Program Book - 27th Fungal Genetics Conference

Asilomar Conference GroundsMarch 12 – 17, 2013Scientific Program Chairs:Katharine Borkovich, University of California, RiversideFrancis Martin, INRA, Nancy, FranceFungal Policy CommitteeBarbara Howlett, Chair (2007-2013)University of MelbourneNeil Gow (2007-2013)Univ. of Abderdeen Institute of Medical SciencesNicholas Talbot (2007 – 2013)Washington Singer LaboratoriesFrancine Govers (2009-2015)Wageningen UniversityBarry Scott (2009-2015)Institute of Molecular BioSciences, MasseyUniversityEric U Selker (2009-2015)University of OregonNick Read (2011-2017)University of EdinburghFrances Trail (2011-2017)Michigan State UniversityLinda Kohn (2011-2017)University of TorontoMarc Orbach (FGC Grant Coordinator)University of ArizonaEx officio, Mike PlamannDirector, Fungal Genetics Stock CenterEx officio, Kevin McCluskeyCurator, Fungal Genetics Stock CenterThe 2013 Fungal Conference logo was created by Xiaoping Li and Mona Pokharel, New Mexico StateUniversity.

Thursday, March 147:30 am - 8:30 am Breakfast Crocker Hall8:30 am – 5:00 pm Registration Surf and Sand8:30 am - 12:00 noon Plenary Session II Merrill Hall and ChapelOrganismic Molecular Interactions Chair: Nick TalbotYong-Hwan LeeAntonio Di PietroPeter DoddsFrancine GoversAlexandra C. BrandLarge-scale Biology for Fungal Pathogenicity in Magnaporthe oryzaeMAPK-mediated control of infectious growth in Fusarium oxysporumAnalysis of effector proteins from flax rust and wheat stem rustDissecting Phytophthora blight; making sense out ofsignalling, effectors and host targetsUnderstanding directional growth in fungi12:00 noon - 1:00 pm Lunch – Crocker HallBox lunches will be available on a first come, first served basis for meeting attendees on the deck outside of theAdministration Building.Following lunch, the morning speakers will be available on the benches outside the administration building tomeet with students. Please allow time for students. In the event of rain, please go inside the AdministrationBuilding.12:15 pm - 1:15 pm GSA Careers Luncheon Crocker HallAd hoc Workshops Box lunches will be available for those attending the sessions.12:15 pm - 1:30 pm Neurospora Business Meeting Chapel12:30 pm - 2:00 pm JGI Sequencing and Analysis Tools and Initiatives Merrill Hall3:00 pm - 6:00 pm Concurrent Sessions IICool Tools for Fungal BiologyFungi and Evolutionary TheoryCytoskeleton, Motors, and IntracellularTransportNucleic Acid-Protein Interactions that ImpactTranscription and TranslationInteractions between Fungi and AnimalsFungal Volatiles and Organic Compounds asSignaling AgentsGenomics and Biochemistry of Degradationof Complex Molecules in the EnvironmentMiguel Penalva and Kevin McCluskeyHanna Johannesson and Duur AanenSamara Reck-Peterson and PingWangMichael Freitag and Mark CaddickNeil Gow and Clarissa NobileJoan Bennett and Richard SplivalloJonathan Walton and Dan CullenMerrill HallChapelHeatherFred FarrForumKilnNautilusScripps6:00 pm - 7:00 pm Dinner Crocker Hall7:30 pm - 10:30 pm Poster Session II and ExhibitsODD numbered posters from 7:30 – 8:30 and EVEN numbered posters from 8:30 – 9:30FiresidePavilionPoster NumberTopic221-350 Comparative and Functional Genomics354-475 Gene Regulation2

SCHEDULE OF EVENTSFriday, March 157:30 am - 8:30 am Breakfast Crocker Hall8:30 am – 5:00 pm Registration Surf and Sand8:30 am - 12:00 noon Plenary Session III Merrill Hall and ChapelSensing, Cell Biology and Development Chair: Michelle MomanyMeritxell RiquelmeJoseph HeitmanMichael BrunnerStephen OsmaniGregory JeddThe illuminated Spitzenkörper of Neurospora crassa: tracking and tracingsecretory vesiclesEvolution of sexual reproduction: A view from the Fungal KingdomMetabolic compensation of the Neurospora clock by a glucose-dependentfeedback of the circadian repressor CSP1 on the core oscillatorIntegration of the fungal cell cycle with growth and developmentA Neurospora cell-free system reconstitutes peroxisome membrane proteinsynthesis and organelle-specific targeting12:00 noon - 1:00 pm Lunch – Crocker HallBox lunches will be available on a first come, first served basis for meeting attendees on the deck outside of theAdministration Building.Following lunch, the morning speakers will be available on the benches outside the administration building tomeet with students. Please allow time for students. In the event of rain, please go inside the AdministrationBuilding.Ad hoc Workshops Box lunches will be available for those attending the sessions.12:15 pm - 1:30 pm FungiDB Kiln12:15 pm - 1:30 pm Magnaporthe Comparative Genomics Chapel3:00 pm - 6:00 pm Concurrent Sessions IIIPathogenic Signaling via Effector ProteinsCell Wall, Polarity and Hyphal Tip GrowthSexual Regulation and Evolution in the FungiOxidative Stress, ROS Signaling andAdaptation to HypoxiaPhylogenomicsSynthetic BiologyFungicides and Antifungal CompoundsBrett Tyler and Sebastien DuplessisStephan Seiler andErnestina Castro-LongoriaFrances Trail and Nicolas CorradiGeraldine Butler and Barry ScottJason Stajich and Joey SpataforaNancy Keller and Peter PuntDaniele Debieu and Paul VerweijMerrill HallChapelHeatherFred FarrForumKilnNautilusScripps6:00 pm - 7:00 pm Dinner Crocker Hall7:00 pm - 8:00 pm GSA Education Special Interest Group Mixer Surf and Sand7:30 pm - 10:30 pm Poster Session III and ExhibitsODD numbered posters 7:30 – 8:30 and EVEN numbered posters 8:30 – 9:30.FiresidePavilionPoster NumberTopic476 – 639 Pathogenic and Mutualistic Interactions640 - 688 Population and Evolutionary Genetics714 - 741 Other Topics27th Fungal Genetics Conference | 3

Saturday, March 167:30 am - 8:30 am Breakfast Crocker Hall9:00 am – 12:00 noon Registration Surf and Sand8:30 am - 12:00 noon Plenary Session IVFunctional Ecology and Fungal Communities Chair: Jim AndersonMerrill Halland ChapelTatiana GiraudMechanisms allowing the formation of new fungalpathogenic species on novel hosts, causing emergingdiseasesB. D. Lindahl The decisive role of mycorrhizal fungi as regulators ofcarbon sequestration in boreal forest ecosystemsEdward J. LouisPopulation Genomics of Saccharomyces Yeasts:Ecology and AdaptationMarc-André Selosse The mycorrhizal symbiosis as a network linking plantsEva H. Stukenbrock Unraveling speciation and specialization processes inplant pathogenic fungi using comparative populationgenomics12:00 noon - 1:00 pm Lunch – Crocker HallBox lunches will be available on a first come, first served basis for meeting attendees on the deck outside of theAdministration Building.Following lunch, the morning speakers will be available on the benches outside the administration building tomeet with students. Please allow time for students. In the event of rain, please go inside the AdministrationBuilding.2:00 pm - 5:00 pm Concurrent Session IVParallels between Fungal Pathogens of Plantsand AnimalsSecondary MetabolismLight Sensing and Circadian RhythmsFungal Evo-DevoEnvironmental MetagenomicsDimorphic TransitionsTropic Growth and FusionBarbara Howlett and Axel BrakhageGillian Turgeon and Bettina TudzynskiLuis Larrondo and Reinhard FischerSteve Harris and Brian ShawChris Schadt and Betsy ArnoldAnne Dranginis and AlexAndrianopoulosAndre Fleissner and Nick ReadMerrill HallChapelHeatherFred FarrForumKilnNautilusScripps5:30 pm - 5:45 pm Poster Awards Merrill Hall andChapel5:45 pm - 6:30 pm Perkins/Metzenberg Lecture:Regine Kahmann, Max Planck Institute for TerrestrialMerrill Hall andChapelMicrobiology6:30 pm - 8:30 pm Closing Banquet Crocker Hall8:30 pm - 12:00 am Closing Party featuring The Amplified DNA Band Merrill Hall8:30 pm - 12:00 am Quiet Alternative Surf and SandSunday, March 177:30 am - 8:30 am Breakfast Crocker Hall4

EXHIBITSThe following companies have contributed to the support of this meeting.Registrants are encouraged to visit the exhibits.Bayer SAS – Bayer CropScience14 Impasse Pierre BaizetLyon, France 69009Tel: 33 472 85 23 43Email: marco.busch@bayer.comBayer CropScience, the subgroup of Bayer AGresponsible for the agricultural business, has annualsales of EUR 7.255 billion (2011) and is one of theworld’s leading innovative crop science companiesin the areas of seeds, crop protection and nonagriculturalpest control. The company offers anoutstanding range of products including high valueseeds, innovative crop protection solutions basedon chemical and biological modes of action as wellas an extensive service backup for modern,sustainable agriculture. In the area of nonagriculturalapplications, Bayer CropScience has abroad portfolio of products and services to controlpests from home and garden to forestryapplications. The company has a global workforceof 21,000 and is represented in more than 120countries.ElsevierRadarweg 291043 NXAmsterdam, The NetherlandsTel: 31 20 4853835Email: a.helsloot@elsevier.comWebsite: www.elsevier.comElsevier is a world-leading provider of scientific,technical and medical information products andservices. The company works in partnershipwith the global science and health communitiesto publish more than 2,000 journals, includingFungal Genetics and Biology, Fungal Biologyand Fungal Ecology. All articles are availableonline through ScienceDirect.Union Biometrica, Inc.Tel: 508-893-3115E-mail: dstrack@unionbio.comWebsite: www.unionbio.comUnion Biometrica provides flow cytometry forobjects that are too large for traditionalcytometers, such as fungal pellets, and offersan alternative to manual sorting. Theseinstruments analyze and dispense objectsbased on size and fluorescent parameters.Automating this process offers increased speed,sensitivity, quantification, and repeatability ofexperiments.MO BIO Laboratories2746 Loker AvenueCarlsbad, CA 92010Tel: 760-929-9911Email: customercare@mobio.comWebsite: www.mobio.comMO BIO Laboratories, Inc. is a global leader insolutions for nucleic acid purification, offeringinnovative tools for research in plant biology.Our patented Inhibitor Removal Technologyensures isolation of high quality, inhibitor-freenucleic acids from even the toughest plantsamples, removing phenolics, polysaccharidesand other PCR inhibiting substances.6

CONCURRENT SESSIONS SCHEDULESWednesday, March 13 3:00 PM–6:00 PMMerrill HallCell Signaling Involved in Fungal Developmentand PathogenesisCo-chairs: Naweed Naqvi and Stefanie PöggelerAbstracts for this session begin on page 29.3:00 - 3:20Alexander V. MichkovStability of a G protein alpha subunit in genetic backgrounds lackingthe G beta subunit or a cytosolic guanine nucleotide exchange factor.3:20 - 3:40Jae-Hyuk YuThe Putative Guanine Nucleotide Exchange Factor RicA MediatesUpstream Signaling for Growth and Development in Aspergillus.3:40 - 4:00Oezguer BayramThe Aspergillus nidulans MAPK module AnSte11-Ste50-Ste7-Fus3controls development and secondary metabolism.4:00 - 4:20Ines TeichertThe developmental PRO40/SOFT protein participates in signaling viathe MIK1/MEK1/MAK1 module in Sordaria macrospora.4:20 - 4:40 Break4:40 - 5:00Linqi WangA Fungal Adhesin Guides Community Behaviors by Autoinduction andParacrinal Signaling.5:00 - 5:20JinRong XuSurface recognition and appressorium morphogenesis inMagnaporthe oryzae.5:20 - 5:40Marie NishimuraPlant cues promote stealth infection in fungal plant pathogens.5:40 - 6:00Andrea HerrmannUnravelling the GTPase polarity complex in Claviceps purpurea.Wednesday, March 13 3:00 PM–6:00 PMChapelGenetics and Genomics of Interactions withBacteria, Insects and PlantsCo-chairs: Nemat Keyhani and Christian HertweckAbstracts for this session begin on page 32.3:00 - 3:20M. J. BidochkaEndophytic insect parasitic fungi feed insect-derived nitrogen toplants.3:20 - 3:40Rusty J. RodriguezGenotype-Environment Interactions and the Interplay BetweenClimate Change and Plant-Fungal Symbioses.3:40 - 4:00Kirstin ScherlachChemical mediators of pathogenic and mutualistic bacterial-fungalinteractions.4:00 - 4:20Chengshu WangComparative genomic analysis of entomopathogenic fungi.4:20 - 4:40 Break4:40 - 5:00Morten SchiøttSynergistic interactions between leaf-cutting ants and their fungalsymbiont facilitate degradation of plant substrate.5:00 - 5:20Charissa de BekkerUnraveling the metabolome: how zombie ant fungi heterogeneouslycontrol ant brains.5:20 - 5:40Artemio MendozaTrichoderma rhizosphere’s competency, endophytism and plantcommunication: A molecular approach.5:40 - 6:00Markus KünzlerEffector proteins in fungal defense against fungivorous nematodes:Targets and functional significance.27th Fungal Genetics Conference | 7

CONCURRENT SESSIONS SCHEDULESWednesday, March 13 3:00 PM–6:00 PMHeatherMembrane Trafficking and MolecularOrganizationCo-chairs: Vicky Sophianopoulou and Gero SteinbergAbstracts for this session begin on page 35.3:00 - 3:20Barbara ValentDistinct secretion systems operate during biotrophic invasion by therice blast fungus, Magnaporthe oryzae.3:20 - 3:40Yujiro HiguchiThe cellular role of early endosome motility in Ustilago maydis.3:40 - 4:00George DiallinasThe arrestin-like protein ArtA is essential for ubiquitylation andendocytosis of the UapA transporter in response to both broad-rangeand specific signals.4:00 - 4:20Guido GrossmannEscaping the hustle - zones of differential protein turnover in theyeast plasma membrane.4:20 - 4:40 Break5:00 - 5:20Samara Reck-PetersonWhole-genome sequencing identifies novel alleles of genes requiredfor organelle distribution and motility in Aspergillus nidulans.5:20 - 5:40Rosa R. Mouriño-PérezDynamics of exocytic markers and cell wall alterations in anendocytosis mutant of Neurospora crassa.5:40 - 6:00Barry J. Bowman“The vacuole” of Neurospora crassa may be composed of multiplecompartments with different structures and functions.Wednesday, March 13 3:00 PM–6:00 PMFred Farr ForumGenome Defense, Epigenetics and RNAiCo-chairs: Patrick Shiu and Sven SaupeAbstracts for this session begin on page 37.3:00 - 3:20Patrick K. T. ShiuMeiotic silencing by unpaired DNA in Neurospora.3:20 - 3:40Zhenyu ZhangMechanism of quelling, a small RNA-mediated gene silencingpathway.3:40 - 4:00Xuying WangSIS, a sex genome defense mechanism operating in Cryptococcusneoformans.4:00 - 4:20Asen DaskalovFungi use prion folds for signal transduction processes involvingSTAND proteins.4:20 - 4:40 Break4:40 - 5:00Haoping LiuRegulation of white and opaque cell-type formation in Candidaalbicans by H3K56 acetylation and nucleosome assembly factors CAF-1 and HIR.5:00 - 5:20Matthew Z. AndersonEpigenetic Regulation of Subtelomeric Gene Noise in Candidaalbicans.5:20 - 5:40Zachary A. LewisChromatin regulation of genome stability.5:40 - 6:00Shinji HondaOpposing activities of the HCHC and DMM complexes maintain properDNA methylation in Neurospora crassa.8

CONCURRENT SESSIONS SCHEDULESWednesday, March 13 3:00 PM–6:00 PMKilnGenomics and MycorrhizaeCo-chairs:Abstracts for this session begin on page 39.3:00 - 3:20A. KohlerThe mycorrhizal genome initiative (MGI): Identification of symbiosisregulatedgenes by using RNA-Seq.3:20 - 3:40Jaqueline HessTransposable element dynamics in the Amanita: insights on theevolution of genome architecture accompanying the transition fromsaprotrophic to ectomycorrhizal ecologies.3:40 - 4:00Alga ZuccaroBroad compatibility in the root endophyte Piriformospora indica isassociated with host-adapted colonization strategies.4:00 - 4:20Anders P. V. TunlidExamining the saprotrophic ability of ectomycorrhizal fungi usinggenomics, transcriptomics and spectroscopy.4:20 - 4:40 Break4:40 - 5:00Nils OS HögbergInteraction between the saprotrophic fungus Serpula lacrymans andliving pine roots.5:00 - 5:20Stephen J. MondoUncovering the evolutionary pressures shaping the Glomeromycota-Glomeribacter endosymbiosis.5:20 - 5:40Alija MujicA draft genome of the ectomycorrhizal fungus Rhizopogonvesiculosus: Characterization of mating system and heterozygositywithin the dikaryon.5:40 - 6:00H.-L. LiaoMetatranscriptomic analysis of ectomycorrhizal root clusters in Pinustaeda: new methodologies for assessing functional gene expression insitu.Wednesday, March 13 3:00 PM–6:00 PMNautilusRegulation and Comparative Genomics ofCarbon and Nitrogen MetabolismCo-chairs: Richard Wilson and Ronald de VriesAbstracts for this session begin on page 42.3:00 - 3:20Carl R. FellbaumThe role of carbon in fungal nutrient uptake and transport:implications for resource exchange in the arbuscular mycorrhiza.3:20 - 3:40Jessie FernandezMechanisms of adaptation to host rice cells by the blast fungus.3:40 - 4:00Sylvia KlaubaufSimilar is not the same: Differences in the function of the (hemi-)cellulolytic regulator XlnR (Xlr1/Xyr1) in filamentous fungi.4:00 - 4:20Richard B. ToddRegulating the Aspergillus nidulans global nitrogen transcriptionfactor AreA.4:20 - 4:40 Break4:40 - 5:00Miia R. MäkeläTranscriptional analysis of oxalate degradation in the white rotbasidiomycete Dichomitus squalens.5:00 - 5:20Gesabel Y. Navarro VelascoTOR-mediated control of virulence functions in the trans-kingdompathogen Fusarium oxysporum.5:20 - 5:40Firoz ShahTranscriptional regulation of peptidases and nitrogen transportersduring the assimilation of organic nitrogen by the ectomycorrhizalfungi Paxillus involutus.5:40 - 6:00Michael HynesRegulation of glycolysis and gluconeogenesis by antisensetranscription in Aspergillus nidulans?27th Fungal Genetics Conference | 9

CONCURRENT SESSIONS SCHEDULESWednesday, March 13 3:00 PM–6:00 PMScrippsEducation, Outreach, and ProfessionalDevelopmentCo-chairs: Steven Denison and Mimi ZolanAbstracts for this session begin on page 45.3:00 - 3:20Michael KoonceCentrosome-Nuclear Disconnect Creates Mitotic Chaos in a ClosedMitosis System.3:20 - 3:40Claire BurnsUsing Fungal Barcoding to Introduce Non-science Majors to ScientificResearch.3:40 - 4:00Andrea GargasComGen Authentic Research Experiences (C-ARE): Fungal geneticanalysis.4:00 - 4:20Patricia J. PukkilaWearing two hats: Tips for combining commitments to research andto university-wide initiatives in education.4:20 - 4:40 Break4:40 - 5:00Virginia K. HenchFacilitating an Interdisciplinary Learning Community AmongstUndergraduate Research Fellows By Emphasizing Scientific Inquiry asthe Unifying Thread.5:00 - 5:20Relly BrandmanMOOCs: Education for Everyone.5:20 - 5:40Break into groups to discuss promises and pitfalls of online courses.5:40 - 6:00Panelists Relly Brandman and Heaher Hallen-Adams respond toquestions and comments from the working groups.Thursday, March 14 3:00 PM–6:00 PMMerrill HallCool Tools for Fungal BiologyCo-chairs: Miguel Penalva and Kevin McCluskeyAbstracts for this session begin on page 47.3:00 - 3:20S. E. BakerThe Environmental Molecular Sciences Laboratory molecular analysiscapabilities for fungal biology.3:20 - 3:40Aric E. WiestDevelopment and utilization of arrayed mutant sets for yeasts andfilamentous fungi.3:40 - 4:00Minou NowrousianSequencing-based solutions to identify and characterize fungaldevelopmental genes.4:00 - 4:20Susan KaminskyjAspergillus nidulans as an experimental system to identify novel cellwall growth and maintenance genes through identification of antifungaldrug resistance mutations.4:20 - 4:40 Break4:40 - 5:00David L. JolyIllumina-based genetic linkage map for wheat leaf rust.5:00 - 5:20Miguel PenalvaPeering into the secret-ory life of Aspergillus nidulans with a littlehelp from classical genetics.5:20 - 5:40Patricia J. PukkilaDomains of meiotic DNA recombination and gene conversion inCoprinopsis cinerea (Coprinus cinereus).5:40 - 6:00Xin XiangA Hook protein is critical for dynein-mediated early endosomemovement in Aspergillus nidulans.10

CONCURRENT SESSIONS SCHEDULESThursday, March 14 3:00 PM–6:00 PMFred Farr ForumNucleic Acid-Protein Interactions that ImpactTranscription and TranslationCo-chairs: Michael Freitag and Mark CaddickAbstracts for this session begin on page 53.3:00 - 3:20Koon Ho WongChIP-seq: an inexpensive and powerful method for studying genomewidechromatin remodeling and transcription regulation in fungi.3:20 - 3:40Jay C. DunlapRegulatory Networks Governing Global Responses to Changes in Lightand Time.3:40 - 4:00L. F. LarrondoProtein Binding Microarrays and high-throughput real-time reportersstudies: Building a four-dimensional understanding of transcriptionalnetworks in Neurospora crassa.4:00 - 4:20Ane SesmaEnding messages: alternative polyadenylation in filamentous fungi.4:20 - 4:40 Break4:40 - 5:00Amanda L. Misener BloomPost-transcriptional gene regulation contributes to host temperatureadaptation and virulence in Cryptococcus neoformans.5:00 - 5:20Johannes FreitagDual targeting of glycolytic enzymes by alternative splicing andtranslational read-through.5:20 - 5:40Mian ZhouNon-optimal codon usage determines the expression level, structureand function of the circadian clock protein FREQUENCY.Thursday, March 14 3:00 PM–6:00 PMKilnInteractions between Fungi and AnimalsCo-chairs: Neil Gow and Clarissa NobileAbstracts for this session begin on page 55.3:00 - 3:20Elaine M. BignellElicitation of host damage occurs in a temporally programmedmanner during Aspergillus fumigatus infections.3:20 - 3:40Stuart LevitzExploiting innate recognition of fungi for vaccine development.3:40 - 4:00Jose C. PerezRegulatory circuits governing Candida albicans proliferation in amammalian host.4:00 - 4:20Judith BermanDramatic ploidy change as an adaptive strategy in Candida albicans...4:20 - 4:40 Break4:40 - 5:00Yen-Ping HsuehNematode-trapping fungi eavesdrop on nematode pheromones.5:00 - 5:20Xiaorong LinA morphogenesis regulator controls cryptococcal neurotropism.5:20 - 5:40M. BrockSit and wait: Special features of Aspergillus terreus in macrophageinteractions and virulence.5:40 - 6:00Dawn ThompsonThe mutational landscape of gradual acquisition of drug resistance inclinical isolates of Candida albicans.5:40 - 6:00Michael FeldbruggeA transcriptome-wide view on microtubule-dependent mRNAtransport.12

CONCURRENT SESSIONS SCHEDULESThursday, March 14 3:00 PM–6:00 PMNautilusFungal Volatiles and Organic Compounds asSignaling AgentsCo-chairs: Joan Bennett and Richard SplivalloAbstracts for this session begin on page 58.3:00 - 3:20Birgit PiechullaFungi reacting to rhizobacterial volatiles.3:20 - 3:40Seogchan KangEnhancement of plant growth and stress resistance by Fusariumvolatile organic compounds: A novel mechanism mediating plantfungalinteractions.3:40 - 4:00Jessica C. HargartenThe Role of Quorum-sensing Molecules in Interactions betweenCandida albicans and its Host.4:00 - 4:20Vong shian Simon Ip ChoInnate Immunity in Fusarium graminearum.4:20 - 4:40 Break4:40 - 5:00Lea AtanasovaThe Trichoderma reesei polyketide synthase gene pks1 is necessaryfor yellow-green pigmentation of conidia and is involved in theestablishment of environmental fitness.5:00 - 5:20Richard HungSemiochemicals and signaling: plant responses to Trichodermavolatile organic compounds.5:20 - 5:40El Ghalid MennatIdentification of chemoattractant compounds from tomato rootexudate that trigger chemotropism in Fusarium oxysporum.5:40 - 6:00Richard SplivalloThe mixed fungal and bacterial origin of truffle aroma.Thursday, March 14 3:00 PM–6:00 PMScrippsGenomics and Biochemistry of Degradation ofComplex Molecules in the EnvironmentCo-chairs: Jonathan Walton and Dan CullenAbstracts for this session begin on page 61.3:00 - 3:20K. IgarashiFungal transcriptome as database for proteome and refinement toolof gene annotation.3:20 - 3:40Irina S. DruzhininaDevelopmental regulation and cellulase gene expression inTrichoderma reesei.3:40 - 4:00D. FloudasParallel losses of genes associated with saprotrophy inectomycorrhizal Agaricomycotina lineages.4:00 - 4:20Emma MasterCo-expression analysis of Phanerochaete carnosa during growth onhardwood and softwood species to predict proteins with unknownfunction relevant to biomass conversion.4:20 - 4:40 Break4:40 - 5:00Yitzhak HadarFunctional Analysis of the Pleurotus ostreatus Manganese-PeroxidaseGene Family.5:00 - 5:20Monika SchmollCarbon source and light dependent regulation of gene clusters inTrichoderma reesei (Hypocrea jecorina).5:20 - 5:40Chiaki HoriGenome-wide analysis of eleven white- and brown-rot Polyporalesprovides insight into mechanisms of wood decay.5:40 - 6:00Alex LichiusTranscription factor shuttling during cellulase induction inTrichoderma reesei.27th Fungal Genetics Conference | 13

CONCURRENT SESSIONS SCHEDULESFriday, March 15 3:00 PM–6:00 PMMerrill HallPathogenic Signaling via Effector ProteinsCo-chairs: Brett Tyler and Sebastien DuplessisAbstracts for this session begin on page 64.3:00 - 3:20Marie-Cecile CaillaudDissecting nuclear immunity using Arabidopsis downy mildew effectoras probes.3:20 - 3:40Claire Veneault-FourreyThe mutualistic fungus Laccaria bicolor uses the effector proteinMiSSP7 to alter host jasmonate signaling and establish symbiosis.3:40 - 4:00Shiv D. KaleIdentification and characterization of an RXLR-like effector familyfrom medically relevant fungi.4:00 - 4:20Yuanchao WangIdentification and functional assay of Phytophthora sojae avirulenceeffectors.4:20 - 4:40 Break4:40 - 5:00Gregory J. FischerFungal lipoxygenases: a novel instigator of asthma?5:00 - 5:20Martha C. GiraldoMagnaporthe oryzae has evolved two distinct mechanisms of effectorsecretion for biotrophic invasion of rice.5:20 - 5:40Anupama GhoshDomains for plant uptake of Ustilago maydis secreted effectors.5:40 - 6:00Edouard EvangelistiPenetration-specific effectors from Phytophthora parasitica favourplant infection.Friday, March 15 3:00 PM–6:00 PMChapelCell Wall, Polarity and Hyphal Tip GrowthCo-chairs: Stephan Seiler and Ernestina Castro-LongoriaAbstracts for this session begin on page 67.3:00 - 3:20Michael BölkerThe function of Rho type small GTPases for cell polarity in Ustilagomaydis.3:20 - 3:40Peter SudberyA quantitative model of hyphal tip growth based on the spatialdistribution of exocyst subunits in the human fungal pathogenCandida albicans.3:40 - 4:00Johannes WagenerCell wall integrity signaling in Aspergillus fumigatus.4:00 - 4:20Roland Wedlich-SoldnerOptimization of polarity establishment through coupling of multiplefeedback loops.4:20 - 4:40 Break4:40 - 5:00Vincent BuloneCell wall structure and biosynthesis in oomycetes and true fungi: acomparative analysis.5:00 - 5:20Lakshmi Preethi YerraCellular morphogenesis of Aspergillus nidulans conidiophores: asystematic survey of protein kinase and phosphatase function.5:20 - 5:40Diego Delgado-ÁlvarezSeptum formation starts with the establishment of a septal actintangle (SAT) at future septation sites.5:40 - 6:00Norio TakeshitaVisualization of apical membrane domains in Aspergillus nidulans byPhotoactivated Localization Microscopy (PALM).14

CONCURRENT SESSIONS SCHEDULESFriday, March 15 3:00 PM–6:00 PMHeatherSexual Regulation and Evolution in the FungiCo-chairs: Frances Trail and Nicolas CorradiAbstracts for this session begin on page 69.3:00 - 3:20Ignazio CarboneClonality and sex impact aflatoxigenicity in Aspergillus populations.3:20 - 3:40Nicolas CorradiToolkit for sexual reproduction in the genome of Glomus spp; asupposedly ancient asexual lineage.3:40 - 4:00Frances TrailComparative transcriptomics identifies new genes for peritheciumdevelopment.4:00 - 4:20Hanna JohannessonRapid evolution of female-biased genes: a novel example from theeukaryotic model organism Neurospora crassa.4:20 - 4:40 Break4:40 - 5:00Katherine A. BorkovichSelf-attraction can not bypass the requirement for two mating typegenes during sexual reproduction in Neurospora crassa.5:00 - 5:20Céline M. O'GormanFertility in Aspergillus fumigatus and the identification of anadditional ‘supermater’ pair.5:20 - 5:40Julia BöhmSexual reproduction and mating type function in the penicillinproducing fungus Penicillium chrysogenum.5:40 - 6:00Patrik InderbitzinThe Sclerotinia sclerotiorum mating type locus (MAT) contains a 3.6-kb region that is inverted in every generation.Friday, March 15 3:00 PM–6:00 PMFred Farr ForumOxidative Stress, ROS Signaling andAdaptation to HypoxiaCo-chairs: Geraldine Butler and Barry ScottAbstracts for this session begin on page 723:00 - 3:20A. NantelTranscriptional regulatory networks controlling the early hypoxicresponse in Candida albicans.3:20 - 3:40Olaf KniemeyerProteomic analysis of the hypoxic response of the human-pathogenicfungus Aspergillus fumigatus.3:40 - 4:00N. PontsFgap1-mediated response to oxidative stress in trichotheceneproducingFusarium graminearum.4:00 - 4:20Nallely Cano-DominguezThe role of NADPH oxidases in Neurospora crassa cell fusion.4:20 - 4:40 Break4:40 - 5:00Elizabeth A. VealPeroxiredoxins in ROS responses -Why evolve peroxidases that areinactivated by peroxides?5:00 - 5:20Lauren S. RyderNADPH oxidases regulate septin-mediated cytoskeletal re-modelingduring plant infection by the rice blast fungus Magnaporthe oryzae.5:20 - 5:40Gemma M. CartwrightRedox regulation of an AP-1-like transcription factor, YapA, in thefungal symbiont Epichloë festucae.5:40 - 6:00Benjamin A. HorwitzInteraction between phenolic and oxidant signaling in Cochliobolusheterostrophus.27th Fungal Genetics Conference | 15

CONCURRENT SESSIONS SCHEDULESFriday, March 15 3:00 PM–6:00 PMScrippsFungicides and Antifungal CompoundsCo-chairs: Daniele Debieu and Paul VerweijAbstracts for this session begin on page 81.3:00 - 3:20D. A. MacdonaldChemically Induced Haploinsufficiency Screens to Identify DrugMechanism of Action in Aspergillus Fumigatus.3:20 - 3:40Branka KorosecInhibition of benzoate 4-monooxygenase (CYP53A15) fromCohliobolus lunatus by cinnamic acid derivatives.3:40 - 4:00Marcelo HS RamadaSecretome analysis of Trichoderma harzianum cultivated in thepresence of Fusarium solani cell wall or glucose.4:00 - 4:20Carol E. DavisMetabolic adaptation of the oomycete Phytophthora infestans duringcolonization of plants and tubers.4:20 - 4:40 Break4:40 - 5:00Paul E. VerweijThe fungi strike back: multidrug resistance in Aspergillus fumigatusand agricultural use of fungicides.5:00 - 5:20D. SanglardEffect of antifungal resistance on virulence of Candida spp.5:20 - 5:40Sabine FillingerFrom enzyme to fungal development or how sdhB mutations impactrespiration, fungicide resistance and fitness in the grey mold agentBotrytis cinerea.5:40 - 6:00Gabriel ScallietDeciphering fungicide resistance mechanisms in phytopatogenic fungi,towards an assessment of resistance risk in new active ingredientresearch.Saturday, March 16 2:00 PM–5:00 PMMerrill HallParallels between Fungal Pathogens of Plantsand AnimalsCo-chairs: Barbara Howlett and Axel BrakhageAbstracts for this session begin on page 84.2:00 - 2:20Sarah J. GurrEmerging fungal (and Oomycete) threats to plant and ecosystemhealth.2:20 - 2:40Axel A. BrakhageMelanin as virulence determinant of human and plant pathogenicfungi.2:40 - 3:00Joanna PotrykusNutrient immunity and systemic readjustment of metal homeostasismodulate fungal iron availability during the development of renalinfections.3:00 - 3:20A. SharonCommon strategies in plant and human "necrotrophic" pathogens:role of PCD.3:20 - 3:40 Break3:40 - 4:00Nick J. TalbotSeptin-mediated plant tissue invasion by the rice blast fungusMagnaporthe oryzae.4:00 - 4:20Katja SchaeferComponents of the urease complex govern virulence of Fusariumoxysporum on plant and animal hosts.4:20 - 4:40Anja KombrinkThe role of LysM effectors in fungal fitness.4:40 - 5:00Harshini C. WeerasingheGenes important for in vivo survival of the human pathogenPenicillium marneffei.27th Fungal Genetics Conference | 17

CONCURRENT SESSIONS SCHEDULESSaturday, March 16 2:00 PM–5:00 PMChapelSecondary MetabolismCo-chairs: Gillian Turgeon and Bettina TudzynskiAbstracts for this session begin on page 87.2:00 - 2:20B. CondonGenomic profiles of secondary metabolism genes in Cochlioboluspathogens.2:20 - 2:40Candace ElliottA biosynthetic gene cluster for the antifungal metabolite phomenoicacid in the plant pathogenic fungus, Leptosphaeria maculans.2:40 - 3:00Eva-Maria NiehausFusarin C biosynthesis in Fusarium fujikuroi: the fusarin C gene cluster,their function and regulation.3:00 - 3:20H. Corby KistlerCellular development integrating primary and induced secondarymetabolism in the filamentous fungus Fusarium graminearum.3:20 - 3:40 Break3:40 - 4:00Nancy KellerLaeA sleuthing reveals cryptic gene clusters in pathogenic Aspergilli.4:00 - 4:20Kristina M. SmithThe KMT6 Histone H3 K27 Methyltransferase Regulates Expression ofSecondary Metabolites and Development in Fusarium graminearum.4:20 - 4:40M. ViaudSecondary metabolism in Botrytis cinerea: the grey and pink sides of apathogen.4:40 - 5:00Frank KempkenIs fungal secondary metabolism regulated by competing insects?Saturday, March 16 2:00 PM–5:00 PMHeatherLight Sensing and Circadian RhythmsCo-chairs: Luis Larrondo and Reinhard FischerAbstracts for this session begin on page 90.2:00 - 2:20Martha W. MerrowCircadian rhythms in gene expression in Aspergillus nidulans.2:20 - 2:40C. HongCircadian clock-gated cell division cycles in Neurospora crassa.2:40 - 3:00Kevin K. FullerLight regulates growth, stress resistance and metabolism in the fungalpathogen Aspergillus fumigatus.3:00 - 3:20Paulo CanessaShedding light on Botrytis biology: characterization of the WC1photoreceptor and FRQ homologues in the necrotrophic plantpathogen Botrytis cinerea.3:20 - 3:40 Break3:40 - 4:00Carmen Ruger-HerrerosThe transcription factor FL is phosphorylated and interacts with atrehalose related protein in Neurospora crassa.4:00 - 4:20Alfredo H. Herrera-EstrellaRegulation of gene expression in response to light in Trichodermaatroviride.4:20 - 4:40Victoriano GarreGenome-wide analysis of light responses in Mucor circinelloides.4:40 - 5:00Phillipp WiemannShedding light on secondary metabolite cluster gene expression,sporulation, UV-damage repair and carotenogenesis in the ricepathogen Fusarium fujikuroi.18

CONCURRENT SESSIONS SCHEDULESSaturday, March 16 2:00 PM–5:00 PMFred Farr ForumFungal Evo-DevoCo-chairs: Steve Harris and Brian ShawAbstracts for this session begin on page 93.2:00 - 2:20Antonis RokasThe Molecular Foundations of the Fungal Lifestyle.2:20 - 2:40Daniel J. EbboleGene expression and regulation during conidial morphogenesis inNeurospora crassa.2:40 - 3:00David S. HibbettComparative developmental morphology in lentinoid mushrooms:toward a new fungal evo-devo?3:00 - 3:20Steven D. HarrisThe Cdc42 GTPase module and the evolution of conidiophorearchitecture in Aspergillus.3:20 - 3:40 Break3:40 - 4:00Audrey M. V. Ah-FongCdc14 association with basal bodies in the oomycete Phytophthorainfestans indicates potential new role for this protein phosphatase.4:00 - 4:20Jurgen W. WendlandMolecular Determinants of Sporulation in Ashbya gossypii.4:20 - 4:40Heesoo ParkTHE velvet regulators in Aspergilli.4:40 - 5:00R. DebuchyA network of HMG-box transcription factors regulates sexual cycle inthe fungus Podospora anserina.Saturday, March 16 2:00 PM–5:00 PMKilnEnvironmental MetagenomicsCo-chairs: Chris Schadt and Betsy ArnoldAbstracts for this session begin on page 95.2:00 - 2:20Donald R. ZakMicrobial Responses to a Changing Climate: Implications for theFuture Functioning of Terrestrial Ecosystems.2:20 - 2:40Mizue NaitoThe Interaction of Mycoplasma-related Endobacteria with theirArbuscular Mycorrhizal Fungal Host.2:40 - 3:00Ning ZhangMetagenomic analysis reveals hidden fungal diversity in grassrhizosphere and tree foliage.3:00 - 3:20Weiguo FangHost-to-pathogen gene transfer facilitated infection of insects by apathogenic fungus.3:20 - 3:40 Break3:40 - 4:00Kabir PeayStructure and function of soil fungal communities across NorthAmerican pine forests.4:00 - 4:20Gregory BonitoGenomic analysis of Mortierella elongata and its endosymbioticbacterium.4:20 - 4:40Richard C. HamelinIntegrative genomics of poplar-fungal pathogen interactions.4:40 - 5:00M.-S. BenitezFungal pathogen and endophyte genetics within the context of forestcommunity dynamics.27th Fungal Genetics Conference | 19

CONCURRENT SESSIONS SCHEDULESSaturday, March 16 2:00 PM–5:00 PMNautilusDimorphic TransitionsCo-chairs: Anne Dranginis and Alex AndrianopoulosAbstracts for this session begin on page 97.2:00 - 2:20Richard BennettEpigenetic Switching Regulates the Yeast-Hyphal Transition in Candidaalbicans.2:20 - 2:40Linqi WangExtracellular and intracellular signaling orchestrates morphotypetransitionand virulence in human pathogen Cryptococcusneoformans.2:40 - 3:00Chad A. RappleyeHistoplasma strain variations and differences in pathogenic-phasetranscriptomes.3:00 - 3:20Hayley E. BugejaThe C 2H 2 transcription factor HgrA promotes hyphal growth in thedimorphic pathogen Penicillium marneffei.3:20 - 3:40 Break3:40 - 4:00Joerg T. KaemperA conserved splicing factor is required for vesicle transport in Ustilagomaydis.4:00 - 4:20Sarah A. GilmoreN-acetylglucosamine (GlcNAc) Triggers a Morphogenetic Program inSystemic Dimorphic Fungi.4:20 - 4:40Gregory M. GauthierA GATA transcription factor encoded by SREB functions as a globalregulator of transcription in Blastomyces dermatitidis.4:40 - 5:00Bridget M. BarkerFunctional Analysis of Genes in Regions of Introgression inCoccidioides.Saturday, March 16 2:00 PM–5:00 PMScrippsTropic Growth and FusionCo-chairs: Andre Fleissner and Nick ReadAbstracts for this session begin on page 100.2:00 - 2:20Carla J. EatonRole of the cell fusion gene idcA in fungal mutualism.2:20 - 2:40Pablo S. AguilarRole of extracellular calcium in budding yeast cell fusion.2:40 - 3:00Chia-Chen ChangThe role of calcium and calmodulin during cell fusion and colonyinitiation in Neurospora crassa.3:00 - 3:20Javier Palma-GuerreroLFD-1 is a component of the membrane merger machinery during cellcellfusion in Neurospora crassa.3:20 - 3:40 Break3:40 - 4:00Martin WeichertSpecific Structural Features of Sterols Affect Cell-Cell Signaling andFusion in Neurospora crassa.4:00 - 4:20David TurraCo-option of a sex pheromone receptor and MAPK signalling pathwayfor chemotropism of Fusarium oxysporum towards plant hostcompounds.4:20 - 4:40Britta HerzogCharacterization of new STRIPAK complex interaction partners in thefilamentous ascomycete Sordaria macrospora.4:40 - 5:00Darren ThomsonCharacterisation of contact-dependant tip re-orientation in Candidaalbicans hyphae.20

PLENARY SESSION ABSTRACTSCarbohydrate-active enzymes in fungal genomes. Bernard Henrissat. AFMB, CNRS and Aix-Marseille University, Marseille, France.We term carbohydrate-active enzymes (CAZymes) the enzymes that assemble and breakdown complex carbohydrates and carbohydrate polymers. Assuch carbohydrates are crucial for fungi as carbon sources but also for cell wall synthesis/remodelling, host pathogen interactions, energy storage etc.Unlike many other classes of enzymes which carry limited informative power, the peculiarities of CAZymes and of their substrates turn these enzymes intoextremely powerful probes to examine genomes and explain the lifestyle of living organisms and fungi in particular. Over the last few years we haveexplored the CAZyme content of over 200 fungal genomes and we will review how evolution shapes the CAZyme profiles of fungi.Suggested reading :- Cantarel et al. (2009) The Carbohydrate-Active EnZymes database (CAZy): an expert resource for Glycogenomics. Nucleic Acids Res. 37: D233-D238- Ohm et al. (2012) Diverse lifestyles and strategies of plant pathogenesis encoded in the genomes of eighteen Dothideomycetes fungi. PLoS Pathogens,8(12): e1003037.- O’Connell et al.(2012) Life-style transitions in plant pathogenic Colletotrichum fungi defined by genome and transcriptome analyses. Nature Genetics44, 1060-1065- Floudas et al. (2012) The Paleozoic origin of white rot wood decay reconstructed using 31 fungal genomes. Science, 336, 1715-1719- Ma et al. (2010) Comparative genomics reveals mobile pathogenicity chromosomes in Fusarium oxysporum. Nature 464, 367-373- Martin et al. (2010) Périgord black truffle genome uncovers evolutionary origins and mechanisms of symbiosis. Nature 464, 1033-1038.Genome-wide approaches to identify and characterize lignocellulolytic enzymes. Adrian Tsang. Biol, Concordia Univ, Montreal, Canada.Lignocellulosic material is both the most abundant source of biomass on the planet and an enormous storehouse of sugars. Yet the sugars in cellulosicmaterial are remarkably recalcitrant. The ability to detect new enzymes, to produce them in large quantities, and to understand how they work will lay thegroundwork for the development of more efficient and economical processes for lignocellulosic biomass. We are particularly interested in harnessing theligocellulolytyic ability of thermophilic fungi as they are potential reservoirs of thermostable enzymes for industrial applications. So far, fewer than 50fungal species have been described as thermophiles. We have sequenced over 20 species of thermophilic fungi, see www.fungalgenomics.ca. Most ofthese thermophiles belong to the orders Sordariales and Eurotiales, three species belong to the Mucorales and one to Onygenales. We have developedcomputational tools to improve the identification genes in fungal genomes in general, and genes encoding extracellular proteins in particular becausebiomass-degrading enzymes are predominantly extracellular proteins. In addition to using informatics tools to identify orthologues of lignocellulolyticenzymes, we have analyzed the transcriptomes and exo-proteomes of the thermophilic fungi when cultured in a variety of agricultural straws to reveal thestrategies used by different fungi in the decomposition of lignocellulose as well as identifying novel extracellular proteins that may play a role in biomassdecomposition. Over 2000 genes encoding potential lignocellulolytic proteins have been identified. The Sordariales possess a larger repertoire oflignocellulolytic enzymes than the thermophiles from other orders. The genes predicted to encode lignocellulolytic proteins have been cloned andtransformed into Aspergillus niger for the production of recombinant enzymes. Biochemical characterization of the recombinant enzymes show that inaddition to producing enzymes that are thermostable, the thermophiles also produce enzymes that have temperature optimum in the 40-50°C range.22

PLENARY SESSION ABSTRACTSThursday, March 14 8:30 AM–12:00 NOONMerrill Hall and ChapelPlenary Session II: Organismic Molecular InteractionsChair: Nick TalbotLarge-scale Biology for Fungal Pathogenicity in Magnaporthe oryzae. Yong-Hwan Lee 1,2 . 1) Department of Agricultural Biotechnology, Seoul NationalUniv, Seoul, 151-921, Korea; 2) Center for Fungal Genetic Resources, and Center for Fungal Pathogenesis, Seoul National University, Seoul 151-921, Korea.Rice blast is a compelling model system for studying host-parasite interactions due to its socioeconomic impact and the availability of both the rice andfungal genomic sequences. In an attempt to understand the molecular mechanisms of rice blast, we have been taking both forward and reverse geneticsapproaches. Our researches using reverse genetics approach focus on identifying and characterizing the genes involved in signal transduction pathwaysleading to appressorium formation, genes encoding transcription factors, and genes that are required for post-penetration stages. For forward geneticsstudies, we carried out a large-scale insertional mutagenesis of the Magnaporthe oryzae strain KJ201 via Agrobacterium tumefaciens-mediatedtransformation, generating over 25,000 mutants. We also developed high throughput phenotype screening system that enables rapid and robust assay ofmutant phenotypes. In addition to our endeavor to functional and comparative genomics, we built a cyber-infrastructure for storage of heterogeneousdata and analysis of such data in multiple contexts. The whole genome sequence information of M. oryzae as well as most of the results from experimentalbiology is housed in our customized database. Our comprehensive and integrative approaches coupled with a web-based Laboratory InformationManagement System would provide a novel platform for systems biology initiatives for fungal pathogenesis.MAPK-mediated control of infectious growth in Fusarium oxysporum. Antonio Di Pietro. Departamento de Genetica, Universidad de Cordoba, 14071Cordoba, Spain.In fungal pathogens, contact with the host triggers a developmental and metabolic transition towards infectious growth. What exactly defines infectiousgrowth and how it is controlled by environmental and host-derived stimuli is not fully understood. We study infectious growth in Fusarium oxysporum, asoilborne fungus that causes vascular wilt disease on a wide range of plant species and opportunistic infections in immunocompromised humans. One ofthe key players in pathogenicity is Fmk1, a conserved mitogen-activated protein kinase (MAPK) that is essential for infection-related processes such aschemotropism, host adhesion, penetration and invasive growth in the plant tissue. Most Fmk1-dependent virulence functions require the homeodomaintranscription factor Ste12, and are repressed in the presence of the preferred nitrogen source ammonium through a mechanism that requires thetransporter MepB and the bZIP factor MeaB. Recent data suggest that ammonium repression is mediated by a shift in extracellular pH, which results inrapid changes in the phosphorylation pattern of different MAPKs. Our current research addresses the mechanisms through which pH controls invasivegrowth of F. oxysporum by reprogramming the activation status of cellular MAPK signalling cascades.Analysis of effector proteins from flax rust and wheat stem rust. Peter Dodds 1 , narayana Upadhyaya 1 , Ann-Maree Catanzariti 2 , Markus Koeck 1 , Adnanenemri 1 , Rohit Mago 1 , Simon Williams 3 , Thomas Ve 3 , Maryam Rafiqi 4 , Wenjie Wu 2 , Adrienne Hardham 2 , David Jones 2 , Jeff Ellis 1 , Bostjan Kobe 3 , Robert Park 5 .1) Plant industry, CSIRO, Canberra, ACT, Australia; 2) Australian National University, Research School of Biology,; 3) University of Queensland, School ofChemistry and Molecular Biosciences; 4) Justus Liebig University, Giessen, Germany; 5) University of Sydney, Camden.Rust fungi cause economically important diseases of cereal crops worldwide, with stem rust caused by the fungus Puccinia graminis tritici one of themost serious diseases in wheat. Because of the ability of the fungus to evolve increased virulence towards previously resistant varieties, continuousbreeding and the identification of new sources of resistance is necessary to keep apace of the threat of rust epidemics. We have been studying how theplant immune system can recognise and respond to rust pathogens using the flax rust model system. Rusts are obligate parasites of plants, and produce aspecialised infection structure called the haustorium which directly penetrates an infected cell and is the main site of nutrient extraction for the fungus. Asuite of disease effector proteins are secreted from haustoria and enter the host cells where they may allow the rust to commandeer host cell biology. It isthese translocated effector proteins that are recognised by host immune receptors, known as resistance (R) proteins. We have been exploring thestructure and function of host-translocated effectors from flax rust and also searching for effector candidates from stem rust that are recgonised by knownwheat R genes.Using genome and transcriptome sequencing we have predicted, carefully curated and analysed the transcription of 400 candidate effectorgenes from the Australian stem rust strain 21-0. To screen for effectors recognized by wheat R genes, we have developed a bacterial Type III SecretionSystem (TTSS)-based delivery assay from the non-pathogen Pseudomonas fluorescens strain Pfo. We are screening candidate effectors on a set of 18wheat cultivars carrying 22 different R genes. Thus far we have identified on effector protein induces a rapid cell death response specifically on a wheatgenotype carrying Sr22. We are also analyzing sequence variation in effector candidates between clonal field isolates that have mutated to overcome theresistance genes that have been deployed in agriculture.27th Fungal Genetics Conference | 23

PLENARY SESSION ABSTRACTSDissecting Phytophthora blight; making sense out of signalling, effectors and host targets. Francine Govers. Lab. of Phytopathology, WageningenUniversity, Wageningen, Netherlands.The plant pathogen Phytophthora infestans causes late blight, the disease that was responsible for the Irish potato famine in the mid-nineteenth century.This oomycete has a hemibiotrophic life style, a narrow host range and a large genome of ~ 240 Mb. Comparative genomics revealed features illuminatingits success as a pathogen, such as rapid turnover and massive expansion of families encoding secreted proteins, and peculiar gene innovations resulting inproteins with oomycete-specific domain combinations. An example of a novel protein family is the GPCR-PIPK family. Its twelve members all have a N-terminal 7-transmembrane domain typical for G-protein coupled receptors (GPCRs) combined with a phosphatidylinositol phosphate kinase (PIPK) domainat the C-terminus. This domain structure suggests that GPCR-PIPKs use GPCRs to directly feed extracellular signals into phospholipid signalling pathways.Their differential expression and localization point to distinct roles in various cellular processes. For one GPCR-PIPK we could demonstrate a role in asexualdevelopment, including spore germination, hyphal elongation and sporangia cleavage, whereas inactivation of another GPCR-PIPK disturbs sexualdevelopment. For successful infection Phytophthora secretes a variety of proteins including a large number of effectors that share the host-cell targetingmotif RXLR. Inside host cells these RXLR effectors promote virulence by manipulating the cell machinery via interaction with host targets therebysuppressing host defence. However, in plants carrying matching resistance genes RXLR effectors trigger defence and thus act as avirulence factors. Here Iwill focus on an RXLR effector that interacts with an exocyst component and show how the interplay between this effector and its host target influencesthe host-pathogen interaction.Understanding directional growth in fungi. Alexandra C. Brand. Aberdeen Fungal Group, Univ Aberdeen, Scotland, United Kingdom.Fungal hyphae are programmed to explore their surroundings in search of nutrients and, for pathogens, success can depend on locating and identifyingsuitable host penetration sites. Fungi have therefore evolved mechanisms that link the sensing of environmental cues with an appropriate growthresponse. The intracellular components involved in polarised growth in fungi are generally well-conserved and have been studied in model organisms suchas Saccharomyces cerevisiae, Neurospora crassa and Aspergillus spp. However, how environmental signals interact with the molecular machinery of hyphaltip growth is less well-understood. Candida albicans is an opportunistic pathogen that exhibits pre-programmed, or tropic, growth responses to specificstimuli. This makes it a useful model for dissection of the regulatory pathways that control hyphal tip behaviour. A variety of external stimuli, includingelectric fields, surface modification and nanofabrication techniques, have been used to examine the physical properties of apical growth, such asdirectional memory, asymmetric tip organisation and hyphal tip force. In addition, these methods have been coupled with reverse genetics, fluorescenceprotein-tagging and live-cell imaging to identify cell-polarity components that can enhance, or even reverse, the direction of hyphal growth. The evidenceto date suggests that the direction of hyphal growth reflects the net output from overlapping positional determinants. In addition, there is a strongassociation between proper hyphal tip regulation and the ability of a fungus to invade and damage host tissue.24

PLENARY SESSION ABSTRACTSFriday, March 15 8:30 AM–12:00 NOONMerrill Hall and ChapelPlenary Session III: Sensing, Cell Biology and DevelopmentChair: Michelle MomanyThe illuminated Spitzenkörper of Neurospora crassa: tracking and tracing secretory vesicles. Meritxell Riquelme 1 , Eddy Sánchez-León 1 , Rosa Fajardo-Somera 1 , Erin L. Bredeweg 2 , Olga Callejas-Negrete 1 , Robert W. Roberson 3 , Salomon Bartnicki-García 1 , Michael Freitag 2 . 1) Dept Microbiology, Center forScientific Research and Higher Education of Ensenada CICESE, Ensenada, Baja California 22860, Mexico; 2) Center for Genome Research and Biocomputing,Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR 97331, U.S.A; 3) School of Life Sciences,Arizona State University,Tempe, AZ 85287, U.S.A.Tip growth in fungal hyphae is maintained by the vectorial traffic of secretory vesicles to the apex, where they accumulate at the Spitzenkörper (Spk),before fusing with the apical plasma membrane (PM) to provide the enzymatic machinery necessary for cell expansion. Confocal microscopy ofNeurospora crassa strains expressing fluorescently tagged proteins that are predicted to participate in cell wall synthesis revealed that the Spk vesiclescontain different enzymatic activities. Microvesicles (chitosomes) at the core of the Spk contained chitin synthases CHS-1, -3, -4, -5 and -6, whereasmacrovesicles at the outer layer carried glucan synthase (GS). The coordinated action of coats, tethers, motors, Rabs and SNAREs allows the multiplevesicular carriers and their cargoes to traffic between organelles and to be delivered at their final destinations. While it remains to be elucidated whatregulates the spatial stratification of the Spk, our most recent analyses show the differential co-localization of Rab GTPases YPT-1 and SEC-4 with microand macrovesicles, respectively; suggesting different biogenesis for these vesicles. Prior to v-SNARE and t-SNARE recognition and fusion with the PM,secretory vesicles are presumably tethered to their target acceptor membrane in a process mediated by the exocyst, an octameric complex. In N. crassa anintact exocyst complex is required for formation of the Spk and the maintenance of regular hyphal growth. Two distinct localization patterns of the exocystsubunits were observed at the hyphal tip. EXO-70 and EXO-84 accumulated at the frontal part of the Spk external layer, coinciding partially with themacrovesicular layer, whereas the exocyst components SEC-3, 5, 6, 8 and 15 formed a delimited crescent at the apical PM. This suggests the formation oftwo distinct exocyst subcomplexes that may unite during vesicle tethering in post-Spk traffic steps. Collectively our results prove the direct involvement ofthe Spk in cell wall synthesis and confirm that the region of exocyst-mediated vesicle fusion in the hyphal apex coincides with the exocytotic gradientpredicted by the Vesicle Supply Center (VSC) model for fungal morphogenesis, with a maximum at the pole and vanishing gradually in the subapex.Evolution of sexual reproduction: A view from the Fungal Kingdom. Joseph Heitman. Department of Molecular Genetics and Microbiology, DukeUniversity, heitm001@duke.edu.Sex is nearly universal in eukaryotes, and thought to have evolved once. Sex promotes genetic diversity and evolution, yet also confers costs. Bothmechanisms of sex determination and mechanics of sexual reproduction are remarkably diverse. Fungi are exceptional models to analyze these processes,and their study reveals surprising insight into both sex and its impact. We focus on how mating-type identity is specified and modes and roles of sexualreproduction in generating diversity. Many fungi have bipolar sexual cycles with two opposite mating types and a bi-allelic mating type locus. In theBasidiomycota many species have a more complex tetrapolar sexual cycle with two unlinked multi-allelic mating type loci, resulting in thousands of matingtypes and enhanced outcrossing but restricted inbreeding. Our studies reveal how transitions from ancestral tetrapolar to derived bipolar systems haveoccurred in pathogenic species embedded within saprobic sibling taxa. The tetrapolar-bipolar transition has occurred repeatedly in pathogens of plantsand animals, suggesting it might be selected during host adaptation. Pathogenic Cryptococcus species have taken this transition further to a unipolarsexual cycle. These species have global largely unisexual populations and reproduce via an unusual homothallic unipolar sexual cycle involving only onemating type (same-sex mating, unisexual reproduction). Like a-a opposite sex mating, a-a unisexual mating can admix parental diversity in the progeny.However, in other cases solo a-a unisex involves selfing of identical genomes with no genetic diversity to exchange. Why organisms engage in selfingchallenges conventional views on the roles of sex. We find unisex generates genetic diversity de novo, preserving well-adapted genomic configurationswhile generating more limited genetic diversity for selection to act upon. Discovery that other fungi and eukaryotic parasite pathogens also reproduceunisexually generalizes these findings, and suggests unisex may have evolved because it mitigates costs of sex. Studies of fungal sex and its evolution andimpact illustrate general principles by which diversity is generated and maintained with implications for saprobic and pathogenic microbes andmulticellular eukaryotes.Metabolic compensation of the Neurospora clock by a glucose-dependent feedback of the circadian repressor CSP1 on the core oscillator. GencerSancar, Cigdem Sancar, Francois Cesbron, Michael Brunner. Dpet Biochemistry, Univ Heidelberg, Heidelberg, Germany.CSP1 is a global morning specific transcription repressor of Neurospora that modulates expression of about 800 genes. Expression of CSP1 is stimulatedby glucose and under circadian control of the white-collar complex (WCC). In csp1 mutant strains the circadian period length decreases with increasingglucose concentrations due to increased expression of WC1. In contrast, in wild-type strains the period is compensated for changes in glucoseconcentration and WC1 levels are independent of glucose. CSP1 contributes to metabolic compensation of the circadian clock by glucose-dependentrepression of wc1 transcription, which counterbalances the glucose-dependent translation efficiency of wc1 RNA. Forced over-expression of CSP1 reducesof WC1 expression and results in dampening of the circadian clock. Many target genes of CSP1 are rhythmically expressed with an evening specific phasewhile target genes of the WCC are morning specific.Integration of the fungal cell cycle with growth and development. Meera Govindaraghavan, Kuo-Fang Shen, Stephen Osmani. Dept Molec Gen, OhioState Univ, Columbus, OH.A universally important aspect of growth and development is the integration of mitosis with cell division. This helps ensure that cells maintain theirnormal size, shape and nuclear number, which in the fungi can vary considerably. For example, the highly polarized mode of growth of the filamentousfungi is subject to complex developmental regulation yielding diverse cell types containing from one to dozens of nuclei. How fungi integrate theregulation of the developmental axis involving mitosis, cytokinesis, and morphogenesis to maintain their defined cellular shapes, with distinctive numbers27th Fungal Genetics Conference | 25

PLENARY SESSION ABSTRACTSof nuclei, remains a mystery. However, recent studies of the mitotic NIMA kinase indicates it plays additional non-mitotic cytoplasmic functions inAspergillus nidulans that impinge on fungal development. These insights were derived initially from defining the interphase subcellular locations of NIMAwhich revealed it locates to both forming and mature septa and additionally locates to tips of growing interphase cells. Subsequent studies revealed thatseptal pores are subject to cell cycle regulation which prevents cytoplasmic movement between mitotic cell compartments and their adjacent interphasepartners. We further find that NIMA markedly affects the regulation of cell tip dominance and morphology via a mechanism involving NIMA location atmicrotubule +ends and the modulation of interphase cytoskeletal functions. Collectively the findings indicate that the mitotic NIMA kinase has roles toregulate communication between adjacent hyphal cells as well as cytoskeletal functions important for normal tip cell growth. Thus NIMA has the potentialto help integrate nuclear division with cell division and morphogenesis.A Neurospora cell-free system reconstitutes peroxisome membrane protein synthesis and organelle-specific targeting. Gregory Jedd. Temasek LifeSciences Laboratory, Singapore, Singapore.A central problem faced by eukaryotic cells is how to ensure that membrane proteins are localized to the appropriate organelle. Peroxisomes areubiquitous eukaryotic organelles that proliferate through growth and division, and can also arise de novo from endoplasmic reticulum (ER)-derivedprecursors. Two distinct views for the biogenesis of peroxisome membrane proteins (PMPs) are currently entertained. In the direct targeting model, PEX19recognizes PMPs in the cytosol and ferries them to the peroxisome where interaction with PEX3 prompts PMP release and membrane integration. In thesecond model, nascent PMPs are integrated to the ER membrane first, and then traffic to the peroxisome membrane. In this case, PEX19 functions as asorting receptor to package PMPs into ER-derived vesicles. My talk will focus on development of a cell-free system that reconstitutes PMP synthesis andtargeting to the peroxisome membrane. I will discuss how distinct chaperones and sequences associated with transmembrane domains distinguish directER and peroxisome targeting pathways.26

PLENARY SESSION ABSTRACTSSaturday, March 16 8:30 AM–12:00 NOONMerrill Hall and ChapelPlenary Session IV: Functional Ecology and Fungal CommunitiesChair: Jim AndersonMechanisms allowing the formation of new fungal pathogenic species on novel hosts, causing emerging diseases. Tatiana Giraud. ESE, Univ Paris 11,Orsay, France.We have studied different mechanisms allowing the formation of novel fungal pathogenic species on novel hosts. A theoretical model combined with aliterature survey have confirmed that mating within hosts in pathogenic ascomycetes may allow the rapid formation of new species by host adaptationalone, without requiring the evolution of mate choice. We also present studies on mechanisms of speciation in Microbotryum a fungal complex causingthe sterilizing anther-smut disease in many species of Caryophyllaceae. Multiple gene phylogenies and measures of hybrid inviability and sterility haverevealed the existence of multiple cryptic species, each being specific to one or a few plant host. We investigated the evolution of reproductive isolation inthe species complex, and showed that hybrid inviability and sterility increased with the genetic distance between the species. We show that selfing is themain barrier to gene flow between close species and that hybrid sterility arise because of karyotypic rearrangements. Cophylogeny analyses showed thatMicrobotryum species have evolved through frequent host shifts to moderately distant hosts. Current geographic distribution of host species seemed tobe of little relevance for understanding the putative historical host shifts, because most fungal clades had overlapping ranges. We did detect someecological similarities between host species that were diseased by closely related anther smut species, including pollinator or habitat similarities. However,genetics underlying the host-parasite interactions appeared to be the most important factor influencing host-shifts and specialization: multi-host speciesparasitized closely related plant species and related species in the Microbotryum phylogeny were associated with members of the same host clade. Weperformed a cross-inoculation experiment and showed that both host and pathogen phylogenies were indeed significant predictors of host range, with atleast partly independent effects. We investigated whether some Microbotryum species have arisen via hybridization.We also detected hybrids in natureand underwent a population genomic study to unravel the genomic architecture of introgression. Anther smut fungi appear as excellent models to unravelthe mechanisms of formation of new fungal species onto novel hosts.The decisive role of mycorrhizal fungi as regulators of carbon sequestration in boreal forest ecosystems. B.D. Lindahl, K.E. Clemmensen, I.T.M. Bödeker,E. Sterkenburg. Dept. Forest Mycol. & Path., SLU, Uppsala, Sweden.Boreal forest soils represent a significant global sink for carbon, but poor knowledge about the mechanisms that regulate the dynamics of soil carbonpools hampers the development of predictive ecosystem models. Such models are urgently needed to guide proper management of forest land, in orderto mitigate increasing atmospheric CO2 levels. By analysing the natural abundance of rare isotopes ( 14 C, 13 C and 15 N), we found that the mycelium ofmycorrhizal fungi represents a major source of soil carbon. Up to 50-70% of stored carbon was estimated to enter the soil via plant roots. Ratios betweenfungal biochemical markers (ergosterol and chitin) indicate that a rapid turnover of fungal mycelium minimizes carbon storage and favours efficientnitrogen recycling, whereas slow mycelial turnover favours carbon sequestration and immobilisation of nitrogen. In order to relate taxonomic andfunctional diversity to carbon dynamics, we used 454-sequencing of ITS2 amplicons to analyse fungal communities in environmental samples. In forestsoils with low nitrogen availability we found high abundance of ectomycorrhizal genera with differentiated extra-radical mycelium (cord formers). Memberof this group also correlated with high activities of classII peroxidases, known to facilitate break down of complex organic matter. Further evidence thatthese fungi act as “mycorrhizal white rotters” were obtained by amplification of peroxidase mRNA from soil extracts. The mRNA could be connected tomycorrhizal species by sequence homology. We propose that conditions of low nutrient availability favour the establishment of mycorrhizal species thatare adapted to minimize immobilisation of nitrogen in stable organic pools. As their own mycelium represents a major sink for soil-derived nutrients, thesespecies have to re-cycle their own biomass rapidly, in order to enable efficient delivery of nutrients to their host plants. They also possess potent oxidativeenzymes that may be used to increase mobility of organic nutrients. As a side-effect of their highly efficient nutrient recycling, presence of these fungi alsominimizes long term carbon sequestration in soils.Population Genomics of Saccharomyces Yeasts: Ecology and Adaptation. Edward J. Louis. Ctr Genetics and Genomics, Univ Nottingham, Nottingham,United Kingdom.The budding yeast, Saccharomyces cerevisiae, along with its close relatives, have only recently become reasonable models for the study of populationgenetics and evolution. This has been due to the lack of understanding of their natural history and ecology. Now that some understanding of budding yeastin nature is in hand, we can apply the powerful genetic and molecular tools available to questions of evolution through adaptation to ecological niches.The combination of population genomics and quantitative trait analysis has led to some understanding of the genetic architecture underlying traits whichmay be relevant to adaptation to particular environments in different yeast populations/species.27th Fungal Genetics Conference | 27

PLENARY SESSION ABSTRACTSThe mycorrhizal symbiosis as a network linking plants. Marc-André Selosse. Centre d'Ecologie Fonctionelle et Evolutive, Montpellier, France.Most plants form mycorrhizal symbioses with soil fungi, which turn out to form networks between plants. Indeed, fungal individuals are large enough tocolonize several root systems, and most mycorrhizal fungi are host-generalists that can link different plant species. The most dramatic evidence for suchnetworks is the repeated emergence of mycoheterotrophy (MH) in plant evolution. MH plants are achlorophyllous and receive carbon (C) fromsurrounding green plants by way of shared mycorrhizal fungi. They recently made strong achievements due to two tools: molecular barcoding identifiedthe fungi, and natural isotopic abundances ( 13 C, 15 N) confirmed which fungal mycorrhizal guild provides C. Temperate MHs belong to orchids andMonotropoideae (an Ericaceae subfamily) associate often with high specificity to basidiomycetes that form themselves ectomycorrhizae with surroundingtrees. Intermediate evolutionary steps exist, in green orchids and Montropoideae that use C from mycorrhizal fungi in addition to their photosynthesis.This so-called mixotrophic nutrition depends on ectomycorrhizal fungi, and thus also on mycorrhizal networks. Phylogenies support that mixotrophypredisposed to evolution of MHs. In some green mixotrophic orchids, the survival of rare achlorophyllous variants further supports MH abilities, but thelow fitness of these variants suggests that mixotrophy can be evolutionarily metastable. By contrast, tropical MH plants belong to diverse families, displaylower specificities, and often associate to arbuscular mycorrhizal (AM) fungi. The isotopic fractionation for 15 N and 13 C along the [green plant-AM fungi-MHplants] continuum shows differences as compared to the same continuum for temperate MHs, which associate with ectomycorrhizal basidiomycetes. Thissupports different exchange mechanisms. Moreover, MHs associated to basidiomycetes have higher total N and 15 N content than autotrophic plants, whileAM-associated MHs do not. I hypothesize that AM-associated MHs evolved mainly to support C nutrition, under selection of shaded, N-rich tropicalforests. Conversely, basidiomycetes-associated MHs may have been first selected for N acquisition in N-limited, but less shaded, temperate forests. Thus,the convergent exploitation of mycorrhizal networks may result from different evolutionary pathways that depend on the biome.Unraveling speciation and specialization processes in plant pathogenic fungi using comparative population genomics. Eva H Stukenbrock 1 , Freddy BChristiansen 2 , Julien Y Dutheil 1 , Bruce A McDonald 3 , Thomas Bataillon 2 , Mikkel H. Schierup 2 . 1) Fungal Biodiversity, Max Planck Institute for TerrestrialMicrobiology, Marburg, Germany; 2) Bioinformatics Research Center, Aarhus Univeristy, C.F. Moellers Alle 1110, 8000 Aarhus, Denmark; 3) PlantPathology, ETH Zurich, LFW B16, 8092 Zurich, Switzerland.The emergence of new fungal pathogens in managed ecosystems is an urgent matter of consideration. Studies have documented that naturalecosystems serve as reservoirs and sources of new crop infecting pathogens. We know, however, very little about the ecology and diversity of fungalpathogens in natural ecosystems. A goal of our research is to infer diversification and speciation processes of plant pathogens in natural and managedecosystems. We study a species complex of plant pathogenic fungi including the wheat pathogen Zymoseptoria tritici (synonym Mycosphaerellagraminicola). Speciation of Z. tritici was associated with wheat domestication and dates back to 10-12000 ya. Several closely related species of Z. triticiexist in natural grasslands in Iran. These wild grass pathogens co-exist and have over-lapping host ranges. In spite of their close relatedness, ecological orspatial factors have allowed speciation to occur. We have taken a comparative population genomics approach to study the underlying evolutionaryprocesses that drive Zymoseptoria diversification. Our study includes full genome sequences from 52 fungal isolates representing four Zymoseptoriaspecies. We perform population genomics analyses and document recent speciation times in the Zymoseptoria complex and present day small effectivepopulation sizes. Using within and between species rates of non-synonymous and synonymous variation we show a strong impact of natural selection ingenome evolution of Zymoseptoria spp. This is at odds with the small effective population sizes estimated and suggests that population sizes werehistorically large but unstable. A significant finding is that speciation of Z. tritici did not entail an apparent loss of variation in spite of the homogenousagro-ecosystem where it has evolved. In contrast, we observe a dramatic loss of variation in the closest wild relative, Z. pseudotritici. The mosaic genomepatterns in Z. pseudotritici are consistent with a very recent hybrid speciation event resulting from a cross between two divergent haploid individuals. Weestimate that the hybridization occurred ~500 sexual generations ago between closely related, but isolated species. Based on the comparative populationgenomic analyses we reveal rapid evolution and distinct patterns of species evolution in natural and managed ecosystems.28

CONCURRENT SESSION ABSTRACTSWednesday, March 13 3:00 PM–6:00 PMMerrill HallCell Signaling Involved in Fungal Development and PathogenesisCo-chairs: Naweed Naqvi and Stefanie PöggelerStability of a G protein alpha subunit in genetic backgrounds lacking the G beta subunit or a cytosolic guanine nucleotide exchange factor. Alexander V.Michkov, Katherine A. Borkovich. Plant Pathology and Microbiology, University of California, Riverside, Riverside, CA.Heterotrimeric G proteins consist of alpha, beta and gamma subunits. Regulation is accomplished through the alternation between binding of GDP(inactive form) and GTP (active form) by the alpha subunit and dissociation of the alpha subunit and beta-gamma dimer. GDP/GTP exchange is facilitatedby both cell surface G protein coupled receptors and cytosolic guanine nucleotide exchange factors (GEFs), such as RIC8. Neurospora crassa has three Galpha subunits (GNA-1, GNA-2 and GNA-3), one G beta (GNB-1), and one G gamma (GNG-1). Interestingly, mutants lacking gnb-1 or the cytosolic GEF ric8exhibit some defects in common with the gna-1 deletion mutant, which may be explained by the reduced GNA-1 protein levels observed in these mutants.Previous studies in our laboratory showed that levels of gna-1 mRNA are similar in wild type and mutants lacking gnb-1 or ric8, consistent with a posttranscriptionalmechanism. Using genetic and biochemical approaches, this study investigated the mechanism underlying regulation of GNA-1 stability inregards to GTP/GDP bound state and amount of protein (normal or overexpressed). The results demonstrate that levels of GNA-1 protein are not visiblyreduced over 36 hours in a wild-type background after halting translation using cycloheximide, suggesting GNA-1 is very stable in wild type. To checkstability of GDP or GTP bound GNA-1 in different backgrounds, we transformed mutants lacking the gna-1 gene and gnb-1 or ric8 with a wild type (gna-1 WT ) or constitutively active, GTPase-deficient gna-1 allele (gna-1 Q204L ). Overexpressing gna-1 WT (GDP bound) in a wild-type background increased the levelof GNA-1 protein ~ 3 fold, while overexpression in a gnb-1 mutant gave a nominal increase (~ 1.6x). Overexpressing gna-1 Q204L (GTP bound) in the Dgnb-1or Dric8 backgrounds led to ~ 2 fold higher levels of GNA-1 compared to wild type. In summary, GNA-1 is very stable in wild type, but stability decreasesdramatically in gnb-1 and ric8 deletion mutants. The GTP-bound G alpha protein is more stable in a gnb-1 mutant background than GDP-bound GNA-1protein.The Putative Guanine Nucleotide Exchange Factor RicA Mediates Upstream Signaling for Growth and Development in Aspergillus. Nak-Jung Kwon 1 , HeeSoo Park 2 , Seunho Jung 3 , Sun Chang Kim 4 , Jae-Hyuk Yu 1,2 . 1) Dept Bacteriology, University of Wisconsin, Madison, WI. USA; 2) Molecular and EnvironmentalToxicology Center, University of Wisconsin, Madison, WI, USA,; 3) Department of Bioscience and Biotechnology, and Center for Biotechnology Research inUBITA, Konkuk University, Seoul, Republic of Korea; 4) Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Dae-Jon,Republic of Korea.Heterotrimeric G proteins (G proteins) govern growth, development, and secondary metabolism in various fungi. Here, we characterized ricA, whichencodes a putative GDP/GTP exchange factor for G proteins in the model fungus Aspergillus nidulans and the opportunistic human pathogen Aspergillusfumigatus. In both species, ricA mRNA accumulates during vegetative growth and early developmental phases, but it is not present in spores. The deletionof ricA results in severely impaired colony growth and the total (for A. nidulans) or near (for A. fumigatus) absence of asexual sporulation (conidiation). Theoverexpression (OE) of the A. fumigatus ricA gene (AfricA) restores growth and conidiation in the DAnricA mutant to some extent, indicating partialconservation of RicA function in Aspergillus. A series of double mutant analyses revealed that the removal of RgsA (an RGS protein of the GanB Gasubunit), but not sfgA, flbA, rgsB, or rgsC, restored vegetative growth and conidiation in AnricA. Furthermore, we found that RicA can physically interactwith GanB in yeast and in vitro. Moreover, the presence of two copies or OE of pkaA suppresses the profound defects caused by DAnricA, indicating thatRicA-mediated growth and developmental signaling is primarily through GanB and PkaA in A. nidulans. Despite the lack of conidiation, brlA and vosAmRNAs accumulated to normal levels in the ricA mutant. In addition, mutants overexpressing fluG or brlA (OEfluG or OEbrlA) failed to restore developmentin the AnricA mutant. These findings suggest that the commencement of asexual development requires unknown RicA-mediated signaling input in A.nidulans.The Aspergillus nidulans MAPK module AnSte11-Ste50-Ste7-Fus3 controls development and secondary metabolism. Oezguer Bayram 1* , Oezlem SarikayaBayram 1 , Yasar Luqman Ahmed 2 , Jun-Ichi Maruyama 1,4 , Oliver Valerius 1 , Silvio Rizzoli 3 , Ralf Ficner 2 , Stefan Irniger 1 , Gerhard Braus 1 . 1) Institute ofMicrobiology & Genetics, Department of Molecular Microbiology and Genetics, Georg-August-Universität, Grisebachstr. 8, D 37077 Goettingen, Germany;2) Department of Molecular Structural Biology, Institute for Microbiology and Genetics, Georg-August-Universität, Goettingen; 3) European NeuroscienceInstitute, Deutsche Forschungsgemeinschaft Center for Molecular Physiology of the Brain/Excellence Cluster 171, 37077 Göttingen; 4) Department ofBiotechnology, The University of Tokyo, Tokyo, Japan.The sexual Fus3 MAP kinase module of yeast is highly conserved in eukaryotes and transmits external signals from the plasma membrane to the nucleus.We show here that the module of the filamentous fungus Aspergillus nidulans (An) consists of the AnFus3 MAP kinase, the upstream kinases AnSte7 andAnSte11, and the AnSte50 adaptor. The fungal MAPK module controls the coordination of fungal development and secondary metabolite production. Itlacks the membrane docking yeast Ste5 scaffold homolog but similar to yeast the entire MAPK module interacts with each other at the plasma membrane.AnFus3 is the only subunit with the potential to enter the nucleus from the nuclear envelope. AnFus3 interacts with the conserved nuclear transcriptionfactor AnSte12 to initiate sexual development and phosphorylates VeA which is a major regulatory protein required for sexual development andcoordinated secondary metabolite production. Our data suggest that not only Fus3 but even the entire MAPK module complex of four physicallyinteracting proteins can migrate from plasma membrane to nuclear envelope.27th Fungal Genetics Conference | 29

CONCURRENT SESSION ABSTRACTSThe developmental PRO40/SOFT protein participates in signaling via the MIK1/MEK1/MAK1 module in Sordaria macrospora. Ines Teichert 1 , EvaSteffens 1 , Nicole Schnab 1 , Benjamin Fränzel 2 , Christoph Krisp 2 , Dirk A. Wolters 2 , Ulrich Kück 1 . 1) General & Molecular Botany, Ruhr University Bochum,Bochum, Germany; 2) Analytical Chemistry, Ruhr University Bochum, Bochum, Germany.Filamentous fungi are able to differentiate multicellular structures like conidiophores and fruiting bodies. Using the homothallic ascomycete Sordariamacrospora as a model system, we have identified a number of developmental proteins essential for perithecium formation. One is PRO40 [1], thehomolog of Neurospora crassa SOFT, and this protein was employed for protein-protein interaction studies to gain insights into its molecular function.Data from yeast two hybrid experiments with PRO40 as bait show an interaction of PRO40 with the MAP kinase kinase (MAPKK) MEK1. MEK1 is a memberof the cell wall integrity (CWI) pathway, one of three MAP kinase modules present in S. macrospora. The S. macrospora CWI pathway consists of MAPkinase kinase kinase (MAPKKK) MIK1, MAPKK MEK1 and MAP kinase (MAPK) MAK1, with additional upstream components, protein kinase C (PKC1) andRHO GTPase RHO1. Data from tandem affinity purification - MS experiments with PRO40 and MEK1 as bait indicate that PRO40 forms a complex withcomponents of the CWI pathway. Analysis of single and double knockout mutants shows that PRO40, MIK1, MEK1 and MAK1 are involved in the transitionfrom protoperithecia to perithecia, hyphal fusion, vegetative growth, and cell wall stress response. Differential phosphorylation of MAPKs in a pro40knockout strain was detected by Western analysis. We propose that PRO40 modulates signaling through the CWI module in a development-dependentmanner. Further interaction studies and complementation analyses with PRO40 derivatives provide mechanistic insight into the function of PRO40domains during fungal development. [1] Engh et al. (2007) Eukaryot Cell 6:831-843.A Fungal Adhesin Guides Community Behaviors by Autoinduction and Paracrinal Signaling. Linqi Wang, Xunyun Tian, Rachana Gyawali, Xiaorong Lin.Biology, Texas A&M University, College Station, TX.Microbes live mostly in a social community rather than in a planktonic state. Such communities have complex spatiotemporal patterns that requireintercellular communication to coordinate gene expression. Here, we demonstrate that Cryptococcus neoformans, a model eukaryotic pathogen, respondsto an extracellular signal in constructing its colony morphology. The signal that directs this community behavior is not a molecule of low molecular weightlike pheromones or quorum sensing molecules, but a secreted protein. We successfully identified this protein as the conserved adhesin Cfl1 in theextracellular matrix. The released Cfl1 acts as an auto-induction signal to stimulate neighboring cells to phenocopy Cfl1-expressing cells. We propose thatsuch adhesin/matrix-initiated communication system exists in divergent microbes and our work represents the first adhesin/matrix-mediated signalingmechanism in simple eukaryotes.Surface recognition and appressorium morphogenesis in Magnaporthe oryzae. JinRong Xu. Dept Botany & Plant Pathology, Purdue Univ, West Lafayette,IN 47906.Appressorium formation and penetration play critical roles in plant infection in the rice blast fungus and other foliar pathogens. In Magnaporthe oryzae,the cAMP signaling and Pmk1 MAP kinase pathways are known to regulate surface recognition, appressorium formation, penetration, and invasive growth.Like other filamentous ascomycetes, M. oryzae contains two genes encoding catalytic subunits of PKA. Whereas the cpkA mutant was delayed inappressorium formation and reduced in virulence, the cpk2 mutant had no detectable phenotypes except a slight reduction in conidiation. However, thecpkA cpk2 double mutant recently identified in our lab had distinct defects in growth, conidiation, appressorium formation, and plant infection. Detailedcharacterization of its phenotype is under the way and will be helpful to better understand the relationship between the cAMP signaling and Pmk1pathways. For upstream signals, MSB2 functions as a surface sensor upstream from the Pmk1 pathway for regulating appressorium formation andpenetration. In addition to its mucin and transmembrane domains, the cleavage domain and C-terminal cytoplasmic tail are important for Msb2 functions.Results from experiments aiming to determine cleavage and intracellular signaling of Msb2 will be presented. We also have used the msb2 or msb2 sho1mutant to generate the double or triple mutants with CBP1 and PTH11, two other putative surface sensor genes. Phenotype characterization of thesemutants will be used to determine the functional relationship of MSB2 with other surface sensors invovled in appressorium morphogenesis.Plant cues promote stealth infection in fungal plant pathogens. Marie Nishimura. Plant-Microb Interact Unit, Natl Inst Agrobiol Sciences, Ibaraki, Japan.Fungal cell wall, mainly composed of polysaccharides, is a major source of microbe-associated molecular patterns (MAMPs) which are recognized by hostinnate immune receptors. Although recognition of the fungal cell wall MAMPs, such as chitin oligomers, activates defense responses in plants, fungal plantpathogens invade the hosts likely by evading the host innate immunity. We have found that the ascomycete rice pathogen Magnaporthe oryzaeaccumulates a-1,3-glucan on the cell wall during infection. The accumulation of a-1,3-glucan was dependent on the cell wall integrity MAP kinase (Mps1)pathway, which was activated by a plant wax component. a-1,3-glucan was not essential for formation of infectious structures but was required for thesuccessful infection by protecting the cells from plants’ antifungal enzymes and by delaying the host defense responses. Furthermore, histocytochemicalobservation have revealed that the ascomycete Cochlioborus miyabeanus and the basidiomycete Rhizoctonia solani have also accumulate a-1,3-glucan onthe cell walls specifically during plant invasion. Thus, plant cues appear to trigger surface accumulation of a-1,3-glucan in these fungi. In addition, riceplants secreting bacterial a-1,3-glucanase rapidly induced defense responses against these pathogens and showed multiple fungal disease resistance.Considering that a-1,3-glucan is non-degradable in many plants, our study suggested that masking cell wall surfaces with a-1,3-glucan is a stealth infectionstrategy commonly used by fungal plant pathogens. Our study also indicated that recognition of plant cues play an important role in promoting stealthinfection in fungal plant pathogens.30

CONCURRENT SESSION ABSTRACTSUnravelling the GTPase polarity complex in Claviceps purpurea. Andrea Herrmann 1 , Janine Schürmann 1 , Britta Tillmann 2 , Michael Bölker 2 , Paul Tudzynski 1 .1) IBBP, WWU Muenster, Schlossplatz 8, 48143 Muenster, Germany; 2) Philipps-Universität, Karl-von-Frisch-Strasse 8, 35032 Marburg, Germany.Claviceps purpurea is a plant pathogen infamous for its production of toxic alkaloids on infected host plants like barley. Consumption of infected grainsleads to severe symptoms up to the death of the patient. Infection patterns are complex and the topic of intensive research. One interesting aspect is thestrict polarity of the hyphal growth during the first infection stage which seems to be crucial for the non-recognition of C. purpurea as a pathogen by thehost. To address the question of the importance of polarity the structure and dynamics of the polarity complex are the focus of this work. The guaninenucleotide exchange factors (GEFs) Cdc24 and Dock180 belong to different families, Cdc24 being a member of the Dbl GEF family and Dock180 a CZH GEF.Cdc24-GFP localises cytosolically and to hyphal tips whereas Dock180-GFP is present in small vesicles in the hypha, though concentrated at the tip region,too. Cdc24 DHPH domains are able to activate the small GTPases Rac and Cdc42 of C. purpurea and U. maydis In vitro, whereas the catalytic domain ofDock180 only activates Rac in both organisms. Despite the proven activation Cdc24 does not interact with any GTPase in yeast two hybrid assays. Dock180shows a weak interaction with Rac and the two p21-activated kinases (PAKs) Ste20 and Cla4. Thus, both GEFs do not share many characteristics apart fromtheir GEF activity. The PAKs Ste20 and Cla4 and the scaffold protein Bem1 are involved in the polarity complex, too. Ste20 localises to hyphal tips andinteracts with Cdc42 in a loading status dependent manner, whereas Cla4 is the main partner of Rac. Other interactions of Ste20 with Dock180 and Cla4could also be shown. Bem1 is present in the cytosol - concentrated at the hyphal tip - and links most of the proteins of the polarity complex as interactionswith Cdc24, Cla4, Ste20 and Dock180 have been detected. Taken together we postulate at least two different polarity complexes, the Rac complex and theCdc42 complex. Both are gathered by Bem1, but Cla4 is the main partner of Rac, whereas Ste20 plays a similar role for Cdc42. Dock180 is mainly linked toRac, Cdc24 can be active in both complexes. We are interested in the spatial and temporal formation and regulation of these complexes and its influenceon polarity and virulence which will be the subject of further studies.27th Fungal Genetics Conference | 31

CONCURRENT SESSION ABSTRACTSWednesday, March 13 3:00 PM–6:00 PMChapelGenetics and Genomics of Interactions with Bacteria, Insects and PlantsCo-chairs: Nemat Keyhani and Christian HertweckEndophytic insect parasitic fungi feed insect-derived nitrogen to plants. M.J. Bidochka, S.W. Behie. Brock University, St. Catharines, ON, Canada.Metarhizium is a fungus with a bifunctional lifestyle: it is a common plant endophyte and is also a pathogen to a large number of insects, which are asource of nitrogen. It is possible that the endophytic capability and insect pathogenicity of Metarhizium are coupled to provide an active method ofnitrogen transfer to plant hosts via fungal mycelia. We used soil microcosms to show the ability of M. robertsii to translocate insect-derived nitrogen toplants. Insects were injected with 15N-labeled nitrogen, and we tracked the incorporation of 15N into amino acids in two plant species. We alsoinvestigated the exchange of plant carbon for insect-derived nitrogen in this symbiosis. Unlike mycorrhizal fungi, Metarhizium, is not a fastidious fungusand is easily genetically manipulated. We also performed gene knockout experiments on carbon and nitrogen transporters to test if the exchange ofcarbon and nitrogen is reciprocal in this fungus-plant symbiosis.Genotype-Environment Interactions and the Interplay Between Climate Change and Plant-Fungal Symbioses. Rusty J. Rodriguez 1,2,3 , Yong Ok Kim 3 , ClaireWoodward 3 , Leesa Wright 2 , Regna Redman 2,3 . 1) Symbiogenics, Seattle, WA; 2) Adaptive Symbiotic Technologies, Seattle, WA; 3) University of Washington,Biology, Seattle, WA.Symbiotic associations span a continuum from parasitism to mutualism and the outcome of specific associations is context driven based on intergenomicinteractions and environmental factors. These factors will determine the ability plants and animals to adapt to a changing climate. For example,plants in natural ecosystems adapt to abiotic stress by forming symbiotic associations with fungal endophytes that confer stress tolerance. Without theendophytes, the plants are not stress tolerant and do not survive in the habitats to which they are adapted. Symbiotically conferred stress tolerancetypically occurs in a habitat-specific manner, a phenomenon we designate Habitat Adapted Symbiosis (HAS). Although several biochemical processes havebeen correlated to plant stress tolerance, few processes correlate with symbiotically conferred stress tolerance. Symbiotically conferred stress toleranceinvolves altered plant gene regulation, increased metabolic efficiency, and an increased ability to manage reactive oxygen species. I will describe howfungal endophytes adapt plants across environmental gradients and present a working model for symbiotically conferred stress tolerance.Chemical mediators of pathogenic and mutualistic bacterial-fungal interactions. Kirstin Scherlach. Biomolecular Chemistry, Leibniz Institute for NaturalProduct Research and Infection Biology, HKI Beutenbergstr. 11a 07745 Jena, Germany.Natural products play a key role in symbiotic interactions between microorganisms and higher organisms. Their function may range from signalingcompounds in mutualism to virulence factors and toxins in pathogenic relationships. In many cases the chemical basis of such interactions is still unknown.By combining genome mining, bioinformatic analyses and chemical analytical techniques we uncovered the biosynthesis of a number of virulence factorsand toxins from bacterial-fungal associations that impact agriculture, medicine and biotechnology. We elucidated an unprecedented case of symbioticcooperation where a fungus tailors a pathogenicity factor provided by its endosymbiotic bacteria to increase its phytotoxic potency. This toxin (rhizoxin)causes rice seedling blight, a plant disease accounting for severe losses in agriculture. Another bacterium, Janthinobacterium agaricidamnosum, initiatessoft rot of cultured mushrooms. With the help of imaging mass spectrometry we discovered a peptide toxin that not only contributes to pathogenicity butalso displays potent antifungal activity against major human pathogens. Furthermore, we identified the molecular basis for the biosynthesis of a highlytoxic polyketide produced by Burkholderia gladioli, a common contaminant of the food fermentation fungus Rhizopus oligosporus. Bongkrekic acid is arespiratory toxin that efficiently inhibits adenine nucleotide translocase. Through sequencing of the bacterial genome and functional analyses of thebiosynthetic genes new insights into the process of polyketide assembly were gained. The discovery of these secondary metabolites as mediators ofbacterial-fungal interaction and their biogenetic origins not only facilitates the understanding of complex ecological processes but also opens avenues tothe development of new drug candidates and potential bio-control agents against crop diseases.Comparative genomic analysis of entomopathogenic fungi. Guohua Xiao, Qiang Gao, Peng Zheng, Xiao Hu, Chengshu Wang. Institute of Plant Physiologyand Ecology, Shanghai Institutes for Biological Sciences, CAS, Shanghai, China.There are about one thousand of fungal species capable of infecting and killing insects, spiders and mites. The species like Metarhizium anisopliae, M.robertsii, M. acridum and Beauveria bassiana has been developed as environmentally friendly biological control agents against different insect pests. Thecaterpillar pupae specific fungus Cordyceps militaris, however, has been used a traditional Chinese Medicine for hundreds of years. We sequenced thegenome of model entomopathogenic fungi Metarhizium spp., B. bassiana and C. militaris and conducted comparative genomic studies. We found that theinsect pathogens have a strikingly larger proportion of genes encoding secreted proteins, particularly proteases, than other sequenced fungi. Aphylogenomic analysis confirmed that fungal entomopathogenicity has evolved multiple times so similar expansion of families of proteases and chitinasesreflects a convergent evolution. Like that of C. militaris which can readily performs a sexual life, a single mating type locus identified in clonallyreproductive species Metarhizium and Beauveria indicated that the later are sexually heterothallic. High throughput transcriptome analysis indicated thatthe pathogens could regulate transcriptional responses with fine-tuned gene sets for host recognition, development and adaptation to different niches.The information from our studies advanced the understanding of the evolution of fungal entomopathogenicity and will benefit future molecular studies offungus-host interactions and thereby facilitate the development of cost-effective mycoinsecticides.32

CONCURRENT SESSION ABSTRACTSSynergistic interactions between leaf-cutting ants and their fungal symbiont facilitate degradation of plant substrate. Morten Schiøtt 1 , Henrik H. de FineLicht 2 , Adelina Rogowska-Wrzesinska 3 , Pepijn Kooij 1 , Peter Roepstorff 3 , Jacobus J. Boomsma 1 . 1) Department of Biology, University of Copenhagen,Copenhagen, Denmark; 2) Department of Mycology, Lund University, Lund, Sweden; 3) Department of Biochemistry and Molecular Biology, University ofSouthern Denmark, Odense, Denmark.About 50 million years ago a single ancestor of today’s more than 230 species of fungus-growing ants committed herself irreversibly to farming fungi forfood instead of being a hunter-gatherer as most other ants. However, the white-rot litter decomposing Leucocoprini (Agaricales) that were domesticatedremained mostly uncommitted to the symbiosis until a single lineage became an obligate symbiont of a derived clade of these ants - the so called higherattines. Coevolution of ants and fungi has subsequently produced specific adaptations in both partners, including the development of special hyphal tips(gongylidia) of the fungus on which the ants feed. Recent work has shown that many fungal enzymes pass through the ant digestive system unharmed tobe mixed (as fecal fluid) with the fresh leaf pulp that the ants deposit on top of their gardens. To understand the function of this form of fungal enzymetransfer, we have used state of the art proteomics and high-throughput genome sequencing to identify the proteins found in the ant fecal fluid. Fecalproteins of Acromyrmex leafcutter ants were separated with SDS-PAGE followed by tandem mass spectrometry, and the resulting peptide tags were usedas queries to Blast-search a low coverage genome sequence of the fungal symbiont. Using this strategy we identified 34 protein sequences encoded by thefungal genome. Enzyme assays for selected fecal proteins showed that they functionally disappeared from the fecal droplets when the ants were deprivedof their fungal symbiont. We further used qPCR to establish that many of these proteins are more highly expressed in gongylidia than in mycelium,suggesting that they have been actively selected to be ingested by the ants. A substantial fraction of the fecal proteins are enzymes that are widely used byplant-pathogens to break down cell walls to access the easily degradable nutrients inside living cells. Of special interest is the finding of a polyphenoloxidizinglaccase enzyme that shows signs of positive selection in the higher attine ant symbionts, and may be an important prerequisite for the ability tocope with the polyphenols present in plant tissues. The results indicate that the leafcutter ants and their fungal symbionts have evolved traits-syndromesthat are partially convergent with those found in plant-pathogenic fungi.Unraveling the metabolome: how zombie ant fungi heterogeneously control ant brains. Charissa de Bekker, David Hughes. Biology and Entomology,Center for Infectious Disease Dynamics,Pennsylvania State University, State College, PA.Fungal entomopathogens rely on cellular heterogeneity during the different stages of insect host infection. Their pathogenicity is exhibited through thesecretion of secondary metabolites. Infection strategies of this group of environmentally important fungi can thus be studied by analyzing theirmetabolome. Next to generalists such as Beauveria bassiana and Metarhizium anisopliae, specialist species exist that are able to control host behavior.One of the most dramatic examples is the death grip of ants infected by Ophiocordyceps unilateralis, where ants are being used as a vehicle and finally biteinto vegetation before dying, aiding fungal spore dispersal after death. To establish this the fungus must not only overcome the immune system of thehost, but also manipulate the brain and atrophy the muscles. To date, most work on manipulation of host behavior has described the ant’s behavior,leaving the molecular processes from the fungal point of view unresolved. To start unraveling the mechanisms underlying this phenomenon we arecombining metabolite profiling with an ex vivo insect tissue culturing system that allows us to study fungal metabolites secreted in different parts withinthe host. Using this technique we established that B. bassiana and M. anisopliae, and O. unilateralis heterogeneously react to brain and muscle tissue bysecreting a significantly different array of metabolites. The combination of these approaches with a concrete understanding of the host-parasiteinteraction in nature is allowing us to understand both the diversity of secondary metabolites as well as make discoveries regarding the temporal dynamicsthese fungi employ when releasing metabolites that affect the host. This project is financed by the Marie Curie International Outgoing Fellowships andPenn State University .Trichoderma rhizosphere’s competency, endophytism and plant communication: A molecular approach. Artemio Mendoza 1 , Johanna Steyaert 1 , NataliaGuazzone 1 , Maria Fernanda Nieto-Jacobo 1 , Mark Braithwaite 1 , Robert Lawry 1 , Damian Bienkowski 1 , Christopher Brown 2 , Kirstin MacLean 1 , Robert Hill 1 ,Alison Stewart 1 . 1) Bio-Protection Research Centre, Lincoln University, Lincoln, New Zealand; 2) Biochemistry Department and Genetics Otago, Universityof Otago, New Zealand.Establishment of root symbiosis is one the key drivers of biocontrol success for members of the fungal genus Trichoderma. This root symbiosis isdescribed as a two-step process, whereby Trichoderma species colonise the soil surrounding the root (rhizosphere) and then penetrate the root tissue andestablish an endophytic relationship. The ability to colonise and then proliferate over time within the rhizosphere is termed rhizosphere competence (RC).There have been numerous reports of Trichoderma biocontrol strains which persist within the rhizosphere for the growing season of the crop plant. Ourresults strongly suggest that RC is widespread among members of the genus Trichoderma and that RC interactions are strain and host plant specific. Forendophytes and their host plants to maintain a mutualistic relationship requires a constant molecular dialogue between the organisms involved. Forexample, the fungal-derived phytohormone, indole acetic acid (IAA), plays an important role in signalling between Trichoderma and the model plantArabidopsis thaliana. There are however, additional, currently unknown, chemical signals which may be even more important for a positive interactionbetween Trichoderma and plants. By using a soil-maize-Trichoderma as a model system in in situ sterile conditions we are currently analysing the RC andendophytism transcriptomes of two Trichoderma species: T. virens and T. atroviride. Using a combination of bioinformatics, quantitative RT-PCR (for stagespecific genetic markers from Trichoderma) and fluoro-labelled Trichoderma strains we are currently identifying and analysing promising Trichodermacandidates involved in endophytism and RC. A comprehensive panorama of the Trichoderma-soil-plant interaction will be discussed in this conference.27th Fungal Genetics Conference | 33

CONCURRENT SESSION ABSTRACTSEffector proteins in fungal defense against fungivorous nematodes: Targets and functional significance. Therese Wohlschlager 1 , Stefanie Schmieder 1 ,Alex Butschi 2 , Paola Grassi 3 , Alexander Titz 4 , Stuart Haslam 3 , Michael Hengartner 2 , Markus Aebi 1 , Markus Künzler 1 . 1) Institute of Microbiology, ETH Zürich,Switzerland; 2) Institute of Molecular Life Sciences, University of Zürich, Switzerland; 3) Division of Molecular Biosciences, Imperial College, London, UnitedKingdom; 4) Department of Chemistry, University of Konstanz, Germany.The defense of fungi against fungivores is largely based on the production of intracellular toxins. A significant proportion of these toxins are peptides andproteins that are synthesized by the ribosome and stored in the cytoplasm. Protein toxins include lectins that target specific glycoepitopes in the intestineof the fungivore upon ingestion and kill the fungivore by a yet unknown mechanism. In our laboratory, we focus on the functional characterization offungal protein toxins that are directed against nematodes. We use the model nematode Caenorhabditis elegans to identify the targets and to study thetoxicity mechanism of these fungal defense effector proteins in the nematode. In addition, we employ the fungivorous nematodes Aphelenchus avenaeand Bursaphelenchus willibaldi to study the diversity, the functional significance and the transcriptional regulation of these proteins in the fungus.Recently, we identified a nematotoxic lectin from the mushroom Laccaria bicolor that is homologous to animal lectins involved in innate immunity againstbacteria. We found that the nematotoxicity of the lectin is based on its specific binding to methylated fucose residues on nematode N-glycans. Amonganimals, this epitope is only present in worms and molluscs but not in insects or vertebrates. We performed affinity chromatography of C. elegans wholeworm protein extracts using the L. bicolor lectin and other nematotoxic fungal lectins recognizing protein-bound glycans. The results of this analysissuggest that these lectins target the same set of glycoproteins in the nematode intestine and may confer toxicity by a common mechanism. In order toaddress the functional significance of these proteins for fungal defense against fungivorous nematodes, we expressed some of the fungal proteinsdisplaying toxicity towards C. elegans, in the filamentous ascomycete Ashbya gossypii. These transformants were fed to A. avenae and the propagation ofthe fungivorous nematode on the various transformants was determined. Expression of some effector proteins significantly inhibited propagation of thenematode suggesting that these proteins have a role in fungal defense against these organisms. Experiments addressing the relative fitness of the variousA. gossypii transformants upon selective pressure of feeding by A. avenae are under way.34

CONCURRENT SESSION ABSTRACTSWednesday, March 13 3:00 PM–6:00 PMHeatherMembrane Trafficking and Molecular OrganizationCo-chairs: Vicky Sophianopoulou and Gero SteinbergDistinct secretion systems operate during biotrophic invasion by the rice blast fungus, Magnaporthe oryzae. Barbara Valent 1 , Martha Giraldo 1 , ChangHyun Khang 1,4 , Yasin Dagdas 2 , Yogesh Gupta 2 , Thomas Mentlak 2,5 , Mihwa Yi 1 , Melinda Dalby 1 , Hiromasa Saitoh 3 , Ryohei Terauchi 3 , Nicholas Talbot 2 . 1) DeptPlant Pathology, Kansas State Univ, Manhattan, KS; 2) School of Biosciences, Univ of Exeter, Exeter, U.K; 3) Iwate Biotechnology Research Center, Kitakami,Iwate, Japan; 4) Current Address: Dept of Plant Biology, Univ of Georgia, Athens, GA; 5) Current Address: Cambridge Consultants Ltd, Cambridge, U.K.During biotrophic invasion, Magnaporthe oryzae secretes cytoplasmic effectors, which preferentially accumulate in biotrophic interfacial complexes(BICs) and are translocated into the cytoplasm of the rice cells, and apoplastic effectors, which remain in the extracellular space between the fungal cellwall and the rice plasma membrane. BICs localize in front of the tips of filamentous hyphae that enter rice cells, and remain subapically beside the firstbulbous invasive hyphal cells after hyphal differentiation. In contrast, secreted apoplastic effectors uniformly outline the entire bulbous invasive hypha.We have determined that cytoplasmic effector genes were highly up-regulated in the BIC-associated cells at early invasion stages, and that effectorpromoters played the major role in determining preferential BIC localization of cytoplasmic effectors. Subapical BIC-associated hyphal cells continued toexpress protein secretion machinery components while invasive hyphae grew elsewhere in the host cell, suggesting that these subapical invasive hyphalcells are involved in active secretion. Disruption of the conventional ER-Golgi secretion pathway by Brefeldin A treatment blocked secretion of apoplasticeffectors, but not secretion of cytoplasmic effectors. Pathogen mutants that failed to express exocyst complex components or a t-SNARE were defective insecretion of cytoplasmic effectors, as well as defective in pathogenicity. In contrast, secretion of apoplastic effectors was not impaired in these mutants.Our data suggest that M. oryzae possesses distinct secretory mechanisms for targeting cytoplasmic and apoplastic effectors during rice invasion.The cellular role of early endosome motility in Ustilago maydis. Yujiro Higuchi 1 , Peter Ashwin 2 , Gero Steinberg 1 . 1) Biosciences, University of Exeter,Exeter EX4 4QD, UK; 2) Mathematics Research Institute, University of Exeter, Exeter EX4 4QF, UK.Early endosomes (EEs) are dynamic organelles that move along microtubules, which is mediated by the motor proteins kinesin-3 and dynein. Despite ourgrowing knowledge about the mechanistics of motion, the physiological significance of EE motility remains elusive. A recent study suggested that RNAbindingproteins travel on EEs, which might support local protein translation at the hyphal tip. However, evidence for apical translation is missing. Here, weinvestigate the distribution of ribosomes, using native levels of ribosomal proteins. We will summarize our findings on protein translation and will discussthe role of EE-dependent transport of RNA-binding proteins in the light of our findings.The arrestin-like protein ArtA is essential for ubiquitylation and endocytosis of the UapA transporter in response to both broad-range and specificsignals. George Diallinas, Mayia Karachaliou, Sotiris Amillis, Minos Evangelinos, Alexandros Kokotos. Faculty of Biology, University of Athens, Athens,Greece.We investigated the role of all arrestin-like proteins of Aspergillus nidulans in respect to growth, morphology, sensitivity to drugs and specifically for theendocytosis and turnover of the uric acid-xanthine transporter UapA. All arrestin null mutants are viable showing wild-type growth and morphology,except one which is affected in conidiospore production, but several have modified profiles in respect to N or C source utilization and drug sensitivity. Asingle arrestin, ArtA, is essential for HulARsp5-dependent ubiquitination and endocytosis of UapA in response to ammonium or substrates. Geneticanalysis further showed that residues 545-561 of the UapA C-tail, which includes a critical di-acidic motif, is required for efficient UapA endocytosis.Mutational analysis of ArtA shows that the N-terminal region (2-123) and both PY elements are essential for its function. ArtA undergoes HulA-dependentubiquitination at residue Lys343 and this modification is critical for the efficiency of UapA ubiquitination and endocytosis, especially in response toammonium. Lastly, we show that ArtA is essential for vacuolar turnover of transporters specific for purines (AzgA) or L-proline (PrnB), but not for anaspartate/glutamate transporter (AgtA). Our results are discussed within the frame of recently proposed mechanisms on how arrestins are activated andrecruited for ubiquitination of transporters in response to broad range signals, but also put the basis for understanding how arrestins, such as ArtA,regulate the turnover of a specific transporter in the presence of its substrates.Escaping the hustle - zones of differential protein turnover in the yeast plasma membrane. Guido Grossmann 1,2 , Vendula Stradalova 3 , MichaelaBlazikova 3 , Miroslava Opekarová 4 , Jan Malinsky 3 , Widmar Tanner 5 . 1) Center for Organismal Studies, University of Heidelberg, Heidelberg, Germany; 2)Department for Plant Biology, Carnegie Institution for Science, Stanford, CA; 3) Institute of Experimental Medicine, Academy of Sciences of the CzechRepublic, Prague, Czech Republic; 4) Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic; 5) Cell biology andplant biochemistry, University of Regensburg, Regensburg, Germany.The plasma membrane (PM) consists of specialized domains that differ in their protein distribution, lipid composition and structure, and that areessential for PM functions such as membrane transport or signal perception and transduction. The mechanisms that establish and maintain thisheterogeneity are still largely unknown but involve interactions between membrane constituents, local modification of the membrane structure, tetheringto other cellular components like ECM/cell wall or cytoskeleton, and polarized exo- and endocytosis. In the PM of bakers yeast, stable rod-shapedmembrane invaginations exist, called membrane compartment of Can1 (MCC), that exhibit a specific composition of lipids and proteins, and are stabilizedby a protein structure called eisosome. Chemical and genetic screens, revealed important roles of the membrane potential, lipid composition, and proteinscaffolds in organizing the PM into specialized domains. The distribution of MCC domains further determines the distribution of the PM-associated corticalER that dynamically covers large areas of the PM. Mapping of endocytic events revealed very low rates of clathrin-mediated endocytosis in PM areascovered by cER and within MCC. The formation of such "quiet zones" provides a mechanism for membrane domain formation through local confinementof membrane turnover.27th Fungal Genetics Conference | 35

CONCURRENT SESSION ABSTRACTSWhole-genome sequencing identifies novel alleles of genes required for organelle distribution and motility in Aspergillus nidulans. Kaeling Tan, AnthonyRoberts, Martin Egan, Mark Chonofsky, Samara Reck-Peterson. Cell Biology, Harvard Med Sch, Boston, MA.Many organelles are transported long distances along microtubules in eukaryotic organisms by dynein and kinesin motors. To identify novel alleles andgenes required for microtubule-based transport, we performed a genetic screen in the filamentous fungus, Aspergillus nidulans. We fluorescently-labeledthree different organelle populations known to be cargo of dynein and kinesin in Aspergillus: nuclei, endosomes, and peroxisomes. We then used afluorescence microscopy-based screen to identify mutants with defects in the distribution or motility of these organelles. Using whole-genomesequencing, we found a number of single nucleotide polymorphisms (SNPs) that resulted in misdistribution of peroxisomes, endosomes, or nuclei. Some ofthese SNPs were novel alleles of cytoplasmic dynein/ nudA, Arp1/ nudK (dynactin), Lis1/ nudF, and kinesin-1/ kinA. Here, we characterize the in vivotransport defects in these novel mutants and analyze the single molecule in vitro motility properties of purified mutant motor proteins. We also describeour methods for using whole genome sequencing as a tool in mutagenesis studies in A. nidulans.Dynamics of exocytic markers and cell wall alterations in an endocytosis mutant of Neurospora crassa. Rosa R. Mouriño-Pérez, Ramón O. Echauri-Espinosa, Arianne Ramírez-del Villar, Salomón Bartnicki-García. Microbiology Department, CICESE, Ensenada, B.C., Mexico.Morphogenesis in filamentous fungi depends principally on the establishment and maintenance of polarized growth. This is accomplished by the orderlymigration and discharge of exocytic vesicles carrying cell wall components. We have been searching for evidence that endocytosis, an opposite process,could also play a role in morphogenesis. Previously, we found that coronin deletion (Neurospora crassa mutant, Dcrn-1) causes a decrease in endocytosis(measured by the rate of uptake of FM4-64) together with marked alterations in normal hyphal growth and morphogenesis accompanied by irregularitiesin cell wall thickness. The absence of coronin destabilizes the cytoskeleton and leads to interspersed periods of polarized and isotropic growth of thehyphae. We used CRIB fused to GFP as an exocytic reporter of activated Cdc-42 and Rac-1. By confocal microscopy, we found that CRIB-GFP was present Inwild-type hyphae as a thin hemispherical cap under the apical dome, i. e. when growing in a polarized fashion and with regular hyphoid morphology. In theDcrn-1 mutant, the location of CRIB-GFP shifted between the periods of polarized and isotropic growth, it migrated to the subapical region and appearedas localized patches. Significantly, cell growth occurred in the places where the CRIB-GFP reporter accumulated, thus the erratic location of the reporter inthe Dcrn-1 mutant correlated with the morphological irregularity of the hyphae. We found that the Dcrn-1 mutant had a higher proportion of chitin thanthe WT strain (14.1% and 9.1% respectively). We also compared the relative cell wall area (TEM images) and we found a different ratio wall/cytoplasmbetween the Dcrn-1 mutant and the WT strain. In conclusion, we have found that the mutant affected in endocytosis has an an altered pattern ofexocytosis as evidenced by its distorted morphology and displaced exocytic markers. A direct cause-effect relationship between endocytosis andexocytosis remains to be established.“The vacuole” of Neurospora crassa may be composed of multiple compartments with different structures and functions. Barry J. Bowman 1 , Emma JeanBowman 1 , Robert Schnittker 2 , Michael Plamann 2 . 1) MCD Biology, University of California, Santa Cruz, CA; 2) Department of Biology, University of Missouri,Kansas City, KA.The structure of the “vacuole” in Neurospora crassa and other filamentous fungi is highly variable with cell type and position in the hypha. Largespherical vacuoles are typically observed in older hyphal compartments, but approximately 100 microns behind the hyphal tip, vacuolar markers are seenin a dynamic network of thin tubules. At the edge of this network nearest the tip, a few distinct round organelles of relatively uniform size (2-3 microns)have been observed (Bowman et al. Eukaryotic Cell 10:654 ). The function of these round organelles is unknown, although the vacuolar ATPase and avacuolar calcium transporter are strongly localized there. To help identify organelles we have tagged SNARE proteins and Rab GTPases with GFP and RFP.Several of these tagged proteins (sec-22, rab-7, rab-8) appear in the tubular vacuolar network and in the membrane of the round organelles. A uniqueaspect of the round organelles is their association with dynein and dynactin (Sivagurunathan et al. Cytoskeleton, 69:613). In strains with mutations in thetail domain of the dynein heavy chain the dynein is often seen in clumps. This aggregated dynein appears to be tightly associated with (and possibly inside)the round organelles, but not in the tubular vacuolar network. Further analysis of the location of SNARE and Rab proteins may help to identify the functionof the round organelles.36

CONCURRENT SESSION ABSTRACTSWednesday, March 13 3:00 PM–6:00 PMFred Farr ForumGenome Defense, Epigenetics and RNAiCo-chairs: Patrick Shiu and Sven SaupeMeiotic silencing by unpaired DNA in Neurospora. Thomas M. Hammond 1 , Hua Xiao 2 , Erin C. Boone 2 , Logan M. Decker 2 , David G. Rehard 2 , Seung A. Lee 2 ,Tony D. Perdue 3 , Patricia J. Pukkila 3 , Patrick K. T. Shiu 2 . 1) School of Biological Sciences, Illinois State University, Normal, IL; 2) Division of BiologicalSciences, University of Missouri, Columbia, MO; 3) Department of Biology, University of North Carolina, Chapel Hill, NC.Neurospora crassa has a cytoplasm that is shared by the entire hyphal network, making it particularly vulnerable to attack by repetitive elements.Accordingly, several surveillance mechanisms are in place to protect the genome integrity of the fungus. For example, if a gene is lacking a partner duringhomolog pairing in meiosis, all copies of this gene are silenced by a process known as meiotic silencing by unpaired DNA (MSUD). MSUD requires commonRNAi proteins (e.g., RNA-directed RNA polymerase, Dicer, and Argonaute) and may work as follows: an unpaired gene triggers the production of anaberrant RNA molecule, which is made double-stranded (by SAD-1) and processed into siRNAs (by DCL-1). These siRNAs are subsequently used to destroycomplementary mRNAs (by SMS-2). The aforementioned proteins colocalize in the perinuclear region, possibly forming a silencing complex that inspectsand processes RNA molecules as they exit the nucleus. We will discuss the recent advances in our understanding of this unique silencing pathway.Mechanism of quelling, a small RNA-mediated gene silencing pathway. Zhenyu Zhang, Shwu-Shin Chang, Yi Liu. Dept Physiology, Univ Texas SW Med Ctr,Dallas, TX.RNAi is a conserved gene silencing mechanism from fungi to mammals. Quelling is an RNAi-related phenomenon that post-transcriptionally silencesrepetitive DNA and transposon in Neurospora. We previously identified a type of DNA damage-induced small RNA called qiRNAs that originate fromribosomal DNA. To understand how small RNAs are generated from repetitive DNA, we carried out a genetic screen to identify genes required for qiRNAbiogenesis. Factors directly involved in homologous recombination (HR) and chromatin remodeling factors required for HR are essential for qiRNAproduction. HR is also required for quelling, and quelling is also the result of DNA damage, indicating that quelling and qiRNA production share a commonmechanism. Together, our results suggest that DNA damage triggered HR-based recombination allows the RNAi pathway to recognize repetitive DNA toproduce small RNA.SIS, a sex genome defense mechanism operating in Cryptococcus neoformans. Xuying Wang, Sabrina Darwiche, Joseph Heitman. Department ofMolecular Genetics and Microbiology, Duke University Medical Center, Durham, NC.Cryptococcus neoformans is a human fungal pathogen that undergoes a dimorphic transition from yeast to hyphae during a-a opposite-sex mating and a-a unisexual reproduction (same-sex mating). Infectious spores are generated during both processes. We previously identified a sex induced silencing (SIS)pathway in the C. neoformans serotype A var. grubii lineage, in which tandem transgene arrays trigger RNAi-dependent gene silencing at a high frequencyduring a-a opposite-sex mating, but at an ~250-fold lower frequency during asexual mitotic vegetative growth. Here we report that SIS also operatesduring a-a unisexual reproduction. A self-fertile strain containing either SXI2a-URA5 or NEO-URA5 transgene arrays exhibited an elevated silencingfrequency during solo and unisexual mating compared with mitotic vegetative growth. We also found that SIS operates at a similar efficiency on transgenearrays of the same copy number during either a-a unisexual reproduction or a-a opposite-sex mating. URA5-derived small RNAs were detected in thesilenced progeny of a-a unisexual reproduction and RNAi core components were required, providing evidence that SIS induced by same-sex mating is alsomediated by RNAi via sequence-specific small RNAs. This study, together with our previous finding of SIS in a-a opposite-sex mating of the C. neoformansserotype A var. grubii lineage, demonstrates that SIS is a conserved process between the divergent C. neoformans serotype A and serotype D siblingspecies. In each case, our data show that the SIS RNAi pathway operates to defend the genome via squelching transposon activity during the sexual cycles.Thus, our discovery of SIS brings a fresh perspective to meiotic silencing involving the upregulation of RNAi pathways as a strategy to guard genomicintegrity during sex. More importantly, the presence of SIS in both a-a unisexual reproduction and a-a opposite-sex mating indicate that SIS may betriggered by the shared pheromone sensing Cpk1 MAPK signal transduction cascade. Ongoing studies focus on defining at a mechanistic level how the SISRNAi pathway is initiated, including identifying new components involved in SIS.Fungi use prion folds for signal transduction processes involving STAND proteins. Asen Daskalov, Khalid Salamat, Sven J. Saupe. CNRS, IBGC UMR5095,BORDEAUX, AQUITAINE, France.Prions are proteins embedding genetic information into their structural state. Generally, those proteins exist in a soluble state and sporadically asinfectious amyloid aggregates. Podospora anserina’s [Het-s] is one of the best characterized fungal prions with a remarkably high prevalence in wildpopulations. [Het-s] functions in vegetative incompatibility - a biological process occurring during anastomosis between two genetically incompatiblestrains. The HET-s protein exists in a soluble state - [Het-s*] - or can switch to an aggregated amyloid state - [Het-s] - the prion form. When an [Het-s] prioninfected strain fuses with a strain expressing the alternative allelic variant of the het-s locus - het-S - a cell death reaction of the heterokaryon occurs.Recent studies shed light on the mechanism of [Het-s]/HET-S incompatibility reaction. Differing by 13 amino acids both proteins shares a two domainarchitecture; a globular N-terminal domain called HeLo and a C-terminal Prion Forming Domain (PFD). The latter is able to adopt a b-sheet richconformation with a specific b-solenoid fold. It has been demonstrated that in presence of [Het-s] amyloid fibers HET-S turns into a pore-forming toxin:transconformation of the HET-S PFD by [Het-s] fibers triggers the refolding of the HET-S HeLo domain, inducing the cell death reaction. In an attempt tobetter characterize the conserved features of the [Het-s] b-solenoid fold and identify new distant homologues of HET-S/s, we have generated a minimalconsensus sequence motif of it. Surprisingly, the second best hit in a BLASTp search is in the N-terminal region (3-23) of the product encoded by nwd2, theimmediately adjacent gene to het-S. NWD2 is a STAND protein. STAND proteins form signal transducing hubs through oligomerization upon ligandrecognition. That in mind and several bioinformatics observations led us to propose that HET-S and NWD2 are functional partners in various filamentousfungal species using the amyloid fold in a signal transducing pathway. We will present experimental evidence that NWD2 is able to trigger HET-S toxicity in27th Fungal Genetics Conference | 37

CONCURRENT SESSION ABSTRACTSmuch the same way as [Het-s] does. Further in silico analysis identify a number of these STAND/prion-like gene pairs and suggest that signal transductionthrough an amyloidal prion-like fold is a general widespread mechanism in fungi.Regulation of white and opaque cell-type formation in Candida albicans by H3K56 acetylation and nucleosome assembly factors CAF-1 and HIR. John S.Stevenson, Haoping Liu. Department of Biological Chemistry, University of California, Irvine, Irvine, CA.CAF-1 and HIR are highly conserved histone chaperone protein complexes that function in the assembly of nucleosomes onto chromatin. CAF-1 ischaracterized as having replication-coupled nucleosome activity whereas the HIR complex can assemble nucleosomes independent of replication. HistoneH3K56 acetylation, controlled by the acetyltransferase Rtt109 and deacetylase Hst3, also plays a significant role in nucleosome assembly. How differentcell types with the same genotype are formed and heritability maintained is a fundamental question in biology. We utilized white-opaque switching inCandida albicans as a system to study mechanisms of cell-type formation and maintenance. Opaque cell specification is under the control of interlockingtranscriptional feedback loops, with Wor1 being the master regulator. We showed that H3K56 acetylation plays an important role in the regulation ofwhite-opaque switching. The rtt109D/D mutant is defective in stochastic and environmentally stimulated white-opaque switching and cannot maintainopaque cell type. Inhibition of Hst3 by nicotinamide induces opaque cell formation in Rtt109 dependent manner. The Hst3 level is down-regulated in thepresence of genotoxins and ectopic expression of HST3 blocks genotoxin induced switching, providing a pathway for genotoxin induced white-opaqueswitching. We now show that CAF-1 and HIR modulate white-opaque switching frequencies in a H3K56 acetylation associated manner. Unique to C.albicans, the cac2D/D mutant shows increased sensitivity to the Hst3 inhibitor nicotinamide, while the rtt109D/D cac2D/D and hir1D/D cac2D/D mutantsare resistant to nicotinamide. CAF-1 plays a major role in maintaining cell types as the cac2D/D mutant exhibited increased switching frequencies in bothdirections, and switches at a high frequency to opaque in response to nicotinamide. Like the rtt109D/D mutant, the hir1D/D cac2D/D double mutant isdefective in maintaining the opaque cell fate, blocks nicotinamide induced opaque formation, and the defects are suppressed by ectopic expression of themaster white-opaque regulator Wor1, suggesting an overlapping function of CAF-1 and HIR in epigenetic regulation cell fate determination in a H3K56acetylation dependent manner.Epigenetic Regulation of Subtelomeric Gene Noise in Candida albicans. Matthew Z Anderson, Joshua A Baller, Lauren J Wigen, Judith Berman. Genetics,Cell Biology and Development, Univeristy of Minnesota, St Paul, MN.Candida albicans grows within a wide range of fluctuating host niches, and the ability to rapidly adapt enhances its success as a commensal and as apathogen. The recently expanded telomere-associated (TLO) gene family consists of fourteen expressed members in C. albicans. Each TLO gene encodes aparalog of a single Mediator complex component. Thirteen expressed TLOs are located at the chromosome ends as the most telomere-proximal openreading frame. Individual TLO expression at both the transcript and protein level was extremely noisy. Noise originated from single cell variability in TLOexpression due to intrinsic factors. Deletion of chromatin modifying enzymes that function in subtelomeric silencing abolished TLO noise, as did ectopicallyexpressing a TLO from an internal locus. Conversely, transcriptional variation of a low noise gene increased significantly when ectopically expressed in thesubtelomere. Interestingly, deletion of the Mediator component MED3, which inhibits Tlo from incorporating into Mediator, also drastically reduced TLOnoise and supports an autoregulatory mechanism for TLO noise. These data suggest subtelomeric chromatin structure regulates TLO gene noise throughthe action of chromatin modifiers and Mediator. We propose that TLO noise is beneficial to C. albicans by producing heterogeneous cell populations thatincorporate different Tlo proteins in Mediator, producing a range of transcriptional profiles in the population that allows some cells to survive in alteredenvironmental conditions.Chromatin regulation of genome stability. Zachary A. Lewis. Department of Microbiology, University of Georgia, Athens, GA.Genome instability results from defective DNA replication or repair and is associated with human diseases such as cancer. Chromatin structure impactsvirtually all DNA-templated processes in the nucleus, including replication and repair. To identify new chromatin factors that are required for genomestability, we screened the Neurospora knockout collection for strains that are sensitive to the DNA damaging agent methyl methanesulfonate (MMS). Theprimary screen uncovered over 500 MMS-sensitive knockout strains, including knockouts of putative regulators of chromatin structure. We are currentlytesting this group of knock out strains for sensitivity to other agents that induce DNA damage. We have also initiated molecular analyses of newlyidentified regulators of genome stability. Our current progress will be summarized.Opposing activities of the HCHC and DMM complexes maintain proper DNA methylation in Neurospora crassa. Shinji Honda 1,2 , Eun Yu 1 , Eric Selker 1 . 1)University of Oregon, Institute of Molecular Biology, Eugene, OR; 2) University of Fukui, Life Science Unite, Fukui, Japan.Proper regulation of heterochromatin and DNA methylation is critical for the normal function of cells. We show that heterochromatin and DNAmethylation are faithfully controlled in Neurospora by opposing activities of the silencing complex HCHC and the anti-silencing complex DMM. Theworkings of these two complexes were investigated. HCHC consists of four proteins, the two chromo domain proteins HP1 and CDP-2, the histonedeacetylase HDA-1 and the AT-hook motif protein CHAP. We found that histone deacetylase activity is critical for HCHC function but the H3K9me3 bindingactivity of the CDP-2 chromo domain is not. Instead, CDP-2 serves as an essential bridge between HP1 and HDA-1. CHAP interacts directly with HDA-1,binds in a methylation-independent way to the A:T-rich DNA that forms the cores of methylated regions and is important for stable association of HDA-1with chromatin. HCHC is involved in the spreading of DNA methylation in dmm mutants. The DMM complex consists of a presumed histone demethylase,DMM-1, plus DMM-2, which is characterized by a fungal-specific Zn(II) 2Cys 6 DNA-binding domain (“Zn-Cys”). We found that DMM-2 strongly binds to DNAfrom euchromatin/heterochromatin junctions, thereby promoting the stable association of DMM-1 at the edge of heterochromatin domains to preventaberrant spreading of DNA methylation.38

CONCURRENT SESSION ABSTRACTSWednesday, March 13 3:00 PM–6:00 PMKilnGenomics and MycorrhizaeCo-chairs: Anders Tunlid and Tom BrunsThe mycorrhizal genome initiative (MGI): Identification of symbiosis-regulated genes by using RNA-Seq. A. Kohler 1 , E. Tisserant 1 , E. Morin 1 , C. Veneault-Fourrey 1 , S. Abba 2 , F. Buscot 3 , J. Doré 4 , G. Gay 4 , M. Girlanda 2 , S. Herrmann 3 , T. Johansson 5 , U. Lahrmann 6 , E. Martino 2 , S. Perotto 2 , M. Tarrka 3 , A. Tunlid 5 , A.Zuccaro 6 , I. Grigoriev 7 , F. Martin 1 . 1) Lab of Excellence ARBRE, Tree-Microbes Department, INRA-Nancy, Champenoux, France; 2) Dipartimento di Scienzedella Vita e Biologia dei Sistemi, Università di Torino,Torino, Italy; 3) Department Soil Ecology, UFZ Centre for Environmental Research Leipzig-Halle Ltd.,Halle, Germany; 4) Ecologie Microbienne UMR CNRS 5557, USC INRA 1193, Universite Claude-Bernard LYON 1, Villeurbanne, France; 5) Microbial Ecology,Lunds University, Lund, Sweden; 6) Max-Planck Insitute for Terrestrial Microbiology, Marburg, Germany; 7) DOE Joint Genome Institute, Walnut Creek,California, USA.Genome and transcriptome analyses of Laccaria bicolor and Tuber melanosporum (Martin et al., 2008, 2010) revealed that the ectomycorrhizal symbiosisprobably developed several times during evolution by generating different ‘symbiosis molecular toolkits’. In L. bicolor a large set of small-secreted proteinsacts as putative effectors but not in T. melanosporum, while the up-regulation of transporter-coding genes seems to be a common feature of bothinteractions. To better understand the evolutionary origin of mycorrhizal symbiosis and to elucidate the molecular mechanisms involved, a largesequencing project of species from different taxa, phylogenetic clades and symbiotic lifestyles (ectomycorrhizae, ericoid and orchid mycorrhizae) wasstarted in 2011 by the Joint Genome Institute and the mycorrhizal genome initiative. To identify and to compare symbiosis-regulated genes large scaleIllumina transcriptome sequencing of mycelium and mycorrhizal roots from Paxillus involutus, Piloderma croceum, Hebeloma cylindrosporum, Sebacinavermifera, Tulasnella calospora and Oidiodendron maius was performed. Small-secreted proteins, transporters, CAZymes but also many lineage specificproteins were among the highly up-regulated transcripts.Martin, F., Aerts, A., Ahrén, D., Brun, A., Duchaussoy, F., Kohler, A., et al. 2008. The genome sequence of the basidiomycete fungus Laccaria bicolorprovides insights inot the mycorrhizal symbiosis. Nature 452 :88-92Martin, F., Kohler, A., Murat, C., Balestrini, R., Coutinho, P.M., Jaillon, O., Montanini, B., et al. 2010. Périgord black truffle genome uncovers evolutionaryorigins and mechanisms of symbiosis. Nature 464 :1033-1038.Transposable element dynamics in the Amanita: insights on the evolution of genome architecture accompanying the transition from saprotrophic toectomycorrhizal ecologies. Jaqueline Hess 1 , Inger Skrede 2 , Anne Pringle 1 . 1) Organismic and Evolutionary Biology, Harvard University, Cambridge, MA; 2)Microbial Evolution Research Group, Department of Biology, University of Oslo, Oslo, Norway.Transposable elements (TEs) form an integral structural part of the genomes of many higher Eukaryotes. Their ability to proliferate independently andinto a large number of copies can lead to extensive amounts of repetitive DNA that is of no obvious benefit to the host. At first thought to be relativelyunderrepresented in Fungi, genome sequencing over the last decade has led to the discovery of many fungal genomes that are densely populated withTEs. Among those are the genomes of the ectomycorrhizal (ECM) fungi Laccaria bicolor (around 30% TE) and Tuber melanosporum (around 60% TE) as wellas a number of fungal pathogens, including Puccinia graminis and Melamspora larici-populina (both around 45% TE). The high TE content in these species,especially when compared to saprotrophic fungal species, suggests an association between symbiotic ecology, both mutualistic and antagonistic, and theability of TEs to invade and persist in their genomes. However, the mechanisms for this are currently not well understood. In order to assess whether highTE content is a feature of other ECM species and to get a more detailed picture of TE content changes around the transition from free-living to ECMecology, we have sequenced the genomes of five members of the genus Amanita: three ECM species and two saprotrophs, as well as the saprotrophicoutgroup Volvariella volvacea. Using the draft genome assemblies, we have developed methodology to estimate TE content from short-read data andexamine changes therein within quantitative and phylogenetic frameworks. Overall, we find no direct relationship between ECM status and increased TEcontent in the Amanita but instead discover patterns that suggest population genetics to be a strong driver of TE content. We will discuss our findings withrespect to the influence of TEs in the evolution of genome architecture around the origin of ECM symbiosis.Broad compatibility in the root endophyte Piriformospora indica is associated with host-adapted colonization strategies. Urs Lahrmann, Yi Ding, AlgaZuccaro. Organismic Interactions, MPI Marburg, Marburg, Germany.Their host range defines plant associated fungi as either specialists, which are adapted to one or few distinct hosts, or generalists who are able to thrivein highly variable host environments. Specialists and their hosts are in an evolutionary arms race that leads to the development of weapons perfectlytailored to the respective host. Conversely, broad-host range species must evolve adaptations to cope with a plethora of different host-associated signalsand host-specific defense mechanisms. The evolutionary force, in this case, drives the expansion and diversification of the fungal arsenal and the hostadaptedgene expression to better suite different plants. The mechanisms underpinning broad compatibility in root symbiosis are largely unexplored. Thegeneralist root endophyte Piriformospora indica that stimulates growth, alleviates salt stress and induces systemic resistance to pathogens in differenthosts can establish a long lasting interaction with the roots of barley and Arabidopsis, two morphologically and biochemically very distinct plants. We showhere that in these two hosts, root colonization proceeds very differently. While in Arabidopsis the fungus establishes and maintains biotrophic nutritionwithin living epidermal cells, in barley the symbiont undergoes a nutritional switch to saprotrophy that is associated with the production of secondarythinner hyphae (SH) in dead cortex cells. Consistent with a diversified trophic behavior, genome-wide expression profiling revealed a strong induction ofgenes encoding cell wall degrading enzymes and nutrient transporters in barley but not in Arabidopsis at a late colonization stage. In particular smallsecreted proteins (SSPs < 300 amino acids) known as effectors have been shown to facilitate colonization by manipulating host defense andreprogramming plant metabolism during symbiosis. Expression of P. indica genes encoding SSPs was induced in both hosts at different symbiotic stage, butthe majority of these SSPs were either Arabidopsis or barley responsive with the larger number expressed during biotrophy in Arabidopsis and duringsaprotrophy in barley. Our study reveals that broad compatibility in root endophytes requires strong phenotypic plasticity and the expression ofalternative lifestyle strategies in a host-dependent way.27th Fungal Genetics Conference | 39

CONCURRENT SESSION ABSTRACTSExamining the saprotrophic ability of ectomycorrhizal fungi using genomics, transcriptomics and spectroscopy. Anders P V Tunlid. Dept MicrobialEcology, Ecology, Lund, Sweden.A large part of the nitrogen in forest soils is found in recalcitrant organic matter- protein complexes. Ectomycorrhizal fungi are thought to have a key rolein the decomposition and mobilization of nitrogen from such complexes. The knowledge on the functional mechanisms of these processes, and how theyare regulated by carbon from the host plant and the availability of more easily available forms of nitrogen sources are limited. We examined how the ECMfungus Paxillus involutus degrade organic litter material using spectroscopy and transcriptome profiling. The fungus partially degraded polysaccharides andmodified the structure of polyphenols. The observed chemical changes and the expressed transcriptome were consistent with a hydroxyl radical attack,involving Fenton chemistry similar to that of brown-rot fungi. The set of enzymes expressed by P. involutus during the degradation of the organic matterwas similar to the set of enzymes involved in the oxidative degradation of wood by brown-rot fungi. However, P. involutus lacked transcripts encodingextracellular enzymes needed for metabolizing the released carbon. Further experiments have shown that the decomposition and assimilation of nitrogenfrom organic litter material are triggered by adding glucose. Addition of easily available forms of nitrogen (i.e. ammonium) had minute effects on theseprocesses. Experiments and comparative genomics demonstrate that the saprotrophic activity of P. involutus has been reduced to a radical-basedbiodegradation system that can efficiently disrupt the organic matter-protein complexes and thereby mobilize the entrapped nutrients.Interaction between the saprotrophic fungus Serpula lacrymans and living pine roots. Nils OS Högberg 1 , Anna Rosling 1 , Annegret Kohler 2 , Martin Francis 2 ,Stenlid Jan 1 . 1) Department of Forest Mycology, BioCenter, SLU, Uppsala, Sweden; 2) INRA, Nancy, France.Recently it has been shown, with a Comparative genomic perspective, that brown rot and mycorrhiza fungi have evolved from white rot ancestors. Woodis a composite material composed of lignin, cellulose and hemicellulose. White rot fungi are able to degrade all of these components with a combination ofcarbohydrate active and oxidative enzymes. During the course of evolution brown rot and mycorrhiza have lost most of the genes in these gene families.Nevertheless, brown rot fungi are efficient wood decomposers that degrade cellulose and hemicellulose by means of hydroxyl radical production andremaining carbohydrate active enzymes. The family Boletales includes both brown rot fungi and mycorrhiza and it is tentative to speculate that there hasbeen a parallel evolution of these ecological strategies. Here we test the effect of infecting pine roots with the brown rot fungus Serpula lacrymans. Theinteraction was neutral since plant growth was not stimulated but not reduced either. The fungus formed a mantle around the pine roots but not theHartig net that is typical for ectomycorrhiza. Fungal gene expression was compared with the wood decay transcriptome. 1250 genes were more than twofoldupregulated compared to a glucose medium control. A large proportion of the upregulated genes (62 %) are unknown. Carbohydrate active genesrepresent only 3% of this gene set and genes with oxidoreductase activity, including monoxygenases represent 4% of the upregulated genes. This isconsiderably lower compared to saprotrophic growth on wood where carbohydrate active enzymes accounted for 26% and oxidative enzymes for 19%which dominated the gene expression on wood. Gene expression for genes involved in transportation was about the same, around 10% in this experimentand under wood decomposition. Several genes that indicate an interaction with a host were also upregulated. In conclusion, gene expression wasmarkedly different between a glucose medium, wood decomposition and growth on pine roots. This may be a signal of symbiosis, the effect on pineseedling growth was neutral. Thus we cannot conclude if the interaction is beneficial or negative to the host.Uncovering the evolutionary pressures shaping the Glomeromycota-Glomeribacter endosymbiosis. Stephen J. Mondo, Teresa E. Pawlowska. PlantPathology, Cornell University, Ithaca, NY.Many eukaryotes interact with heritable endobacteria to satisfy diverse metabolic needs. Of the characterized fungal-bacterial endosymbioses, theassociation between Gigasporaceae (Glomeromycota) and Ca. Glomeribacter is one of the best described. Glomeribacter is a member of the Burkholderialineage of b-proteobacteria, and was shown previously to represent one of the few cases of an ancient, long-term non-essential endosymbiont. In order tofurther explore what adaptations have taken place to shape this unique bacterial lifestyle, we have sequenced three Glomeribacter genomes anddeveloped a computational pipeline to compare across bacteria engaging in different lifestyles using genome wide patterns of mutation accumulation. Weused PAML to identify gene orthologs that exhibited both over-accumulation and under-accumulation of amino acid substitutions and then used thesedata to compare across taxa at the level of functional gene categories. We found that bacteria can be grouped by lifestyle using this approach.Glomeribacter, as expected, appears most similar to other potentially long-term non-essential endosymbionts. Therefore, we were able to exploit thedifferences in mutation accumulation patterns between these taxa to identify processes, which may be relevant within the particular interaction betweenGlomeribacter and its host. While several of these processes, including vitamin synthesis and amino acid transport, have been identified previously, weadditionally discovered features related to lipid biosynthesis and energy metabolism to be of potential importance for this symbiosis. Interestingly, genesexhibiting an under-accumulation of nonsynonymous substitutions (indicative of purifying selection) in Glomeribacter tend to be involved inrecombination, cell division, and ribosome maintenance. While these processes are typically fast evolving in endosymbiotic organisms, they may representfeatures that increase the stability of Glomeribacter in their fungal host population and increase their resilience to genetic drift. We speculate that theseprocesses are unique to the Glomeribacter-Glomeromycota symbiosis and could partially explain why Glomeribacter has been successful as a nonessentialendosymbiont for over 400 million years.40

CONCURRENT SESSION ABSTRACTSA draft genome of the ectomycorrhizal fungus Rhizopogon vesiculosus: Characterization of mating system and heterozygosity within the dikaryon. AlijaMujic, Joseph Spatafora. Botany and Plant Pathology, Oregon State University, Corvallis, OR.Species of Rhizopogon are EM symbionts of trees in family Pinaceae and produce basidiospores within hypogeous false truffles that are dispersed bymycophagous mammals. All known members of R. subgenus Villosuli form obligate EM relationships with Pseudotsuga spp. (Douglas Fir) and are the onlymembers of the genus known to possess this host association. R. vesiculosus, along with its cryptic sister species R. vinicolor, possess a sympatricdistribution where sampled within the range of their host tree, P. menziesii. While the sporocarp and EM morphology of these fungi may be highly similar;they possess striking life history differences with R. vesiculosus producing larger vegetative genets and displaying greater population structure at bothlocal and landscape scales. We have sequenced the genome of R. vesiculosus using dikaryotic tissue and a whole genome shotgun sequencing approach onthe Illumina HiSeq platform. De novo assembly of the genome was performed using VELVET 1.19 and gene predictions were made using AUGUSTUS withLaccaria bicolor as a training model. The draft genome assembled to a total length of 46 Mb in 6700 contigs with an N50 of 26,783, a maximum contig sizeof 446,818 bp, and 12,604 predicted genes. Here we characterize the mating system of R. vesiculosus, which possesses both an A-locus encoding aheterodimer transcription factor, as well a B-locus encoding transmembrane pheromone receptors and pheromone precursor genes. We presentcomparisons of the mating system of R. vinicolor and its similarities to other members of Boletales (e.g., Serpula) and differences with Agaricales (e.g.,Laccaria). Due to the dikaryotic nature of the genome sequence produced for R. vesiculosus, single nucleotide polymorphisms (SNPs) can be observed andused to characterize allelic variation. SNPs observed in protein coding regions of both MAT loci indicate that R. vesisculosus is likely heterothallic. We havealso characterized heterozygosity across the whole genome in order to identify hypervariable regions. This genome will allow for comparative analysis ofgene content, mating type system with other Basidiomycota and, ultimately, for population/species-level genomic studies within Rhizopogon.Metatranscriptomic analysis of ectomycorrhizal root clusters in Pinus taeda: new methodologies for assessing functional gene expression in situ. H.-L.Liao 1 , Y. Chen 2 , T. D. Bruns 3 , K. G. Peay 4 , J. W. Taylor 3 , S. Branco 3 , J. M. Talbot 4 , R. Vilgalys 1 . 1) Department of Biology Duke University, Durham, NC; 2)School of Medicine, Duke University, Durham, NC; 3) Department of Plant and Microbial Biology, UC-Berkeley, Berkeley, CA; 4) Department of Biology,Stanford University, Stanford, CA.A highly diverse community of ectomycorrhizal (ECM) fungi are known to associate with members of the genus Pinus. Less is known about how diversefungal communities affect functional diversity within ECM roots. Here we present an optimized method for metatranscriptomic analysis of the ECM-pineroot interaction in a natural system. RNA was purified using a CTAB method from individual ECM root clusters collected at varying spatial scales across thedistribution range of P. taeda, and sequenced using Illumina HiSeq technology. About 35 million qualified reads were obtained. Sequences were initiallyassembled using reference based mapping (Bowtie) to sort the reads that represent rRNA from fungal and bacterial species. Reads from divergent regions(D1-D2) of fungal LSU rRNA were used to identify dominant ECM and other fungal community members. Subsequently, P. taeda genes and functionalgenes of dominant fungal species were sorted using public cDNA databases. The Trinity package was used for de novo assembly of un-mapped reads(mostly fungal genes). Blastx and Go packages were used for gene annotation. A typical ECM root cluster was found 45% P. taeda genes, 3% fungal rRNA,0.05% bacterial 16S rRNA, 30% fungal functional genes, 10% unknown sequences, and 12% unassembled reads. Analysis of D1-D2 LSU sequencesconfirmed that a single ECM fungal species usually dominates individual root clusters. De novo assemblies of fungal genes yielded 120 thousand contigsfrom 10 million reads representing 90 thousand unique genes with highly similarity to known ECM fungi. Functional analysis revealed that most of thetranscripts recovered were involved with translation, protein degradation, heat shock, superoxide metabolism, electron transfer, signaling, and C/Nmetabolism. Highly expressed transcripts recovered from Piloderma, which was abundant in our samples, included genes encoding a wide array ofmetabolic enzymes: chitosanase, phosphatase, glutamine synthetase, terpene synthases, b-glucanase; transporters for P+ and oligopeptides; cell signaling:calmodulin, cAMP-regulated phosphoprotein (Igo1); C/N related genes: lectin, cross-pathway control (cpc1); as well as several genes with unknownfunction. Future studies will seek to address how ECM metatranscriptomes change in response to different Pinus hosts and across different spatial scales.27th Fungal Genetics Conference | 41

CONCURRENT SESSION ABSTRACTSWednesday, March 13 3:00 PM–6:00 PMNautilusRegulation and Comparative Genomics of Carbon and Nitrogen MetabolismCo-chairs: Richard Wilson and Ronald de VriesThe role of carbon in fungal nutrient uptake and transport: implications for resource exchange in the arbuscular mycorrhiza. Carl R. Fellbaum 1 , Emma W.Gachomo 1 , Gary D. Strahan 2 , Philip E. Pfeffer 2 , E. Toby Kiers 3 , Heike Bücking 1 . 1) Biology and Microbiology, South Dakota State University, Brookings, SD; 2)Agricultural Research Service, Eastern Regional Research Center, US Department of Agriculture, Wyndmoor, PA; 3) Department of Ecological Science, VrijeUniversiteit, Amsterdam, The Netherlands.Arbuscular mycorrhizal (AM) fungi can substantially contribute to host plant nitrogen (N) nutrition in exchange for carbon (C). We studied the effect of Csupply on fungal N uptake and transport in the AM symbiosis via 15 N labeling, enzymatic assays and qPCR analysis of fungal genes putatively involved in Nmetabolism. We found that an increase in C supply stimulated 15 N transport and increased the enzymatic activity of arginase and urease in the intraradicalmycelium (IRM). The fungus responded to an increase in the C supply with an upregulation of genes involved in N assimilation and arginine biosynthesis,but with a downregulation of a fungal urease in the extraradical mycelium (ERM). The effect on fungal gene expression in the IRM was relatively small, butgenes involved in arginine biosynthesis were downregulated by an increase in C availability. The results indicate that C from the host triggers N uptake bythe AM fungus, the conversion of N into arginine in the ERM, the transport of arginine to the IRM and subsequent breakdown of arginine via the catabolicarm of the urea cycle. When the fungus had access to a C supply independent from the host 15 N transport was reduced and a change in the geneexpression pattern indicated that the fungus changed its nutrient allocation strategy when the fungus was less dependent on the host for its C supply. In acommon mycelial network, the AM fungus Glomus aggregatum transported more N to a more photosynthetically active plant when given the choicebetween a shaded versus a non-shaded plant host. The results indicate that AM fungi are able to distinguish between hosts differing in their carbon supplyand that carbon is an important trigger for fungal nitrogen uptake and transport in the AM symbiosis.Mechanisms of adaptation to host rice cells by the blast fungus. Jessie Fernandez, Richard A. Wilson. Plant Pathology, University of Nebraska-Lincoln,Lincoln, NE 68516, USA.To infect rice, the devastating blast fungus Magnaporthe oryzea has distinct morphogenetic stages that allow it to breach the surface of the host leaf andinvade the plant tissue. How the fungus monitors the transition from the nutrient-free surface to the nutrient-rich interior of the leaf, what controls thegenetic reprogramming necessary to produce infectious hyphae, and how it acquires nutrient during biotrophic in planta growth is poorly understood. M.oryzae’s trehalose-6-phosphate synthase 1 (Tps1) enzyme integrates carbon and nitrogen metabolism in the fungal cell to regulate virulence via a novelNADPH-dependent genetic switch. Loss of Tps1 function results in Dtps1 strains that can form functional appressoria and penetrate the rice surface but failto grow beyond the first infected cell. Impaired invasive growth of Dtps1 strains is due to loss of glucose sensing, inactivation of the NADPH-dependentgenetic switch, and altered carbon assimilation. Moreover, NADPH-requiring antioxidation systems are shut down in Dtps1 strains, rendering themhypersensitive to oxidative stress. Taken together, we discuss here how, using classical and high-throughput reverse genetics, we are exploring thedynamics of this critical NADPH-dependent genetic switch to understand how M. oryzae controls infectious hyphal development during biotrophy, how itresponds to and acquires nutrient from the host, and how these processes are integrated to allow successful colonization of rice cells.Similar is not the same: Differences in the function of the (hemi-) cellulolytic regulator XlnR (Xlr1/Xyr1) in filamentous fungi. Sylvia Klaubauf 1* , HariMander Narang 1 , Evy Battaglia 2 , Tetsuo Kobayashi 3 , Kurt Brunner 4 , Astrid R. Mach Aigner 4 , Robert L. Mach 4 , Ronald P. de Vries 1,2 . 1) Fungal Physiology, CBS-KNAW Fungal Biodiversity Centre, Utrecht, Netherlands; 2) Microbiology, Utrecht University, Utrecht, Netherlands; 3) Department of BiologicalMechanisms and Functions, Graduate School of Bioagricultural sciences, Nagoya University, Furo-cho, Chikusa-ku, Nagoya-shi, Aichi, Japan; 4) Institue ofChemical Engineering, Research Area Biotechnology and Microbiology, Working Group Gene Technology, Vienna, Austria.The (hemi-) cellulolytic transcriptional activator XlnR (Xlr1/Xyr1) is a major regulator in fungal xylan and cellulose degradation as well as in the utilizationof D-xylose via the pentose catabolic pathway. XlnR homologs are commonly found in filamentous ascomycetes and often assumed to have the samefunction in different fungi. However, a comparison of the saprobe Aspergillus niger and the plant pathogen Magnaporthe oryzae showed differentphenotypes for deletion strains of XlnR. In this study wild type and xlnR/xlr1/xyr1 mutants of six fungi were compared: Fusarium graminearum, M. oryzae,Trichoderma reesei, A. niger, Aspergillus nidulans and Aspergillus oryzae. The comparison included growth profiling on relevant substrates and detailedanalysis of protein profiles of extracellular enzymes as well as extracellular enzyme activities. The resulting data demonstrated significant differences in theinfluence of XlnR and its orthologs on plant polysaccharide degradation by these fungi. For example, in A. niger cellulolytic enzymes, such ascellobiohydrolase and b-glucosidase are strongly down-regulated in the mutant strain, whereas this is not the case for the other two Aspergillus species.Moreover, in A. oryzae the L-arabinose releasing enzyme a-arabinofuranosidase is clearly regulated by AoXlnR, whereas this enzyme is known to be undercontrol of another regulator, AraR, in A. niger and not affected by XlnR. In contrast, M. oryzae Xlr1 does not significantly affect enzyme activities in thisstudy. Based on extracellular protein profiles, disruption of Xyr1 results in the disappearance of only some bands in F. graminearum, while nearly all bandsdisappear in T. reesei Dxyr1. This comparison emphasizes the functional diversity of a fine-tuned (hemi-) cellulolytic regulatory system in filamentous fungi,which might be related to the adaptation of fungi to their specific biotopes.42

CONCURRENT SESSION ABSTRACTSRegulating the Aspergillus nidulans global nitrogen transcription factor AreA. Richard B. Todd. Department of Plant Pathology, Kansas State University,Manhattan, KS.Nitrogen nutrient utilization genes are regulated in Aspergillus nidulans by the GATA DNA-binding transcription activator AreA. The transcriptionalactivity of AreA is highly regulated by multiple mechanisms including autogenous transcriptional regulation, differential areA transcript stability,interaction of AreA with the corepressor NmrA, and repression by the negative-acting GATA factor AreB. In addition, AreA shows regulated nuclearaccumulation. AreA accumulates in the nucleus specifically during nitrogen starvation, and is rapidly exported to the cytoplasm upon addition of nitrogennutrients to nitrogen-starved cells. I will focus on recent developments in our understanding of AreA nuclear import and nuclear export, the key controlpoints of regulated AreA nuclear accumulation. We have shown that the six conserved nuclear localization signals (NLSs) in AreA show redundancy andcollaborate to mediate nuclear import. In contrast, a single CrmA exportin-dependent nuclear export signal (NES) in AreA is required for nuclear export.We have shown that fusion of the AreA NES to a constitutively nuclear protein confers nucleocytoplasmic localization and a loss of function phenotype.We have exploited this phenotype to select mutants defective in the AreA-CrmA interaction.Transcriptional analysis of oxalate degradation in the white rot basidiomycete Dichomitus squalens. Miia R. Mäkelä, Johanna Rytioja, Outi-Maaria Sietiö,Sari Timonen, Annele Hatakka, Kristiina Hildén. Department of Food and Environmental Sciences, University of Helsinki, Helsinki, Finland.Basidiomycetous white rot fungi are the most efficient degraders of lignocellulose with a unique ability to mineralize the recalcitrant lignin polymer.Lignocellulose decay involves a complex enzymatic system, but is also suggested to be promoted by the fungal secretion of oxalic acid. White rot fungisynthesize oxalate as a metabolic waste compound and typically secrete it to their environment in millimolar quantities. As oxalate is a toxic compound,regulation of its intra- and extracellular concentration is extremely crucial for fungi and also for lignocellulose degradation since high oxalate levels areshown to inhibit the decomposition reactions. Therefore, specific oxalate-converting enzymes, namely oxalate decarboxylases (ODCs) that work inconjunction with formate-degrading formate dehydrogenases (FDHs), are recognized as key fungal enzymes in lignocellulose decay. Dichomitus squalens isa white rot fungus that degrades effectively all the wood polymers, i.e. cellulose, hemicelluloses and lignin, and secretes oxalic acid during its growth onwood. The genome of D. squalens harbours 5 putative ODC and 3 putative FDH encoding genes, while these numbers differ in other fungi based oncomparative genomics. In order to enlighten the roles of the multiple oxalic-acid catabolising enzymes of D. squalens, the expression of the odc and fdhgenes was followed with quantitative real-time RT-PCR when the fungus was grown on its natural substrate, i.e. Norway spruce (Picea abies) wood. Inaddition, the effect of organic acid (oxalic acid) and inorganic acid (HCl) supplementation on the relative transcript levels of the oxalate-catabolizing geneswas examined in the submerged liquid cultures of D. squalens. The results show for the first time the sequential action of ODC and FDH at the transcriptlevel in a white rot fungal species. The constitutive expression of odc1 suggests the pivotal role of the corresponding enzyme during the growth of D.squalens on wood. In addition, the strong upregulation of the transcription of odc2 in oxalic-acid amended cultures indicates the distinct roles of individualODC isoenzymes.TOR-mediated control of virulence functions in the trans-kingdom pathogen Fusarium oxysporum. Gesabel Y. Navarro Velasco, Antonio Di Pietro.Departamento de Genética, Universidad de Córdoba, 14071 Córdoba, Spain.Infectious growth of fungal pathogens is controlled by environmental cues, including nutrient status. The soilborne fungus Fusarium oxysporum producesvascular wilt disease in more than a hundred different crop species and can cause lethal systemic infections in immunodepressed humans. Previous workshowed that the preferred nitrogen source ammonium causes repression of infection-related processes in F. oxysporum that could be reversed byrapamycin, a specific inhibitor of the conserved protein kinase TOR. Here we generated mutations in upstream components that should result inconstitutive activation of TOR, including null mutants in tuberous sclerosis complex 2 (TSC2), a small GTPase that represses TOR activity, as well as strainsexpressing a dominant activating allele of the small GTPase Rag (ragA Q86L ), an activator of TOR. The Dtsc2 mutants and, to a minor extent, the ragA Q86Lstrains showed defects in hyphal growth and colony morphology on several amino acids, as well as decreased efficiency in cellophane penetration andvegetative hyphal fusion. These phenotypes were exacerbated in Dtsc2ragA Q86L double mutants and could be reversed by rapamycin, suggesting that theyare caused by hyperactivation of TOR. The mutants caused significantly lower mortality on tomato plants and on larvae of the animal model host Galleriamellonella. These results suggest that TOR functions as a negative regulator of fungal virulence on plant and animal hosts.Transcriptional regulation of peptidases and nitrogen transporters during the assimilation of organic nitrogen by the ectomycorrhizal fungi Paxillusinvolutus. Firoz Shah 1 , Francois Rineau 2 , Tomas Johansson 1 , Anders Tunlid 1 . 1) Microbial Ecology Group, Department of Biology, Lund University, SE-22362,Lund, Sweden; 2) Centre for Environmental Sciences, Hasselt University, Building D, Agoralaan, 3590 Diepenbeek, Limburg, Belgium.Proteins and amino acids form a major part of the organic nitrogen (N) sources in soils. Though a poorly characterized process, this N is mobilized andbecomes available to plants due to the activity of ectomycorrhizal (ECM) fungi. We have examined the role of ectomycorrhizal extracellular peptidases andamino acid transporters in the degradation, uptake and transfer of various protein sources (BSA, Gliadin and pollen) as well as of plant litter material usingthe ECM model fungus Paxillus involutus. During N-deprived conditions, all substrates induced secretion of peptidase activities. The activity had acidic pHoptimum (2.3-3.0), and it was mainly due to aspartic peptidases and with minor contribution of metallo and serine peptidases. The activity was partly andtemporarily repressed by low concentrations of ammonium (1mg/L). Transcriptional analysis showed that P. involutus expressed a large array of proteinsand enzymes involved in the assimilation of organic N including peptidases, N-transporters and enzymes of the N-metabolism. Extensive in-silico analysisrevealed the presence of genes encoding 312 peptidases, 129 N transporters and 284 enzymes involved in amino acid metabolism. Out of these, 89peptidases and 37 N-transporters and 109 amino acid metabolism enzymes encoding genes were significantly upregulated during organic N assimilation.The genes were encoding a variety of secreted (23) and non-secreted (20) peptidases which were differentially expressed depending on the medium withthe highest expression of the aspartic and metallo peptidases. Apart from the YAAH/ATO family, upregulated genes were found in all the other families oftransporters for amino acids, oligopeptides, ammonium, urea and allantoate/allantoin. The results shows that the expression levels of peptidases andtransporters in P. involutus are coordinately regulated during the assimilation of organic N sources.27th Fungal Genetics Conference | 43

CONCURRENT SESSION ABSTRACTSRegulation of glycolysis and gluconeogenesis by antisense transcription in Aspergillus nidulans? Michael Hynes 1 , Koon Ho Wong 2 , Sandra Murray 1 . 1)Dept Gen, Univ Melbourne, Parkville, Victoria, Australia; 2) Dept. of Biological Chemistry & Molecular Pharmacology, Harvard Medical School, Boston, MA.The last step in glycolysis is carried out by pyruvate kinase, encoded by pkiA, converting phospho-enol-pyruvate to pyruvate for metabolism tooxaloacetate and acetyl-CoA. The key step in gluconeogenesis is conversion of oxaloacetate to phospho-enol-pyruvate by PEP carboxykinase, encoded byacuF. Simultaneous activity of these enzymes would generate a nasty futile cycle. A number of observations suggests that control of the expression ofthese two genes involves activation of sense transcription and negative control by activation of antisense transcription. For pkiA, ChIP studies have foundbinding of the gluconeogenic activators AcuK and AcuM and of the acetate dependent FacB activator in the downstream region. Cognate binding sites areconserved in filamentous ascomycetes. RNA Seq, polII ChIP and RT-PCR analysis indicates antisense transcription during growth on acetate or proline ascarbon sources. Old data (de Graaf, van den Broek, Visser; Cur. Genetics 13: 315, 1988) showed that transformation of a construct lacking these 3’ sitesresulted in inappropriate pkiA expression on acetate. In response to growth on gluconeogenic carbon sources, the acuF gene is activated by AcuK andAcuM binding to sites in the 5’ upstream region. Studies with an acuF-lacZ gene fusion indicate positive control by AcuK and AcuM but a loss of the glucoserepression observed in Northerns suggesting negative regulation acting via 3’ sequences in response to growth on glycolytic carbon sources. Support forthis is provided by transcription studies. Modulation of the balance between the opposing activities of these two gene products is proposed to result fromtranscriptional interference involving collision of RNA polymerase molecules.44

CONCURRENT SESSION ABSTRACTSWednesday, March 13 3:00 PM–6:00 PMScrippsEducation, Outreach, and Professional DevelopmentCo-chairs: Steven Denison and Mimi ZolanCentrosome-Nuclear Disconnect Creates Mitotic Chaos in a Closed Mitosis System. Michael Koonce, Irina Tikhonenko. Translational Medicine,Wadsworth Center, NYS Department of Health, Albany, NY.In many fungi, protists, and unicellular algae, cells divide via a mitotic mechanism that functions within a closed nuclear compartment. Closed mitosisrequires tight coordination between the centrosome and nucleus to ensure a smooth transition from cytoplasmic to nuclear activities. In many cases, thecentrosome/spindle pole body is directly embedded into the nuclear envelope, an arrangement that facilitates this transition. However in other organisms(e.g. Cryptococcus, S. Pombe, Dictyostelium), the centrosome is simply attached to the cytoplasmic side of the nucleus during interphase, gaining access tothe intranuclear volume only during mitosis when this organelle builds the spindle. We have identified a kinesin motor protein (Kif9) that is responsible formaintaining the centrosome-nucleus interaction in Dictyostelium. In the absence of Kif9, centrosomes separate from nuclei and the two organelles moveindependently. We used this null mutant to examine what happens if centrosomes fail to engage nuclei when cells enter mitosis and to examineinteractions in multi-nucleated arrangements. We find that centrosomes replicate and separate in the cytosol, but do not form a visible spindle apparatus.Nuclei that lack centrosomes import tubulin, but also fail to develop a spindle apparatus. Replicated daughter centrosomes are able to integrate into thenuclear envelope and can go on to form monopolar spindles. All centrosomes (nuclear or cytoplasmic) trigger cytokinesis activity. Moreover, the nuclearenvelope is promiscuous and can dock multiple centrosomes if they are available. In these cases, multipolar or multiple independent spindles can arisewhich lead to aneuploid nuclear products. Our work illustrates the significance of maintaining a one centrosome-one nucleus relationship to ensure properchromosome segregation. This is particularly important in multinucleated syncytia where unpaired activities would result in multiple combinations ofcentrosome-nuclear engagements. In addition, a firm coupling between these two organelles links nuclei to microtubule force generating machinery thatis crucial for nuclear transport and positioning. We will further present a model for how the Kif9 kinesin functions to maintain centrosome-nuclear paring.For our work, we gratefully acknowledge support from the NSF (MCB-1051612).Using Fungal Barcoding to Introduce Non-science Majors to Scientific Research. Claire Burns. Washington & Jefferson College Washington, PA.Non-scientists, even those with an interest in biology, tend to have a poor understanding of fungi; one example is the perception that mushrooms are“some kind of weird plant”. This lack of awareness regarding fungi and their importance allows non-majors students to enter the classroom withoutpreconceptions or expectations. Fungi provide an excellent starting point to introduce students to broader biological themes such as cell biology, diversityof life, evolution, conservation, and molecular biology. In addition, the relative paucity of species classification for fungi when compared with otherkingdoms provides an opportunity for non-majors students to engage in primary research projects. In a liberal arts college class entitled “’Shrooms”,students collect mushrooms in the field, cultivate the fungi, extract DNA, and identify the species using DNA barcoding. Fungal identifications andsequences can contribute to ongoing fungal barcoding efforts. In future classes, students will expand this research to include identification of soil-bornefungi at a field station located close to gas-fracking operations.ComGen Authentic Research Experiences (C-ARE): Fungal genetic analysis. Gita Bangera 1 , Andrea Gargas 2 . 1) Bellevue College, Bellevue, WA, USA; 2)Symbiology LLC, Middleton, WI, USA.ComGen (Community College Genomics Research Initiative) teaches students the skills of self-directed learning, critical thinking, and analysis.Community college students in this program receive a mini-graduate school experience, following a single requisite course in cell biology. Students workon original research projects, learn to troubleshoot their experiments, organize lab meetings and student journal clubs, and network within the scientificcommunity. In one research track students work with DNA from described fungal collections, learning DNA-based techniques including PCR amplification,DNA sequencing and sequence analysis. Student-gathered sequence information is used to advance identification and phylogenetic results for thesecollections. With NSF Award DUE #1225857 ComGen (C-ARE): Dissemination, Enrichment and Expansion Project the project will be expanding tocommunity college partners throughout the Seattle/Tacoma region of Washington State.Wearing two hats: Tips for combining commitments to research and to university-wide initiatives in education. Patricia J. Pukkila. Dept Biol, Univ NorthCarolina, Chapel Hill, NC.In an editorial which appeared in the New York Times, Gary Gutting argued that the primary role of universities is to “nourish a world of intellectualculture; that is, a world of ideas, dedicated to what we can know scientifically, understand humanistically, or express artistically”(http://opinionator.blogs.nytimes.com/2011/12/14/what-is-college-for/). At research universities, faculty are expected to make substantial contributionsto their disciplines, to society, and to educating students. It is important for faculty to seek pan-university roles, and making acknowledged contributionsto changing your campus culture can be deeply satisfying. This session will encourage you to consider how you might productively multitask in ways thatcan actually benefit your research productivity in addition to improving the intellectual climate on your campus. Supported in part by the HHMI throughthe Precollege and Undergraduate Science Education Program.27th Fungal Genetics Conference | 45

CONCURRENT SESSION ABSTRACTSFacilitating an Interdisciplinary Learning Community Amongst Undergraduate Research Fellows By Emphasizing Scientific Inquiry as the UnifyingThread. Virginia K. Hench 1,2 , Patricia J. Pukkila 1,2 . 1) Department of Biology, University of North Carolina at Chapel Hill, NC 27599; 2) Office forUndergraduate Research, University of North Carolina at Chapel Hill, NC 27599.The HHMI-Future Scientists and Clinicians (HHMI-FSC) fellowship is 1 of 3 components of the HHMI Science Learning Communities program at UNCChapel Hill. The HHMI-FSC program was designed to foster an intellectual community that empowers high-ability students from low-incomebackgrounds to engage in biomedical research for 2 summers. Each year, 12 new fellows are matched with mentors in labs spanning a range of biomedicalareas. They work fulltime in labs on their own research project and meet weekly as a group to engage in interactive programming that targets skills criticalfor success in science beyond the bench. One area of emphasis has been the process of inquiry itself. The goal is for students to transition from being apair of hands executing protocols to active learners invested in their own projects and able to speak with authority about why experiments are performedin particular ways and what conclusions can be drawn from data generated. This starts with coaching students to state the questions that they are tryingto answer and think through whether an experimental setup is consistent with what they say they are trying to find out. Assignments and feedback aredesigned to reinforce this principle. One of the most satisfying aspects of doing science is getting to follow one’s own instinctive curiosities and developthe methodologies needed to navigate new terrains. Undergraduates are usually still trying to define their own specific curiosities. Pushing students todescribe what they are curious and passionate about is one feasible strategy that can help students identify pursuits that fit their interests and talents.Another successful strategy has been to require returning second year fellows to share science learning experiences via 15-30 minute long talks for theirpeers. Some took the opportunity to become more immersed in their lab’s focus, while others branched into questions like what motivates scientists towork in foreign countries and what has genomic anthropology told us about human evolution. Project aims were developed through conversationsbetween the fellow and instructor. The one constraint was for fellows to organize their presentations around questions. Feedback indicated thatpresenters benefited from having to give presentations and others enjoyed learning about a broader array of topics.MOOCs: Education for Everyone. Relly Brandman. Course Operations, Coursera.46

CONCURRENT SESSION ABSTRACTSThursday, March 14 3:00 PM–6:00 PMMerrill HallCool Tools for Fungal BiologyCo-chairs: Miguel Penalva and Kevin McCluskeyThe Environmental Molecular Sciences Laboratory molecular analysis capabilities for fungal biology. S. E. Baker. Environmental Molecular SciencesLaboratory, Pacific Northwest Natl Lab, Richland, WA.Tools for analysis of classical and reverse genetic mutants play an important role in fungal biology research. The Environmental Molecular SciencesLaboratory (EMSL) at the Pacific Northwest National Laboratory is a US Department of Energy national user facility. EMSL develops and utilizes cuttingedge mass spectrometry, NMR, imaging and computational capabilities to accelerate research in a number of areas. We have used EMSL’s massspectrometry capabilities to characterize glycosylation of secreted proteins of Aspergillus niger. In addition, we have explored the use of laser ablation andnano-DESI mass spectrometry for spatial localization of molecules associated with Trichoderma reesei mycelium. Finally, spores from wildtype and albinostrains of Aspergillus carbonarius were characterized using helium ion microscopy. As a national user facility, the EMSL is open to the fungal biologycommunity through a competitive, peer-reviewed proposal process.Development and utilization of arrayed mutant sets for yeasts and filamentous fungi. Aric E Wiest, Kevin McCluskey. Fungal Genetics Stock Center,Kansas City, MO.Advancements in high throughput functional genomics has allowed the generation of vastly increasing numbers of strains carrying single gene deletions.For some organisms these include mutations distributed across the genome. The FGSC has generated or acquired sets of arrayed mutants for severaldifferent yeast or filamentous fungal species including Neurospora crassa, Magnaporthe grisea, Cryptococcus neoformans, Candida albicans, Aspergilusnidulans, and Pichia pastoris. These arrayed sets allow rapid screening for desired traits across a broad number of gene deletions. Details of construction,replication and manipulation of these arrayed sets will be presented. Custom arraying, construction of functional sets, and cryopreservation will also bediscussed.Sequencing-based solutions to identify and characterize fungal developmental genes. Minou Nowrousian, Ines Teichert, Gabriele Wolff, Ulrich Kück.Dept. of General & Molecular Botany, Ruhr University Bochum, Bochum, Germany.During sexual development, filamentous ascomycetes form complex, three-dimensional fruiting bodies for the protection and dispersal of spores. We areusing a combination of classical genetics, next generation sequencing, molecular and microscopic methods to learn more about this differentiation processin the model organism Sordaria macrospora, and here we present data on the identification/characterization of transcription factors and signalingmolecules that are involved in development. Whole genome sequencing of mutant pro44 was used to identify the mutation that causes sterility in themutant strain. For Illumina/Solexa sequencing, pooled DNA from progeny of crosses of the mutant with the wild type was used, and we were able topinpoint the causative mutation in the mutant strain through bioinformatics analysis. pro44 carries a mutation in a GATA-type transcription factor, andfertility can be restored by transformation with the wild-type allele. In a second approach, we used laser microdissection to isolate young fruiting bodies(protoperithecia) of the wild type and mutant pro1 that carries a deletion of another transcription factor gene essential for sexual development. Linearamplification of RNA from microdissected protoperithecia yielded enough material for RNA-seq analysis. A comparison with total mycelium revealedsignificant differences in in gene expression between protoperithecia and non-reproductive mycelia. Among the genes strongly up-regulated inprotoperithecia were the pheromone precursor genes ppg1 and ppg2. This was confirmed by fluorescence microscopy of egfp expression under control ofppg1 regulatory sequences. In protoperithecia, many genes are under control of the transcription factor PRO1; thus, by combining laser microdissectionand RNA-seq, we can now perform genome-wide analyses of genes that are dependent on a development-specific transcription factor for correctexpression in a defined developmental structure in fungi. Among the genes that are dependent on PRO1 for correct expression in protoperithecia is pro44,which is among the 500 most strongly expressed genes in wild-type, but not pro1 protoperitheica. In summary, our data indicate that PRO1 and PRO44 aremembers of a transcription factor network that regulates gene expression and cell differentiation in developing fruiting bodies.Aspergillus nidulans as an experimental system to identify novel cell wall growth and maintenance genes through identification of anti-fungal drugresistance mutations. Xiaoxiao Sean He, Shengnan Jill Li, Susan Kaminskyj. Biology, University of Saskatchewan, Saskatoon, Saskatchewan, Canada.Systemic fungal infections are estimated to contribute to ~10% of hospital deaths. Systemic fungal infections are most dangerous for the young, the old,and the already sick, since their immune systems are less vigorous. Most antifungal drugs in current clinical use target ergosterol (polyenes) or theergosterol biosynthetic pathway (azoles and allylamines). Drugs against beta-glucan synthesis (echinocandins) are effective against aspergillosis andcandidaisis. The use of compounds that target fungal enzymes inevitably leads to the development and natural selection of drug resistant fungal strains.Not only are the anti-fungal drugs in current clinical use losing efficacy in some situations, but in addition the high level of conservation between animaland fungal physiology leaves relatively few relevant targets to explore. However, it is likely that for any drug-enzyme combination there will be relativelyfew mutations that could increase drug resistance while still maintaining enzyme function. We are using Aspergillus nidulans as an experimental modelsystem to assess the number and identity of mutations that lead to drug resistance. As proof of concept, we grew wild type A. nidulans on replicate platescontaining a sub-lethal concentration of Calcofluor. These developed fast-growing sectors beginning at ~ 5 d (70 rounds of mitosis). Preliminary resultsshow that many of these sectors harboured heritable, single-gene mutations. To date, mutated genes that confer robust, heritable resistance to Calcofluorthat were identified by next generation sequencing have roles in cell wall synthesis, cell wall integrity regulation, or drug detoxification. We suggest thisstrategy will be useful for predicting genetically-mediated anti fungal resistance adaptation and help us to be ahead in the drug-resistance arms race.27th Fungal Genetics Conference | 47

CONCURRENT SESSION ABSTRACTSIllumina-based genetic linkage map for wheat leaf rust. David L. Joly 1,2 , Barbara Mulock 3 , Christina A. Cuomo 4 , Barry J. Saville 2 , Brent D. McCallum 3 , GuusBakkeren 2 . 1) Pacific Agri-Food Research Centre, Agriculutre and Agri-Food Canada, Summerland, British Columbia, Canada; 2) Forensic Science Programand Environmental & Life Sciences Graduate Program, Trent University, Peterborough, ON, Canada; 3) Cereal Research Centre, Agriculture and Agri-FoodCanada, Winnipeg, MB, Canada; 4) Broad Institute of Massachusetts Institute of Technology and Harvard University, Cambridge, MA 02142.Few genetic maps have been made for rust fungi; yet they are useful in identifying candidate loci for phenotypic traits or in unravelling chromosomalarrangements. This lack of maps is, in part, due to the obligate biotrophic nature of rusts and the difficulties in manipulating their life cycle in a way thatenables controlled crosses. Recently, the genome sequence of a wheat leaf rust (Puccinia triticina) isolate was determined and this prompted thesequencing of additional isolates using next-generation sequencing technologies. This has dramatically increased the amount of sequence informationavailable at a substantially decreased per base cost. Fifty-seven F2 progeny of a wheat leaf rust sexual cross between race 9 (SBDG) and race 161 (FBDJ)were sequenced using Illumina. In order to generate a high-resolution genetic linkage map, genome-wide single-nucleotide polymorphisms (SNPs) wereidentified. Employing the genome sequence information from the two parents and the F1 isolate, more than 25,000 SNPs were selected and used togenerate a genetic linkage map. Although they were obtained from different isolates, the genetic map and the reference genome were integrated,allowing the creation of pseudomolecules. Those represent a strong improvement over the currently fragmented status of the reference genome.Moreover, at least 9 seedling and 2 adult-plant avirulence genes were shown to segregate in this F2 population and candidate genes identified using thegenetic map are currently being investigated.Peering into the secret-ory life of Aspergillus nidulans with a little help from classical genetics. Miguel Penalva 1 , Areti Pantazopoulou 1 , Mario Pinar 1 ,Herbert N. Arst, Jr. 2 . 1) Cellular and Molecular Biology, Centro de Investigaciones Biologicas CSIC, Madrid, Spain; 2) Department of Microbiology, ImperialCollege, London, UK.Model fungi have survived the revolution of modern biology partly through their amenability to classical genetic analysis. Unquestionably, classicalgenetics lay at the root of the unmatched success of the yeast Saccharomyces cerevisiae, that exotic fungal visitor so pleasantly accepted into the parlourof true eukaryotic cells and in the conservatory of gene regulation that dominated the fungal community at the end of last millennium. Formerlyfashionable, classical genetics became nearly extinct with the advent of the ‘omics era’, their demise confirmed with each of the uncountabledevelopments of low-cost sequencing. However, we shall illustrate how extraordinarily powerful classical genetics can be, used in combination withsequencing techniques, to address general questions on the organization of the Golgi in eukaryotic cells. The Golgi is essential for secretion, and therefore,for hyphal growth. Thus, we begin with a sequenced, well-characterized heat-sensitive X ts mutation in an A. nidulans Golgi gene. An X ts strain ismutagenised to isolate suX suppressor mutations, reversing the absence of growth resulting from X ts at the restrictive temperature. Less interestingintragenic reversion/pseudo-reversion events are identified by the inability of any given suX X ts strain to produce single mutant X ts progeny when crossedto a wild-type. These mutations are next sequenced and archived. The remaining extragenic suppressors are allocated to one of the eight A. nidulanschromosomes by parasexual analysis, exploiting the rarity of mitotic recombination. Next, meiotic crosses between the suX X ts strain and a panel ofparental strains carrying markers in the suX chromosome are analysed to detect genetic linkage. Once linkage is detected, suX is further mapped to thesmallest feasible chromosomal interval. Candidate genes in the annotated genome interval, hopefully conspicuous at this stage to the educated eye, or, asa last resort, the whole interval between the genetic boundaries, are sequenced to identify the suppressor. The combination of gene mapping withsequencing eliminates the cumbersome identification of a single causative mutation (aka ‘a needle in a haystack’) hidden amongst the genetic variability ofthe mutant and parental strains, inherent to whole genome sequencing approaches.Domains of meiotic DNA recombination and gene conversion in Coprinopsis cinerea (Coprinus cinereus). Patricia J. Pukkila 1 , Wendy Schackwitz 2 . 1) DeptBiol, Univ North Carolina, Chapel Hill, NC, USA; 2) US DOE Joint Genome Institute, Walnut Creek, CA, USA.We have shown previously that rates of meiotic recombination are highly non-uniform along the assembled chromosomes of C. cinerea (Stajich et al.PNAS 107: 11889-11894, 2010). That study revealed an over-representation of paralogous multicopy genes in regions with elevated levels of meioticexchange. In addition, retrotransposon-related sequences were not found in large segments of the genome with low levels of meiotic exchange. However,the study was limited by the available markers, and only 31 Mb of the 36 Mb genome could be mapped. More recently, we have resequenced 45 meioticsegregants and 4 complete tetrads. We developed a simple script to detect crossover and gene conversion events involving over 75,000 SNPs spanning 35Mb. The data were analyzed using MSTmap (Wu et al. PLoS Genetics 4: e1000212, 2008). The new dataset revealed sub-telomeric recombination hotspotsat every chromosome end, and 36% of the crossovers were associated with uninterrupted tracts of gene conversion. The conversion tracts (2-8 SNPs) werequite short (8-219 nt), and the median distance between the flanking SNP markers was also small (500 nt). Since these subtelomeric hotspots correspondto sites of synaptic initiation in C. cinerea (Holm et al. Carlberg Res. Commun. 46: 305-346, 1981), these data may contribute to our understanding of howhomologous chromosome pairing and synapsis are coordinated with meiotic recombination. Supported by the U.S. Department of Energy Joint GenomeInstitute Community Sequencing Program. The work conducted by the U.S. DOE JGI is supported by the Office of Science of the U.S. Department of Energyunder Contract No. DE-AC02-05CH11231.A Hook protein is critical for dynein-mediated early endosome movement in Aspergillus nidulans. Jun Zhang 1 , Rongde Qiu 1 , Herbert Arst 2 , MiguelPeñalva 3 , Xin Xiang 1 . 1) Department of Biochemistry and Molecular Biology, Uniformed Services University, Bethesda, Maryland, USA; 2) Department ofMedicine, Imperial College London, London, UK; 3) Department of Molecular and Cellular Medicine, Centro de Investigaciones Biológicas CSIC, Ramiro deMaeztu 9, Madrid, Spain.It has been hypothesized that cytoplasmic linker proteins such as CLIP-170 facilitate motor-driven organelle transport by serving as an additional linkerbetween the organelle and the microtubule track. However, mammalian and fungal cells lacking CLIP-170 do not exhibit any apparent defects in vesicletransport. We recently found that in the filamentous fungus Aspergillus nidulans, the HooK protein ortholog, HookA, is critical for dynein-mediatedtransport of early endosomes. HookA mutants were obtained from a genetic screen for mutants defective in dynein-mediated early endosome movement,and the HookA gene was identified by a combination of classical genetic and whole-genome-sequencing approaches. The HookA protein is homologous tohuman Hook proteins containing a N-terminal microtubule-binding domain, a coiled-coil domain and a C-terminal cargo-binding domain, an organizationsimilar to that of CLIP-170. Both the N- and C-terminal domains of HookA are required for dynein-mediated early endosome transport, and HookAassociates with early endosomes via its C-terminal domain in a dynein-independent manner. Importantly, HookA physically interacts with dynein/dynactin,and this interaction is independent of the C-terminal early-endosome-binding domain but dependent upon the N-terminal microtubule-binding domain.Together, our results suggest that HookA may facilitate cargo-motor-track interactions during dynein-mediated transport of early endosomes.48

CONCURRENT SESSION ABSTRACTSThursday, March 14 3:00 PM–6:00 PMChapelFungi and Evolutionary TheoryCo-chairs: Hanna Johannesson and Duur AanenReaching the wind: the fluid mechanics of spore discharge, and potential for dispersal mechanisms to shape the evolution of sporocarp and sporemorphologies. Anne Pringle 1 , Michael Brenner 2 , Joerg Fritz 2 , Marcus Roper 3 , Agnese Seminara 2 . 1) Organismic & Evolutionary Biology, Harvard University,Cambridge, MA; 2) School of Engineering and Applied Sciences, Harvard University, Cambridge, MA; 3) Department of Mathematics, University ofCalifornia, Los Angeles, CA.Fungi play critical roles in human agriculture and Earth’s biogeochemistry, but mechanisms of fungal dispersal are poorly understood. Thinking hasfocused on the passive spread of spores by air and water, and neglected the biomechanics used by fungi to actively move spores to new habitats. In thistalk we focus on terrestrial ascomycetes, a group including plant and animal pathogens, mycorrhizal fungi, and lichens. We build theory to catalog andexplain the morphological features used by ascomycetes to shoot spores and facilitate the crossing of the boundary layer, a sheath of nearly still airsurrounding sporocarps. Crossing the boundary layer is critically important to the fitness of a spore: a spore that cannot escape will fall back on the parentfungus, where probabilities of germination and survival are low. But after crossing the boundary layer, a spore must also travel in wind, and by explicitlymodeling discharge and dispersal we identify a previously unsuspected trade-off constraining the sizes of spores. Large spores cross boundary layers moreeffectively, while small spores are more easily carried by wind. Spore dispersal shapes the epidemiology of disease, and will mediate range shifts inresponse to global change; understanding how and how quickly fungi move across landscapes will enable both management and conservation.Neurospora tetrasperma mating-type chromosomes: Testing hypotheses on the effects of degeneration and introgression on performance. Jennifer L.Anderson, Yu Sun, Pádraic Corcoran, Hanna Johannesson. Department of Evolutionary Biology, Uppsala University, Uppsala, Sweden.Following hybridization between species, parts of one species’ genome can be incorporated into the genome of the other. This transfer of geneticmaterial, introgressive hybridization, is a well-known driver of speciation, diversification, and adaptive evolution. Introgression has occurred repeatedly inthe fungus Neurospora tetrasperma and has resulted in the presence of large regions of DNA (< 4 Mbp tracts) from other species of Neurospora on themating-type (mat) chromosomes of N. tetrasperma. The mat chromosomes of N. tetrasperma also contain large regions of suppressed recombination thatare associated with the accumulation of mutations and possibly a reduction in biological fitness. It has been proposed that introgressions of DNA fromother taxa, with freely recombining mat chromosomes, onto the mat chromosomes of N. tetrasperma could counteract the deleterious effects of mutationaccumulation and “reinvigorate” fitness. Alternatively, interspecific introgression into N. tetrasperma mat chromosomes could be either neutral ordeleterious to fitness, but are maintained due to lack of recombination between mat chromosomes. To test these hypotheses we have quantifiedphysiological performance (linear growth rate, LGR) in homokaryons from eight strains of N. tetrasperma with mat chromosomes that differ inintrogression history (e.g. introgressions from different species) and degree of degeneration. Differences in LGR between mating types and chromosometypes (introgressed or degenerate) will inform our understanding how hybridization and chromosomal structure and content effect physiologicalperformance and possibly fitness.Nuclear arms races: sexual selection for masculine mushrooms. Bart Nieuwenhuis, Duur Aanen. Laboratory of Genetics, Wageningen University,Wageningen, Netherlands.When many gametes compete to fertilize a limited number of compatible gametes, sexual selection will favor those traits that increase competitiveadvantage during mating. In animals and plants, sperm and pollen competition have yielded many interesting adaptations for improved mating success. Infungi, similar processes have not been directly shown yet. We test the hypothesis that sexual selection can increase competitive fitness during mating,using experimental evolution in the mushroom fungus Schizophyllum commune. Mating in S. commune occurs by donation of nuclei to a mycelium. Thesefertilizing ‘male’ nuclei migrate through the receiving ‘female’ mycelium. In our setup, an evolving population of nuclei was serially mated with a nonevolvingfemale mycelium for 20 sexual generations. Four of the 12 tested strains had significantly increased competitive fitness and one had decreasedfitness. The main characteristic that explained fitness change was the relative success in colonization of the female mycelium. In most cases, no trade-offswere found with other fitness components. Our results show that sexual selection can act in mushroom fungi and that sexual selection can lead toincreased competitive ability during mating.Genome-wide mutation dynamic within a long-lived individual of Armillaria. James B. Anderson. Deptartment of Biology,, Univ Toronto, Mississauga,Ontario, Canada.Mutation is the ultimate source of all genetic variation in populations and yet the events themselves remain unobservable and buried in the past. Longlivedindividuals of Armillaria gallica, a common opportunistic fungal pathogen of tree roots in temperate forests of the northern hemisphere, provide aspatial context for the mutational dynamic. Each individual of A. gallica arises in a single mating event between two haploid gametes and the resultingdiploid genotype then grows vegetatively to occupy a discrete spatial territory including many adjacent tree root systems. In effect, this leaves a spatialrecord of growth over time within which mutations can be pinpointed. To identify mutations, the entire genomes of three spatially separated samples ofone individual of A. gallica approximately 200 by 60 m in size were sequenced and compared. In this comparison, mutations and chromosomal regions ofloss of heterozygosity (LOH) were identified and then assayed in another 22 isolates from the same individual.by conventional PCR and Sanger sequencing.The genotype network of all 10 mutations and two LOH events in the 90 MB genome assembly was without internal conflict. Further, the spatialdistribution of genotypes was non-random and appeared to reflect the vegetative expansion leading to the present-day individual. I will discuss theimplications of the whole-genome data in estimating mutation rates and cellular generation times.27th Fungal Genetics Conference | 49

CONCURRENT SESSION ABSTRACTSRapid genetic change and plasticity in arbuscular mycorrhizal fungi is caused by a host shift and enhanced by segregation. C. Angelard, I. Sanders.University of Lausanne, Biophore, 1015 Lausanne, Switzerland.Arbuscular mycorrhizal fungi (AMF) are among the most abundant symbionts of plants, improving plant productivity and diversity. They are clonal; a traitassumed to limit adaptability. However, AMF harbour genetically different nuclei. We hypothesized that AMF can respond rapidly to a change ofenvironment through changes in the frequency of nuclei and by making genetically novel offspring. We subjected AMF parents and offspring to a hostshift. We observed genetic changes in all AMF lines. Genetic and phenotypic responses were different among offspring and some displayed higher fitnessthan their parents. Our results demonstrate that AMF rapidly undergo genetic change in response to the environment and that nucleotype frequency playsa role in how they perform in the new environment. Even though clonal, AMF offspring display greater genetic change and plasticity in response to hostshift. Such genetic and phenotypic flexibility is likely to be key to their ecological success.Meiotic Drive: A Single Gene Conferring Killing and Resistance in Fungal Spore Killer. Pierre Grognet 1,2* , Fabienne Malagnac 1,2 , Hervé Lalucque 1,2 , PhilippeSilar 1,2 . 1) Univ Paris Diderot, Sorbonne Paris Cité, Laboratoire Interdisciplinaire des Energies de Demain, 75205 Paris CEDEX 13 France; 2) Univ Paris Sud,Institut de Génétique et Microbiologie, Bât. 400, 91405 Orsay cedex, France.Meiotic drives (MD) are nuclear genetic loci ubiquitous in eukaryotic genomes that cheat the Mendel laws by distorting segregation in their favor. Allknown MD are composed of at least two linked genes, the distorter that acts as a toxin by disrupting the formation of gametes, and the responder thatacts as an antitoxin and protects from the deleterious distorter effects. In fungi, MDs are known as Spore Killers (SK). In the model ascomycete Podosporaanserina, MD has been associated with deleterious effect during ascospore formation of the Het-s prion and in Neurospora crassa a resistance gene(responder) to the Sk-2 and Sk-3 distorters has been identified. MDs are easily studied in P. anserina thanks to the ascus structure as SKs are identified bythe presence of 2-spored asci in crosses between strains. Here, we identify and characterize by targeted deletion in P. anserina Spok1 and Spok2, two MDelements. We show that they are related genes with both spore-killing distorter and spore-protecting responder activities carried out by the same allele,unlike other known MD. These alleles act as autonomous elements and exert their effects in any region of the genome. Moreover, Spok1 acts as aresistance factor to Spok2 killing. As Spok1 and Spok2 belong to a multigene family, these Spore Killer genes represent a novel kind of selfish genes thatproliferate in population through meiotic distortion.Cryptic population subdivision, sympatric coexistence and the genetic basis of local adaptation in Neurospora discreta. Pierre Gladieux, David Kowbel,Christopher Hann-Soden, John Taylor. Department of Plant and Microbial Biology, University of California, Berkeley, CA.Identifying the genes for ecologically relevant traits is a central challenge in empirical population genetics. Species distributed across strongenvironmental gradients are excellent models to discover and identify the genetic targets of local selection as they are more likely to experience spatiallyheterogeneous selection pressures leading to local adaptation of ecologically important traits. We studied the origin of ecological differentiation in N.discreta phylogenetic species 4 (PS4), a species with a broad latitudinal distribution. We Illumina-sequenced the complete genomes of 52 individualsrepresenting 8 collections sites in Alaska, New Mexico, Washington, California, and Western Europe (average sequencing depth: 52X). Reads were mappedto the N. discreta PS4 reference genomes, and analyses were based on a final set of ca. 1.2 million high-quality SNPs. Phylogenetic analyses identified fourwell-supported clades. Papua New-Guinea individuals formed the most basal clade. Individuals from Alaska and Europe on the one hand, and from NewMexico on the other hand grouped into sister clades, and individuals from California were basal to these two clades. Individuals from Washington, sampledwithin the same site, grouped with either the New Mexico individuals, or the California individuals, indicating the coexistence in sympatry of two divergentpopulations. The observed pattern of population subdivision is being used as a reference to identify genes departing from the genome-wide background,and showing increased divergence consistent with divergent selective pressures, or decreased divergence consistent with gene-flow. Our findingsemphasize the need to continue exploration to uncover divergent populations of Neurospora, and place N. discreta, along with N. crassa, among thehandful of species that have the attributes to serve as outstanding evolutionary and ecological model organisms.Ecological context in symbioses: when is your enemy also your friend? Georgiana May 1 , Paul Nelson 2 . 1) Dept Ecol, Evol, Behavior,#100, Univ Minnesota,St Paul, MN; 2) EEB graduate program University of Minnesota St. Paul MN.Most plants are rife with fungal symbiotic partners with many of these having little apparent effect on the host's health and fitness. In this work, weexplore the degree to which the outcome of interactions between an endophytic fungus, pathogen and plant host depend on ecological context. Inparticular, we ask whether interactions between the endophyte of maize, Fusarium verticilliodes, with the pathogen Ustilago maydis, depend on hostresistance to the pathogen. In the case of a host susceptible to the pathogen, the two fungal species should meet frequently, and compete over hostresources, potentially driving greater virulence to the host in one or the other fungal species. In the case of a host resistant to the pathogen, theendophyte might be a "bystander" to the pathogen, because the two meet too infrequently to drive their co-evolutionary interaction. We show evidencethat the two fungal species have evolved stronger antagonistic interactions in maize susceptible to the pathogen, and further, that this might beassociated with greater virulence by the pathogen. Results of modeling will also be presented from which we predict longer term evolutionary trajectoriesfor this 3-way interaction.50

CONCURRENT SESSION ABSTRACTSThursday, March 14 3:00 PM–6:00 PMHeatherCytoskeleton, Motors, and Intracellular TransportCo-chairs: Samara Reck-Peterson and Ping WangThe molecular basis of extended dynein run-length. Sreedhar Kilaru, Martin Schuster, Gero Steinberg. School of Bioscinces, Univ Exeter, EX4 4QD Exeter,UK.Dynein is a minus-end directed motor that utilises ATP to transport organelles along microtubules. In fungi, a major "cargo" of dynein are earlyendosomes that are taken over long distance from the plus-ends near the growing apex to the central part of the hyphal cell. In cell-free assays it wasshown that single dynein motors can only overcome 1 micrometer, and long-distance motility of organelles requires binding of several dynein motors thatcooperate to extend the transport distance. We recently showed that this does not apply to the fungus Ustilago maydis. Here, single dynein motors moveover 30 micrometers, raising the question of the underlying molecular mechanism for this extraordinary motor performance. This talk will provide acomprehensive explanation for this phenomenon.The role of microtubule-based motors in the spatiotemporal control of autophagy. Martin Egan, Mark McClintock, Samara Reck-Peterson. Cell Biology,Harvard Medical School, Boston, MA.Autophagy is a highly conserved eukaryotic process in which components of the cytoplasm, including damaged organelles and misfolded proteins, aresequestered into double membrane-bound vesicles called autophagosomes that are subsequently delivered to the vacuole for recycling. In fungi,autophagy is linked to cellular remodeling and differentiation, while in mammals dysfunction in the autophagy pathway has been implicated in cancer andneurodegenerative diseases. Here we explore the role of microtubule-based motors in the spatiotemporal control of autophagy in the model filamentousfungus Aspergillus nidulans. Using a molecular genetic and live-cell imaging approach, we identify the motors responsible for autophagosome motility, anddissect their role in the delivery and fusion of autophagic vesicles with the vacuolar system. Furthermore, we examine the role of microtubule-basedmotors in the clearance of aggregation-prone proteins associated with motor neuron disease, and determine the effect of these aggregates on normalmicrotubule-based transport processes.Microtubule-dependent co-transport of mRNPs and endosomes. Sebastian Baumann 1,2 , Thomas Pohlmann 1,2 , Andreas Brachmann 2,3 , MichaelFeldbrügge 1,2 . 1) Heinrich-Heine University Düsseldorf, Institute for Microbiology, 40204 Düsseldorf, Germany; 2) Max Planck Institute for TerrestrialMicrobiolgy, Department of Organismic Interactions, Karl-von-Frisch-Str. 10, 45043 Marburg, Germany; 3) Biocenter of the Ludwigs Maximilians UniversityMunich, Genetics Section, Grosshaderner Str. 2-4, 82152 Planegg-Martinsried, Germany.Long-distance transport of mRNAs is important in determining polarity in eukaryotes. Molecular motors shuttle large messenger ribonucleoproteincomplexes (mRNPs) containing mRNAs, RNA-binding proteins and associated factors along microtubules. However, precise mechanisms including theinterplay of molecular motors and a potential connection to membrane trafficking remain elusive. In recent studies we identified the RNA-binding proteinRrm4 as the key player in microtubule-dependent mRNA transport in Ustilago maydis. Combining in vivo CLIP and RNA-live imaging revealed a subset ofmRNAs that are bound by Rrm4 and transported processively throughout the hyphae. Studying the molecular motors revealed that shuttling is mediatedby Kin3 and Dyn1/2. The same set of motors acts in endosome trafficking and indeed, studying the SNARE Yup1 and the small GTPase Rab5 we found cotransportwith endosomes as a novel mechanism for mRNP transport. Currently, we address the link between mRNAs and endosomes.Role of tea1 and tea4 homologs in cell morphogenesis in Ustilago maydis. Flora Banuett, Woraratanadharm Tad, Lu Ching-yu, Valinluck Michael.Biological Sciences, California State University, Long Beach, CA.We are interested in understanding the molecular mechanisms that govern cell morphogenesis in Ustilago maydis. This fungus is a member of theBasidiomycota and exhibits a yeast-like and a filamentous form. The latter induces tumor formation in maize (Zea mays) and teosinte (Zea mays subsp.parviglumis and subsp. mexicana). We used a genetic screen to isolate mutants with altered cell morphology and defects in nuclear position. One of themutants led to identification of tea4. Tea4 was first identified in Schizosaccharomyces pombe, where it interacts with Tea1 and other proteins thatdetermine the axis of polarized growth. Tea4 recruits a formin (For3), which nucleates actin cables towards the site of growth, and thus, polarizessecretion (Martin et al., 2005). Tea1 and Tea4 have been characterized in Aspergillus nidulans and Magnaporthe oryzae (Higashitsuji et al., 2009; Patkar etal., 2010; Takeshita et al., 2008; Yasin et al., 2012). Here we report the characterization for the first time of the Tea4 and Tea1 homologs in theBasidiomycota. The U. maydis tea4 ORF has coding information for a protein of 1684 amino acid residues that contains a Src homology (SH3) domain, aRAS-associating domain, a phosphatase binding domain, a putative NLS, and a conserved domain of unknown function. All Tea4 homologs in theBasidiomycota contain a RA domain. This domain is absent in Tea4 homologs in the Ascomycota, suggesting that Tea4 performs additional functions in theBasidiomycota. We also identified the Umtea1 homolog, which codes for a putative protein of 1698 amino acid residues. It contains three Kelch repeats.The Tea1 homologs in the Ascomycota and Basidiomycota contain variable numbers of Kelch repeats. The Kelch repeat is a protein domain involved inprotein-protein interactions. The tea1 gene was first identified in S. pombe and is a key determinant of directionality of polarized growth (Mata and Nurse,1997). To understand the function of tea1 and tea4 in several cellular processes in U. maydis, we generated null mutations. We demonstrate that tea4 andtea1 are necessary for the axis of polarized growth, cell polarity, normal septum positioning, and organization of the microbutubule cytoskeleton. We alsodetermined the subcellular localization of Tea1::GFP and Tea4::GFP in the yeast-like and filamentous forms.27th Fungal Genetics Conference | 51

CONCURRENT SESSION ABSTRACTSAspergillus nidulans septin interactions and post-translational modifications. Yainitza Hernandez-Rodriguez 1 , Shunsuke Masuo 2 , Darryl Johnson 3 , RonOrlando 3,4 , Michelle Momany 1 . 1) Plant Biology, University of Georgia, Athens, GA, US; 2) Laboratory of Advanced Research A515, Graduate School of Lifeand Environmental Sciences, University of Tsukuba, Tennodai, Tsukuba, Ibaraki, JP; 3) Department of Chemistry, University of Georgia, Athens, GA, US; 4)Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, US.Septins are cytoskeletal elements found in fungi, animals, and some algae, but absent in higher plants. These evolutionarily conserved GTP bindingproteins form heteroligomeric complexes that seem to be key for the diverse cellular functions and processes that septins carry out. Here we usedAspergillus nidulans, a model filamentous fungus with well defined vegetative growth stages to investigate septin-septin interactions. A. nidulans has fiveseptins: AspA/Cdc11, AspB/Cdc3, AspC/Cdc12 and AspD/Cdc10 are orthologs of the “core-filament forming-septins” in S. cerevisiae; while AspE is onlyfound in filamentous fungi. Using S-tag affinity purification assays and mass spectrometry we found that AspA, AspB, AspC and AspD strongly interact inearly unicellular and multicellular vegetative growth. In contrast, AspE appeared to have little or no interactions with core septins in unicellular stagesbefore septation. However, after septation AspE interacted with other septins, for which we postulate an accessory role. AspE localized to the cortex ofactively growing areas and to septa, and localizations are dependent on other septin partners. Interestingly, core septin localizations can also depend onaccessory septin AspE, particularly post-septation. In addition, LC-MS/MS showed acetylation of lysine residues in AspA before septation and AspC afterseptation. Western blot analysis using an anti-acetylated lysine antibody showed that AspC is highly acetylated in all stages examined, while other septinsshowed acetylation post-septation. Though LC-MS analysis failed to detect phosphorylation of septins, this modification has been widely reported in fungalseptins. Using phosphatase treatments and Western Bloting, we found phosphorylation of AspD, but no other septins. This is interesting because AspDbelongs to a special group of septins that lack a C-terminal coiled-coil found in other septins. However, septin localization is not affected by the absence ofAspD/Cdc10, but by the absence of filamentous fungi specific septin AspE. Our data suggests that septin interactions and modifications change duringdevelopment and growth in A. nidulans, and that some modifications are septin specific.Altered Ras1 trafficking impairs the pathogencity of Cryptococcus neoformans. Connie B. Nichols, Teresa O'Meara, Kyla Selvig, Sandra Breeding, J.Andrew Alspaugh. Dept. of Medicine, Duke University Medical Center, Durham, NC, USA.Cryptococcus neoformans is an opportunistic human fungal pathogen. The ability to cause disease is linked to several different determinants, one ofwhich is the ability to grow at high temperature. Previously we found that one branch of the Ras1 signaling cascade mediates cell morphology andcytokinesis in response to mild stress, such as growth at high temperature. Inactivation of Ras1 and other components of this signaling branch negativelyimpacts C. neoformans pathogenicity. Additionally, this branch of the Ras1 signaling cascade is dependent on the trafficking of Ras1 from theendomembranes to the plasma membrane and is mediated by palmitoylation of the Ras1 protein. We have identified and characterized several C.neoformans protein acyltransferases (PATs), the enzymes responsible for palmitoylation, to further understand the role of palmitoylation and traffickingon Ras1 function and activity during high temperature growth and pathogenesis. Although there is some degree of functional redundancy in this proteinfamily, we identified individual PATs that are required for stress response and virulence in models of cryptococcal disease.Quantification of the thigmotropic response of Neurospora crassa to microfabricated slides with ridges of defined height and topography. KarenStephenson 1 , Fordyce Davidson 2 , Neil Gow 3 , Geoffrey Gadd 1 . 1) Division of Molecular Microbiology, College of Life Sciences, University of Dundee, Dundee,United Kingdom; 2) Division of Mathematics, University of Dundee, Dundee, United Kingdom; 3) Institute of Medical Sciences, University of Aberdeen,Aberdee, United Kingdom.Thigmotropism is the ability of an organism to exhibit an orientation response to a mechanical stimulus. We have quantified the thigmotropic responseof Neurospora crassa to microfabricated slides with ridges of defined height and topography. We show that mutants that lack the formin BNI-1 and theRho-GTPase CDC-42, an activator of BNI-1, had an attenuated thigmotropic response. In contrast, null mutants that lacked cell end-marker protein TEA-1and KIP-A, the kinesin responsible for its localisation, exhibited significantly increased thigmotropism. These results indicate that vesicle delivery to thehyphal tip via the actin cytoskeleton is critical for thigmotropism. Disruption of actin in the region of the hyphal tip which contacts obstacles such as ridgeson microfabricated slides may lead to a bias in vesicle delivery to one area of the tip and therefore a change in hyphal growth orientation. This mechanismmay differ to that reported in Candida albicans in so far as it does not seem to be dependent on the mechanosensitive calcium channel protein Mid1. TheN. crassa Dmid-1 mutant was not affected in its thigmotropic response. Although it was found that depletion of exogenous calcium did not affect thethigmotropic response, deletion of the spray gene, which encodes an intracellular calcium channel with a role in maintenance of the tip-high calciumgradient, resulted in a decrease in the thigmotropic response of N. crassa. This predicts a role for calcium in the thigmotropic response. Our findingssuggest that thigmotropism in C. albicans and N. crassa are similar in being dependent on the regulation of the vectorial supply of secretory vesicles, butdifferent in the extent to which this process is dependent on local calcium-ion gradients.Dynein drives oscillatory nuclear movements in the phytopathogenic fungus Ashbya gossypii and prevents nuclear clustering. S. Grava, M. Keller, S.Voegeli, S. Seger, C. Lang, P. Philippsen. Biozentrum, Molecular Microbiology, University of Basel, CH 4056 Basel, Switzerland.In the yeast Saccharomyces cerevisiae the dynein pathway has a specific cellular function. It acts together with the Kar9 pathway to position the nucleusat the bud neck and to direct the pulling of one daughter nucleus into the bud. Nuclei in the closely related multinucleated filamentous fungus Ashbyagossypii are in continuous motion and nuclear positioning or spindle orientation is not an issue. A. gossypii expresses homologues of all components of theKar9/Dyn1 pathway, which apparently have adapted novel functions. Previous studies with A. gossypii revealed autonomous nuclear divisions and,emanating from each MTOC, an autonomous cytoplasmic microtubule (cMT) cytoskeleton responsible for pulling of nuclei in both directions of the hyphalgrowth axis. We now show that dynein is the sole motor for bidirectional movements. Surprisingly, deletion of Kar9 shows no phenotype. Dyn1, thedynactin component Jnm1, the accessory proteins Dyn2 and Ndl1, and the potential dynein cortical anchor Num1 are involved in the dynamic distributionof nuclei. In their absence, nuclei aggregate to different degrees, whereby the mutants with dense nuclear clusters grow extremely long cMTs. Like inbudding yeast, we found that dynein is delivered to cMT +ends, and its activity or processivity is probably controlled by dynactin and Num1. Together withits role in powering nuclear movements, we propose that dynein also plays (directly or indirectly) a role in the control of cMT length. Those combineddynein actions prevent nuclear clustering in A. gossypii and thus reveal a novel cellular role for dynein.52

CONCURRENT SESSION ABSTRACTSThursday, March 14 3:00 PM–6:00 PMFred Farr ForumNucleic Acid-Protein Interactions that Impact Transcription and TranslationCo-chairs: Michael Freitag and Mark CaddickChIP-seq: an inexpensive and powerful method for studying genome-wide chromatin remodeling and transcription regulation in fungi. Koon Ho Wong,Kevin Struhl. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA.Chromatin Immuno-precipitation (ChIP) is a commonly used technique for studying protein-DNA interactions. When coupled with the Next GenerationSequencing (NGS) technology, ChIP-seq can map and measure genome-wide locations and occupancies of any protein-of-interest at very high resolution,and is an invaluable technique for studying chromatin-associated processes including transcription regulation. However, owing to the fact that NGSexperiments are expensive, this powerful technique has yet been widely applied to fungal studies. The output of current sequencing technologies vastlyexceeds the sequencing depth requirement of ChIP-seq experiments as well as many NGS applications in fungi. We have developed a multiplex sequencingmethod that allows up to 96 different samples to be included in a single sequencing reaction, providing a means to obtain whole-genome data at a highlyaffordable cost. Using multiplex sequencing, ChIP-seq and a technique called Anchor-Away for conditional depletion of proteins from the nucleus, we havegained important insights into different aspects of transcription regulation including the repression mechanism of the Cyc8-Tup1 co-repressor complex inSaccharomyces cerevisiae. Examples on how ChIP-seq applications may be broadly applied to address common questions regarding transcriptionregulation will also be presented.Regulatory Networks Governing Global Responses to Changes in Light and Time. Jay C. Dunlap, Jennifer J. Loros, & the P01 Consortium**"FunctionalAnalysis and Systems Biology of Model Filamentous Fungi". coordinated from Dept Gen, Geisel School of Medicine at Dartmouth, Hanover, NH.**including PIs Deb Bell-Pedersen, Michael Freitag, James Galagan, Matthew Sachs, Eric Selker, Jeff Townsend, and members of their labs at institutionsnot listed here. Free-living fungi live in a profoundly rhythmic environment characterized by daily changes in light intensity and temperature. Some fungihave well described systems for anticipation of temporal change, circadian systems, and nearly all fungi can respond acutely to changes in light intensity.The nuts and bolts of the regulatory structures underlying circadian regulation and responses to blue light are well known in Neurospora. The circadianclock comprises a negative feedback loop wherein a heterodimer of proteins, WC-1 and WC-2, acts as a transcription factor (TF) to drive expression of frq.FRQ stably interacts with a putative RNA helicase (FRH) and with casein kinase 1, and the complex down-regulates the White Collar Complex (WCC). Withappropriate phosphorylation mediated delays, this feedback loop oscillates once per day (Baker, Loros, & Dunlap, FEMS Microbiol. Reviews 36: 95-106,2012). In turn, blue light is detected by FAD stably bound by WC-1, eliciting photochemistry that drives a conformational change in the WCC resulting inactivation of gene expression from promoters bound by the WCC (Chen, Dunlap & Loros, FGB 47, 922-9, 2010). With this as context, the consortium teamlisted above is using the tools of next generation sequencing, recombineering and luciferase reporters to see how the initial simple steps of clock controland light perception ramify via regulatory networks to elicit development in response to the cues of light and time. Interestingly, the same players andnetworks appear to be involved in many places. For instance, the circadian feedback loop yields rhythmic activation of WCC that regulates many genesincluding transcription factors (TFs). Genes encoding TFs that do not affect the circadian feedback loop itself provide circadian output. In this manner theseTFs act as second order regulators, transducing regulation from light responses or from the core circadian oscillator, to banks of output clock-controlledgenes (ccgs), some of which are in turn other TFs. Assembling the global regulatory networks governing light and clock regulation is now a feasible goal.Protein Binding Microarrays and high-throughput real-time reporters studies: Building a four-dimensional understanding of transcriptional networks inNeurospora crassa. A. Montenegro-Montero 1 , A. Goity 1 , C. Olivares-Yañez 1 , A. Stevens-Lagos 1 , M. Weirauch 2 , A. Yang 3 , T. Hughes 3 , L. F. Larrondo 1 . 1)Genética Molecular y Microbiología, Pontificia Universidad Católica de Chile, Santiago, Chile; 2) CAGE, Cincinnati Children`’s Hospital Medical Center,University of Cincinnati. U.S.A; 3) Banting and Best Department of Medical Research, University of Toronto, Canada.It has been suggested that ~20% of the Neurospora-transcriptome may be under circadian control. Nevertheless, there is scarce information regardingthe regulators that are involved in the rhythmic expression of clock-controlled genes (ccgs). We are using a high-throughput platform, based on variouscodon-optimized luciferase transcriptional- and translational-reporters, to monitor time-of-day-specific gene expression and to identify key elementsmediating circadian transcriptional control. Thus, we have identified transcription factors -such as SUB-1- that affect the expression of known and novelccgs, among which there are transcriptional regulators that give access to a group of third-tier ccgs. In addition, we are characterizing several rhythmicbZIP-coding genes as potential nodes of circadian regulation. In order to characterize regulatory networks in which these and all Neurospora transcriptionfactors participate, we are using double-stranded DNA microarrays containing all possible 10-base pair sequences to examine their binding specificities andin that way, predict possible targets on a genome-wide manner. Currently, these Protein Binding Microarray studies have provided DNA-bindingspecificities for over 120 Neurospora transcription factors granting an unprecedented and powerful tool for transcriptional network studies. Finally, wehave generated graphic tools to explore the spatial differences observed in the temporal control of gene expression. Funding: Conicyt/Fondecyt/regular1090513.Ending messages: alternative polyadenylation in filamentous fungi. Julio Rodriguez-Romero, Ane Sesma. CBGP/ Univ Politécnica de Madrid, Pozuelo deAlarcón, Madrid, Spain.The 3' end polyadenylation of pre-mRNAs is a two-step process. First, pre-mRNAs are cleaved at their 3' end. The second step involves the addition of thepolyA tail by RNA polymerases. Presence of multiple potential 3' end cleavage sites is common in eukaryotic genes, and the selection of the right site isregulated during development and in response to cellular cues. This mechanism of alternative (or non-canonical) polyadenylation generates mRNAisoforms with different exon content or 3' UTR lengths and regulates the presence of cis elements in the mRNA. Proteins involved in alternativepolyadenylation (APA) include Cleavage Factor I in metazoans (CFIm), Hrp1 in yeast and Rbp35 in filamentous fungi. The cis elements present in the 3'UTRs such as miRNA target sites modulate gene expression by affecting cytoplasmic polyadenylation, subcellular localization, stability, translation and/ordecay of the mRNA. Therefore, the selection of a proper 3' end cleavage site represents an important step of regulation of gene expression. Using DirectRNA Sequencing (DRS), we are carrying out in the rice blast fungus Magnaporthe oryzae a comprehensive map of genome wide polyadenylation sites and27th Fungal Genetics Conference | 53

CONCURRENT SESSION ABSTRACTSquantifying their usage under different nutritional conditions (rich and minimal media, carbon and nitrogen starvation) in the wild type and the Drbp35mutant. Results of these polyadenylation maps will be presented, including candidate APA targets, sequence motifs present in long 3' UTRs, Rbp35-dependent mRNA isoforms, and conservation of significant mRNA isoforms in other filamentous fungi.Post-transcriptional gene regulation contributes to host temperature adaptation and virulence in Cryptococcus neoformans. Amanda L. MisenerBloom 1,2 , Kurtis Downey 1 , Nathan K. Wool 1 , John C. Panepinto 1,2 . 1) Microbiology/Immunology, SUNY University at Buffalo, Buffalo, NY; 2) Witebsky Centerfor Microbial Pathogenesis and Immunology, SUNY University at Buffalo, Buffalo, NY.In response to the hostile host environment, pathogens must undergo rapid reprogramming of gene expression to adapt to the stresses they encounter.Upon exposure to host temperature, Ribosomal protein (RP) transcripts are rapidly repressed in C. neoformans. We are interested in investigating specificmechanisms involved in this response, as this repression may be a critical process in host temperature adaptation. Using a mutant null of the majordeadenylase, Ccr4, we have discovered that this repression is in part due to enhanced degradation of RP-transcripts. Ccr4 lacks a nucleic acid bindingdomain and therefore must be recruited to mRNA targets via RNA binding proteins. Using MEME analysis and chromatographic techniques, we haveidentified a shared cis element in the 3’UTR of RP transcripts that is recognized by the zinc knuckle protein, Gis2. We are currently investigating theimportance of this protein-RNA interaction in the expression of RP genes.Host temperature-induced enhanced degradation of RP transcripts is also dependent on the dissociable RNA polymerase II subunit, Rpb4. Specifically, wedemonstrated that in an rpb4D mutant, RP-transcript deadenylation is impaired, suggesting that Rpb4 may be required for Ccr4-targeted degradation. Inaddition, we observed that upon a shift to 37°C, Rpb4 travels from the nucleus to the cytoplasm, supporting a role for Rpb4 in coupling transcription anddegradation. Interestingly, this coupling is not restricted to the RP transcripts, as Rpb4 is also involved in enhanced decay of ER stress transcripts followingtheir peak induction, one hour after a shift to host temperature. We have demonstrated that signaling through PKH enhances the degradation of the RPtranscriptsin response to host temperature, but not the ER stress transcripts, highlighting the complexity of this system. We report that whentranscription and degradation are uncoupled by the loss of Rpb4, growth at host temperature is impaired and virulence in a mouse model of disseminatedcryptococcosis is attenuated. Our data suggests that coupling of transcription and degradation via Rpb4 allows the cell to control the intensity andduration of different responses at specific times following exposure to host temperature, contributing to the ability of C. neoformans to adapt to thisstress.Dual targeting of glycolytic enzymes by alternative splicing and translational read-through. Johannes Freitag, Julia Ast, Alina Stiebler, Michael Bölker.Department of Biology, Philipps-Universität Marburg, Marburg, Germany.Processing of mRNA is a highly conserved process in eukaryotes involving three major steps. Nascent transcripts are capped at their 5’end, introns areremoved by splicing and the 3’end is cleaved and polyadenylated. In the plant pathogenic fungus Ustilago maydis, several genes show hallmarks ofdifferential splicing and alternative polyadenylation resulting in the production of C-terminally extended proteins. We detected that this process leads togeneration of an extended glyceraldehyde-3-phosphate dehydrogenase (GAPDH) isoform harboring a C-terminal peroxisomal targeting sequence (PTS1).We could also detect peroxisomal isoforms of two further glycolytic enzymes, phosphoglycerate kinase (PGK) and triosephosphate isomerase (TPI).Remarkably, peroxisomal isoforms of PGK and TPI are generated by translational read-through in U. maydis. Further analysis revealed that dual targetingof glycolytic enzymes to peroxisomes and the cytoplasm is not restricted to U. maydis but occurs in a variety of fungal species. Interestingly, in differentspecies variable mechanisms to generate extended peroxisomal isoforms of glycolytic enzymes are operating. In the ascomycete Aspergillus nidulans thePTS1-motif of PGK is derived from alternative splicing and polyadenylation, while translational read-through is used to generate a peroxisomal isoform ofGAPDH. We could also show that some enzymes are partially targeted to peroxisomes by means of weak peroxisomal targeting signals. Dual localization ofglycolytic enzymes to peroxisomes and the cytoplasm appears to be widespread in fungi. This indicates that fungal peroxisomes are endowed with a morecomplex metabolism than previously assumed. Thus, the consideration of alternative splicing and translational read-through will be of importance infuture proteomic and metabolomic studies of organelles.Non-optimal codon usage determines the expression level, structure and function of the circadian clock protein FREQUENCY. Mian Zhou 1 , Jinhu Guo 5 ,Joonseok Cha 1 , Michael Chae 1 , She Chen 2 , Jose Barral 3 , Matthew Sachs 4 , Yi Liu 1 . 1) Department of Physiology, UT Southwestern Medical Center, Dallas, TX;2) National Institute of Biological Sciences, Beijing, China; 3) Departments of Neuroscience and Cell Biology and Biochemistry and Molecular Biology, TheUniversity of Texas Medical Branch, Galveston, TX; 4) Departments of Biology, Texas A&M University, College Station, TX; 5) School of Life Sciences, SunYat-sen University, Guangzhou, China.Codon usage bias has been observed in the genomes of almost all organisms and is thought to result from selection for efficient and accurate translationof highly expressed genes 1-3. In addition, codon usage is also implicated in the control of transcription, splicing and RNA structure 4-6. Many genes,however, exhibit little codon usage bias. The lack of codon bias for a gene is thought to be due to lack of selection for mRNA translation. Alternatively,however, non-optimal codon usage may also have biological significance. The rhythmic expression and the proper function of the Neurospora FREQUENCY(FRQ) protein are essential for circadian clock function. Here, we show that, unlike most genes in Neurospora, frq exhibits non-optimal codon usage acrossits entire open reading frame. Optimization of frq codon usage results in the abolition of both overt and molecular circadian rhythms. Codon optimizationnot only increases FRQ expression level but surprisingly, also results in conformational changes in FRQ protein, impaired FRQ phosphorylation, andimpaired functions in the circadian feedback loops. These results indicate that non-optimal codon usage of frq is essential for its circadian clock function.Our study provides an example of how non-optimal codon usage is used to regulate protein expression levels and to achieve optimal protein structure andfunction.A transcriptome-wide view on microtubule-dependent mRNA transport. Carl Haag 1 , Julian Konig 2 , Kathi Zarnack 3 , Michael Feldbrugge 1 . 1) Institut forMicrobiology, Heinrich-Heine University, Düsseldorf, NRW, Germany; 2) MRC LMB Cambridge, UK; 3) EBI Hinxton, UK.Long distance transport of mRNAs regulates spatio-temporal gene expression during polar growth. In filaments of U. maydis, for example, microtubuledependentshuttling of mRNAs is crucial to determine the axis of polarity. The key component of this transport system is the RNA-binding protein Rrm4that binds a distinct set of target mRNAs. Recently, we discovered a novel mechanism for mRNA transport, namely the co-transport of Rrm4 andassociated mRNAs with endosomes. Here, new insights on mRNA transport will be presented using the improved in vivo UV-crosslinking technique: iCLIP.This technique allows identification of target mRNAs at the transcriptome-wide level with single nucleotide resolution.54

CONCURRENT SESSION ABSTRACTSThursday, March 14 3:00 PM–6:00 PMKilnInteractions between Fungi and AnimalsCo-chairs: Neil Gow and Clarissa NobileElicitation of host damage occurs in a temporally programmed manner during Aspergillus fumigatus infections. Elaine M. Bignell. Microbiology Section,Imperial College London, London, United Kingdom.Background: In tissue-invasive lung infections caused by the mould Aspergillus fumigatus the molecular basis of host damage remains unclear. It has longbeen hypothesised that the secretion of proteolytic enzymes by invading A. fumigatus hyphae provides a mechanism by which epithelial damage ismediated. However, in whole animal studies of disease it has not been possible to substantiate an important role of fungal proteases since A. fumigatusmutants lacking individual or multiple enzyme functions retain the ability to cause fatal infections. One of the first cellular lines of defence against A.fumigatus infection is the monolayer of epithelial cells which line the mammalian airway. Epithelial cells provide a physical barrier against endothelialinvasion and initiate an inflammatory immune response upon contact with A. fumigatus spores. Here we show that the A. fumigatus pH-responsivetranscription factor, PacC, which governs expression of secreted proteases and secondary metabolism genes, is required for invasion of the murinepulmonary epithelium, and pathogenicity. Results: We determined, via murine and epithelial infection assays, that DpacC mutants are defective inelicitation of early-phase host damage which occurs, in wild type isolates, via a novel contact-dependent mechanism. Transcriptomic analyses of murineaspergillosis revealed aberrant cell wall biosynthesis in infecting DpacC isolates, suggesting a novel role for the A. fumigatus cell wall in pathogen-mediatedhost damage. Concordant with these findings PacC null mutants were shown to have signficiantly heightened chitin content in the fungal cell wall andwere hypersensitive to cell wall perturbing agents, including caspofungin. The mechanistic relevance of cell wall-mediated host damage was verified bycomparative analysis of damage elicited by cell wall extracts and heat-killed hyphae from wild type and DpacC isolates. Conclusion: A. fumigatus elicitshost damage in a biphasic manner, initally via a novel contact-dependent mechanism involving cell wall components, and later via soluble mediators. A.fumigatus mutants deficient in the pH-responsive transcription factor PacC suffer deficits in both mechanisms. On the basis of this functionaltranscriptomic analysis we propose a new model of biphasic host damage during A. fumigatus infections.Exploiting innate recognition of fungi for vaccine development. Stuart Levitz. Medicine, University of Massachusetts, Worcester, MA.Most licensed vaccines work by promoting protective antibody responses. However, some populations, such as the elderly and theimmunocompromised, generally have poor antibody responses to conventional vaccines. Moreover, for many infectious and neoplastic diseases, vaccinesthat arm adaptive T cell responses appear necessary. Thus, a major challenge in vaccinology is the development of platforms and adjuvants that effectivelypromote protective T cell and antibody responses. The immune system has evolved to innately recognize components of the fungal cell wall, particularly b-glucans. Research in my laboratory, in collaboration with Gary Ostroff, has focused on how this innate recognition of the fungal cell wall can be exploitedfor vaccine development. To achieve this aim, we have used glucan particles (GPs) as a novel vaccine platform. GPs are hollow, highly purifiedmicrocapsules prepared from Saccharomyces cerevisiae cell walls. GPs are composed predominantly of b-1,3-glucan and are recognized by b-glucanreceptors (particularly Dectin-1) on dendritic cells and other phagocytes. GPs also potently activate complement, resulting in opsonization and recognitionby complement receptors. GPs can be loaded with antigens and immunomodulators such that the “payload” is released following phagocytosis. We havedemonstrated robust and long-lasting antigen-specific T cell (Th1- and Th17-biased) and antibody responses following immunization of mice with GPs“encapsulated” with antibody. Moreover, vaccination of mice with GPs loaded with fungal antigens can protect mice against lethal challenges with thepathogenic fungi Cryptococcus neoformans and Histoplasma capsulatum.Regulatory circuits governing Candida albicans proliferation in a mammalian host. Jose C. Perez 1 , Carol A. Kumamoto 2 , Alexander D. Johnson 1 . 1)Microbiology and Immunology, UCSF, San Francisco, CA; 2) Molecular Biology and Microbiology, Tufts University, Boston, MA.The fungus Candida albicans resides in the gastrointestinal tract of most, if not all, human adults and is also a leading cause of life-threatening fungalinfections in immunocompromised individuals. C. albicans has no known environmental reservoir suggesting that it has extensively co-evolved to thrive inits host. To uncover the C. albicans gene circuits governing its proliferation in a host, we used mouse models of intestinal colonization and systemicinfection to screen a set of ~75 transcription regulator deletion strains. These mutant strains were chosen because they showed no gross phenotypeswhen cultured under a variety of laboratory growth conditions. We identified eight transcription regulators that play roles in intestinal colonization,systemic infection or both. Through genome-wide chromatin immunoprecipitation and transcriptional profiling experiments, we determined the targetgenes and the general circuitry controlled by these regulators. Our results reveal multiple biological functions necessary for C. albicans to inhabit amammalian host, the acquisition of carbon and nitrogen sources being prominent among them. These findings highlight common challenges faced bybacterial and eukaryotic (fungal) species when colonizing the mammalian intestine and illustrate how evolution has tinkered with the C. albicansregulatory circuitry to meet these demands.Dramatic ploidy change as an adaptive strategy in Candida albicans... Meleah A. Hickman, Ben Harrison, Darren Abbey, Anja Forche, Carsten Paulson,Kathleen Matter, Judith Berman. Dept Gen, Cell Biol & Dev, Univ Minnesota, Minneapolis, MN.For over 100 years, Candida albicans has been considered an obligate diploid, although it clearly tolerates single chromosome aneuploidy as well as longtracts of homozygosity. We recently identified tetraploid, triploid as well as intriguing reductions to below diploid C. albicans cells, some from the clinic,others from a mouse host and others following stress exposure in vitro. Tetraploidy arises either through parasex (mating between diploid cells) or defectsin mitosis. Stress conditions, including exposure to the antifungal drug flucanozole, increase the frequency of tetraploid formation. The polyploid state isrelatively unstable even under standard laboratory conditions and loss of a heterozygous marker increases by an order of magnitude as compared todiploid populations. A small subset of tetraploid cells return to a near diploid state very rapidly even without exposure to the stresses usually used toinduce concerted chromosome loss. The diploid derivatives of polyploid cells exhibit a wide range of chromosome aneuploidies and homozygosities, thusgenerating a wide range of genetic diversity within a single population. Evolution experiments with fluconazole suggest that diploid cells undergo transientpolyploidization in response to fluconazole and that polyploid cells adapt to stress conditions more rapidly.27th Fungal Genetics Conference | 55

CONCURRENT SESSION ABSTRACTSNematode-trapping fungi eavesdrop on nematode pheromones. Yen-Ping Hsueh 1 , Parag Mahanti 2 , Frank Schroeder 2 , Paul Sternberg 1 . 1) Howard HughesMedical Institute and Division of Biology, California Inst of Technology, Pasadena, CA; 2) Boyce Thompson Institute and Department of Chemistry andChemical Biology, Cornell University, Ithaca, NY.The recognition of molecular patterns associated with specific pathogens or food sources is fundamental to ecology and plays a major role in theevolution of predator-prey relationships. Recent studies showed that nematodes produce an evolutionarily highly conserved family of small molecules, theascarosides, which serve essential functions in regulating nematode development and behavior. Here we show that nematophagous fungi, naturalpredators of soil-dwelling nematodes, can detect and respond to ascarosides. Nematophagous fungi use specialized trapping devices to catch andconsume nematodes, and previous studies demonstrated that most fungal species do not produce traps constitutively but rather initiate trap-formation inresponse to their prey. We found that ascarosides, which are constitutively secreted by many species of soil-dwelling nematodes, represent a conservedmolecular pattern used by nematophagous fungi to detect prey and trigger trap formation. Ascaroside-induced morphogenesis is conserved in severalclosely related species of nematophagous fungi and occurs only under nutrient-deprived condition. Our results demonstrate that microbial predatorseavesdrop on chemical communication among their metazoan prey to regulate morphogenesis, providing a striking example of predator-prey coevolution.We anticipate that these findings will have broader implications for understanding other inter-kingdom interactions involving nematodes, whichare found in almost any ecological niche on Earth.A morphogenesis regulator controls cryptococcal neurotropism. Xiaorong Lin 1 , Bing Zhai 1 , Karen Wozniak 2 , Srijana Upadhyay 1 , Linqi Wang 1 , ShupingZhang 3 , Floyd Wormley 2 . 1) Biology, Texas A&M University, TAMU-3258, TX; 2) Biology, the University of Texas at San Antonio, San Antonio, Texas, USA; 3)Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, Texas, USA.Cryptococcus neoformans is the major causative agent of cryptococcal meningitis, a disease that is responsible for more than 600,000 deaths each year.This ubiquitous environmental pathogen enters host lungs through inhalation and typically establishes asymptomatic latent infections. However,extrapulmonary dissemination often occurs in individuals with weakened immunity and Cryptococcus has a predilection to infect the brain. Braininfections are fatal and formidable to treat due to the poor penetration of most antifungals to the brain. Unfortunately, little is known about cryptococcalfactors that control its neurotropism. Here we report that a morphogenesis regulator Znf2 controls the tissue tropism of cryptococcal infection. Inparticular, activation of Znf2 abolishes Cryptococcus extrapulmonary dissemination and consequently leads to the absence of fatal brain infections in theinhalation infection model. Although Znf2 overexpression strains are avirulent in this animal model, these strains are capable of proliferating in the animallungs during the early stages of infections. Histological examinations and cytokine profiling revealed that the Znf2 overexpression strain causes enhancedmonocyte infiltration in the animal lungs. Consistently, the Znf2 overexpression strain stimulates pro-inflammatory host responses while suppressesdeleterious Th2 host responses during early stage of infection in the pulmonary infection model. Such protective host defense responses might haveprevented the extrapulmonary dissemination of Cryptococcus. In the intravenous infection model where the lung infection was bypassed and there wasuniform hematogenous dissemination, the Znf2 overexpression strain showed a specific defect in the brain infection. Taken together, our data indicatethat Znf2 helps polarize the host immunity towards protection and that it mediates cryptococcal tissue tropism during infection.Sit and wait: Special features of Aspergillus terreus in macrophage interactions and virulence. M. Brock 1 , I.D. Jacobsen 2 . 1) MicrobialBiochemistry/Physiology, Friedrich Schiller University and Hans Knoell Institute, Jena, Germany; 2) Molecular Pathogenicity Mechanisms, Hans KnoellInstitute Jena, Germany.While Aspergillus fumigatus is known as the main cause of invasive pulmonary aspergillosis in immunocompromised patients, Aspergillus terreus is anemerging pathogen prevalent in some local hot spots. When tested in embryonated egg or murine infection models A. terreus required substantiallyhigher infectious doses compared to A. fumigatus to cause high mortality rates. Furthermore, when A. fumigatus and A. terreus infections were followedby in vivo imaging using bioluminescent reporter strains, germination and tissue invasion of A. terreus was significantly delayed. To elucidate differences inmore detail, the interaction of A. terreus and A. fumigatus with macrophages was compared. A. terreus was phagocytosed significantly faster, whichappears mainly due to higher exposure of galactomannan and glucans on the surface of conidia. Additionally, although phagocytosis of both speciesresulted in phagolysosome maturation, A. fumigatus efficiently inhibited acidification, which was not the case for A. terreus. However, within this acidicenvironment of phagolysosomes A. terreus showed long-term persistence without significant inactivation of conidia. Further analyses revealed thatinefficient blocking of acidification by A. terreus was due to differences in the spore colour pigment of both species. Recombinant production of anaphthopyrone synthase from Aspergillus nidulans enabled A. terreus to inhibit the acidification to a similar extent as observed for A. fumigatus. Thisalteration of the phagolysosomal environment resulted in an increased escape from macrophages and was accompanied by increased virulence in amurine infection model. We speculate that the long-term persistence of A. terreus wild-type strains in acidified phagolysosomes might be responsible forhigh dissemination rates observed in infected human patients, because A. terreus might hitchhike inside immune effector cells to reach secondary sites ofinfection.56

CONCURRENT SESSION ABSTRACTSThe mutational landscape of gradual acquisition of drug resistance in clinical isolates of Candida albicans. Jason Funt 1 , Darren Abbey 7 , Luca Issi 5 , BrianOliver 3 , Theodore White 4 , Reeta Rao 5 , Judith Berman 6 , Dawn Thompson 1 , Aviv Regev 1,2 . 1) Broad Institute of MIT and Harvard, 7 Cambridge Center,Cambridge, MA 02142; 2) Howard Hughes Medical Institute, Department of Biology, Massachusetts Institute of Technology, 77 Masscahusetts Ave,Camridge, MA 02140; 3) Seattle Biomedical Research Institute, Seattle, WA; 4) School of Biological Sciences, University of Missouri at Kansas City, MS; 5)Worcester Polytechnic Institute, Department of Biology and Biotechnology, 100 Institute Road, Worcester MA 01609; 6) Tel Aviv University, Ramat Aviv,69978 Israel; 7) University of Minnesota, Minneapolis MN 55455 USA.Candida albicans is both a member of the healthy human microbiome and a major pathogen in immunocompromised individuals1. Infections are mostcommonly treated with azole inhibitors of ergosterol biosynthesis. Prophylactic treatment in immuncompromised patients2,3 often leads to thedevelopment of drug resistance. Since C. albicans is diploid and lacks a complete sexual cycle, conventional genetic analysis is challenging. An alternativeapproach is to study the mutations that arise naturally during the evolution of drug resistance in vivo, using isolates sampled consecutively from the samepatient. Studies in evolved isolates have implicated multiple mechanisms in drug resistance, but have focused on large-scale aberrations or candidategenes, and do not comprehensively chart the genetic basis of adaptation5. Here, we leveraged next-generation sequencing to systematically analyze 43isolates from 11 oral candidiasis patients, collected sequentially at two to 16 time points per patient. Because most isolates from an individual patientwere clonal, we could detect newly acquired mutations, including single-nucleotide polymorphisms (SNPs), copy-number variations and loss ofheterozygosity (LOH) events. Focusing on new mutations that were both persistent within a patient and recurrent across patients, we found that LOHevents were commonly associated with acquired resistance, and that persistent and recurrent point mutations in over 150 genes may be related to thecomplex process of adaptation to the host. Conversely, most aneuploidies were transient and did not directly correlate with changes in drug resistance.Our work sheds new light on the molecular mechanisms underlying the evolution of drug resistance and host adaptation.27th Fungal Genetics Conference | 57

CONCURRENT SESSION ABSTRACTSThursday, March 14 3:00 PM–6:00 PMNautilusFungal Volatiles and Organic Compounds as Signaling AgentsCo-chairs: Joan Bennett and Richard SplivalloFungi reacting to rhizobacterial volatiles. Birgit Piechulla, Piyali Das, Uta Effmert. University of Rostock, Rostock, Germany.Microorganisms, similar as other organisms are able to synthesize and release volatile organic compounds (VOCs), which are responsible forcharacteristic blends or aromas of for example foodstuff such as wine and cheese as well as spoiled meat. The capability of microorganisms to emitcomplex volatile mixtures is tremendous. More than 800 volatiles are presently known that are emitted by microorganisms (database of volatiles ofmicroorganisms DOVE-MO). Beside the wealth of volatile emissions, to date not much is known about the biological functions of these compounds. Tostudy volatile-mediated interactions of plant associated bacteria and fungi, various rhizobacteria and phytopathogenic fungi were co-cultivated in bipartitePetri dishes, which allow only volatiles to traverse from one to the other compartment. The volatiles of Serratia, Stenotrophomonas, Pseudomonas,Burkholderia and Staphylococcus inhibited the growth of Aspergillus, Fusarium, Microdochium, Neurospora, Rhizoctonia, Phaecilomyces, Penicillium,Phoma, Sclerotinia, Trichoderma and Verticillium in species specific manner. The reactions of Sclerotinia scleotiorum to Serratia sp. 4Rx13 volatiles werestudied in more detail, e.g. radial growth, biomass formation, catalase activity and lipid peroxidation. Furthermore, the volatile mixture of Serratia sp.4Rx13 was studied using headspace collection systems and GCMS analysis. Ca. 100 volatiles were separated, some of them were identified, most of themremain unknowns or structures have to be elucidated. References: Kai et al. (2007) Arch. Microbiol. 187:351-360 Vespermann et al. (2007) Appl. Environ.Microbiol. 73:5639-5641 Kai et al. (2010) Appl. Microbiol. Biotechnol. 88:965-976 Effmert et al. (2012) Chem. Ecol. 38:665-703.Enhancement of plant growth and stress resistance by Fusarium volatile organic compounds: A novel mechanism mediating plant-fungal interactions.Seogchan Kang 1,3 , Vasileios Bitas 1,3 , Nate McCartney 2,3 , Jim Tumlinson 2,3 . 1) Plant Pathology & Environmental Microbiology, Pennsylvania State Univ,University Park, PA; 2) Entomology, Pennsylvania State Univ, University Park, PA; 3) Center for Chemical Ecology, Pennsylvania State Univ, University Park,PA.Every organism employs an elaborate network of signaling pathways for sensing stimuli from surrounding environments and neighboring organisms andtranslating them into specific molecular and cellular responses. Production and perception of a vast array of secreted proteins and metabolites plays keyroles in this mechanism. A group of secreted molecules that are ubiquitous but often overlooked is volatile organic compounds (VOCs). VOCs can travel farfrom their point of production through the atmosphere as well as porous soils, making them ideal signaling molecules for mediating organismalinteractions without physical contact. Roles of animal- and plant-derived VOCs in directing animal behaviors and roles of plant VOCs in chatters of “talkingtrees” are well known and serve critical roles in diverse ecological processes. In contrast, the available knowledge of microbial VOCs as semiochemicals islimited and mostly circumstantial. Multiple isolates of Fusarium oxysporum, a soil-borne, cosmopolitan fungus that often resides in the rhizosphere ofmany plants, produce unknown VOCs that drastically enhance the growth and stress resistance of Arabidopsis thaliana. Other Fusarium species alsopromoted Arabidopsis growth. Molecular and cellular changes underpinning the Fusarium VOC-mediated signaling will be discussed. Given the vastdiversity of fungi in nature and the critical importance of fungal communities for the ecology and fitness of plants, VOC-mediated signaling is a mostlyuncharted frontier, waiting for systematic exploration.The Role of Quorum-sensing Molecules in Interactions between Candida albicans and its Host. Jessica C. Hargarten 1 , Thomas M. Petro 2 , Kenneth W.Nickerson 1 , Audrey L. Atkin 1 . 1) School of Biological Sciences, University of Nebraska, Lincoln, Lincoln, NE; 2) Department of Oral Biology, University ofNebraska Medical Center, Lincoln, NE.Candida albicans is a polymorphic fungus that is capable of causing the life threatening disease Candidiasis once it reaches the bloodstream of asusceptible host. The capability to switch between morphologies, and its ability to synthesize and secrete the quorum sensing molecule (QSM) farnesol areknown virulence factor. Previously, we showed that C. albicans mutants that produced less farnesol are less pathogenic to mice than their parental strainin a tail vein assay. Also, oral administration of farnesol to the mice prior to infection increased mortality. In contrast, farnesol blocks the yeast to myceliatransition in vitro, which should have a protective effect. These observations pose the dilemma of finding a mechanism whereby a molecule which blocksthe yeast to mycelia transition can also act as a virulence factor. We hypothesize that farnesol functions as a virulence factor by modulating the hostinnate immune response. Distinct Candida morphologies elicit different host immune responses. Both white and opaque cells stimulate leukocytemovement, but only white cells secrete a small molecular weight chemoattractant that draws the leukocyte directly towards the white cell and stimulatesengulfment by mouse macrophages. The white cells are also less susceptible to killing by human macrophages and neutrophils than opaque cells, possiblydue to their increased capabilities of escape once phagocytosed. The chemical identity of this chemoattractant is currently unknown, but the reasonbehind its continued secretion by white cells is intriguing. One likely candidate is farnesol because opaque cells, unlike white cells, do not accumulatedetectable levels of farnesol. Macrophages are capable of detecting and responding to exogenous farnesol. Earlier our group reported that farnesolstimulates the expression of both pro-inflammatory and regulatory cytokines by mouse macrophage. The production of these warning signals is animportant indicator of how the body ultimately hopes to clear the infection. Others have shown that farnesol suppresses the anti-Candida activity ofmacrophages through its cytotoxic effects, thus making it all the more difficult to eliminate the fungus early in infection. Here we report the in vitro role offarnesol and other known QSM in macrophage chemotaxis and relative phagocytosis of C. albicans.58

CONCURRENT SESSION ABSTRACTSInnate Immunity in Fusarium graminearum. Vong shian Simon Ip Cho 1,2 , Gitte Erbs 3 , Thomas Sundelin 3 , Peter Busk 4 , Mari-Anne Newman 3 , Stefan Olsson 1 .1) Genetics and Microbiology, University of Copenhagen, Copenhagen, Denmark; 2) USDA-ARS Cereal Disease Laboratory, University of Minnesota, SaintPaul, MN, USA; 3) Transport Biology, University of Copenhagen, Copenhagen, Denmark; 4) Dept. Biotechnology, Aalborg University, Copenhagen,Denmark.Fungi are often mostly recognized as plant pathogens that cause harm to important economical plants. In nature however, fungi are frequently victims ofbacterial parasitism but little is known about fungal defense mechanisms. The potential existence of fungal innate immunity was studied using Fusariumgraminearum as model organism and bacterial flagellin to mimic the presence of bacteria in an in vitro environment. The presence of flagellin triggered aninitial mitochondrial and cell membrane hyperpolarization which was detected using the florescent dye DiOC 7(3). This was followed by the production ofthe secondary signalling molecule Nitric Oxide (NO), common to innate immunity signalling in other eukaryotes. NO was monitored using the fluorescentdye DAF-FM. NO appears to be produced by an inducible enzyme that is regulated by complex mechanisms but centrally modulated byCalcium/Calmodulin. Inhibition studies suggest the presence of a Nitric Oxide Synthase (NOS), but no typical arginine utilizing NOS was identified withinthe F. graminearum’s genome by homology search. Various genes bearing resemblance to the archetypal NOS, as well as argininosuccinate lyase weredeleted. However, the mutants still produced NO. The presence of alternative pathways contributing towards the production of NO was investigated byadding a variety of potential substrates to challenged cultures. Various reactions were observed suggesting that several pathways are present. Inconclusion, F. graminearum reacts strongly to the presence of the bacterial Microbial Associated Molecular Pattern (MAMP) flagellin with an up-regulationof NO production showing the presence of innate immunity-like responses also in fungi.The Trichoderma reesei polyketide synthase gene pks1 is necessary for yellow-green pigmentation of conidia and is involved in the establishment ofenvironmental fitness. Lea Atanasova 1 , Benjamin P. Knox 2 , Christian P. Kubicek 1 , Scott E. Baker 2 , Irina S. Druzhinina 1 . 1) Microbiology Group, Research AreaBiotechnology and Microbiology, Institute of Chemical Engineering, Vienna University of Technology, 1060 Vienna, Austria; 2) Chemical and BiologicalProcess Development Group, Pacific Northwest National Laboratory, Richland, WA, USA.The economically important genus Trichoderma (Hypocreales, Ascomycota, Dikarya) is well known for its mycotrophic lifestyle and for the broad range ofbiotrophic interactions with plants and animals. Moreover it contains several cosmopolitan species characterized by their outstanding environmentalopportunism. These properties have given rise to the use of several species in agriculture as biopesticides and biofertilizers while T. reesei is applied forproduction of bioenergy-related enzymes. The molecular basis of the opportunistic success of Trichoderma is not yet well understood. While there is someevidence for a role of secreted enzymes and proteins, less is known about a possible role of secondary metabolites. Recently it was predicted that the PKSencoding gene pks1 from T. reesei and its orthologues are most likely responsible for the characteristic yellow-green pigmentation of conidia. To reveal thefull function of the gene we deleted it from the wild-type strain QM 6a what resulted in complete loss of the green coloration of conidia. Theecophysiological profiling of Dpks1 showed that the gene is also involved in multiple functions at different stages of the T. reesei life cycle. Testing theantagonistic antifungal potential of the T. reesei Dpks1 mutant against several host/prey fungi suggested that the loss of pks1 reduced the ability tocombat them by means of both mechanisms: the pre-contact inhibition and direct overgrowth. However the overall analysis of mycoparasitic interactionssuggests that the gene is most likely involved in protection against other fungi rather than in attacking them. Interestingly, we noticed the increasedproduction of volatile compounds by the Dpks1 strains. The phenotype microarrays showed that PKS1 encoding gene restricts T. reesei from conidiation ona number of the best utilized carbon sources but does not influence the sexual development except the alteration of stromata pigmentation. The data fortranscriptional response of genes putatively involved in above mentioned processes will be presented.Semiochemicals and signaling: plant responses to Trichoderma volatile organic compounds. Richard Hung. Plant Biology, Rutgers, The State University ofNew Jersey, New Brunswick, NJ.Volatile organic compounds (VOCs) produced by Trichoderma viride have recently been shown to have plant growth promoting effects on Arabidopsisthaliana. This finding adds a new facet to the multiple methods which fungi in the genus Trichoderma promote plant health and are beneficial to humans.Both above and below ground growth was greater in A. thaliana exposed to naturally produced T. viride VOCs as compared to controls. The average rootmass of control plants was 0.36g and the average mass of VOC exposed plants was 0.77g showing a 113% increase in plant mass. In addition there was a60% increase in chlorophyll concentration (5.5mg/g control, 8.8mg/g test). GCMS analysis of the VOCs produced by T. viride has resulted in 51 identifiedcompounds. Several compounds from the GCMS data were chosen to determine the effects of individual compounds on the health of A. thaliana. Thecompound trans-2-octenol at concentrations of 1ppm caused decreased dry weight (14% less than control) and extended root length (16% longer thancontrol), indicative of stress. At 1 and 10ppm, the compound 2,5-dimethylfuran, which has been reported to be produced by Trichoderma but was notfound in the aforementioned GCMS analysis, caused only visual differences. The exposed A. thaliana had extended stems as compared to controls but noother differences. In summary, the individual compounds of the T. viride volatile profile that were tested, did not promote plant growth.Identification of chemoattractant compounds from tomato root exudate that trigger chemotropism in Fusarium oxysporum. El Ghalid Mennat, DavidTurra, Antonio Di Pietro. Departamento de Genética, Universidad de Córdoba, 14071 Córdoba, Spain.Fusarium oxysporum is a soilborne pathogen that causes vascular wilt disease on a wide range of plant species, including tomato (Solanum lycopersicum).The host signals that trigger fungal infection are currently unknown. A chemotropic response of F. oxysporum towards tomato root exudate was observedusing a plate assay that measures directed growth of fungal germ tubes towards chemoattractants. To purifiy the chemoattractant coumpound(s) fromtomato root exudate, we applied a series of purification methods including extraction with organic and inorganic solvents, fractionation by size exclusionand ion exchange chromatography. The compound(s) showing chemoattractant activity were found in the hydophilic fraction, had a molecular weightbetween 30 and 50 kDa and were sensitive to boiling and treatment with proteinase K, suggesting that they correspond to one or several secreted tomatoproteins. Polyacrylamide gel electrophoresis of the active fraction revealed multiple protein bands of the expected size, two of which displayedchemoattractant activity when eluted from the gel. Identification of the active protein(s) by LC-ESI-MS is currently ongoing. Identification of the secretedchemoattractant(s) from tomato roots will advance our understanding of the molecular events that trigger fungus-root interactions.27th Fungal Genetics Conference | 59

CONCURRENT SESSION ABSTRACTSThe mixed fungal and bacterial origin of truffle aroma. Richard Splivallo 1 , Aurélie Deveau 2 , Nayuf Valdez 1 , Nina Kirchhoff 1 , Pascale Frey-Klett 2 , PetrKarlovsky 1 . 1) Molecular Phytopathology and Mycotoxin Research, Georg-August University of Goettingen, Grisebachstrasse 6, Germany; 2) UMR1136INRA Université de Lorraine "Interactions Arbres/Micro-organismes", Labex ARBRE, IFR110 EFABA, Centre INRA of Nancy, 54280 Champenoux, France.Truffles are symbiotic ectomycorrhizal fungi which develop on plant roots. Their fruiting bodies are highly appreciated by humans for their aroma, whichtypically comprises twenty to fifty volatiles per truffle species. The biosynthetic routes leading to characteristic truffle volatiles have not yet been fullycharacterized. By similarity to yeasts, volatile synthesis in truffles most likely involves amino acid and fatty acid catabolism. Truffle fruiting bodies furthercontain a diverse microbial community which might be able to generate volatiles or biotransform volatile-precursors on its own. Our aim was toinvestigate the formation of sulphur containing volatiles in truffles, because sulphur volatiles play a major role in the truffle ecology and are determinantof their quality (for humans). We demonstrate that sulphur volatiles characteristic of the white truffle T. borchii are actually produced by bacteriacolonizing truffle fruiting bodies. Under laboratory bioassays, sulphur containing compounds (thiophenes volatiles) resulted from the transformation bybacteria of non-volatile precursor(s) into volatiles. Interestingly in our assays thiophene volatiles were detectable only from bacteria and not from trufflemycelium, while other compounds such as dimethyl disulfide were detectable from both organisms. This indicates that some volatiles might be producedby both truffle mycelium and bacteria, but thiophene volatiles most likely originate from bacteria only. Characterization of the bacterial population byFluorescence In Situ Hybridization highlighted that the concentration of thiophene volatiles correlated with the bacterial density inside fruiting bodies. Thisgives further ground to the bacterial origin of thiophene volatiles. Additionally the production of thiophene volatiles was suppressed upon treating trufflefruiting bodies with antibacterial or antifungal agents, suggesting that the precursors of thiophene volatiles might be synthesized by both fungi andbacteria. These unexpected results disprove the earlier belief that truffles were able to synthesize their aroma on their own. They add a new dimension toplant-fungal interactions by highlighting the importance of the bacterial community associated to truffle fruiting bodies.60

CONCURRENT SESSION ABSTRACTSThursday, March 14 3:00 PM–6:00 PMScrippsGenomics and Biochemistry of Degradation of Complex Molecules in the EnvironmentCo-chairs: Jonathan Walton and Dan CullenFungal transcriptome as database for proteome and refinement tool of gene annotation. K. Igarashi 1 , C. Hori 1 , M. Ishiguro 1 , Y. Uemura 2 , A. K. Takeda 2 , S.Kaneko 3 , M. Samejima 1 . 1) University of Tokyo, Tokyo, Japan; 2) Genaris, Inc., Kanagawa, Japan; 3) National Food Research Institute, Ibaraki, Japan.So far, wood-rotting basidiomycetes, such as white-rot and brown-rot fungi, are the organisms known to grown on wood. They produce various enzymesto outside of their cell, extracellular part of the mycelia, in order to degrade major components of plant cell wall such as cellulose, hemicellulose andlignin. There are many enzymes, which can be utilized for the biomass conversion, in those fungi, as well as the proteins helping and/or accelerating thedegradation of the plant cell wall. Therefore, combination of correct annotation of these genes and the proteome analysis of the extracellular enzymes arequite important for biomass utilization. In the present study, we have cultivated the white-rot basidiomycetes Flammulina velutipes (Enoki-take, wintermushroom) and Phanerochaete chrysosporium in various biomass-degrading culture, and the transcriptome databases were constructed by sequencing ofthe cDNA library using 454 sequencer. In F. velutipes, we identified 19 novel biomass-degrading enzymes including 12 carbohydrate-active enzymes(CAZymes) by 2-dimentional gel electrophoresis of extracellular proteins from cellulose-grown culture, using the transcriptome data as a referencesequence. In the case of P. chrysosporium, the transcriptome sequence data was also used to improve the gene annotation, and more than 1,000 genesare newly annotated by the algorithms refined by cDNA sequences. The improvement of gene annotation caused accurate prediction of introns andshowed unique monodispersed distribution of intron length in this fungus.Developmental regulation and cellulase gene expression in Trichoderma reesei. Irina S. Druzhinina 1,2 , Razieh Karimi-Aghcheh 1 , Lea Atanasova 1 , ChristianP. Kubicek 1,2 . 1) Microbiology Group, Institute of Chemical Engineering, Vienna, Austria; 2) Austrian Center of Industrial Biotechnology, c/o Institute ofChemical Engineering, Vienna University of Technology, Vienna, Austria.We have recently shown that expression of cellulase and hemicellulase encoding genes in Trichoderma reesei (teleomorph Hypocrea jecorina) isobligatorily dependent on the function of the protein methyltransferase LAE1. Its orthologue in Aspergillus nidulans, LaeA, is a part of the VELVET proteincomplex consisting of LaeA, VeA and VelB that regulates secondary metabolism and sexual reproduction. Here we have investigated a possible role ofVEL1, the T. reesei orthologue of A. nidulans VeA, in expression cellulase genes and the development of the fungus. The T. reesei vel1 gene is not expressedin the darkness and is expressed at a relatively low level under illumination. Deletion of the vel1 locus causes a complete loss of conidiation and essentialalteration in sexual development such as loss of formation of perithecia. Overexpression of vel1 under the constitutive expression signals of tef1 did notenhance conidiation in light or darkness. However it led to irregular formation of infertile perithecia in the darkness. Deletion of vel1 did not affectcellulase gene expression, but vel1 overexpression strongly enhanced it. Consistent findings were also obtained for expression of xylanase and b-xylosidasegenes. The stimulation of cellulase gene expression by overexpressing vel1 was dependent on a functional lae1 gene. Our data show that VEL1 controlsphotoinduced sexual development and light-independent conidiation. In addition, while vel1 overexpression stimulates cellulase gene expression, isdispensable for this process and its action is therefore different from that of LAE1.Parallel losses of genes associated with saprotrophy in ectomycorrhizal Agaricomycotina lineages. D. Floudas 1 , L. Nagy 1 , A. Kohler 2 , A. Kuo 3 , I. Grigoriev 3 ,F. Martin 2 , D. Hibbett 1 . 1) Biology, Clark Univ, Worcester, MA; 2) Lab of Excellence ARBRE, Tree-Microbes Department, INRA-Nancy, 54280 Champenoux,France; 3) DOE Joint Genome Institute, Walnut Creek, CA.Mushroom forming fungi (Agaricomycotina) play pivotal roles in the cycling of nutrients in terrestrial ecosystems. Agaricomycotina exhibit diverselifestyles including saprotrophs and symbionts, such as mutualistic ectomycorrhizas. Previously, as part of the Saprotrophic Agaricomycotina Project (SAP),we performed analyses of fungal genomes focusing on wood decayers, which suggested that white rot is the plesiomorphic nutritional strategy ofAgaricomycetes and emerged 300 million years ago at the end of Carboniferous era. Our analyses also suggested that the brown rot mechanism and themycorrhizal lifestyle of Laccaria bicolor have emerged from white rot ancestors. The transitions from white rot to brown rot have taken place several timesin Agaricomycotina and were accompanied by losses of genes encoding enzymes involved in lignin and crystalline cellulose degradation. A similar patternwas reconstructed for the transition from a saprotrophic towards a mycorrhizal lifestyle in L. bicolor, which was the first mycorrhizal species in theAgaricomycotina to have its genome sequenced. However, L. bicolor represents only one of many ectomycorrhizal lineages recognized acrossAgaricomycotina. Here, we present data from eleven newly sequenced mycorrhizal genomes of Agaricomycotina, generated under the auspices of theMycorrhizal Genomes Initiative (MGI), in addition to 8 new genomes of decayers from the SAP. With these new genomes in hand, we are able to explorehow the emergence of mycorrhizal lifestyles is associated with changes in numbers of genes encoding enzymes involved in degradation of plantbiopolymers. The results suggest that ectomycorrhizal lifestyles have emerged multiple times from both white rot and brown rot ancestors inAgaricomycotina. The transitions to the ectomycorrhizal lifestyle show parallelism in gene losses between L. bicolor and other mycorrhizalAgaricomycotina lineages. However, patterns of retention of genes encoding lignocellulolytic enzymes vary across ectomycorrhizal lineages. For example,cellobiohydrolases, which are involved in the degradation of crystalline cellulose, have been retained in several mycorrhizal lineages. The results suggestthat the emergence of ectomycorrhizal lineages in Agaricomycotina has been associated with different degrees of reduction of their saprotrophic ability.27th Fungal Genetics Conference | 61

CONCURRENT SESSION ABSTRACTSCo-expression analysis of Phanerochaete carnosa during growth on hardwood and softwood species to predict proteins with unknown functionrelevant to biomass conversion. Hitoshi Suzuki 1 , Chi Yip Ho 2 , Kin Chan 2 , Philip Wong 1 , Yunchen Gong 1 , Elisabeth Tillier 1 , Emma Master 1 . 1) University ofToronto, Toronto, Ontario, Canada; 2) Mount Sinai Hospital, Toronto, Ontario, Canada.Softwood is the predominant form of land plant biomass in the Northern hemisphere and is among the most recalcitrant biomass resource to bioprocesstechnologies. The white rot fungus Phanerochaete carnosa has been isolated almost exclusively from softwoods, while most known white-rot species,including the model fungus Phanerochaete chrysosporium, were mainly isolated from hardwoods. Growth studies of P. carnosa and P. chrysosporium onsapwood and heartwood from deciduous and coniferous species revealed comparable growth of P.carnosa on all wood samples, while P. chrysosporiumgrew poorly on heartwood from conifers. A contributing factor to growth on extractive-rich heartwood samples could be the comparatively high numberof P450 monooxygenases encoded by P. carnosa. Notably, genome sequencing revealed that P. carnosa possesses one of the largest P450 contingents(239 P450s) among the sequenced and annotated wood-rotting basidiomycetes. However, like most sequencing efforts, a significant fraction of the P.carnosa genome comprises genes that encode proteins with unknown function. Moreover, transcripts from several of these genes were identified inmycelia collected at a single time point from P. carnosa cultivations growing on woody biomass. Accordingly, the aim of the current study was to analyzeco-expression patterns of known and unknown genes to identify those with unknown function that might be most relevant to biomass conversion. Ourapproach was to separately cultivate P. carnosa on ball-milled trembling aspen (Populus tremuloides) and ball-milled white spruce (Picea glauca) and tocollect mycelia at five time points over a one-month cultivation period. RNA collected from all cultures at each time point was sequenced separately usingthe Illumina HiSeq platform. Co-expression patterns will be described and used to predict new gene products that are particularly interesting to target fordetailed biochemical characterization.Functional Analysis of the Pleurotus ostreatus Manganese-Peroxidase Gene Family. Tomer Salame, Doriv Knop, Dana Levinson, Oded Yarden, YitzhakHadar. Microbiology and Plat Pathology, Hebrew Unversity, Rehovot, Israel.Mn amendment to P. ostreatus cultures enhances degradation of recalcitrant aromatic compounds. Manganese peroxidase (MnP) isoenzymes are keyplayers in these processes. The MnP gene family is comprised of five Mn -dependent peroxidases (mnp3, 6, 7, 8 and 9) and four versatile-peroxidases(mnp1, 2, 4 and 5; VPs). In liquid medium, Mn amendment resulted in a drastic up-regulation of the predominantly expressed mnp3 and mnp9, and downregulationof mnp4. To obtain direct evidence for the role of these enzymes, we produced genetically-modified (knockout, knockdown and/or overexpression)strains in mnps and studied their degradation capacity. The compounds studied were: azo-dyes such as orange II and reactive black,recalcitrant pharmaceutical compounds found in treated waste water such as Carbamazepine and lignocellulosic agricultural waste. We engineered atransformant, constitutively expressing mnp4 a VP naturally repressed by Mn (designated OEmnp4) under the control of the b-tubulin promoter. Now,despite the presence of Mn in the medium, OEmnp4 produced mnp4 transcript as well as VP activity as soon as four days after inoculation. OEmnp4decolorized the azo-dyes two days earlier relative to the wild type in Mn amended medium. RNAi silencing targeting mnp3 resulted in a delay in thedecolorization capacity which occurred concomitantly along with a marked reduction of the expression level of all mnps, particularly mnp3 and mnp9. Thisobservation supported the conclusion that MnPs are involved in the process but could not determine the specific contribution of the different genes to theoutcome. Therefore we produced a Dku80 strain, exhibiting a 100% homologous DNA recombination rate, to enable specific gene replacement.Subsequently, homokaryon mnp2, 3, 4 and 9 knockout strains were produced. In Mn amended GP, orange II decolorization was not significantly inhibitedby any of these strains, indicating on functional redundancy. In Mn deficient GP, inactivation of mnp4 proved that it encodes the key VP responsible forMn dependent and Mn independent peroxidase activity, as well as resulted in reduction of the azo dye reactive black 5 decolorization capacity. The toolsand protocols developed increase the amenability of P. ostreatus to genetic manipulations and expand options for gene function analyses.Carbon source and light dependent regulation of gene clusters in Trichoderma reesei (Hypocrea jecorina). Doris Tisch 2 , Monika Schmoll 1 . 1) Health andEnvironment, Bioresources, Austrian Institute of Technology AIT, Tulln, Austria; 2) Vienna University of Technology, Institute of Chemical Engineering,Vienna, Austria.Trichoderma reesei (anamorph of Hypocrea jecorina) is one of the most prolific producers of plant cell wall degrading enzymes. Regulation of the genesencoding these enzymes occurs in response to the nutrient sources available in the environment and many of them are responsive to light as well.Cellulose as the natural substrate induces the most complete enzyme set, while induction of cellulases also occurs on sophorose and lactose. In contrast,no cellulases are induced on glycerol and the respective genes are repressed on glucose. We therefore investigated the transcriptome on these five carbonsources in light and darkness and aimed to identify genes specifically expressed under cellulase inducing conditions. These conditions are characterized bya significant enrichment of genes involved in C-compound and carbohydrate degradation and transport among the upregulated gene set. Genes downregulatedunder inducing conditions show a significant enrichment in amino acid metabolism and energy metabolism. We were further interested whetherlight dependent regulation is clustered in the genome and if the carbon source is relevant for activation of light dependent clusters. We found that lightdependent clustering predominantly occurs upon growth on cellulose, with the most significant regulation in a gene cluster comprising env1. This clusterappears on glucose as well, but is not down regulated in mutants of blr1 or blr2. Also cbh2, the arabinofuranosidase gene abf2 and the histoneacetyltransferase gene gcn5 are part of light dependent clusters. Hierarchical clustering of gene expression patterns was performed to reveal functionaldivergence of gene regulation with respect to light response or carbon specific regulation. Glycoside hydrolase genes follow the whole transcriptomepattern with carbon source being superior to light in terms of regulation. ENV1 in in part the G-protein beta subunit GNB1 were found to be crucial forcarbon source specific regulation of G-protein coupled receptors, genes involved in secretion, sulphur metabolism and oxidative processes as well astransporters. We conclude that clustered regulation of light responsive genes preferentially occurs upon growth cellulose and that ENV1 and to a lesserextent GNB1 play a role in carbon source dependent regulation of specific gene groups in light.62

CONCURRENT SESSION ABSTRACTSGenome-wide analysis of eleven white- and brown-rot Polyporales provides insight into mechanisms of wood decay. Chiaki Hori 1,2 , Kiyohiko Igarashi 1 ,David Hibbett 3 , Bernard Henrissat 4 , Masahiro Samejima 1 , Dan Cullen 2 . 1) Graduate School of Agricultural and Life sciences, University of Tokyo, Tokyo,Japan; 2) Forest Products Laboratory, USDA, Madison, WI; 3) Biology Department, Clark University, Worcester, MA; 4) CNRS, Marseille, France.Many efficient wood decay fungi belong to the Polyporales, and these can be categorized as white-rot fungi or brown-rot fungi, based on decay patterns.White-rot fungi degrade cell wall polysaccharides such as cellulose and hemicellulose as well as the more recalcitrant phenylpropanoid polymer, lignin. Incontrast, brown-rot fungi depolymerize the polysaccharides but the modified lignin remains in the wood. Comparative analysis of white- and brown-rotgene repertoires and expression profiles have revealed substantial variation but considerable uncertainty persists with respect to precise mechanisms.Addressing this issue, we performed genome-wide analysis of carbohydrate-active enzymes (CAZy) and some oxidative enzymes related to polysaccharidesdegradation in eleven white- and brown-rot fungi. This analysis included classifying and enumerating genes from three recently sequenced polyporalesBjerkandera adusta, Ganoderma sp. and Phlebia brevispora. Furthermore, comparative secretomic analysis of seven Polyporales grown on wood culturewere conducted. Summarizing, the average number of genes coding CAZy in the genomes of white-rot fungi was 373, significantly more than the 283observed in brown-rot fungi. Notably, white-rot fungi have genes encoding cellulase and hemicellulase such as those belonging to glycoside hydrolase (GH)families 6, 7, 9 and 74, whereas these are lacking in genomes of brown-rot polyporales. White-rot genes encoding oxidative enzymes potentially related tocellulose degradation such as cellobiose dehydrogenase (CDH), polysaccharides monooxygenase (PMO, formerly GH61), cytochrome b562 with cellulosebindingmodule, are also increased relative to brown-rot fungi. Indeed, secretomic analysis identified GH6, GH7, CDH and PMO peptides only in white-rotfungi. Overall, these results show that, relative to brown rot fungi, white rot polyporales maintain greater enzymatic diversity supporting lignocelluloseattack.Transcription factor shuttling during cellulase induction in Trichoderma reesei. Alex Lichius, Christian P. Kubicek, Verena Seidl-Seiboth. Institute ofChemical Engineering, Vienna University of Technology, Vienna, Austria.For economically feasible production of liquid fuels and other value-added compounds from lignocellulosic plant material, strategies are required toboost cellulolytic and hemicellulolytic enzyme production by industrially relevant fungi. One promising approach is to modulate the transcriptional controlmediating release from carbon catabolite repression (CCR) and induction of cellulase, hemicellulase and xylanase gene expression. To better understandthe underlying molecular dynamics during induction, we characterized nucleo-cytoplamic shuttling of the two transcription factors carbon cataboliterepressor 1 (CRE1) and xylanase regulator 1 (XYR1) of Trichoderma reesei by means of live-cell imaging. In submerged cultures, nuclear import and exportof CRE1 upon repression and induction, respectively, occurred within minutes and therefore was generally faster than shuttling of XYR1. Under CCRconditions XYR1 expression levels were very low, and its nuclear signal required up to one hour to significantly increase upon replacement into an inducingcarbon source. Cultured directly under inducing conditions, nuclear accumulation of XYR1 was detectable after about 20h post inoculation, and stronglyincreased within the following 24 hours. CRE1 under the same conditions was localized exclusively to the cytoplasm. In plate cultures, nuclear recruitmentof CRE1 and XYR1 differed within the central area, the subperiphery and the periphery of the colony depending on the provided carbon source. Mostinterestingly, under inducing conditions we found evidence for increased nuclear recruitment of CRE1 in the central area, correlating with strong nuclearimport of XYR1 in the same region. Notably, the cytoplasmic signal of CRE1 was usually elevated in leading hyphae, whereas XYR1 was never significantlyrecruited to the colony periphery. Taken together our data provide the first temporal resolution of transcription factor shuttling during the induction ofcellulase gene expression in Trichoderma reesei, and reveal some interesting differences between the subcellular localization of CRE1 and XYR1 insubmerged and plate cultures, respectively. These differences indicate that the mycelial organization during fungal growth might be another importantregulatory element to consider for the industrial scale production of cellulolytic enzymes.27th Fungal Genetics Conference | 63

CONCURRENT SESSION ABSTRACTSFriday, March 15 3:00 PM–6:00 PMMerrill HallPathogenic Signaling via Effector ProteinsCo-chairs: Brett Tyler and Sebastien DuplessisDissecting nuclear immunity using Arabidopsis downy mildew effector as probes. Marie-Cecile Caillaud 1 , Lennart Wirthmueller 1,2 , Shuta Asai 1 , SophiePiquerez 1 , Georgina Fabro 1,3 , Jonathan Jones 1 . 1) The Sainsbury Laboratory, Norwich, United Kingdom; 2) John Innes Centre, Norwich, United Kingdom; 3)Present address: CIQUIBIC-CONICET, Universidad Nacional de Cordoba, Argentina.An important role in plant defence has been attributed to nuclear dynamics, since a growing number of reports reveal that the nuclear localization of keycomponents of plant immunity is essential for disease resistance. Recent studies suggest that effectors may manipulate host transcription or other nuclearprocess for the benefit of the pathogen. However, the specific mechanisms by which these effectors promote susceptibility remain unclear. Theinteraction between Arabidopsis and Hyaloperonospora arabidopsidis (Hpa) has been studied intensively during the past twenty years, and it has becomeone of the most well-understood model systems to help us understand pathogen effector biology and the plant immune system. The recent identificationof 15 nuclear-localized Hpa effectors (HaRxLs) provides a powerful tool to dissect plant nuclear immunity. When stably expressed in planta, nuclear-HaRxLs cause diverse developmental phenotypes which highlight their interferences with fundamental plant regulatory mechanisms. Remarkably, nuclearHaRxLs-plant targets are often transcriptional regulators, which may act in complex with immunity co-factors. Here, we report recent insights into ourunderstanding of the arms race between obligate pathogen and its host.The mutualistic fungus Laccaria bicolor uses the effector protein MiSSP7 to alter host jasmonate signaling and establish symbiosis. Claire Veneault-Fourrey 1 , Jonathan Plett 1,3 , Yohann Daguerre 1 , Aurélie Deveau 1 , Annegret Kohler 1 , Jennifer Morrell-Falvey 2 , Annick Brun 1 , Francis Martin 1 . 1) UMR1136IaM_INRA/UHP, Lorraine Univ / INRA, Lab of Excellence ARBRE, Nancy, France; 2) Oak Ridge National Laboratory, Oak Ridge, TN 37831-6422, USA; 3)Hawkesbury Institute for the Environment, University of Western Sydney, Australia.Roots of most trees form a nutrient-acquiring symbiosis with mutualistic fungi. Mycorrhiza-induced Smal Secreted protein MiSSP7, a fungal effectorprotein necessary for the mutualistic interaction between of the ectomycorrhizal fungus Laccaria bicolor and Populus spp. host trees, is secreted by thefungus in contact with plant tissues and is taken up via endocytosis into plant cells where it localizes to the nucleus and targets plant transcription throughan unknown mechanism Here we demonstrate that MiSSP7 interacts with the jasmonic acid receptorJAZ6 of Populus trichocarpa and that PtJAZ6 interactswith a number of other nuclear localized proteins that likely form a DNA binding complex. MiSSP7 is able to block jasmonic acid signaling in both L. bicolorhost and non-host plants, likely through its interaction with a jasmonate receptor. Loss of MiSSP7 expression in L. bicolor can be complemented bytransgenically varying the transcription of PtJAZ6 or through inhibiting jasmonic acid biosynthesis in poplar roots. We conclude that MiSSP7, in contrast toarbuscular mycorrhizal fungi and pathogenic bacteria that promote jasmonate signaling to colonize host tissues, is a novel effector used to promotemutualism by blocking jasmonic acid signaling. In addition to MiSSP7, L. bicolor expresses other MiSSPs to communicate with its host-plant. In particular,we demonstrate that MiSSP8 an apoplastic effector is required for symbiosis.Plett JM, et al. (2011) Curr Biol . 21:1197-1203.Identification and characterization of an RXLR-like effector family from medically relevant fungi. Shiv D. Kale 1* , Kelly C. Drews 1,2 , Helen R. Clark 1,3 , HuaWise 1,4 , Vincenzo Antignani 1 , Tristan A. Hayes 1,2 , Christopher B. Lawrence 1,2 , Brett M. Tyler 4,5 . 1) Virginia Bioinformatics Institute, Virginia Tech., Blacksburg,VA; 2) Department of Biological Sciences, Virginia Tech., Blacksburg, VA; 3) Department of Biochemistry, Virginia Tech., Blacksburg, VA; 4) Center forGenome Research and Biocomputing, Oregon State University, Corvallis, OR; 5) Department of Botany and Plant Pathology, Oregon State University,Corvallis, OR.Fungal infections have become an increasingly significant problem for immunocompromised individuals, transplant recipients, the elderly, several casesinvolving healthy individuals. There is a significant growth in incidences of morbidity and mortality associated with medically important fungi, specificallyAspergillus species. Aspergillus fumigatus virulence has been attributed to production of pigments, adhesins on the surface of the cell wall, secretedproteases, and mycotoxins. Current treatments consist of oral corticosteroids, antifungal medications, and/or surgery to remove aspergillomas. Many ofthese treatments have substantial shortcomings. Detection and diagnosis is also weighty problem as most clinical tests take weeks for results allowing theinfection to proceed. Appropriately, the paradigm for human fungal interactions has been focused on the host deficiencies mediating virulence ofopportunistic pathogenic fungi. There has been substantial progress in identifying and characterizing secreted proteins (effectors) from bacterial,oomycete, and fungal plant pathogens. A subset of these effector proteins are able to enter host cells and modulate host intracellular functions. Using ourbioinformatics pipeline we have been able to identify a family of secreted proteins from A. fumigatus sharing a conserved N-terminal RXLR-like motif. Wefound this family is expanded amongst primary fungal pathogens. The RXLR and RXLR-like motifs from known intracellular effectors of plant pathogenicand mutualistic oomycetes and fungi have been shown to facilitate effector entry into plant cells via binding external phosphatidylinositol-3-phosphate(PI3P). Here we describe AF2, a candidate effector from A. fumigatus that contains a N-terminal RxLR-like motif. Through the use of confocal microscopyand flow cytometry we show AF2 is rapidly able to enter several primary and immortalized mammalian cell lines. Through the use of isothermal titrationcalorimetry and liposome binding assays we show AF2 has nanomolar binding affinity for PI3P, and does not bind other mono or poly-PIPs that we havetested thus far. Based on our bioinformatics and biochemical analysis we postulate AF2 is a secreted effector protein capable of rapidly translocating intomammalian cells. We will present our latest findings on the physiological relevance of AF2.64

CONCURRENT SESSION ABSTRACTSIdentification and functional assay of Phytophthora sojae avirulence effectors. Yuanchao Wang, Suomeng Dong, Weixiao Yin. Plant Pathology Dept,Nanjing Agri Univ, Nanjing, China.Phytophthora sojae is a notorious oomycete pathogen producing a great loss on global soybean production annually. The disease outcome betweensoybean and P. sojae depends on whether hosts could recognize pathogen avirulence effectors. Recently identified oomycete avirulence effectors arecharacterized by N-terminal host entry motif (RxLR motif), sequence and transcriptional polymorphisms between virulent and avirulent strains. Benefitfrom 454 genome sequencing and solexa transcriptome sequencing of P. sojae strains, eight RxLR effectors are bioinformatically identified, geneticmapping suggested that two of them perfectly matched Avr3b and Avr1d phenotype respectively. Transient expression of the ORF from avirulence strainon soybean specifically triggered Rps3b and Rps1d mediated program cell death, respectively. confirming that they encodes avirulence effector Avr3b andAvr1d. Transient expression of Avr3b and Avr1d on Nicotiana benthamiana could promote the infection of Phytophthora capasici, suggesting bothavirulence effectors could suppress plant immunity and contribute to pathogen infection. Silencing of Avr3b impaired the virulence of Phytophthora sojae.Our progress in elucidating the mechanism under the inhibiting plant immunity by these effectors will be presented.Fungal lipoxygenases: a novel instigator of asthma? Gregory J. Fischer 1 , Katharyn Affeldt 3 , Erwin Berthier 2 , Nancy P. Keller 1,2,3 . 1) Department of Genetics,University of Wisconsin-Madison, Madison, WI; 2) Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, WI;3) Department of Bacteriology, University of Wisconsin-Madison, Madison, WI.Statement of Purpose: Fungi have long been associated with asthmatic diseases, yet the exact mechanism(s) by which fungi induce asthma is unknown.We propose that fungal lipoxygenase enzymes and their eicosanoid products are involved in asthmatic diseases. Human 5-lipoxygenase derivedleukotrienes induce inflammation, mucus secretion, vasodilation, and bronchial constriction. We hypothesize that the fungal pathogen Aspergillusfumigatus is capable of secreting a 5-lipoxygenase homolog, LoxB, that participates in eicosanoid production, including leukotrienes. This secretedhomolog is translocated into lung epithelial cells, participates in the production of leukotriene and other eicosanoids, and exacerbates asthmaticresponses, such as bronchoconstriction. Together, this work will help delineate the role fungal products play in asthmatic diseases. Methods: We areassessing fungal interactions with lung epithelial cells using a microfluidic in-vitro platform followed by murine asthma model research. To assess theeffects of LoxB overexpression, mass spectrometry was used to identify eicosanoid oxylipins within culture supernatants. Results: We have identified anAspergillus fumigatus lipoxygenase, LoxB, with high identity to human 5-lipoxygenase. Moreover, we have identified a motif in LoxB that may mediateentry into lung epithelial cells. To fully understand the impact of LoxB in asthma, we have developed an Aspergillus fumigatus strain that overexpressesLoxB. Overexpression of LoxB results in increased levels of various eicosanoids that are known to cause airway hyperresponsiveness and increased mucusproduction. Future work will focus on characterizing the effect these eicosanoid products have on the airway and whether fungal effector translocationresult in increased leukotriene levels.Magnaporthe oryzae has evolved two distinct mechanisms of effector secretion for biotrophic invasion of rice. Martha C. Giraldo 1 , Yasin F. Dagdas 2 ,Yogesh K. Gupta 2 , Thomas A. Mentlak 2,4 , Mihwa Yi 1 , Hiromasa Saitoh 3 , Ryohei Terauchi 3 , Nicholas J. Talbot 2 , Barbara Valent 1 . 1) Plant Pathology, KansasState University, Manhattan, KS. USA; 2) School of Biosciences, University of Exeter, EX4 4QD, UK; 3) Iwate Biotechnology Research Center, Kitakami,Iwate, 024-0003 Japan; 4) Cambridge Consultants Ltd, Cambridge, CB4 0DW, U.K.Pathogens secrete effector proteins into host tissue to suppress immunity and cause disease. Pathogenic bacteria have evolved several distinct secretionsystems to target specific effector proteins during pathogenesis, but it was not previously known if fungal pathogens require different secretorymechanisms. We present evidence that the blast fungus Magnaporthe oryzae possesses distinct secretion systems for delivering effector proteins duringbiotrophic invasion of rice cells. M. oryzae secretes cytoplasmic effectors targeted for delivery inside rice cells and apoplastic effectors targeted to theextracellular space. Cytoplasmic effectors preferentially accumulate in the biotrophic interfacial complex (BIC), a novel in planta structure located besidethe tip of the initially filamentous invasive hypha and then remaining next to the first differentiated bulbous invasive hypha cell. In contrast, apoplasticeffectors remain in the extracellular compartment uniformly surrounding the invasive hypha inside the invaded cell. Disruption of the conventional ER-Golgi secretion pathway by Brefeldin A (BFA) treatment blocked secretion of apoplastic effectors, which were retained in the ER, but not secretion ofcytoplasmic effectors. Fluorescence Recovery After Photobleaching experiments confirmed that cytoplasmic effectors continued to accumulate in BICs inthe presence of BFA. Analysis of mutants showed that the BIC is associated with a novel form of secretion involving exocyst components, Exo70 and Sec5,and the t-SNARE Sso1, which are required for efficient delivery of effectors into plant cells and are critical for pathogenicity. By contrast, effectors whichfunction between the fungal cell wall and plant plasma membrane are secreted from invasive hyphae to the apoplast by the ER-Golgi secretory pathwayconserved in eukaryotes. We propose a model for the distinct secretion systems that the rice blast fungus has evolved to achieve tissue invasion.Domains for plant uptake of Ustilago maydis secreted effectors. Anupama Ghosh, Armin Djamei, Shigeyuki Tanaka, Regine Kahmann. Max PIanckInstitute for Terrestrial Microbiology, Department of Organismic Interactions, Karl-Von-Frisch-Strasse 10, D-35043 Marburg, Germany.The genome of the corn smut fungus Ustilago maydis codes for a large repertoire of secreted effectors. Some of them play crucial roles for virulence andestablishment of the biotrophic phase. The chorismate mutase Cmu1 is one such secreted translocated effector of U. maydis. cmu1 deletion strains areattenuated in virulence that is attributed to higher salicylate levels in plants infected with the mutant strain, most likely through alterations in thechanneling of chorismate from the plastids to the cytosol. Here we identify the motif in Cmu1 that is necessary for the translocation of the protein acrossthe plant plasma membrane and present a mutational analysis of this region. To test for uptake we assayed the ability of mutant proteins to complement acmu1 mutant strain as well as the retained ability to complement the growth defect of a Daro7 strain of S. cerevisiae in minimal medium. By deletionanalysis a region of 20 amino acids adjacent to the signal peptide was shown to be essential for the translocation. Microscopic analysis of maize tissueinfected with U. maydis strains expressing Cmu1-mcherry fusion proteins with or without the probable uptake motif revealed that the 20 amino acid motifallows binding of the protein to an as yet unknown plant plasma membrane component. We hypothesize that the translocation of Cmu1 across the plantplasma membrane is a two step process; initiated by binding followed by translocation across the membrane. In addition, we present results where the 20amino acid motif is substituted by motifs from other effectors.27th Fungal Genetics Conference | 65

CONCURRENT SESSION ABSTRACTSPenetration-specific effectors from Phytophthora parasitica favour plant infection. Edouard Evangelisti 1* , Benjamin Govetto 2 , Naima Minet-Kebdani 1 ,Marie-Line Kuhn 1 , Agnes Attard 1 , Franck Panabieres 1 , Mathieu Gourgues 1 . 1) UMR Institut Sophia Agrobiotech, INRA/CNRS/Université de Nice, SophiaAntipolis, France; 2) Institut Méditerranéen de Biodiversité et d'Écologie marine et continentale (IMBE), CNRS-INEE - IRD -Aix Marseille Université -Université d'Avignon - Institut Pytheas.Oomycetes are major crop pests which cause million dollars losses every year. To date only a few efficient chemicals are available against thesefilamentous microorganisms. A better understanding of the molecular events occuring during plant-oomycete interactions will help to propose newstrategies for crop protection. We performed a transcriptional analysis in order to identify oomycete penetration-specific genes and identified a set ofpenetration-specific effectors (PSE) bearing a RXLR motif. This motif was previously shown to promote effector import into plant cells during the biotrophicstage in feeding structures called haustoria. Here we report the functional analysis of three candidate genes, referred to as PSE1, PSE2 and PSE3. The threeeffectors were able to abolish plant defense responses when transiently expressed in Nicotiana plants. Moreover, constitutive expression of PSE1 andPSE3 in A. thaliana led to an enhanced susceptibility to P. parasitica infection suggesting a role for these proteins in P. parasitica pathogenicity. TransgenicArabidopsis lines accumulating PSE1 protein showed several developmental perturbations that were associated with altered auxin physiology. Rootgrowth inhibition assays showed that auxin signaling pathway is not altered by PSE1 accumulation. Nevertheless, the coiled-root phenotype and theenhanced susceptibility of PSE1-expressing lines to P. parasitica were reverted by synthetic auxin 2,4-D supply, or treatment with the auxin efflux inhibitorTIBA suggesting that a reduced auxin accumulation is responsible for these phenotypes. This hypothesis was confirmed by a reduced activity of the pDR5auxin sensitive promoter at the root apex. The alteration of the expression pattern observed for two auxin efflux carriers, PIN4 and PIN7 suggests that aperturbation of auxin efflux could be responsible for the PSE1 associated defects. We proposed that PSE1 could favour P. parasitica virulence byinterfering with auxin content. Our results show that penetration specific effectors can modulate general plant functions to facilitate plant infection.Perturbation of hormone physiology was previously reported for other plant pathogens, including nematodes and bacteria, supporting the hypothesis thatinfection strategies from distant pathogens species could converge onto a limited set of plant targets.66

CONCURRENT SESSION ABSTRACTSFriday, March 15 3:00 PM–6:00 PMChapelCell Wall, Polarity and Hyphal Tip GrowthCo-chairs: Stephan Seiler and Ernestina Castro-LongoriaThe function of Rho type small GTPases for cell polarity in Ustilago maydis. Britta Tillmann 1 , Michaela Wehr 1 , Sonja Frieser 1 , Kay Oliver Schink 2 , JohannesFreitag 1 , Michael Bölker 1 . 1) Dept Biol, Univ Marburg, Marburg, Germany; 2) Institute for Cancer Research, the Norwegian Radium Hospital, Oslo UniversityHospital, Montebello, 0310 Oslo, Norway.Establishment of cell polarization requires the coordinated transport and localized fusion of secretory vesicles. This process is controlled by Rho-typeGTPases that act as molecular switches. Temporal and spatial activation of Rho-GTPases depends on specific guanine nucleotide exchange factors (GEFs).Inactivation of Rho proteins is achieved via interaction with GTPase activating proteins (GAPs) that stimulate the low intrinsic GTPase activity of Rhoproteins. During its life cycle, U. maydis switches between budding and filamentous growth. The Rho type GTPase Rac1 is the main regulator of thismorphogenic transition. The highly related Cdc42 is required for cell separation after mitosis and for formation of retraction septa during filamentousgrowth. We could show that the activator of Rac1, the Rho-GEF Cdc24 is subject to a negative autoregulatory feedback loop. Active Rac1 triggers Cla4dependent multisite phosphorylation of a C-terminal destruction box. This results in rapid degradation of Cdc24 and release of a ternary complexcontaining active Rac1, the scaffold protein Bem1 and the Rac1 effector kinase Cla4. The active Rac1 is subsequently inactivated by GAPs that localize in aring-like fashion underneath of the tip. Both destruction of Cdc24 and inactivation of Rac1 serve to delimit Rac1 activity to the very tip of the fungal hypha.Sustained polarized growth is further supported by recycling of inactive Rac1 to the hyphal tip. This is achieved either by interaction with the Rho proteinGDP dissociation inhibitor Gdi1 or via endocytosis. Active Rho-GTPases recruit specific effectors that trigger the localized fusion of secretory vesicles at thehyphal tip. We found that both Cdc42 and Rac1 interact with Sec3, a subunit of the multiprotein exocyst complex. We have identified several homologs ofexocyst subunits in U. maydis and tested them for functions during polar growth. We could demonstrate that Rac1 is critical for proper localization of theexocyst landmark protein Sec3. We have identified the U. maydis homolog of Smg-GDS, an unconventional activator of Rho GTPases in mammals. Smg-GDS contains a number of armadillo repeats and interacts with both Cdc42 and Rac1. Deletion of the Smg-GDS gene reduces significantly mating andfilament formation, indicating that it contributes to regulation of cell polarity.A quantitative model of hyphal tip growth based on the spatial distribution of exocyst subunits in the human fungal pathogen Candida albicans. DavidCaballero- Lima, Ilyana Kaneva, Simon Watton, C. Jeremy Craven, Peter Sudbery. Dept Molecular Biol & Biotech, Sheffield Univ, Sheffield, S Yorkshire,United Kingdom.We present a quantitative three dimensional treatment of fungal hyphal growth which adapts previous theoretical treatments in the light of advances inour knowledge of the components of polarised growth and their location as revealed by GFP fusions. The model is based on the proposition that vesiclesfuse with the hyphal tip at a rate determined by the experimentally observable local density of exocyst components. Enzymes such as b-1,3 glucansynthase are embedded in the plasma membrane by this process and continue to synthesize cell wall until they are removed from the membrane byendocytosis. The time development of the spatial distribution of the synthase molecules arises from the model. We test the model in the hyphae of thehuman fungal pathogen Candida albicans by quantitative measurements of the distribution of exocyst components and membrane components such asGFP-Rho1, the regulatory subunit of b1,3 glucan synthase, Rom2-GFP, the GEF for Rho1, and the location of actin cortical patches. We show that thepredicted shape and width of the hyphae are in good agreement with that predicted by the model, provided that endocytosis acts to remove cell wallsynthesizing enzymes at the subapical band of cortical actin patches. Thus the pattern of tip growth of fungal hyphae can be satisfactorily explained by asimple but quantitative model rooted within the known molecular processes of polarized growth. At the same time the model exposes the areas ofuncertainty which need to be addressed by future experimentation.Cell wall integrity signaling in Aspergillus fumigatus. Johannes Wagener, Karl Dichtl, Christoph Helmschrott, Sweta Samantaray, Franziska Dirr, MichaelNeubauer. Max von Pettenkofer-Institut, University of Munich, Munich, Germany.Aspergillus fumigatus is an opportunistic pathogen and the most frequent cause of a severe invasive infection termed invasive aspergillosis. Similar toother fungi, this mold is surrounded by a robust cell wall that defines its shape and protects it from physical stress. We have characterized the cell wallintegrity (CWI) pathway of A. fumigates. It comprises at least three major membrane anchored cell wall stress sensors with partially overlapping functions(Wsc1, Wsc3 and MidA), the guanine nucleotide exchange factor Rom2, a Rho GTPase, protein kinase C and a MAP kinase signaling module. We haveshown that the principal CWI components are well conserved from yeasts to filamentous fungi. Though, the importance of the individual components forthe fungal physiology, e.g., cell polarity and conidiation, may significantly differ. Our data stress the importance of the CWI pathway for the antifungal drugsusceptibility and virulence of this pathogen.Optimization of polarity establishment through coupling of multiple feedback loops. Roland Wedlich-Soldner 1 , Tina Fresisinger 1 , Ben Kluender 2 , NikolaMueller 1 , Gisela Beck 1 , Garwin Pichler 4 , Jared Johnson 3 , Richard Cerione 3 , Erwin Frey 2 . 1) Cellular Dynamics adn Cell Patterning, Max Planck Institute ofBiochemistry, Martinsried, Germany; 2) Arnold Sommerfeld Center for Theoretical Physics, Ludwig Maximilians University Munich, Munich, Germany; 3)Department of Chemistry and Chemical Biology, Baker Laboratory, Cornell University, Ithaca, NY, USA; 4) Department of Biology II, Ludwig MaximiliansUniversity Munich, Martinsried, Germany.Establishment of cell polarity - or symmetry breaking - relies on local accumulation of polarity regulators. While simple positive feedback is sufficient todrive symmetry breaking, it is highly sensitive to stochastic fluctuations typical for living cells. By integrating mathematical modeling with quantitativeexperimental validations we now show that in the yeast Saccharomyces cerevisiae only a combination of actin- and Guanine nucleotide DissociationInhibitor (GDI)-dependent recycling of the central polarity regulator Cdc42 is capable of establishing robust cell polarity at a single site during yeastbudding. The GDI pathway consistently generates a single polarization site, but requires Cdc42 to cycle rapidly between its active and inactive form, and istherefore highly sensitive to perturbations of the GTPase cycle. Conversely, actin-mediated recycling of Cdc42 induces robust symmetry breaking but27th Fungal Genetics Conference | 67

CONCURRENT SESSION ABSTRACTScannot restrict polarization to a single site. Our results demonstrate how cells optimize symmetry-breaking through coupling between multiple feedbackloops.Cell wall structure and biosynthesis in oomycetes and true fungi: a comparative analysis. Vincent Bulone. Sch Biotech, Royal Inst Biotech (KTH),Stockholm, Sweden.Cell wall polysaccharides play a central role in vital processes like the morphogenesis and growth of eukaryotic micro-organisms. Thus, the enzymesresponsible for their biosynthesis represent potential targets of drugs that can be used to control diseases provoked by pathogenic species. One of themost important features that distinguish oomycetes from true fungi is their specific cell wall composition. The cell wall of oomycetes essentially consists of(1®3)-b-glucans, (1®6)-b-glucans and cellulose whereas chitin, a key cell wall component of fungi, occurs in minute amounts in the walls of some oomycetespecies only. Thus, the cell walls of oomycetes share structural features with both plants [cellulose; (1®3)-b-glucans] and true fungi [(1®3)-b-glucans, (1®6)-b-glucans and chitin in some cases]. However, as opposed to the fungal and plant carbohydrate synthases, the oomycete enzymes exhibit specific domaincompositions that may reflect polyfunctionality. In addition to summarizing the major structural differences between oomycete and fungal cell walls, thispresentation will compare the specific properties of the oomycete carbohydrate synthases with the properties of their fungal and plant counterparts, withparticular emphasis on chitin, cellulose and (1®3)-b-glucan synthases. The significance of the association of these carbohydrate synthases with membranemicrodomains similar to lipid rafts in animal cells will be discussed. In addition, distinguishing structural features within the oomycete class will behighlighted with the description of our recent classification of oomycete cell walls in three different major types. Genomic and proteomic analyses ofselected oomycete and fungal species will be correlated with their cell wall structural features and the corresponding biosynthetic pathways.Cellular morphogenesis of Aspergillus nidulans conidiophores: a systematic survey of protein kinase and phosphatase function. Lakshmi Preethi Yerra,Steven Harris. University of Nebraska-Lincoln, Lincoln, NE.In the filamentous fungus Aspergillus nidulans, the transition from hyphal growth to asexual development is associated with dramatic changes inpatterns of cellular morphogenesis and division. These changes enable the formation of airborne conidiophores that culminate in chains of sporesgenerated by repeated budding of phialides. Our objective is to characterize the regulatory modules that mediate these changes and to determine howthey are integrated with the well-characterized network of transcription factors that regulate conidiation in A. nidulans. Because protein phosphorylationis likely to be a key component of these regulatory modules, we have exploited the availability of A. nidulans post-genomic resources to investigate theroles of protein kinases and phosphatases in developmental morphogenesis. We have used the protein kinase and phosphatase deletion mutant librariesmade available by the Fungal Genetics Stock Center to systematically screen for defects in conidiophore morphology and division patterns. Our initialresults implicate ANID_11101.1 (=yeast Hsl1/Gin4) in phialide morphogenesis, and also reveal the importance of ANID_07104.1 (=yeast Yak1) in themaintenance of cell integrity during asexual development. Additional deletion mutants with reproducible defects have been identified and will bedescribed in detail. We will also summarize initial results from double mutant analyses that attempt to place specific protein kinase deletions within theregulatory network that controls conidiation.Septum formation starts with the establishment of a septal actin tangle (SAT) at future septation sites. Diego Delgado-Álvarez 1 , S. Seiler 2 , S. Bartnicki-García 1 , R. Mouriño-Pérez 1 . 1) CICESE, Ensenada, Mexico; 2) Georg August University, Göttingen, Germany.The machinery responsible for cytokinesis and septum formation is well conserved among eukaryotes. Its main components are actin and myosins, whichform a contractile actomyosin ring (CAR). The constriction of the CAR is coupled to the centripetal growth of plasma membrane and deposition of cell wall.In filamentous fungi, such as Neurospora crassa, cytokinesis in vegetative hyphae is incomplete and results in the formation of a centrally perforatedseptum. We have followed the molecular events that precede formation of septa and constructed a timeline that shows that a tangle of actin filaments isthe first element to conspicuously localize at future septation sites. We named this structure the SAT for septal actin tangle. SAT formation seems to bethe first event in CAR formation and precedes the recruitment of the anillin Bud-4, and the formin Bni-1, known to be essential for septum formation.During the transition from SAT to CAR, tropomyosin is recruited to the actin cables. . Constriction of the CAR occurs simultaneously with membraneinternalization and synthesis of the septal cell wall.Visualization of apical membrane domains in Aspergillus nidulans by Photoactivated Localization Microscopy (PALM). Norio Takeshita 1 , Yuji Ishitsuka 2 ,Yiming Li 2 , Ulrich Nienhaus 2 , Reinhard Fischer 1 . 1) Dept. of Microbiology, Karlsruhe Institute of Technology, Karlsruhe, Germany; 2) Institute for AppliedPhysics, Karlsruhe Institute of Technology.Apical sterol-rich plasma membrane domains (SRDs), which can be viewed using the sterol-binding fluorescent dye filipin, are gaining attention for theirimportant roles in polarized growth of filamentous fungi. The size of SRDs is around a few mm, whereas the size of lipid rafts ranges in general between10-200 nm. In recent years, super-resolution microscope techniques have been improving and breaking the diffraction limit of conventional lightmicroscopy whose resolution limit is 250 nm. In this method, a lateral image resolution as high as 20 nm will be a powerful tool to investigate membranemicrodomains. To investigate deeply the relation of lipid membrane domains and protein localization, the distribution of microdomains in SRDs wereanalyzed by super-resolution microscope technique, Photoactivated Localization Microscopy (PALM). Membrane domains were visualized by each markerprotein tagged with photoconvertible fluorescent protein mEosFP for PALM. Size, number, distribution and dynamics of membrane domains, anddynamics of single molecules were investigated. Time-laps analysis revealed the dynamic behavior of exocytosis.68

CONCURRENT SESSION ABSTRACTSFriday, March 15 3:00 PM–6:00 PMHeatherSexual Regulation and Evolution in the FungiCo-chairs: Frances Trail and Nicolas CorradiClonality and sex impact aflatoxigenicity in Aspergillus populations. Ignazio Carbone 1 , Bruce W. Horn 2 , Rodrigo A. Olarte 1 , Geromy G. Moore 3 , Carolyn J.Worthington 1 , James T. Monacell 4,1 , Rakhi Singh 1 , Eric A. Stone 5,4 , Kerstin Hell 6 , Sofia N. Chulze 7 , German Barros 7 , Graeme Wright 8 , Manjunath K. Naik 9 . 1)Department of Plant Pathology, NC State University, Raleigh, NC, USA; 2) National Peanut Research Laboratory, Agricultural Research Service, U.S.Department of Agriculture, Dawson, GA, USA; 3) Southern Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, NewOrleans, LA, USA; 4) Bioinformatics Research Center, NC State University, Raleigh, NC, USA; 5) Department of Genetics, NC State University, Raleigh, NC,USA; 6) International Institute of Tropical Agriculture, Cotonou, Republic of Benin; 7) Departamento de Microbiologia e Inmunologia, Universidad Nacionalde Rio Cuarto, Cordoba, Argentina; 8) Department of Primary Industries, Queensland, Kingaroy, Australia; 9) Department of Plant Pathology, College ofAgriculture, Karnataka, India.Species in Aspergillus section Flavi commonly infect agricultural staples such as corn, peanuts, cottonseed, and tree nuts and produce an array ofmycotoxins, the most potent of which are aflatoxins. Aspergillus flavus is the dominant aflatoxin-producing species in the majority of crops. Populations ofaflatoxin-producing fungi may shift in response to: (1) clonal amplification that results from strong directional selection acting on a nontoxin- or toxinproducingtrait; (2) disruptive selection that maintains a balance of extreme toxigenicities and diverse mycotoxin profiles; (3) sexual reproduction thatresults in continuous distributions of toxigenicity; or (4) female fertility/sterility that impacts the frequency of sexual reproduction. Population shifts thatresult in changes in ploidy or nuclear DNA composition (homokaryon versus heterokaryon) may have immediate effects on fitness and the rate ofadaptation in subsequent fungal generations. We found that A. flavus populations with regular rounds of sexual reproduction maintain higher aflatoxinconcentrations than predominantly clonal populations and that the frequency of mating-type genes is directly correlated with the magnitude ofrecombination in the aflatoxin gene cluster. Genetic exchange within the aflatoxin gene cluster occurs via crossing over between divergent lineages inpopulations and between closely related species. During adaptation, specific toxin genotypes may be favored and swept to fixation or be subjected to driftand frequency-dependent selection in nature. Results from mating experiments in the laboratory indicate that fertility differences among lineages may bedriving genetic and functional diversity. Differences in fertility may be the result of female sterility, changes in heterokaryotic state, DNA methylation, orother epigenetic modifications. The extent to which these processes influence aflatoxigenesis is largely unknown, but is critical to understand for bothfundamental and practical applications, such as biological control. Our work shows that a combination of population genetic processes, especiallyasexual/sexual reproduction and fertility differences coupled with ecological factors, may influence aflatoxigenicity in these agriculturally important fungi.Toolkit for sexual reproduction in the genome of Glomus spp; a supposedly ancient asexual lineage. Nicolas Corradi. Department of Biology, Universityof Ottawa, Ottawa, Ontario, Canada.Arbuscular mycorrhizal fungi (AMF) are involved in a critical symbiosis with the roots of most land plants;the mycorrhizal symbiosis. Despite theirimportance for terrestrial ecosystems worldwide, many aspects of AMF evolution and genetics are still poorly understood, resulting in notorious scientificfrustrations and intense debates; especially regarding the genetic structure of their nuclei (heterokaryosis vs homokayosis) and their mode of propagation(long-term clonality vs cryptic sexuality). This will aim address the latter aspect of their biology - i.e. their mode of reproduction - by cataloguing andhighlighting emerging evidence, based on available genome sequence data, for the presence of a cryptic sexual cycle in the AMF . In particular,investigations along available genome and transcriptome data from several AMF species have unravelled the presence of a battery of genes that arecommonly linked with sexually-related processes in other fungal phyla. These include a gene-set required for the initiation and completion of aconventional meiosis, as well as many other genomic regions that are otherwise found to play a pivotal role in fungal partner recognition. The origin,diversity and functional analysis of some of these sexually-related genes in AMF will be discussed.Comparative transcriptomics identifies new genes for perithecium development. Frances Trail 1 , Usha Sikhakolli 1 , Kayla Fellows 1 , Nina Lehr 2 , JeffreyTownsend 2 . 1) Department of Plant Biology, Michigan State Univ, East Lansing, MI; 2) Department of Ecology and Evolutionary Biology, Yale University,New Haven, CT.In recent years, a plethora of genomic sequences have been released for fungal species, accompanied by functional predictions for genes based onprotein sequence comparisons. However, identification of genes involved in particular processes has been extremely slow, and new methodologies foridentifying genes involved in a particular process have not kept pace with the exponential increase in genome sequence availability. We have performedtranscriptional profiling of five species of Neurospora and Fusarium during six stages of perithecium development. We estimated the ancestraltranscriptional shifts during the developmental process among the species and identified genes whose transcription had substantially and significantlyshifted during the evolutionary process. We then examined phenotypes of knockouts of genes whose expression greatly increased in Fusariumgraminearum perithecium development. In numerous cases, gene disruption resulted in substantial changes in perithecium. These genes were notpreviously identified as candidates for function in perithecium development, illustrating the utility of this method for identification of genes associatedwith specific functional processes.27th Fungal Genetics Conference | 69

CONCURRENT SESSION ABSTRACTSRapid evolution of female-biased genes: a novel example from the eukaryotic model organism Neurospora crassa. Hanna Johannesson, Carrie Whittle.Evolutionary Biology, Uppsala University, Uppsala, Sweden.In animals and plants, sex-biased gene expression plays a major role in gene evolution. In particular, reproductive genes with male-biased expressiontend to exhibit rapid protein evolution and reduced codon bias as compared to female-biased or unbiased genes. Minimal data are available for fungi.Here, we demonstrate that sex-biased expression is associated with gene evolution in the filamentous fungus Neurospora crassa, but in contrast toanimals and plants, the rapid evolution occurs for female-biased genes. Based on analyses of >25,000 expressed sequence tags (ESTs) from male (conidial),female (protoperithecial) and vegetative (mycelial) tissues, we show that reproductive genes with female-biased expression exhibit faster proteinevolution and reduced optimal codon usage than male-biased genes and vegetative genes. Furthermore, our data suggest that female-biased genes arealso more apt to experience selective sweeps. The sex-biased expression effects are observable at the species and population level. We argue that therapid molecular evolution of female-biased genes is best explained by sexual selection via female-female competition, but could also result from matechoiceand/or directional natural selection.Self-attraction can not bypass the requirement for two mating type genes during sexual reproduction in Neurospora crassa. Katherine A. Borkovich,Hyojeong Kim, Sara Wright, Gyungsoon Park, Shouqiang Ouyang, Svetlana Krystofova. Plant Pathology and Microbiology, University of California, Riverside,Riverside, CA.The pheromone receptor PRE-2 is highly expressed in male and female reproductive structures of mat a strains in Neurospora crassa. Trichogynes fromDpre-2 mat a protoperithecia do not respond chemotropically to mat A conidia or form mature fruiting bodies or meiotic progeny. Strains with swappedidentity due to heterologous expression of pre-2 or its cognate pheromone ccg-4 behave normally in crosses with opposite mating-type. Coexpression ofpre-2 and ccg-4 in the mat A background leads to self-attraction and development of barren perithecia that lack ascospores. Further perithecialdevelopment is achieved by inactivation of Sad-1, a gene required for meiotic gene silencing in N. crassa. Results from studies using forced heterokaryonsof opposite mating-type strains show that the presence of one receptor and its compatible pheromone is necessary and sufficient for perithecialdevelopment and ascospore production. Taken together, the results demonstrate that although receptors and pheromones control sexual identity, themating-type genes (mat A and mat a) must be in two different nuclei to allow meiosis and sexual sporulation to occur in N. crassa.Fertility in Aspergillus fumigatus and the identification of an additional ‘supermater’ pair. Céline M. O'Gorman 1 , Sameira S. Swilaiman 1 , Janyce A. Sugui 2 ,Kyung J. Kwon-Chung 2 , Paul S. Dyer 1 . 1) School of Biology, University of Nottingham, University Park, Nottingham NG7 2RD, United Kingdom; 2) MolecularMicrobiology Section, Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health,Bethesda, Maryland, USA.Aspergillus fumigatus is an opportunistic human pathogen that causes a range of allergic and invasive diseases in severely immunocompromisedindividuals, with a very high mortality rate typically in excess of 50%. A functional sexual cycle was discovered in 2009 and a highly fertile ‘supermater’ pair,AFB62 and AfIR928, was later identified from a collection of 50 isolates. Here we describe the results of a larger, worldwide fertility screen and present anadditional ‘supermater’ pair. A set of 126 clinical and environmental A. fumigatus isolates were crossed against two Irish reference strains of each matingtype. A subset of the eight most-fertile strains was then tested in all pairwise combinations. The pairing of isolates 47-169 x 47-154 had consistently highmating efficiency and outcrossing ability after four weeks, therefore it was chosen as an additional ‘supermater’ pair for community use in mating projects.It is important to have alternative tester strains to allow for unexpected mating differences when crossing isolates of diverse genetic origins. This isbecause factors such as heterokaryon incompatibility (het) loci and single nucleotide polymorphisms, can considerably influence sexual compatibility. Theworldwide fertility screen found that approximately 85% of isolates are sexually fertile, indicating that sexual reproduction should be possible in naturewhen suitable environments are present. Next, the plasticity of sexual crossing conditions was tested, to determine whether they could be manipulated toincrease fertility in crosses involving low-fertility strains of interest. A range of environmental and growth conditions were examined, including incubationtemperature, CO 2 level, and oatmeal agar type. Fertility levels were significantly affected by certain parameters. Work is ongoing to integrate these factorsto further optimize fertility in the ‘supermater’ pairs.Sexual reproduction and mating type function in the penicillin producing fungus Penicillium chrysogenum. Julia Böhm 1 , Birgit Hoff 1 , Simon Wolfers 1 ,Céline O'Gorman 2 , Paul Dyer 2 , Stefanie Pöggeler 3 , Ulrich Kück 1 . 1) Christian Doppler Laboratory for Fungal Biotechnology, Ruhr-Universität Bochum,Universitätsstr. 150, 44780 Bochum, Deutschland; 2) School of Biology, University of Nottingham, University Park, Nottingham, NG7 2RD, U.K; 3) AbteilungGenetik eukaryotischer Mikroorganismen, Institut für Mikrobiologie und Genetik, Georg-August Universität Göttingen, 37077 Göttingen, Deutschland.Penicillium chrysogenum is a filamentous fungus of major medical and historical importance, being the original and present day industrial source of theantibiotic penicillin with a world market value of about 600 million € per year. The species has been considered asexual for over 100 years and despiteconcerted efforts it has not been possible to induce sexual reproduction. However, we recently were able to detect mating type loci in different strains,indicating a sexual lifecycle. Isolates, carrying opposite mating types, were found in near-equal proportion in nature and we observed transcriptionalexpression of mating type loci as well as pheromone and pheromone receptor genes [1]. Utilising knowledge of mating-type (MAT) gene organization wenow describe conditions under which a sexual cycle can be induced leading to the production of cleistothecia and meiotic ascospores, which were similarto those described recently for Eupenicillium crustaceum [2]. Evidence of recombination was obtained using both molecular and phenotypic markers. Thenewly identified heterothallic sexual cycle was used for strain development purposes, generating offspring with novel combinations of traits relevant topenicillin production.Furthermore, the MAT1-1-1 mating-type gene, known primarily for a role in governing sexual identity, was also found to control transcription of a widerange of genes including those regulating penicillin production, hyphal morphology and conidial formation, all traits of biotechnological relevance. Forfunctional characterization MAT1-1-1 knockout and overexpression strains were generated and analyzed. These discoveries of a sexual cycle and MATgene function are likely to be of broad relevance for manipulation of other asexual fungi of economic importance.[1] Hoff B, Pöggeler S, Kück U (2008) Eighty years after its discovery, Fleming`s Penicillium strain discloses the secret of its sex. Eukaryot Cell 7: 465-470[2] Pöggeler S, O'Gorman CM, Hoff B, Kück U (2011) Molecular organization of the mating-type loci in the homothallic ascomycete Eupenicilliumcrustaceum. Fungal Biol. 115: 615-624.70

CONCURRENT SESSION ABSTRACTSThe Sclerotinia sclerotiorum mating type locus (MAT) contains a 3.6-kb region that is inverted in every generation. Patrik Inderbitzin 1 , PeriasamyChitrampalam 2 , Karunakaran Maruthachalam 1 , Bo-Ming Wu 3 , Krishna Subbarao 1 . 1) Department of Plant Pathology, University of California-Davis, Davis,CA, USA; 2) Department of Plant Sciences, University of Arizona, Tucson, AZ, USA; 3) Department of Plant Pathology, China Agricultural University, 2 WestYuanmingyuan Rd., Haidian District, Beijing, China.Sclerotinia sclerotiorum is a filamentous ascomycete in the Sclerotiniaceae (Pezizomycotina) and a necrotrophic pathogen of more than 400 hostsworldwide, including many important agricultural crops. In California, the biggest lettuce producer in the United States, S. sclerotiorum is a causal agent oflettuce drop that reduces overall annual lettuce yield by 15%. Little is known about the details of sexual reproduction in S. sclerotiorum, but the structureof the S. sclerotiorum mating type locus MAT, the master regulator of sexual reproduction in ascomycetes, has previously been reported. As in otherhomothallic (self-fertile) ascomycetes, S. sclerotiorum MAT contains both idiomorphs (divergent alleles) fused end-to-end at a single locus. Using 283isolates from lettuce in California and from other states and hosts, we investigated the diversity of S. sclerotiorum MAT, and identified a novel version ofMAT that differed by a 3.6-kb inversion and was designated Inv+, as opposed to the previously known S. sclerotiorum MAT that lacked the inversion andwas Inv-. The inversion affected three of the four MAT genes: MAT1-2-1 and MAT1-2-4 were inverted and MAT1-1-1 was truncated at the 3’-end.Expression of MAT genes differed between Inv+ and Inv- isolates. In Inv+ isolates, only one of the three MAT1-2-1 transcript variants of Inv- isolates wasdetected, and the alpha1 domain of Inv+ MAT1-1-1 transcripts was truncated. Both Inv- and Inv+ isolates were self-fertile, and the inversion segregated ina 1:1 ratio regardless of whether the parent was Inv- or Inv+. This suggested the involvement of a highly regulated process in maintaining equalproportions of Inv- and Inv+, likely associated with the sexual state. The MAT inversion region, defined as the 3.6-kb MAT inversion in Inv+ isolates and thehomologous region of Inv- isolates, was flanked by a 250-bp inverted repeat on either side. The 250-bp inverted repeat was a partial MAT1-1-1 thatthrough mediation of loop formation and crossing over, may be involved in the inversion process. Inv+ isolates were widespread, and in California andNebraska constituted half of the isolates examined. We speculate that a similar inversion region may be involved in mating type switching in thefilamentous ascomycetes Chromocrea spinulosa, Sclerotinia trifoliorum and in certain Ceratocystis species.27th Fungal Genetics Conference | 71

CONCURRENT SESSION ABSTRACTSFriday, March 15 3:00 PM–6:00 PMFred Farr ForumOxidative Stress, ROS Signaling and Adaptation to HypoxiaCo-chairs: Geraldine Butler and Barry ScottTranscriptional regulatory networks controlling the early hypoxic response in Candida albicans. A. Nantel, M. van het Hoog, A. Sellam, C. Beaurepaire, F.Tebbji, M. Whiteway. National Research Council of Canada, Montreal, Quebec, Canada.The ability of Candida albicans to colonize or invade multiple host environments requires that it rapidly adapts to different conditions. Our group hasbeen exploiting ChIP-chip and transcription profiling technologies, together with computer modeling, to provide a better understanding of selecttranscription factor (TF) networks. We used DNA microarrays to measure the changes in transcriptional profiles that occur immediately following thetransfer of C. albicans to hypoxic growth conditions. The impressive speed of this response is not compatible with current models of fungal adaptation tohypoxia that depend on the inhibition of sterol and heme biosynthesis. Functional interpretation of these profiles was achieved using Gene Set EnrichmentAnalysis, a method that determines whether defined groups of genes exhibit a statistically significant bias in their distribution within a ranked gene list.The Sit4p phosphatase, Ccr4p mRNA deacetylase and Sko1p TF were identified as novel regulators of the early hypoxic response. While cells mutated inthese regulators exhibit a delay in their transcriptional responses to hypoxia their ability to grow in the absence of oxygen is not impeded. Promoteroccupancy data on 26 TFs was combined with the profiles of 375 significantly-modulated target genes in a Network Component Analysis (NCA) to producea model of the dynamic and highly interconnected TF network that controls this process. The NCA also allowed us to observe correlations betweentemporal changes in TF activities and the expression of their respective genes, thus allowing us to identify which TFs are potentially subjected to posttranscriptionalmodifications. The TF network is centered on Tye7p and Upc2p which are associated with many of the genes that exhibit the fastest andstrongest up regulations. While Upc2p only associates with downstream promoters, Tye7p is acting as a hub, its own promoter being bound by itself and 7additional TFs. Rap1 and Ahr1 appear to function as master regulators since they bind to a greater proportion of TF gene promoters, including those ofUpc2p and Tye7p. Finally, Cbf1p, Mrr1p and Rap1p show the greatest numbers of unique gene targets. The high connectivity of these models illustratesthe challenges that lie in determining the individual contributions of specific TFs.Proteomic analysis of the hypoxic response of the human-pathogenic fungus Aspergillus fumigatus. Olaf Kniemeyer 1,4,5 , Kristin Kroll 1,5 , Vera Pähtz 1,4,5 ,Martin Vödisch 1,5 , Falk Hillmann 1,5 , Kirstin Scherlach 2 , Martin Roth 3 , Christian Hertweck 2 , Axel A. Brakhage 1,5 . 1) Department of Molecular and AppliedMicrobiology, Leibniz Institute for Natural Product Research and Infection Biology (HKI), Jena, Germany; 2) Department of Biomolecular Chemistry, LeibnizInstitute for Natural Product Research and Infection Biology (HKI), Jena, Germany; 3) Bio Pilot Plant, Leibniz Institute for Natural Product Research andInfection Biology (HKI), Jena, Germany; 4) Integrated Research and Treatment Center, Center for Sepsis Control and Care Jena, University Hospital (CSSC),Jena, Germany; 5) Department of Microbiology and Molecular Biology, Friedrich Schiller University Jena, Beutenbergstr. 11a, 07745 Jena, Germany.Aspergillus fumigatus is a ubiquitous, filamentous fungus which may cause a broad spectrum of disease in the human host, ranging from allergic orlocally restricted infections to invasive mycoses. The most fatal A. fumigatus disease, invasive aspergillosis occurs in patients who are severelyimmunocompromised and is characterized by a high mortality. During the course of the infection A. fumigatus has to cope with several kinds of stressconditions including low oxygen levels (hypoxia). Just recently it was shown that hypoxia adaptation is an important virulence attribute of A. fumigatus. Toidentify novel hypoxia-sensing and adapting pathways we have characterized the changes of the A. fumigatus proteome in response to short (3-24 hours)and long periods (7-10 days) of hypoxia (1% O 2). To maintain reproducible culture conditions, an oxygen-controlled fermenter was used. During long-termcultivation under hypoxia, proteins involved in glycolysis, the pentose phosphate shunt, amino acid biosynthesis, NO-detoxification and respiration showedan increased level. In contrast, proteins involved in sulfate assimilation and acetate activation were down-regulated. Strikingly, hypoxia also inducedbiosynthesis of the secondary metabolite pseurotin A. The proteomic response of A. fumigatus to short periods of hypoxia showed some similarities, butalso marked differences: The level of glycolytic, NO-detoxifying and amino acid biosynthesis enzymes increased under both hypoxic growth conditions.However, the abundance of enzymes of the pentose-phosphate pathway decreased, whereas enzymes involved in ethanol fermentation significantlyincreased. To get a deeper knowledge about the specific role of metabolic pathways in adaptation to hypoxia, we have started to characterize candidategenes for their role in hypoxia by generating deletion mutants. First data will be presented and discussed.Fgap1-mediated response to oxidative stress in trichothecene-producing Fusarium graminearum. M. Montibus, N. Ponts, E. Zehraoui, F. Richard-Forget,C. Barreau. INRA, UR1264-MycSA, BP81, F-33883 Villenave d’Ornon, France.The filamentous fungus Fusarium graminearum infects cereals and corn. It is one of the main causal agent of “Fusarium Head Blight” and “Maize EarRot”. During infection, it produces mycotoxins belonging to the trichothecenes family that accumulate in the grains. Although the biosynthetic pathwayinvolving specific Tri genes has been elucidated, the global regulation of toxin biosynthesis remains enigmatic. It is now established that oxidative stressmodulates the production of toxins by F. graminearum. H 2O 2 added in liquid cultures of this fungus enhances trichothecenes accumulation and increasesTri genes expression. Our working hypothesis is that a transcription factor regulates redox homeostasis, and is involved in Tri genes regulation. In the yeastSaccharomyces cerevisiae, the transcription factor Yap1p mediates response to oxidative stress via nuclear re-localization and activation of genes codingfor detoxification enzymes. In this study, we investigate the role of Yap1p homolog in F. graminearum, Fgap1, in response to oxidative stress and itseventual role in the regulation of trichothecene production. A deleted mutant and a strain expressing a constitutively activated form of the Fgap1 factor inF. graminearum were constructed. We cultured these mutants in GYEP liquid medium supplemented with H 2O 2 to evaluate their sensitivity to oxidativestress and analyse their toxin production. The nuclear localization of constitutively activated Fgap1p as well as wild-type Fgap1p under oxidative stress byH 2O 2 was analyzed. Expression profiles of genes encoding oxidative stress response enzymes potentially controlled by Fgap1p and of genes involved in thebiosynthesis of type B trichothecenes were analyzed by Q-RT-PCR. Trichothecene accumulation is strongly enhanced in the deleted strain, with an increasein Tri genes expression. On the other hand, Tri genes expression and toxin accumulation are drastically repressed in the mutant in which Fgap1p isconstitutively activated. Moreover, the level of expression of two genes encoding catalases is modulated in both mutants. The involvement of Fgap1 inother types of stress has also been investigated. In particular, cadmium and osmotic stress affect growth in the deleted strain.72

CONCURRENT SESSION ABSTRACTSThe role of NADPH oxidases in Neurospora crassa cell fusion. Nallely Cano-Dominguez 1 , Ernestina Casto-Longoria 1 , Jesus Aguirre 2 . 1) Departamento deMicrobiologia, CICESE, Ensenada, Baja California, Mexico; 2) Departamento de Biologia Celular y Desarrollo. Instituto de Fisiologia Celular UNAM, MexicoCity, D.F. Mexico.Hansberg and Aguirre proposed that reactive oxygen species (ROS) play essential roles in cell differentiation in microorganisms. ROS are generatedmainly during mitochondrial electron transport and by the action of certain enzymes. The NADPH oxidases (NOX) are enzymes that catalyze the productionof superoxide by transferring electrons from NADPH to oxygen. Neurospora crassa contains the NADPH oxidases NOX-1 and NOX-2 and a commonregulatory subunit NOR-1. NOX-2 is essential for ascospore germination, while NOX-1 is required for sexual and asexual development, polar growth andcell fusion. NOR-1 is essential for all these NOX functions. We have found that a functional NOR-1::GFP fusion is localized throughout the cytoplasm,enriched at the hyphal tip and sometimes in aggregates. This suggests that the functional NOX complexes are probably not localized at the plasmamembrane. Up to now NOX function in fungi has been evaluated in mutants that completely lack NOX proteins. We generated nox-1 alleles that result inNOX-1 proteins carrying substitutions of proline 382 by histidine or cysteine 542 by arginine, which affect NADPH-binding. Equivalent mutations inphagocytic Nox2/gp91phox do not affect protein stability but completely lack oxidase activity. P382H and C542R mutants did not produce sexual fruitingbodies and showed a decreased growth and differentiation of aerial mycelia, without affecting production of conida. These results indicate that sexualdevelopment depends on ROS production by NOX-1, whereas during asexual differentiation NOX-1 plays an important role independently of its catalyticactivity. Dnox-1, Dnor-1, P382H NOX-1 and C542R NOX-1 mutants were all able to produce some conidial anastomosis tubes (CATs) but they were unableto complete cell-cell fusion. All these mutants are also impaired in vegetative hyphae-hyphae fusion, which might explain the growth defects in Dnox-1 andDnor-1 strains. CATs production is delayed in the presence of antioxidant N- acetyl cystein (NAC) and Dsod-1 strains show an increase in CATs fusions. Theresults suggest that some ROS may be implicated in signaling CATs homing and vegetative fusion.Peroxiredoxins in ROS responses -Why evolve peroxidases that are inactivated by peroxides? Alison M. Day, Jonathon D Brown, Sarah R Taylor, JonathanD Rand, Brian A Morgan, Elizabeth A Veal. Inst Cell & Molecular Biosciences, Newcastle Univ, Newcastle Tyne, United Kingdom.Peroxiredoxins (Prx) are extremely abundant antioxidant enzymes with important roles in protecting against oxidative stress, ageing and cancer.Thethioredoxin peroxidase activity of eukaryotic typical 2-Cys Prx detoxifies hydrogen peroxide but, enigmatically, is highly sensitive to inactivation byperoxide-induced hyperoxidation of a catalytic cysteine residue. It has been proposed that hyperoxidation might allow hydrogen peroxide to act as a signaland/or promote an alternative activity of Prx as a chaperone [1, 2]. However, any advantage to be gained by inhibiting the thioredoxin peroxidase activityand preventing Prx from removing peroxides under oxidative stress conditions has remained obscure. The fission yeast Schizosaccharomyces pombecontains a single 2-Cys Prx, Tpx1. Our previous work has established that, counterintuitively, Tpx1 is vital for adaptive transcriptional responses tohydrogen peroxide due to essential roles in the hydrogen peroxide-induced activation of the p38/JNK/Hog1-related MAPK Sty1 and AP-1-like transcriptionfactor Pap1 [3, 4]. In seeking to understand why the thioredoxin peroxidase activity of Tpx1 should be important for Pap1 activation, we have identifiedthat Tpx1 is the major cellular substrate for thioredoxin. Accordingly, in hydrogen peroxide-treated cells, Tpx1 competitively inhibits the activity ofthioredoxin towards other substrates, including Pap1, and the methionine sulphoxide reductase A, Mxr1. Consequently, we show that the oxidativeinactivation of the thioredoxin peroxidase activity of Tpx1 is important to maintain active Mxr1, repair oxidative protein damage and maintain cell viabilityfollowing exposure to toxic levels of hydrogen peroxide [5]. Based on these discoveries in yeast, we propose that an important function for the reversiblehyperoxidation of eukaryotic 2-Cys Prx is to regulate thioredoxin and thus thioredoxin-mediated signalling and repair processes. I will present further datasupporting this conclusion and discuss its implications for hydrogen peroxide signal transduction.NADPH oxidases regulate septin-mediated cytoskeletal re-modeling during plant infection by the rice blast fungus Magnaporthe oryzae. Lauren S.Ryder 1 , Yasin F. Dagdas 1 , Thomas A. Mentlak 1 , Michael J Kershaw 1 , Martin Schuster 1 , Christopher R Thornton 1 , Jisheng Chen 2 , Zonghua Wang 2 , Nicholas JTalbot 1 . 1) Dept Biosciences, Univ Exeter, Exeter, United Kingdom; 2) Fujian agricultural university.NADPH oxidases (Nox) are flavoenzymes that function by transferring electrons across biological membranes to catalyze reduction of molecular oxygento superoxide. In animal cells, Nox enzymes are implicated in cell proliferation, cell signalling and apoptosis, while in plants Nox are necessary forprogrammed cell death, the response to environmental stresses, pathogen infection, and polarised growth of root hairs. In filamentous fungi, Nox arenecessary for cellular differentiation during sexual reproduction and for developmental processes that involve transitions from non-polarised to polarisedcell growth, such as tissue invasion by mutualistic and pathogenic fungi, and fungal virulence. The underlying function of Nox enzymes in these diversedevelopmental processes remains unclear. The rice blast fungus Magnaporthe oryzae infects plants with a specialized cell called an appressorium, whichuses turgor to drive a rigid penetration peg through the rice leaf cuticle. Here, we show that NADPH oxidases (Nox) are necessary for septin-mediated reorientationof the dynamic F-actin cytoskeleton to facilitate cuticle rupture and plant cell invasion. We report that the Nox2-NoxR complex spatiallyorganises a heteroligomeric septin ring at the appressorium pore, required for assembly of a toroidal F-actin network at the point of penetration pegemergence. Maintenance of the cortical F-actin network during plant infection independently requires Nox1, a second NADPH oxidase, which is necessaryfor penetration hypha elongation. Organisation of F-actin and septins in appressoria are disrupted by application of anti-oxidants, while latrunculinmediateddepolymerisation of appressorial F-actin is competitively inhibited by reactive oxygen species (ROS), providing evidence that regulated synthesisof ROS by fungal NADPH oxidases directly controls septin and F-actin dynamics.27th Fungal Genetics Conference | 73

CONCURRENT SESSION ABSTRACTSRedox regulation of an AP-1-like transcription factor, YapA, in the fungal symbiont Epichloë festucae. Gemma M. Cartwright, Barry Scott, Yvonne Becker.Molec Biosci, Massey Univ, Palmerston Nth, New Zealand.Reactive oxygen species (ROS) are emerging as important regulators required for the successful establishment and maintenance of the mutualisticassociation between the fungal endophyte Epichloë festucae and its grass host Lolium perenne. The generation of reactive oxygen species (ROS) by thefungal NADPH oxidase, NoxA has previously been shown to regulate hyphal growth of E. festucae in planta; a result that has led to the hypothesis thatfungal-produced ROS are key second messengers in the symbiosis. However, the highly reactive nature of these molecules dictates that cells possessefficient sensing mechanisms to maintain ROS homeostasis and prevent oxidative damage to cellular components. The Saccharomyces cerevisiae Gpx3-Yap1 and Schizosaccharomyces pombe Tpx1-Pap1, two-component H 2O 2 sensors, serve as model redox relays for coordinating the cellular response toROS. While proteins related to the Yap1 and Pap1 basic-leucine zipper (bZIP) transcription factors have been identified in a number of filamentous fungi,the components involved in the upstream regulation remain unclear. This study investigated the role of the E. festucae Yap1 homologue, YapA, andputative upstream activators GpxC and TpxA, homologues of Gpx3 and Tpx1, respectively, in responding to ROS. YapA is involved in responding to ROSgenerated at the wound site following inoculation into ryegrass seedlings. However, deletion of yapA did not impair host colonization indicatingredundancy in systems used by E. festucae to sense and respond to plant-produced ROS. In culture, deletion of E. festucae yapA, renders the mutantssensitive to only a subset of ROS and this sensitivity is influenced by the stage of fungal development. In contrast to the H 2O 2-sensitive phenotype widelyreported for fungi lacking the Yap1-like protein, the E. festucae yapA mutant maintains wild-type mycelial resistance to H 2O 2 but conidia of the yapAmutant are very sensitive to H 2O 2. Using a degron-tagged GFP-CL1 as a reporter, we found YapA is required for the expression of the spore specificcatalase, catA. Moreover, YapA is activated by H 2O 2 independently of both GpxC and TpxA, suggesting a novel mechanism of regulation exists in E.festucae. This work provides a comprehensive analysis of the role and regulation of the AP-1 transcription factor pathway in a filamentous fungal species.Interaction between phenolic and oxidant signaling in Cochliobolus heterostrophus. Benjamin A Horwitz 1 , Samer Shalaby 1 , Olga Larkov 1 , MordechaiRonen 2 , Sophie Lev 3 . 1) Department of Biology, Technion - IIT, Haifa, Israel; 2) Department of Plant Science, Tel Aviv University, Ramat Aviv, Israel; 3)Centre for Infectious Diseases and Microbiology, University of Sydney at Westmead Hospital, Westmead, NSW 2145, Australia.The transcription factor ChAP1 is an ortholog of yeast YAP1 in the maize pathogen Cochliobolus heterostrophus. ChAP1 migrates to the nucleus uponexposure to oxidative stress, inducing antioxidant genes such as thioredoxin and glutathione reductase [1]. ChAP1 also localizes to nuclei on contact withthe leaf and during invasive growth. Though reactive oxygen species are encountered on the host, ChAP1 nuclear retention can occur without oxidativestress. One of the signals responsible is provided by phenolic compounds [1-3]. Using a genetically-encoded ratiometric reporter of the redox state, weshowed that leaf extract and phenolics, despite their antioxidant properties, promote nuclear accumulation of ChAP1. To study this dual role of ChAP1 weidentified genes expressed in response to phenolics. Intradiol dioxygenase CCHD1 is rapidly upregulated, independent of ChAP1 [2]. Coumaric acid causedrapid and simultaneous upregulation of most of the b-ketoadipate pathway genes. Deletion of CCHD1 provided genetic evidence that protocatechuic acidis an intermediate in catabolism of many aromatics [3]. The activity of a structure series showed complementary requirements for upregulation of CCHD1and ChAP1 nuclear retention. The ability to metabolize a compound and ChAP1 nuclear retention are inversely correlated. To find additional genesinduced by phenolics, microarrays designed from the predicted coding sequences of the C. heterostrophus genome [4] were hybridized to probes madefrom RNA of cultures exposed to coumaric acid, or controls. Expression of about 90 genes from different pathways primarily for metabolism, for example,the b-ketoadipate, quinic acid and shikimic acid pathways, as well as transporters from different families was altered in response to coumaric acid. Theability to respond to phenolics and detoxify or metabolize them via the b-ketoadipate pathway confers an advantage to plant pathogens, and explains thepresence of at least two response pathways detecting these compounds. [1] Lev et al. (2005) Eukaryot. Cell 4:443-454; [2] Shanmugam et al. (2010) Cell.Microbiol. 12:1421-1434; [3] Shalaby et al. (2012) MPMI 25: 931-940; [4] Ohm et al. (2012) PLoS Pathog 8: e1003037. Supported in part by the IsraelScience Foundation. We thank Michal Levin and Itai Yanai for help with microarray hybridization.74

CONCURRENT SESSION ABSTRACTSFriday, March 15 3:00 PM–6:00 PMKilnPhylogenomicsCo-chairs: Jason Stajich and Joey SpataforaCharacterizing Gene Tree Incongruence on a Genome Scale. Dannie Durand. Biological Sciences, Carnegie Mellon University, Pittsburgh, PA.Gene families evolve through gene duplication and loss, and lateral gene transfer. Reconstructing these events is a powerful approach to understandingthe co-evolution of genes and species and the emergence of novel protein function. Gene duplication, loss, and transfer can all result in a gene tree thatdisagrees with the species tree. This incongruence can be exploited to infer the history of these events, as well as the ancestral lineage in which each eventtook place. This is achieved by fitting the gene family tree to the associated species tree, a process called reconciliation. I will discuss the benefits andchallenges of gene tree reconciliation, with special attention to genome scale analyses. The use of gene tree reconciliation will be compared with nonphylogeneticanalyses of gene family expansion and contraction. The problem of determining whether the observed incongruence is due to geneduplication, lateral transfer, or incomplete lineage sorting will also be discussed. I will present analyses of several large gene tree data sets from wellstudiedspecies lineages, as a practical demonstration of this approach. Our algorithms have been implemented in[http://www.cs.cmu.edu/~durand/Notung], a freely available software tool.Early fungi and their carbohydrate active enzymes. Mary L. Berbee 1* , Satoshi Sekimoto 2 , Joseph Spatafora 3 , Timothy James 4 , Teresita M. Porter 5 , RytasVilgalys 6 . 1) Dept Botany, Univ British Columbia, Vancouver, B.C., Canada; 2) Department Of Biological Sciences, The University Of Alabama, Tuscaloosa, AL;3) Oregon State University, Dept of Botany & Plant Pathology, 2082 Cordley Hall, Corvallis, OR; 4) University of Michigan, Dept of Ecology & Evol Biology,830 N University, Ann Arbor, MI; 5) 16 Yachters Lane, Etobicoke, ON, Canada; 6) Biology Department 130 Science Drive, Biological Sciences Rm 137, DukeUniversity Box 90338, Durham, NC.Early fungi are intermingled with some of the oldest fossils from vascular plants, dated at 400 Ma. However, what the fungi were doing for their nutritionbefore land plants were available has been difficult to reconstruct because in phylogenies of the earliest diverging fungal lineages, saprotrophs andparasites of plants as well as animals are intermingled, and which fungal life style came first is ambiguous. We are using phylogenetic analysis of enzymesinvolved in carbohydrate metabolism to reconstruct the enzymatic capabilities of some of the early terrestrial fungi. Our community sequencing proposalto the US Joint Genome Institute resulted in four new genome sequences for evolutionarily divergent lineages including aquatic fungi, the chytrids andBlastocladiomycota, and zygomycetes. Analysis of the genomes suggests that cellulases and pectinases to degrade plant wall carbohydrates were alreadypresent in the earliest fungal lineages but largely lost from the zygomycetes. This implies that fungi evolved in association with the green algal/green plantlineage. Even with complete genome sequences, the branching order among the aquatic fungi and zygomycetes remains problematical, and branchingorder conflicts from one analysis to another. The conflicts may reflect difficulties involved in modeling evolutionary processes across lineages.Alternatively, the conflicts may indicate that fungi, like animals, underwent a 'Cambrian Explosion' perhaps facilitated by rapid expansion of nutritionalresources offered by radiation of multicellular plants and animals.Better evolution through gene clustering. Jason Slot 1 , Matthew Campbell 2 , Han Zhang 1 , Martijn Staats 3 , Jan van Kan 4 , Antonis Rokas 1 . 1) BiologicalSciences, Vanderbilt University, Nashville, TN; 2) Botany, University of Hawai`i, Manoa, HI; 3) Biosystematics group, Wageningen University, Wageningen,The Netherlands; 4) Laboratory of Phytopathology, Wageningen University, Wageningen, The Netherlands.The recent availability of a large number of fungal genomes has facilitated systematic investigations of metabolic pathway evolution across the kingdom.Through combining phylogenetic and genomic techniques, we have recently examined the evolution of metabolic pathways across a well-sampled fungalphylogeny, and gained new insight into the role of metabolic gene clusters in fungal evolution. The occasional occurrence of horizontal gene transfer ofentire pathways between distantly related fungi via gene clusters suggests that fungal species have access to larger pan-genomes than previously thought.Furthermore, analysis of gene cluster decay suggests these transfers are underestimated by analyses of single strains, and that evolution within clusteredpathways is constrained by natural selection. Increased evolvability in fungi is also implied by the discovery of chromosomal loci that maintain largealternative secondary metabolite gene clusters within recombining lineages. Together, these phylogenomic analyses in fungi illustrate a multi-faceted roleof gene clustering in fungal evolution.Phylogenomics unveils secondary metabolites specific to mycoparasitic lineages in Hypocreales. C. Alisha Owensby, Kathryn E. Bushley, Joseph W.Spatafora. Botany & Plant Pathology, Oregon State University, Corvallis, OR.Hypocreales is an order characterized by a dynamic evolutionary history of interkingdom host jumping, with members that parasitize animals, plants, andother fungi. The monophyly of taxa attacking members of the same kingdom is not supported by molecular phylogenetics, however. For example,Trichoderma spp. and Elaphocordyceps spp. are both mycoparasitic, but are members of different families within Hypocreales, Hypocreaceae andOphiocordycipitaceae, respectively. In fact, both genera are more closely related to insect pathogens, than they are to each other. Multiple species ofTrichoderma have sequenced genomes, and recently genomes of several insect pathogens in Hypocreales have been completed (e.g. Metarhizium spp. andTolypocladium inflatum). The genus Elaphocordyceps represents a unique clade within Hypocreales, because whereas most species in the familyOphiocordycipitaceae are insect pathogens, most Elaphocordyceps parasitize truffles of the ectomycorrhizal genus Elaphomyces [Eurotiales, Ascomycota].To compare genes of a truffle pathogen with hypocrealean insect pathogens and mycoparasites, we sequenced the genome of Elaphocordycepsophioglossoides. Our draft assembly of the E. ophioglossoides genome is ~32 MB and has 10,779 gene models, 36 of which are predicted to producesecondary metabolites. We have identified three very large genes in E. ophioglossoides related to peptaibol producing nonribosomal peptide synthetase(NRPS) genes. Peptaibols, which disrupt osmoregulation by forming ion channels through lipid bilayers, have antibiotic and antifungal activity and are bestdescribed in Trichoderma spp. E. ophioglossoides and its beetle-pathogenic congener, T. inflatum, both possess three putative peptaibol synthetases whichwe identified through analysis of NRPS adenylation domains. Of the three peptaibol-specific domain clades, one is predicted to encode for thenonproteinogenic a-aminoisobutryic acid residues. We also show that, despite being very closely related, E. ophioglossoides and T. inflatum each possess27th Fungal Genetics Conference | 75

CONCURRENT SESSION ABSTRACTSthree different peptaibol-like genes, only two of which appear to be located in syntenic regions. The current distribution of fungi possessing peptaibolgenes is restricted to mycoparasitic lineages of Hypocreales and is generating hypotheses about the role of secondary metabolites in mycoparasitism.Comparative analysis of 35 basidiomycete genomes reveals diversity and uniqueness of the phylum. Robert Riley 1 , Asaf Salamov 1 , Robert Otillar 1 , KirstenFagnan 1 , Bastien Boussau 3 , Daren Brown 4 , Bernard Henrissat 5 , Anthony Levasseur 5 , Benjamin Held 6 , Laszlo Nagy 2 , Dimitris Floudas 2 , Emmanuelle Morin 7 ,Gerard Manning 8 , Scott Baker 9 , Robert Blanchette 6 , Francis Martin 7 , David Hibbett 2 , Igor Grigoriev 1 . 1) Joint Genome Istitute, Lawrence Berkeley NationalLab, Walnut Creek, CA; 2) Clark University, Worcester, MA; 3) UC Berkeley, Berkeley, CA; 4) USDA, Peoria, IL; 5) AFMB, Marseille, France; 6) UMN, St. Paul,MN; 7) INRA, France; 8) Salk Institute, La Jolla, CA; 9) Pacific Northwest National Lab, Richland, WA.Fungi of the phylum Basidiomycota (basidiomycetes), make up some 37% of the described fungi, and are important in forestry, agriculture, medicine,and bioenergy. This diverse phylum includes symbionts, pathogens, and saprobes including wood decaying fungi. To better understand the diversity of thisphylum we compared the genomes of 35 basidiomycete fungi including 6 newly sequenced genomes. The genomes of basidiomycetes span extremes ofgenome size, gene number, and repeat content. A phylogenetic tree of Basidiomycota was generated using the Phyldog software, which uses all availableprotein sequence data to simultaneously infer gene and species trees. Analysis of core genes reveals that some 48% of basidiomycete proteins are uniqueto the phylum with nearly half of those (22%) comprising proteins found in only one organism. Phylogenetic patterns of plant biomass-degrading genessuggest a continuum rather than a sharp dichotomy between the white rot and brown rot modes of wood decay among the members of Agaricomycotinasubphylum. There is a correlation of the profile of certain gene families to nutritional mode in Agaricomycotina. Based on phylogenetically-informed PCAanalysis of such profiles, we predict that that Botryobasidium botryosum and Jaapia argillacea have properties similar to white rot species, althoughneither has liginolytic class II fungal peroxidases. Furthermore, we find that both fungi exhibit wood decay with white rot-like characteristics in growthassays. Analysis of the rate of discovery of proteins with no or few homologs suggests the high value of continued sequencing of basidiomycete fungi.Genome evolution of fungal pathogens from the Magnaporthe oryzae/grisea clade. Helene Chiapello 1,2 , Ludovic Mallet 1,3 , Cyprien Guérin 1 , GabrielaAguileta 4 , François Rodolphe 1 , Annie Gendrault 1 , Jonathan Kreplak 3 , Joelle Amselem 3 , Enrique Ortega-Abboud 5 , Marc-Henri Lebrun 6 , Didier Tharreau 5 ,Elisabeth Fournier 7 . 1) INRA, UR MIG, 78352 Jouy-en-Josas, France; 2) INRA, UR BIA, 31326 Castanet-Tolosan, France; 3) INRA, URGI, 78026 Versailles,France; 4) CRG, Barcelone, Spain; 5) CIRAD, UMR BGPI, TA 54K, 34398 Montpellier, France; 6) INRA, UMR BIOGER, 78850 Thiverval-Grignon, France; 7)INRA, UMR BGPI, TA 54K, 34398 Montpellier, France.The GEMO project aims at characterizing genomic determinants of pathogenicity and evolutionary events involved in adaptation of 9 isolates from theM. oryzae/grisea clade differing in their host specificity. Eight strains from M. oryzae species complex and one of the related species M. grisea have beensequenced and assembled. De novo structural gene annotation was carried out using Eugene (Schiex, 2001) to predict genes and REPET (Flutre, 2011) toannotate Transposable Elements (TEs) in these 9 genomes. Four of them exhibited large supplementary genomic regions potentially issued from anunknown bacterial strain of the Burkholderia genus. An original strategy based on Gotham software (Ménigaud, 2012) was used to accurately quantifythese regions in all the affected genome scaffolds. Functional gene annotations were performed using InterProScan. Databases and interfaces relying onthe GMOD tools (gmod.org) were set up to browse annotations and facilitate further evolutionary analyses. In order to identify gene families, the entireset of the predicted and known proteins of the M. oryzae/grisea genomes were clustered using OrthoMCL (Li, 2003). A total of 20443 clusters (15326assigned to M.oryzae/grisea and 5117 to Burkholderia) were obtained, including 8154 clusters comprising single copy genes shared by all genomes (coregenome) and variable number of genome specific gene families (305-1550). Genes encoding putative Secreted Proteins (SPs) were identified in 2522OrthoMCL clusters (2271 of M. oryzae/grisea and 251 of Burkholderia). Further analyses regarding genome-specific and rice-infecting specific genes andSPs will be presented. OrthoMCL families were processed to infer the phylogenetic reference genealogy of the M. oryzae/grisae complex. We alsoevaluated the ability of individual genes to recover the same topology as that supported by most of the genes by using a recent method based on multipleco-inertia analysis (de Vienne, 2012). Most of the genes exhibit a concordant topology with the reference tree except a small set of ‘outliers’. Furtherinvestigations are currently being performed to determine possible causes for incongruities. Finally, we present preliminary results regarding thecomparison of TE distribution in M. oryzae/grisea species taking into account the reference genealogy of the strains.Leptosphaeria maculans 'brassicae': "Transposable Elements changed my life, I feel different now". Jonathan Grandaubert 1 , Conrad Schoch 2 , HosseinBorhan 3 , Barbara Howlett 4 , Thierry Rouxel 1 . 1) INRA-BIOGER, Thiverval-Grignon, France; 2) NCBI, National Institutes of Health, Bethesda, MD, USA; 3) AAFSaskatoon, Canada; 4) School of Botany, University of Melbourne, Australia.The Dothideomycetes phytopathogens Leptosphaeria maculans and Leptosphaeria biglobosa form a complex of 8 species and putative subspeciessuggested to have diverged “recently”. In 2007, the sequencing of an isolate of Leptosphaeria maculans 'brassicae' (Lmb) provided the first referencegenome for this fungus. The 45-Mb genome has an unusual bipartite structure, alternating large GC-equilibrated and AT-rich regions. These AT-rich regionscomprise one third of the genome and are mainly composed of mosaics of truncated Transposable Elements (TEs) postulated to have “invaded” thegenome 5-10 MYA; they also comprise 5% of the predicted genes of which 20% encode putative effectors. In these regions, both genes and TEs areaffected by Repeat Induced Point mutation (RIP). To investigate when and how genome expansion took place in the evolutionary series, and theconsequences it had on fungal adaptability and pathogenicity, the genomes of five members of the species complex showing contrasted host range andinfection abilities were sequenced. In silico comparison of the reference genome with that of 30-32-Mb genome of L. maculans 'lepidii' (Lml), L. biglobosa'brassicae', L. biglobosa 'thlaspii' and L. biglobosa 'canadensis', showed these species have a much more compact genome with a very low amount of TEs(

CONCURRENT SESSION ABSTRACTSComparing comparative “omics” in Coccidioides spp. Emily A. Whiston, John W. Taylor. Plant & Microbial Biology, U.C. Berkeley, Berkeley, CA.The mammalian pathogens Coccidioides immitis and C. posadasii are the only dimorphic fungal pathogens that form spherules in the host. Furthermore,all of Coccidioides’ closest known relatives are non-pathogenic. In this project, we are interested in genome changes between the Coccidioides lineage andits relatives, and how these changes compare to recently published comparative and population genomics, and transcriptomics studies in Coccidioides.Coccidioides and its closest sequenced relative, Uncinocarpus reesii, are estimated to have diverged 75-80 million years ago. Here, we have sequenced thegenomes of four species more closely related to Coccidioides than U. reesii: Byssoonygena ceratinophila, Chrysosporium queenslandicum, Amauroascusniger and A. mutatus. For each of these four species, we prepared genomic DNA Illumina sequencing libraries; the resulting genome assemblies rangedfrom 23-34Mb, with N50 of 90kb-205kb. Predicted genes were confirmed by RNAseq; the total number of genes ranged from 8,179-9,184. We assessedindividual gene gain/loss, and gene family expansion/contraction in Coccioides using these new genomes and other recently published genomes from theOnygenales order, including the yeast-forming dimorphic pathogens Histoplasma and Paracoccidioides, and the dermatophytes Microsporum andTrichopyton. We have compared these results to genes identified in recently published Coccidioides “omics” studies that show evidence of positiveselection, introgression and/or differential expression.27th Fungal Genetics Conference | 77

CONCURRENT SESSION ABSTRACTSFriday, March 15 3:00 PM–6:00 PMNautilusSynthetic BiologyCo-chairs: Nancy Keller and Peter PuntEngineering Aspergillus oryzae for high level production of L-malic acid. Debbie S Yaver 1 , S. Brown 2 , A. Berry 2 . 1) Expression Technology, Novozymes, Inc.,Davis, CA; 2) Microbial Physiology, Novozymes, Inc., Davis, CA.In the last decade, there has been widespread interest and investment in developing processes for the production of bulk and specialty chemicals fromrenewable feedstocks by fermentation. During this period, Novozymes has successfully developed technology for production of a specialty molecule(hyaluronic acid) by Bacillus fermentation and has been very active in developing technologies for the production of bulk chemicals by metabolicengineering and fermentation using several different microorganisms. An example of the latter is L-malic acid. In the literature it is reported that somewild-type Aspergillus strains produce high levels of malic acid under specific cultivation conditions. Concentrations up to 113 g/L malate (94% w/w fromglucose) reported for A. flavus in fed-batch fermentations (Battat, et. al., 1991. Biotechnol. Bioeng. 37:1108-1116). The goal of our work was to improvemalic acid production in the natural malic acid producing filamentous fungus Aspergillus oryzae NRRL 3488 by overexpression of cloned genes and classicalmutagenesis. More than 75 different recombinant strains were tested containing combinations of overexpression of genes as well as deletions. A highthrough put screen was developed and used to screen mutagenized strains. Combined genetic engineering and mutagenesis/HTS was used to increase themalic acid production rate of A. oryzae NRRL3488 by 4-fold with final C4 acid totals of 340 g/l at 8 days in lab scale fermentations.When synthetic biology meets metabolic engineering: in vivo pathway assembly in Saccharomyces cerevisiae. Niels Kuijpers 1,2 , Daniel Solis Escalante 1,2 ,Jack T. Pronk 1,2,3 , Jean-Marc Daran 1,2,3 , Pascale Daran-Lapujade 1,2 . 1) Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628 BCDelft, The Netherlands; 2) Kluyver Centre for Genomics of Industrial Fermentation, PO Box 5057, 2600 GA Delft, The Netherlands; 3) Platform GreenSynthetic Biology, Julianalaan 67, 2628 BC Delft, The Netherlands.The yeast Saccharomyces cerevisiae is a powerful and versatile workhorse intensively exploited for a wide range of biotechnological applications. Besidesthe large scale production of endogenous products, such as the biofuel ethanol, S. cerevisiae has been genetically engineered to produce manyheterologous compounds, including half of the worldwide insulin market. The past decade has been marked by the conversion of S. cerevisiae into acomplex cell factory with remarkable new capabilities such as the production of the anti-malaria drug precursor artemisinic acid. The ever-increasingdemand for cheap and sustainable production of complex molecules combined with its attractiveness as a host for pathway engineering will inevitablyintensify the exploitation of S. cerevisiae as cell factory in the future. Even in the genetically accessible bakers yeast, expression of dozen of genes is stilllargely based on laborious classical techniques involving successive restrictions and ligations, complemented with the creative application of PCR.However, the increasing size and complexity of today’s constructs in metabolic engineering has made design and construction of plasmids by theseclassical techniques increasingly complicated and time consuming. Although uncovered nearly three decades ago, the high efficiency of S. cerevisiaehomologous recombination is only beginning to reveal its full potential for the assembly of large DNA constructs (Gibson et al., 2008). In vivo assembly inyeast is predicted to have a large impact on laboratory practices, ranging from simple plasmid construction to engineering of complex pathways viaautomated high-throughput strain construction. Despite those promising prospects, in vivo assembly has not yet become a standard technique in mostacademic laboratories. This offers unique possibilities for standardization and, simultaneously, for further optimization. In the present work we describe anapproach designed to improve the efficiency of in vivo assembly and to make a robust, versatile in vivo assembly strategy for multi-component plasmids.As a proof of principle, the method was used to assemble a 21 kb plasmid from 9 overlapping fragments, using only PCR and yeast transformation. GibsonD.G. et al. (2008), Science, 319, 1215-1220.Analysis of the intracellular galactoglycom of Trichoderma reesei grown on lactose. Levente Karaffa 1 , Leon Coulier 2 , Erzsébet Fekete 1 , Karin M.Overkamp 2 , Irina S. Druzhinina 3 , Marianna Mikus 3 , Bernhard Seiboth 3 , Levente Novák 4 , Peter J. Punt 2 , Christian P. Kubicek 3 . 1) Department of BiochemicalEngineering, University of Debrecen, H-4032, Debrecen, Hungary; 2) TNO, P.O. Box 360, 3700 AJ Zeist, The Netherlands; 3) Research Area Biotechnologyand Microbiology, Institute of Chemical Engineering, TU Wien, Gumpendorferstrasse 1a, A-1060 Wien, Austria; 4) Department of Colloid andEnvironmental Chemistry, Faculty of Science and Technology, University of Debrecen, H-4032, Debrecen, Hungary.Lactose (1,4-0-b-D-galactopyranosyl-D-glucose) is used as a soluble carbon source for the production of cellulases and hemicellulases for - among otherpurposes - in the biofuel and biorefinery industries. However, the mechanism how lactose induces cellulase formation in T. reesei is still enigmatic.Previous results raised the hypothesis that intermediates from the two D-galactose catabolic pathway may give rise to the accumulation of intracellularoligogalactosides that could act as inducer. We have therefore used HPAEC-MS to study the intracellular galactoglycome of T. reesei during growth onlactose, in T. reesei mutants impaired in galactose catabolism, and in strains with different cellulase productivity. Lactose, allo-lactose and lactulose weredetected in the highest amounts in all strains, and two trisaccharides (Gal-b-1,6-Gal-b-1,4-Glc/Fru; and Gal-b-1,4-Gal-b-1,4-Glc/Fru) also accumulated tosignificant levels. D-Glucose and D-galactose, as well as two further oligosaccharides (Gal-b-1,3/1,4-Gal; Gal-b-1,2/1,3-Glc) were only detected in minoramounts, In addition, one unknown disaccharide and four trisaccharides were also detected. The unknown hexose disaccharide to correlate with cellulaseformation in the improved mutant strains as well as the galactose pathway mutants, and Gal-b-1,4-Gal-b-1,4-Glc and two other unknown hexosetrisaccharides to correlate with cellulase production only in the pathway mutants, suggesting that these compounds could be involved in cellulaseinduction by lactose.78

CONCURRENT SESSION ABSTRACTSNovel transcriptomics approaches for metabolic pathway engineering target identification in Aspergillus. Peter J. Punt, Martien Caspers, MarvinSteijaert, Eric Schoen, Machtelt Braaksma. Microbiology, TNO, Zeist, Netherlands.Among filamentous fungi Aspergillus sp. are well known production host for several organic acids. These acids, traditionally being food ingredients, morerecently have gained attention as platform or building-block chemicals. These chemicals, currently mostly produced based on petrochemistry, are thestarting point for the production of a wide variety of materials, such as resins, plastics, etc. Production of these compounds via biobased routes will be amajor contribution towards a Biobased Economy. For the production of these bulk compounds robust host organisms are required, suitable for using lowcost lignocellulose-based feedstocks, resistant against adverse conditions due to inhibitory feedstock compounds and capable of coping with high productconcentrations. A. niger was shown to fulfill most of these prerequisites (Rumbold et al., 2009).Based on the extended molecular genetic toolkit systemsbiology approaches were developed for A. niger and other fungi (e.g. Braaksma et al., 2010). These approaches were followed towards production of theseplatform chemicals in A. niger, as demonstrated by the example of itaconic acid (Li et al., 2011, 2012). The recent development of novel high throughputsequence methods has led to new much more efficient transcriptomics approaches such as RNAseq. Combination of these approaches with novelexperimental design and statistical methods for targetgene identification in metabolic pathway engineering will be illustrated. Rumbold, K., van Buijsen,H.J.J., Overkamp, K.M., van Groenestijn, J.W., Punt, P.J., Werf, M.J.V.D. (2009) Microbial production host selection for converting second-generationfeedstocks into bioproducts. Microbial Cell Factories 8, art. no. 64 Braaksma, M., van den Berg, R.A., van der Werf, M.J., Punt, P.J. (2010) A Top-DownSystems Biology Approach for the Identification of Targets for Fungal Strain and Process Development. In: Cellular and Molecular Biology of FilamentousFungi. Eds: K.A. Borkovich & D.J. Ebbole ASM Press, Washington DC. pp. 25-35 Li, A., van Luijk, N., ter Beek, M., Caspers, M., Punt, P., van der Werf, M.(2011) A clone-based transcriptomics approach for the identification of genes relevant for itaconic acid production in Aspergillus. Fungal Genetics andBiology 48 (6), pp. 602-611.A new method for gene mining and enzyme discovery. Y. Huang 1,2,3 , P. Busk 1 , M. Grell 1 , H. Zhao 2,3 , L. Lange 1 . 1) Section for Sustainable Biotechnology,Department of Biotechnology, Chemistry and Environmental Engineering, Aalborg University Copenhagen, Denmark; 2) Environmental Microbiology KeyLaboratory of Sichuan Province, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, Sichuan 610041, PR China; 3) University of theChinese Academy of Sciences, Beijing 100049, PR China.Peptide pattern recognition (PPR) is a non-alignment based sequence analysis principle and methodological approach, which can simultaneouslycompare multiple sequences and find characteristic features. This method has improved the understanding of structure/function relationship for enzymeswithin the CAZY families, which would make it easier to predict the potential function of novel enzymes, creating bigger promises for industrial purposes.Mucor circinelloides, member of the former subdivision Zygomycota, can utilize complex polysaccharides such as wheat bran, corncob, xylan, CMC andavicel as substrate to produce plant cell wall degrading enzymes. Although the genome of M. circinelloides has been sequenced, only few plant cell walldegrading enzymes are annotated in this species. In the present project, PPR was applied to analyze glycoside hydrolase families (GH family) and miningfor new GH genes in M. circinellolides genome. We found 19 different genes encoding GH3, GH5, GH6, GH7, GH9, GH16, GH38, GH43, GH47 and GH125 inthe genome. Of the three GH3 encoding genes found, one was predicted by PPR to encode a b-glucosidase. We expressed this gene in Pichia pastoris andfound that the recombinant protein has high b-glucosidase activity (4884 U/mL). In this work, PPR provided targeted short cut to discovery of enzymeswith a specific activity. Not only could PPR pinpoint genes belonging to different GH families but it did also predict the enzymatic function of the genes.Increased production of fatty acids and triglycerides in Aspergillus oryzae by modifying fatty acid metabolism. Koichi Tamano 1 , Kenneth Bruno 2 , TomokoIshii 1 , Sue Karagiosis 2 , David Culley 2 , Shuang Deng 2 , James Collet 2 , Myco Umemura 1 , Hideaki Koike 1 , Scott Baker 2 , Masayuki Machida 1 . 1) National Instituteof Advanced Industrial Science and Technology (AIST); 2) Pacific Northwest National Laboratory (PNNL).Biofuels are attractive substitutes for petroleum based fuels. Biofuels are considered they do not contribute to global warming in the sense they arecarbon-neutral and do not increase carbons on the globe. Hydrocarbons that are synthesized by microorganisms have potential of being used as biofuelsor the source compounds. In the hydrocarbon compounds synthesized by A. oryzae, fatty acids and triglycerides are the source compounds of biodieselthat is fatty acid methyl ester. We have increased the production by modifying fatty acid metabolism with genetic engineering in A. oryzae. Firstly,enhanced-expression strategy was used for the increase. For four enzyme genes related to the synthesis of palmitic acid [C16:0-fatty acid], the individualenhanced-expression mutants were made. And the fatty acids and triglycerides in cytosol were assayed by enzyme assay kits, respectively. As a result,both fatty acids and triglycerides were most synthesized in the enhanced-expression mutant of fatty acid synthase gene at 2.1-fold and 2.2-fold more thanthe wild-type strain, respectively. Secondly, gene disruption strategy was used for the increase. Disruptants of several enzyme genes related to long-chainfatty acid synthesis were made individually. And one of them showed drastic increase in fatty acid synthesis. In the future, further increase in the synthesisis expected by utilizing genetic engineering in A. oryzae.27th Fungal Genetics Conference | 79

CONCURRENT SESSION ABSTRACTSMolecular biological basis for statin resistance in naturally statin-producing organisms. Ana Rems, Rasmus Frandsen. DTU Systems Biology, TechnicalUniversity of Denmark, Kongens Lyngby, Denmark.Secondary metabolites can be toxic to the organism producing them; therefore gene clusters for biosynthesis of secondary metabolites often includegenes responsible for the organism’s self-resistance to the toxic compounds. One such gene cluster is the compactin (ML-236B) cluster in Penicilliumsolitum. Compactin is an inhibitor of 3-hydroxy-3-methyl-glutaryl-CoA (HMG-CoA) reductase, and is used as a precursor for production of the cholesterolloweringdrug pravastatin. The compactin gene cluster includes two genes encoding proteins that may confer the self-resistance to compactin and itssecretion [1]. The mlcD gene encodes a putative 'HMG-CoA reductase-like protein’, and mlcE encodes a putative efflux pump. However, the function ofthese two putative proteins has not yet been confirmed. We aim to elucidate the biological basis for compactin resistance in the compactin-producingorganism. A codon-optimized version of the mlcD gene was inserted into the Saccharomyces cerevisiae genome. The constructed yeast strain was testedfor sensitivity to lovastatin, a statin structurally similar to compactin, by growing the strain on media containing lovastatin. The strain showed an increasedresistance to lovastatin compared to the wild-type strain. Furthermore, we investigated if MlcD confers the resistance by functional complementation ofthe endogenous HMG-CoA reductases in S. cerevisiae. There are two isozymes of HMG-CoA reductase in yeast, HMG1 and HMG2, both involved in thesterol biosynthetic pathway, which leads to the synthesis of ergosterol. Following deletion of HMG1 and HMG2 genes in S. cerevisiae, we inserted the mlcDgene into the knockout mutants, and tested the resulted strains for sensitivity to lovastatin. The HMG1 and HMG2 knockout mutants were unable to growon minimal media and had an increased sensitivity to lovastatin on rich media. However, insertion of the mlcD gene restored the growth of the yeastmutants and increased their resistance to lovastatin. These results show that MlcD complements the activity of the deleted HMG-CoA reductases, enablingsynthesis of ergosterol in yeast. In addition MlcD confers statin resistance by being insensitive to the inhibiting effects of statins. Reference: [1] Abe Y.,Suzuki T., Ono C., Iwamoto K., Hosobuchi M., Yoshikawa H. Mol Genet Genomics 2002, 267, 5:636-46.Engineering Cyclic Peptide Biosynthesis in Poisonous Mushrooms. Hong Luo, John S. Scott Craig, Robert M. Sgambelluri, Sung-Yong Hong, Jonathan D.Walton. Department of Energy Plant Research Laboratory, Michigan State University, E. Lansing, MI 48824, United States.Ninety percent of fatal mushroom poisonings are caused by alpha-amanitin and related bicyclic peptides found in some species of Amanita, Galerina,Lepiota, and Conocybe. We showed that the amatoxins (mainly amanitins) and related phallotoxins are synthesized on ribosomes in A. bisporigera and theunrelated mushroom G. marginata. The primary gene products are short (34-35 amino acid) proproteins that are initially processed by a dedicated prolyloligopeptidase. A genome survey sequence of A. bisporigera suggested that it has a repertoire of over 40 cyclic peptides, all produced on a singlebiosynthetic scaffold. Members of this extended gene family are characterized by conserved upstream and downstream amino acid sequences, includingtwo invariant proline residues, flanking a six to ten-amino acid “hypervariable” region that encodes the amino acids found in the mature toxins (orpredicted toxins). The evidence indicates that A. bisporigera has evolved a combinatorial strategy that could in principle biosynthesize billions of smallcyclic peptides. In order to study the other steps in amanitin biosynthesis, and to engineer novel cyclic peptides, we have developed a transformationstrategy for the amanitin-producing mushroom G. marginata. This first transformation method uses Agrobacterium-mediated transformation followed byhygromycin selection. Taking advantage of this platform, we are introducing artificial toxin genes that are deliberately designed to provide insights into thepathway. The synthetic genes include those that encode the cyclic octapeptide beta-amanitin, the heptapeptides phalloidin and phallacidin, examples ofthe toxin gene family known from A. bisporigera but not G. marginata, and randomly generated artificial sequences. Currently, thousands of transformantshave been generated through an efficient pipeline and the transformants are being analyzed for production of the expected products. If successful, thenovel peptides will be screened in a number of assays including RNA polymerase (the site of action of alpha-amanitin), membrane ion channels,pathogenic bacteria, and cancer cell lines.80

CONCURRENT SESSION ABSTRACTSFriday, March 15 3:00 PM–6:00 PMScrippsFungicides and Antifungal CompoundsCo-chairs: Daniele Debieu and Paul VerweijChemically Induced Haploinsufficiency Screens to Identify Drug Mechanism of Action in Aspergillus Fumigatus. D. A. Macdonald 1 , A. E. Johns 1 , M.Eberle 2 , P. Bowyer 1 , D. Denning 1 , M. J. Bromley 1 . 1) Institute of Inflammation and Repair, Respiratory & Allergy Centre, University of Manchester,Manchester, United Kingdom; 2) Applied Microbiology, Institute for Applied Life Sciences, University of Karlsruhe, Hertzstrae 16, 76187 Karlsruhe,Germany.Current drugs used to treat Aspergillus infections are limited and suffer from a variety of shortcomings including low efficacy, toxicity and increasingresistance. Despite the discovery of numerous promising drug targets, few lead compounds have been discovered by target based approaches. This can beexplained, in part, by the ‘druggability’ of a target as some compounds which demonstrate promising activity against an enzyme are not active against thewhole cell or are toxic to humans. Consequently most of the antimicrobials presently on the market were originally discovered by random screening ofcompounds against whole cell screens. A solution to this problem is to identify gene targets utilizing compounds that already show antifungal activity andhave clean toxicity profiles.Chemical genetic profiling aids identification of drug mechanism of action as a diploid strain lacking a single copy of a drug’s target is hypersensitive tothat drug. Heterozygote S. cerevisiae and C. albicans libraries have been used to identify the mechanism of action of several promising compounds;however, this has been hindered in A. fumigatus by the complexity in generating an adequate set of heterozygous strains. A high-throughput targetedgene KO method for A. fumigatus has been established by employing fusion-PCR to generate targeted gene disruption cassettes, optimizing the commontransformation protocol for A. fumigatus high-throughput gene disruption, and utilising a diploid Ku80 - /Ku80 - mutant to facilitate more reliablehomologous recombination. Preliminary efforts have produced 46 heterozygous KO strains and subsequently, the feasibility of chemical genetichaploinsufficiency studies in filamentous fungi has been demonstrated with several compounds. High-throughput methods of chemical genetic profiling bypooling multiple heterozygous KO strains into a single culture is currently being validated and preliminary data is promising. This will enable highthroughputmethods for surveying the genome of A. fumigatus for new drug targets and supports unveiling the mechanisms of action of antifungal drugs.Inhibition of benzoate 4-monooxygenase (CYP53A15) from Cohliobolus lunatus by cinnamic acid derivatives. Branka Korosec 1 , Barbara Podobnik 2 , SabinaBerne 3 , Neja Zupanec 1 , Metka Novak 1 , Nada Krasevec 1 , Samo Turk 4 , Matej Sova 4 , Ljerka Lah 1 , Jure Stojan 3 , Stanislav Gobec 4 , Radovan Komel 1,3 . 1) NationalInstitute of Chemistry, Ljubljana, Slovenia; 2) Lek Pharmaceuticals d.d., Verovskova 57, SI-1000 Ljubljana, Slovenia; 3) Institute of Biochemistry, Faculty ofMedicine, University of Ljubljana, Vrazov trg 2, SI-1000 Ljubljana, Slovenia; 4) Chair of Pharmaceutical Chemistry, Faculty of Pharmacy, University ofLjubljana, Askerceva cesta 7, SI-1000 Ljubljana, Slovenia.Fungal infections cause huge economic losses in agriculture. Some of the major phytopathogens also cause serious, and very often lethal, infections inhuman and animals. Plants may be a good source of antifungals since they have to defend themselves by producing numerous secondary metabolites,such as sterols, terpens, polycosanols and phenolic compounds. Successful development of antifungal compounds, based on natural defense molecules,could prove useful in combating infectious and toxin-producing fungi in both agriculture and medicine. In recent years several promising antifungal targetshave been under exploration. One of such is also fungal CYP53, member of the family of highly conserved CYP proteins, involved in detoxification ofbenzoate, a key intermediate in metabolism of aromatic compounds in fungi. High specificity and absence of homologue in higher eukaryotes assignCYP53A15 from the filamentous fungus Cochliobolus lunatus as interesting drug target. In our latest research we explored chemical properties ofisoeugenol for ligand-based similarity searching, and the homology model of CYP53A15 of Cochliobolus lunatus, for structure-based virtual screening of acomposite chemical library. Two cinnamic acid derivatives were amongst the highest scoring compounds. In the past few years, several other reportsabout antifungal activity of cinnamic acid derivatives have been published. In order to investigate the potential inhibitory activity on benzoate 4-monooxygenase (CYP53A15) we analyzed antifungal activity of 9 commercially available, and 10 representative cinnamic acid derivatives from our library.Furthermore, to obtain more information about structure-activity relationship 26 additional cinnamic acid esters and amides were synthesized andincluded in our assays. Among 45 cinnamic acid derivatives tested, 7 compounds have shown antifungal activity against C. lunatus, A. niger and P.ostreatus in in vivo inhibition tests. Compounds with antifungal activity were further evaluated for inhibition of CYP53A15 activity with spectral bindingtitration assay and HPLC. The best two inhibitors of CYP53A15 activity showed 70% inhibition at 600 mM concentration and were selected for furtheroptimization of new lead structures.27th Fungal Genetics Conference | 81

CONCURRENT SESSION ABSTRACTSSecretome analysis of Trichoderma harzianum cultivated in the presence of Fusarium solani cell wall or glucose. Marcelo HS Ramada 1,3 , Andrei SSteindorff 1 , Carlos Bloch Jr. 3 , Cirano J Ulhoa 2 . 1) Brasilia University, Cell Biology Department, Brasilia, DF, Brazil; 2) Federal University of Goias, BiochemistryDepartment, Goiania, GO, Brazil; 3) EMBRAPA CENARGEN, Mass Spectrometry Laboratory, Brasilia, DF, Brazil.Trichoderma harzianum is a fungus well known for its potential as a biocontrol agent of many fungal phytopathogens. The aim of this study was toevaluate the potential of T. harzianum ALL42 to control Fusarium solani, a phytopathogen fungus that causes several losses in common bean and soy cropsin Brazil and to evaluate the secreted proteins of T. harzianum ALL42 when its spores were inoculated and incubated in culture media supplemented (TLE)or not (MM) with nitrogen sources and in the presence or not of F. solani cell walls (FsCW). In the absence of FsCW, the media were supplemented withglucose (GLU). T. harzianum was able to control the phytopathogen growth and started to sporulate in its area after 7 days in a dual culture assay,indicating that it had successfully parasitized the host. T. harzianum was able to grow in TLE+FsCW, MM+FsCW, TLE+GLU, but unable to grow in MM+GLU.Protein quantification showed that TLE+FsCW and MM+FsCW had 45 and 30 fold, respectively, more proteins than TLE+GLU, and this difference wasobserved in the bidimensional gels, as the two supernatants from media supplemented with FsCW had around 200 spots and the one supplemented withglucose only had 18. TLE+FsCW and MM+FsCW had above 80% of spot similarity. A total of 100 proteins were excised from all three conditions andsubmitted to mass spectrometry analysis. 85 out of 100 proteins were identified. The only protein observed in all three conditions is a small protein, calledepl1, involved in eliciting plant-response against phytopathogens. An aspartic protease, previously described as related to mycoparasitism, was only foundwhen T. harzianum was grown with glucose. Gene expression was evaluated and confirmed the gel results. In the media supplemented with FsCW,different hydrolases like chitinases, beta-1,3-glucanases, glucoamylases, alpha-1,3-glucanases, and proteases were identified. Some proteins like a smallcystein-rich, alpha-L-arabinofuranosidase and NPP1, with no known function in mycoparasitism were also identified. T. harzianum ALL42 is able to inhibitthe growth and parasitize F. solani and showed a complex and diverse arsenal of proteins that are secreted in response to the presence of the cell walls,with novel proteins not previously described in mycoparasitism studies.Metabolic adaptation of the oomycete Phytophthora infestans during colonization of plants and tubers. Carol E. Davis, Howard S. Judelson. PlantPathology and Microbiology, University of California, Riverside, CA 92521.Phytophthora infestans is the causative agent of late blight and was responsible for the Irish famine in the 1840’s. Today it still continues to be a globalproblem and in the USA it has been reported that the economic loss on potato crops alone exceeds $6 billion per year. A successful phytopathogenicrelationship depends on the ability of the organism to adapt its metabolism during infection on various nutritional substrates (e.g., plant versus tuber) andat different times throughout infection when nutrients may be limiting. Investigation of this metabolic adaptation is key to understanding how P. infestanssucceeds as a pathogen. To do this, tomato plants and potato tubers were infected with zoospores using a “dipping” method. RNA was extracted at 3 dpiand 6 dpi and subsequently used in library preparation. Following this, the libraries were quality checked by analysis on a Bioanalyzer using a highsensitivity DNA chip. Using Illumina technology (50 bp, paired-end reads) RNA Sequencing was performed. For each sample an average of 262 million readswas obtained. As a reference for the in planta data, RNASeq was also performed on defined and complex media. Mining of the data shows that theexpression profiles of some pathways change, such as glycolysis and gluconeogenesis. Learning how metabolic adaptation occurs will prove useful in thedevelopment of novel control strategies for this plant pathogen.The fungi strike back: multidrug resistance in Aspergillus fumigatus and agricultural use of fungicides. Paul E. Verweij. Medical Microbiology, UMC StRadboud, Nijmegen, Netherlands.Aspergillus fumigatus is a saprophytic mould that causes a range of diseases in humans. The spectrum of diseases includes allergic conditions,aspergilloma and acute and chronic invasive aspergillosis. Acute invasive aspergillosis occurs typically in patients with compromised host defenses such asthose receiving treatment for leukemia. The management of invasive aspergillosis is very difficult as the diagnostic tools often lack sensitivity and thenumber of antifungal agents effective against the infection is limited. The azoles are the most important class of agents used for the treatment andprevention of invasive aspergillosis. Since 1998 resistance to medical triazoles has emerged in the Netherlands in clinical A. fumigatus isolates. Theseisolates commonly showed a multi-azole resistant phenotype and patients with azole-resistant aspergillosis failed to respond to azole therapy. Themortality rate of azole-resistant invasive aspergillosis was 88%. In more than 90% of resistant isolates a combination of changes in the target gene Cyp51Awas found: a substitution at codon 98 and a 34 bp tandem repeat in the promoter region (TR34/L98H). As person-to-person transmission is highly unlikelyin invasive aspergillosis, the dominance of a single resistance mechanism could not be explained by resistance development in azole-treated patients.Surveys in the environment showed that A. fumigatus isolates resistant to medical triazoles could be recovered from the environment, especially fromcultivated soil. The resistance mechanisms in these isolates were identical to those found in clinical isolates. Molecule alignment studies identified 5triazole fungicides that showed highly similar molecule structures to the medical triazoles. These 5 fungicides have been authorized between 1990 and1996, thus preceding the isolation of the first resistant clinical A. fumigatus isolate. TR34/L98H is increasingly reported in European countries, India, Chinaand Iran. Recently a new azole-resistance mechanism was found in Dutch patients and in the environment. The resistance mechanism again consisted of acombination of changes in the Cyp51A-gene: TR46/Y121F/T289A. This new resistance mechanism has spread in addition to TR34/L98H in the Netherlands,and has also been reported in the neighboring country Belgium. The selection of multiple azole resistance mechanisms in the environment posses a threatfor the use of azole drugs in the management of Aspergillus diseases in humans. Research that investigates the selection of azole resistance in A. fumigatusin the environment is urgently warranted.82

CONCURRENT SESSION ABSTRACTSEffect of antifungal resistance on virulence of Candida spp. L. Vale-Silva, A. Lohberger, D. Sanglard. Inst Microbiology, Univ Lausanne and Univ HospCenter, Lausanne, VD, Switzerland.Our laboratory has been involved in the understanding of mechanisms of antifungal resistance in fungal pathogens. While resistance is beneficial to fungiin the presence of antifungals both in vitro and in vivo, resistance mechanisms have some impact on the ability of fungal pathogens to propagate in thehost and cause disease. We have addressed this question with two pathogens, C. albicans (Ca) and C. glabrata (Cg). Our studies have focused on resistanceto azoles, which are widely used compounds. In Cg, azole resistance in clinical isolates is almost exclusively mediated by ABC transporters via gain offunction (GOF) mutations in the transcription factor (TF) CgPDR1. We observed that development of azole resistance in Cg was correlated with gain ofvirulence and fitness in animal models as compared to susceptible isolates. Recent results suggest that ABC transporters (especially CgCDR1) participate tothis phenotype. In further studies on interactions of Cg with murine bone marrow-derived macrophages (BMDM), we found that the GOF mutant was ableto evade phagocytosis as compared to a matched susceptible isolate. This effect was a consequence of impaired adhesion. This suggests that themechanism behind escape from phagocytosis was rather based on decreased recognition and/or adhesion by host macrophages. GOF mutations inCgPDR1 may thus allow Cg to evade the host’s innate immune response, which may in turn contribute to increased virulence. In Ca azole resistance ismediated by mutations in TF and in azole targets. GOF mutations in the three different activators including TAC1, MRR1 and UPC2 regulate specific subclassesof drug resistance genes belonging to ABC-transporters, major facilitators and azole targets, respectively. We have constructed sets of isogenicstrains lacking or carrying each GOF mutations in these TF and addressed their effect on virulence and tissue colonization in animal models. The presenceof GOF mutations in TAC1 and MRR1 had a neutral effect on virulence and ability to colonize host tissues. A UPC2 GOF mutation was negatively affectingvirulence and tissue colonization, which suggests that UPC2 activity generates fitness costs in Ca. Our results therefore highlight that the costs ofresistance mechanisms on virulence are depending on the fungal species and the type of resistance mechanism.From enzyme to fungal development or how sdhB mutations impact respiration, fungicide resistance and fitness in the grey mold agent Botrytiscinerea. Anais Laleve 1 , Anne-Sophie Walker 1 , Stephanie Gamet 2 , Valerie Toquin 2 , Daniele Debieu 1 , Sabine Fillinger 1 . 1) BIOGER, INRA, Thiverval-Grignon,France; 2) BAYER SAS, Bayer CropScience, Lyon, France.Respiration inhibitor fungicides are widely used to control fungal diseases on multiple crops. The succinate dehydrogenase inhibitors (SDHIs) are amongthe latest introduced molecules against the grey mold agent Botrytis cinerea on grapevine. We have recently isolated and characterized B. cinerea fieldstrains resistant to the SDHIs. Most of the strains harbor one single mutation in the succinate dehydrogenase subunit gene, sdhB affecting the ubiquinonebindingpocket. In this study we have introduced these mutations into a B. cinerea wild-type strain (B05.10) in order to evaluate the impact of eachmutation on SDH- and respiratory activity and inhibition by SDHIs from different chemistries. We also analyzed several parameters of B. cinerea’s life cycleto assess the fitness cost associated with the resistance mutation. Our results show a strict correlation between the sdhB mutation and the resistancespectra to SDHIs. These resistances can be fully explained by the affinities of the SDHIs to its modified target enzyme. Four out of the seven sdhB alleles ledto significantly reduced SDH activity and, in three cases (H272L, N230I, P225L), to reduced respiration rates. Concerning the fungal biology, we testedmycelial growth and sclerotia production on different media and temperatures, conidia production and germination, resistance to oxidative stress and ROSproduction, as well as pathogenicity on tomato and bean leaves. All mutants were affected for at least one parameter. However, fitness parameters ofmutants sdhB H272R and sdhB P225L showed the strongest modifications among all strains, e.g., reduced pathogenicity, strongly reduced conidia- and sclerotiaproduction(H272R). A clear correlation between fitness and respiration on one hand, fitness and allele frequency among natural populations on the other,is not yet obvious. However, our results in terms of resistance spectra and fitness parameters should help defining more efficient treatment strategiesagainst grey mold.Deciphering fungicide resistance mechanisms in phytopatogenic fungi, towards an assessment of resistance risk in new active ingredient research.Gabriel Scalliet 1* , Andreas Mosbach 1 , Diana Steinhauer 1 , Edel Dominique 1 , Robert Dietrich 2 . 1) Disease Control, Syngenta Crop Protection Munchwilen AG,Stein, Switzerland; 2) Syngenta Biotechnology, Inc 3054 Cornwallis Rd. Research Triangle Park, NC 27709.Resistance to crop protection fungicides can lead to a rapid loss of efficacy in the field. This is a major threat for the sustainability of our products. Anearly assessment is required in order to define best recommendations in product usage but also guide our choice in developing novel active ingredients. Inaddition to monitoring the situation for marketed fungicides, we are conducting early assessments for new active ingredients which include resistancegeneration, mode of resistance identification and fitness penalty determination. High throughput sequencing has become key for a rapid identification ofresistance mechanisms both for novel fungicides but also to holder chemistries for which the resistance mechanisms were so far not confirmed. A fewexamples will be discussed.27th Fungal Genetics Conference | 83

CONCURRENT SESSION ABSTRACTSSaturday, March 16 2:00 PM–5:00 PMMerrill HallParallels between Fungal Pathogens of Plants and AnimalsCo-chairs: Barbara Howlett and Axel BrakhageEmerging fungal (and Oomycete) threats to plant and ecosystem health. Sarah J. Gurr 1* , Daniel Bebber 1 , Matthew Fisher 2 . 1) Plant Sciences, OxfordUniversity, Oxford, Oxfordshire; 2) Imperial College of Science, Technology and Medicine, London.Fungal diseases have increased in severity and scale since the mid 20th Century and now pose a serious challenge to global food security and ecosystemhealth (Gurr et al., 2011 Fungal Biology Reviews 25 181-188 ). We have demonstrated recently that the threat to plants of fungal infection has nowreached a level that outstrips that posed by bacterial and viral diseases combined (Fisher et al., 2012 Nature 484 185-194) I shall highlight some of themore notable fungal and oomycete plant diseases and will draw attention to the emergence of new pathotypes affecting crop yields and decimating ournatural and managed landscapes. We have calculated that the losses due to persistent disease (that is, non-epidemic) caused by, for example rice blast,wheat stem rust, corn smut, soybean rust and potato late blight, if mitigated, would be sufficient to feed 8.5% of the global population (based on 2000calories per day for 1 year). Moreover, tree losses due to fungal and oomycete diseases such as dutch elm, chestnut blight, sudden oak death, jarrah diebackand pine beetle/blue stain fungus, thus far, have been estimated to account for significant CO2 sequestration losses (Fisher et al., 2012). The spreadof such organisms around the world is facilitated primarily by trade, but there is increasing concern that climate change may allow their establishment inregions hitherto deemed unsuitable. Increasing latitudinal ranges are anticipated under rising temperatures. However, the interactions between climatechange, crops and natural enemies are complex, and the extent to which crop pests and pathogens have altered their latitudinal ranges in response toglobal warming is largely unknown. We can demonstrate, from thousands of observations of hundreds of pests and pathogens, their shift polewards since1950, with a more rapid shift since 1990 (Bebber et al., (under review)). This latitudinal shift is seen in both Hemispheres. Moreover, the rate of movementsince 1950 is identical to that predicted by global climate data. This observed trend cannot be explained by latitudinal variation in technical capacity todetect and report pest incidences.Melanin as virulence determinant of human and plant pathogenic fungi. Axel A. Brakhage, Andreas Thywissen, Juliane Macheleidt, Sophia Keller, VitoValiante, Thorsten Heinekamp. Molec & Appl Microbiology, Leibniz Inst Natural Prod Res Infection Biol-HKI, Jena, Germany.In fungi, melanins are often associated with the cell wall and also contribute to the structural rigidity of spores. In several plant and human pathogenicfungi, melanins contribute to pathogenicity. For example, pigmentless mutants of the plant pathogens Magnaporthe oryzae and Colletotrichumlagenarium, as well as the human-pathogenic fungi Cryptococcus neoformans and Aspergillus fumigatus are less virulent when compared to melaninproducingwild-type strains. In M. oryzae, it was shown that a 1,8-dihydroxynaphthalene (DHN) melanin layer between the cell wall and the cell membraneis essential for turgor generation. The melanin acts as a barrier to the efflux of solute from the appressorium, which occurs as pressure is generated.Cellular turgor is translated into mechanical force of infection hyphae, forcing it through the leaf cuticle (1) . In human-pathogenic fungi, high turgor pressureis not required for penetration of tissue. In these fungi, melanin displays other virulence attributes such as the scavenging of reactive oxygen species. In A.fumigatus, at least two types of melanin are produced: Pyomelanin by polymerization of homogentisic acid, and DHN melanin. Transcription of genesessential for pyomelanin and DHN-melanin biosynthesis is detected during infection of mice. However, pyomelanin seems to be dispensable for fungalvirulence in the murine infection models tested (2,3) . DHN melanin is responsible for the grey-green color of A. fumigatus conidia. The biosynthesis enzymesof DHN melanin are encoded by six genes. Centrally is the polyketide synthase gene pksP, whose deletion results in a mutant strain with drasticallyattenuated virulence. Recent data of our laboratory showed that DHN melanin is essential not only for inhibition of apoptosis of phagocytes by interferingwith the host PI3K/Akt signaling cascade but also for effective inhibition of acidification of conidia-containing phagolysosomes (4,5) . These features allow A.fumigatus to survive in phagocytes and thereby to escape from human immune effector cells and to become an aggressive pathogen. 1) Wilson RA &Talbot NJ (2009) Nat Rev Microbiol. 7: 185-195 2) Keller et al. (2011) PLoS One 6:e26604 3) Schmaler-Ripcke et al. (2009) Appl Environ Microbiol. 75: 493 4)Thywiben et al. (2011) Front Microbiol. 2: 96 5) Volling et al. (2011) Cell Microbiol. 13: 1130.Nutrient immunity and systemic readjustment of metal homeostasis modulate fungal iron availability during the development of renal infections.Joanna Potrykus 1 , David Stead 2 , Dagmar S Urgast 3 , Donna MacCallum 1 , Andrea Raab 3 , Jörg Feldmann 3 , Alistair JP Brown 1 . 1) Aberdeen Fungal Group,University of Aberdeen, Aberdeen, United Kingdom; 2) Aberdeen Proteomics, University of Aberdeen, Aberdeen, United Kingdom; 3) Trace ElementSpeciation Laboratory, University of Aberdeen, Aberdeen, United Kingdom.Iron is a vital micronutrient that can limit the growth and virulence of many microbial pathogens. Here we show, that in the murine model ofdisseminated candidiasis, the dynamics of iron availability are driven by a complex interplay of localized and systemic events. As the infection progresses inthe kidney, Candida albicans responds by broadening its repertoire of iron acquisition strategies from non-heme iron (FTR1-dependent) to heme-ironacquisition (HMX1-dependent), as demonstrated in situ by laser capture microdissection, RNA amplification and qRT-PCR. This suggested changes in ironavailability in the vicinity of fungus. This was confirmed by 56 Fe iron distribution mapping in infected tissues via laser ablation-ICP-MS, which revealeddistinct iron exclusion zones around the lesions. These exclusion zones correlated with the immune infiltrates encircling the fungal mass, and wereassociated with elevated concentrations of murine heme oxygenase (HO-1) circumventing the lesions. Also, MALDI Imaging revealed an increase in hemeand hemoglobin alpha levels in the infected tissue, with their distribution roughly corresponding to that of 56 Fe. Paradoxically, whilst iron was excludedfrom lesions, there was a significant increase in the levels of iron in the kidneys of infected animals. This iron appeared tissue bound, was concentratedaway from the fungal exclusion zones, and was accompanied by increased levels of ferritin and HO-2. This iron accumulation in the kidney correlated withdefects in red pulp macrophage function and red blood cell recycling in the spleen, brought about by the fungal infection. Significantly, this effect could bereplicated by selective chemical ablation of splenic red pulp macrophages by clodronate. Collectively, our data indicate that systemic events shapemicronutrient availability within local tissue environments during fungal infection. The infection attenuates the functionality of splenic red pulpmacrophages leading to elevated renal involvement in systemic iron homeostasis and increased renal iron loading. Simultaneously, localized nutrientimmunity limits iron availability around foci of fungal infection in the kidney. In response, the fungus modulates its iron assimilation strategies.84

CONCURRENT SESSION ABSTRACTSCommon strategies in plant and human "necrotrophic" pathogens: role of PCD. N. Shlezinger 1 , H. Irme 2 , G. Braus 2 , A. Sharon 1 . 1) Tel Aviv University, TelAviv, Israel; 2) Georg-August University Goettingen, 37077 Goettingen, Germany.Botrytis cinerea is a model system to study pathogenicity of necrotrophic fungi. As inferred by the term "necrotrophic", such pathogens must first killhost cells before they can use them as a source of nutrients. To achieve this, B. cinerea promotes apoptotic cell death in infected plants, which facilitateslesion spreading during late infection stage. How the fungus survives the first encounter with living plant tissue remained unclear. We found that hyphaeof B. cinerea undergo massive PCD during early stages of infection, but fully recover upon transition to second phase of infection. Further studies using thefungal and plant mutants showed that survival, and hence pathogenicity of the fungus depended on anti-apoptotic machinery; fungal mutants bearingdefects in the anti-apoptotic gene BcBIR1 had reduced virulence whereas strains over-expressing this gene were hypovirulent. A similar phenomenon wasobserved with another necrotrophic plant pathogen, but not with two hemi-biotrophic pathogens. These results showed that the anti-PCD machinery isessential for pathogenic development necrotrophic plant pathogens. In order to extend these finding to other systems, we generated transgenic strains ofthe human pathogen Aspergillus fumigatus. This fungus shares common stages of infection with B. cinerea and hence might also need the anti-PCDmachinery for infection. Indeed, when conidia of A. fumigates are inoculated on a sensitive tissue, the developing hyphae undergo massive cell death, andsimilar to B. cinerea they fully recover later. Collectively, our results show that certain plant and human fungal pathogen share common strategies andmechanisms when infecting their host. One such mechanism is control of host-induced cell death, through conserved anti-apoptotic machinery.Septin-mediated plant tissue invasion by the rice blast fungus Magnaporthe oryzae. Yasin Dagdas, Lauren Ryder, Michael Kershaw, Nick J. Talbot. DeptBiological Sci, Univ Exeter, Exeter, United Kingdom.Magnaporthe oryzae is the causal agent of rice blast, one of the most serious economic problems affecting rice production. During plant infection, M.oryzae develops a differentiated infection structure called an appressorium. This unicellular, dome-shaped structure generates cellular turgor, that istranslated into mechanical force to cause rupture of the rice cuticle and entry into plant tissue. Development of a functional appressorium requirescompletion of a single round of mitosis shortly after conidial germination, which also leads to initiation of autophagic recycling of the contents of thefungal spore to the appressorium. We have recently shown that a hetero-oligomeric septin GTPase complex is necessary for re-organisation of a toroidal F-actin network at the base of the appressorium which allows re-establishment of polarised fungal growth. Septins scaffold F-actin, via the ezrin-radixinmoesin(ERM) protein, Tea1, and phosphatidylinositide interactions at the appressorium plasma membrane. The septin ring assembles in a Cdc42 andChm1-dependent manner and forms a diffusion barrier to localize the Inverse-Bin-Amphiphysin-RVS (I-BAR)-domain protein, Rvs167, and Wiskott-AldrichSyndrome protein (WASP), Las17 at the point of penetration. This leads to formation of a penetration hyphae that breaches the host cuticle and leads toplant tissue colonization. We present evidence that septin-mediated plant infection is regulated by a specialised NADPH oxidase-tetraspanin complexnecessary for control of F-actin dynamics. We also describe the potential operation of a pressure-mediated checkpoint pathway that leads to initial septinassembly and activation and the re-orientation of the cortical F-actin cytoskeleton to facilitate plant tissue invasion.Components of the urease complex govern virulence of Fusarium oxysporum on plant and animal hosts. Katja Schaefer, Elena Pérez-Nadales, Antonio DiPietro. Departamento de Genética, Universidad de Córdoba, 14071 Cordoba, Spain.In the soilborne pathogen Fusarium oxysporum, a mitogen-activated protein kinase (MAPK) cascade homologous to the yeast filamentous growthpathway controls invasive growth and virulence on tomato plants. Full phosphorylation of Fmk1 requires the transmembrane protein Msb2, a member ofthe family of signalling mucins that have emerged as novel virulence factors in fungal plant pathogens. A yeast two-hybrid screen for proteins interactingwith the Msb2 cytoplasmic tail identified UreG, a component of the urease enzymatic complex. UreG belongs to a set of accessory proteins needed toactivate Apo- urease, which converts urea to yield ammonia and carbon dioxide. The F. oxysporum genome contains two structural urease genes, ure1 andure2. Mutants in ureG or ure1 showed reduced growth on urea as the sole carbon and nitrogen source. Lack of urease activity in the mutants resulted infailure to secrete ammonia and to increase the extracellular pH. The DureG mutants caused significantly reduced mortality on tomato plants and on theanimal model host Galleria melonella, while Dure1 mutants only showed reduced virulence on tomato plants. Real-time qPCR analysis of key genesinvolved in nitrogen uptake and assimilation, as well as in the urea cycle, during infectious growth of F. oxysporum in G. melonella revealed increasedtranscript levels of arginase, which converts arginine to urea. Our results suggest a role for the urease accessory protein UreG in fungal virulence on plantand animal hosts.The role of LysM effectors in fungal fitness. Anja Kombrink 1 , Jason Rudd 2 , Dirk-Jan Valkenburg 1 , Bart Thomma 1 . 1) Phytopathology, WageningenUniversity, Wageningen, Netherlands; 2) Department of Plant Pathology and Microbiology, Rothamsted Research, Harpenden, Hertfordshire, UnitedKingdom.LysM effector genes are found in the genomes of a wide range of fungal species. The encoded LysM effectors are secreted proteins that contain a varyingnumber of LysM domains, which are carbohydrate-binding modules. Ecp6, secreted by tomato leaf mould fungus Cladosporium fulvum, is the firstcharacterized LysM effector. We demonstrated that Ecp6 specifically binds chitin, the major constituent of fungal cell walls that acts as a microbialassociatedmolecular pattern (MAMP) and triggers immune responses upon recognition by the host. Ecp6 outcompetes plant receptors for chitin binding,and thus prevents the activation of immune responses. Many fungal genomes, including saprophytes, carry multiple LysM effector genes that share onlylow sequence conservation and encode a varying number of LysM domains. We speculate that fungal LysM effectors might bind different carbohydratesand exert various functions in fungal fitness. In the fungal wheat pathogen Mycosphaerella graminicola, two LysM effectors were identified. Mg3LysM, butnot Mg1LysM, suppresses chitin-induced immune responses in a similar fashion as Ecp6. Interestingly, unlike Ecp6, both Mg1LysM and Mg3LysM inhibitdegradation of fungal hyphae by plant chitinases, revealing an additional function for LysM effectors in pathogen virulence. We recently observed thatMg1LysM binds to the bacterial cell wall constituent peptidoglycan. Similarly, a LysM effector from the saprophytic fungus Neurospora crassa showedpeptidoglycan binding. We hypothesize that peptidoglycan binding by LysM effectors plays a role in the interaction of fungal species with bacterialcompetitors. The soil-borne fungal pathogen Verticillium dahliae contains seven LysM effectors genes of which one (Vd2LysM) is induced during tomatoinfection. Inoculation with two independent knock-out mutants revealed that Vd2LysM is required for full virulence of V. dahliae. However, Vd2LysM doesnot specifically bind chitin and does not function in a similar fashion as previous characterized LysM effectors. Thus, its function in virulence remainsunclear.27th Fungal Genetics Conference | 85

CONCURRENT SESSION ABSTRACTSGenes important for in vivo survival of the human pathogen Penicillium marneffei. Harshini C. Weerasinghe, Michael J. Payne, Hayley E. Bugeja, AlexAndrianopoulos. Genetics, The University of Melbourne, Parkville, Victoria, Australia.Pathogenic fungi are having an increasing global impact in the areas of health, agriculture and the environment. As such it is essential to understand themechanisms that fungi employ to survive and grow within a host. The emergence of many new “opportunistic fungal pathogens” has to a great extentaltered the traditional view that pathogenicity was solely reliant on the inherent properties of the pathogen. In fact, the ability of a pathogen to causedisease in some hosts but not in others suggests that pathogenic determinants are complex and dynamic, and are largely dependent on specific pathogenhostrelationships. Despite this there are conserved aspects of the interactions between host and pathogen. For example., hosts employ innate immuneresponses as an almost immediate recognition and attack mechanism against invading pathogens. Penicillium marneffei is a temperature dependentdimorphic fungus, growing in a hyphal form producing conidia at 25°C and as a yeast form at 37°C. Despite its importance as an opportunistic pathogen,little is known about the biology and mechanism of infection of P. marneffei. The infectious agents (conidia) are believed to be inhaled, reaching thealveoli of the lungs, where they are phagocytosed by alveolar macrophages for elimination. At this point that P. marneffei switches growth to a pathogenicyeast cell form, and is able to withstand macrophage cytotoxic attacks to cause infection. In order to understand how P. marneffei responds to the host,RNA-seq analysis was used to create a transcriptomic profile of P. marneffei, when infected in murine macrophages. These results were compared to RNAseqdata from hyphal (25°C) and yeast (37°C) cells grown in vitro in order to identify genes that are specifically upregulated during infection. Based on thisanalysis a group of genes of varying functions were chosen for gene deletion studies and tested for defects in pathogenicity. Among these is a group ofPep1-like aspartic endopeptidases which are a uniquely expanded family in P. marneffei and that show reduced virulence in a macrophage model.86

CONCURRENT SESSION ABSTRACTSSaturday, March 16 2:00 PM–5:00 PMChapelSecondary MetabolismCo-chairs: Gillian Turgeon and Bettina TudzynskiGenomic profiles of secondary metabolism genes in Cochliobolus pathogens. B. Condon 1* , D. Wu 1 , K. Bushley 3 , I. Grigoriev 2 , C. Elliott 4 , B. Howlett 4 , B. G.Turgeon 1 . 1) Plant Pathology & Plant-Micro Biology, Cornell University, Ithaca, NY; 2) DOE Joint Genome Institute, Walnut Creek, CA; 3) Dept of PlantPathology, Oregon State University, Corvallis OR; 4) School of Botany, University of Melbourne, Melbourne, AU.The genomes of five Cochliobolus heterostrophus strains, two Cochliobolus sativus strains, and three additional Cochliobolus species (Cochliobolusvictoriae, Cochliobolus carbonum, Cochliobolus miyabeanus) were sequenced at the Joint Genome Institute (JGI). These species are notable pathogens ofeconomically important grasses and many produce a signature secondary metabolite host selective toxin, conferring high virulence to a host of a particulargenotype. We catalogued the suites of secondary metabolism genes [focusing on those encoding backbone nonribosomal peptide synthetases (NRPSs) andpolyketide synthases (PKSs)] in these genomes and performed comparative phylogenomic analyses within the Cochliobolus genus and in a kingdom-widecontext. We found that NPS and PKS genes were broadly conserved, disparately conserved, or species-unique. Distribution patterns for conserved ordisparately conserved genes suggest an evolutionary mechanism involving rapid duplication, loss, and recombination of protein domains. Genescategorized as species unique within our dataset often had lone orthologs in phylogenetically distant species. Expanding genomic resources may revealthat ‘signature’ genes, formerly thought to be species-unique are present broadly, but sporadically, in other fungi. These results have strong implicationsfor understanding evolution of genes for host selective toxins and associated virulence.A biosynthetic gene cluster for the antifungal metabolite phomenoic acid in the plant pathogenic fungus, Leptosphaeria maculans. Candace Elliott 1 ,Damien Callahan 2 , Daniel Schwenk 3 , Markus Nett 4 , Dirk Hoffmeister 3 , Barbara Howlett 1 . 1) School of Botany, University of Melbourne, Melbourne,Australia; 2) Metabolomics Australia, School of Botany, The University of Melbourne, Victoria 3010, Australia; 3) Friedrich-Schiller-Universität, DepartmentPharmaceutical Biology at the Hans-Knöll-Institute, Beutenbergstrasse 11a, 07745 Jena, Germany; 4) Leibniz Institute for Natural Product Research andInfection Biology e.V., Hans-Knöll-Institute, Beutenbergstrasse 11a, 07745 Jena, Germany.Phomenoic acid, a long chain aliphatic carboxylic acid, is a major metabolite produced by Leptosphaeria maculans, a fungal pathogen of Brassica napus(canola). Early biosynthetic studies suggested that the methyl group derived from S-adenosylmethionine (SAM), whereas the incorporation pattern of[13C] acetate suggested a polyketidic origin of the linear portion of phomenoic acid (Devys et al., 1984). We have used domain modelling to predict acandidate polyketide synthase (PKS) for phomenoic acid biosynthesis. Of the 15 predicted polyketide synthases (PKS) in the L. maculans genome, sevenwere reducing with the appropriate domains (KS - keto-synthase; AT - acyltransferase; DH - dehydratase; MT- methyltransferase; ER - enoylreductase; KR -ketoreductase; ACP- acyl carrier protein) for the biosynthesis of phomenoic acid. The most highly expressed of these seven genes, PKS2, was silenced to10% of that of wild type levels and the resultant mutant produced 25 times less phomenoic acid than the wild type did, indicating that PKS2 is involved inphomenoic acid biosynthesis. This gene is part of a cluster and nearby genes are co-regulated. A two-fold reduction in the expression of the adjacenttranscriptional regulator C6TF, led to at least a 20-fold reduction in expression of PKS2, as well as of other genes in the cluster (P450, YogA, RTA1 andMFS), but not of the adjacent ChoK or a hypothetical gene (Hyp). This down-regulated mutant also showed a marked reduction in phomenoic acidproduction. Phomenoic acid is toxic towards another canola pathogen Leptosphaeria biglobosa ‘canadensis’, but L. maculans and to a lesser extent thewheat pathogen, Stagonospora nodorum are more tolerant. Phomenoic acid may play a role in allowing L. maculans to outcompete other fungi in itsenvironmental niche.Fusarin C biosynthesis in Fusarium fujikuroi: the fusarin C gene cluster, their function and regulation. Eva-Maria Niehaus 1 , Karin Kleigrewe 2 , PhilippWiemann 1 , Lena Studt 1,2 , Hans-Ulrich Humpf 2 , Bettina Tudzynski 1 . 1) Institute of Plant Biology and Biotechnology, Schlossplatz 8, 48143 Muenster,Germany; 2) Institute of Food Chemistry, Corrensstr. 45, 48149 Muenster, Germany.The filamentous fungus F. fujikuroi is known to produce a variety of structurally diverse secondary metabolites such as the plant hormones gibberellins,pigments and mycotoxins. In order to reduce the health risk of mycotoxins in food, feed and biotechnologically produced gibberellin preparations,identification of mycotoxin biosynthesis genes is of great importance. The recently sequenced genome of F. fujikuroi contains 17 polyketide synthases(PKS). So far only four of them can be linked to specific products: bikaverin, fusarins, fumonisins and fusarubins. The focus of this work is studying thebiosynthesis and regulation of the mutagenic mycotoxin fusarin C by external signals, such as nitrogen availability and pH. Furthermore the involvement ofpotential transcription factors and global regulators such as AreA, AreB, GS, PacC and three members of the velvet-like complex (Vel1, Vel2, Lae1) wereinvestigated. We show that all nine genes are co-expressed under nitrogen sufficient conditions. Chromatin immunoprecipitation (ChIP) experimentsrevealed a high level of H3K9 acetylation under these favorite conditions. By combination of gene deletion and overexpression of the cluster genes and cocultivationof different mutants, we were able to identify the intermediates and finally unraveled the entire fusarin biosynthetic pathway which we arepresenting in a model.27th Fungal Genetics Conference | 87

CONCURRENT SESSION ABSTRACTSCellular development integrating primary and induced secondary metabolism in the filamentous fungus Fusarium graminearum. Jon Menke 1 , JakobWeber 2 , Karen Broz 3 , H. Corby Kistler 1,3* . 1) Department of Plant Pathology, University of Minnesota, St. Paul, USA; 2) Molekulare Phytopathologie,Universität Hamburg, Germany; 3) USDA ARS Cereal Disease Laboratory, St. Paul, MN, USA.Several species of the filamentous fungus Fusarium colonize plants and produce toxic small molecules that contaminate agricultural products, renderingthem unsuitable for consumption. Among the most destructive of these species is F. graminearum, which causes disease in wheat and barley and oftencontaminates the grain with harmful trichothecene mycotoxins. Induction of these secondary metabolites occurs during plant infection or in culture inresponse to chemical signals. Here we report that trichothecene biosynthesis involves a complex developmental process that includes dynamic changes incell morphology and the biogenesis of novel subcellular structures. Two cytochrome P-450 oxygenases (Tri4p and Tri1p) involved in early and late steps intrichothecene biosynthesis were tagged with fluorescent proteins and shown to co-localize to vesicles we call “toxisomes.” Toxisomes, the inferred site oftrichothecene biosynthesis, dynamically interact with motile vesicles containing a predicted major facilitator superfamily protein (Tri12p) previouslyimplicated in trichothecene export and tolerance. The immediate isoprenoid precursor of trichothecenes is the primary metabolite farnesylpyrophosphate. When cultures are shifted from non-inducing to trichothecene inducing conditions, changes occur in the localization of the isoprenoidbiosynthetic enzyme HMG CoA reductase. Initially localized in the cellular endomembrane system, HMG CoA reductase increasingly is targeted totoxisomes. Metabolic pathways of primary and secondary metabolism thus may be coordinated and co-localized under conditions when trichothecenesynthesis occurs.LaeA sleuthing reveals cryptic gene clusters in pathogenic Aspergilli. Nancy Keller 2 , Wenbing Yin 2 , Saori Amaike 2 , Katharyn Affeld 2 , JinWoo Bok 2 , DanielSchwenk 3 , Dirk Hoffmeister 3 , Joshua Baccile 1 , Ry Forseth 1 , Frank Schroeder 1 . 1) Boyce Thompson Institute and Department of Chemistry and ChemicalBiology, Cornell University, Ithaca, NY 14853, USA; 2) Department of Plant Pathology, Department of Medical Microbiology and Immunology, andDepartment of Bacteriology, University of Wisconsin-Madison, Madison, WI 53706, USA; 3) Department of Pharmaceutical Biology at the Hans-Knöll-Institute, Friedrich-Schiller-Universität, Beutenbergstrabe 11a, 07745 Jena, Germany.The human and plant pathogenic Aspergilli, Aspergillus fumigatus and A. flavus, are known to produce a plethora of secondary metabolites. However,most of these metabolites are not yet characterized although their gene clusters are apparent from genomic sequence. In both species, the nuclearprotein LaeA regulates the expression of many of these uncharacterized gene clusters. Following leads from laeA mutant microarray data, we created genedeletion and overexpression strains and used 2D NMR-based comparative metabolomic analyses to identify previously undescribed metabolites from bothspecies. In A. fumigatus a tryptophan-derived iron(III)-complex, hexadehydro-astechrome (HAS), was found to be the major product of the cryptic has nonribosomalpeptide synthetase (NRPS) cluster. In A. flavus we show that two separate clusters encode enzymes that produce partially overlapping sets ofnovel piperazines, pyrazines, and morpholines. These L-tyrosine metabolites are activated by two NRPS-like proteins, LnaA and LnbA. Loss andoverexpression of these metabolites impacted fungal development in these species.The KMT6 Histone H3 K27 Methyltransferase Regulates Expression of Secondary Metabolites and Development in Fusarium graminearum. Kristina M.Smith, Lanelle R. Connolly, Michael Freitag. Department of Biochemistry and Biophysics, Center for Genome Research and Biocomputing, Oregon StateUniversity, Corvallis, OR 97331.The cereal pathogen Fusarium graminearum produces secondary metabolites toxic to humans and animals, yet coordinated transcriptional regulation ofsecondary metabolite gene clusters remains largely a mystery. By ChIP-sequencing we found that regions of the F. graminearum genome with secondarymetabolite clusters are enriched for a histone modification, trimethylated histone H3 lysine 27 (H3K27me3), associated with gene silencing. Thismodification was found predominantly in regions that lack synteny with other Fusarium species, generally subtelomeric regions. H3K27me3 and di- ortrimethylated H3K4 (H3K4me2/3), modifications associated with gene activity, are found in mutually exclusive regions of the genome. To betterunderstand the role of H3K27me3, we deleted the gene for the putative H3K27 methyltransferase, KMT6, a homolog of Drosophila Enhancer of zeste, E(z).The kmt6 mutant lacks H3K27me3, as shown by western blot and ChIP-sequencing, displays growth defects, is sterile, and produces mycotoxins underconditions where they are not generated in wildtype (WT) strains. RNA-sequencing showed that genes modified by H3K27me3 are most often silent, asabout 75% of the 4,449 silent genes are enriched for H3K27me3. Surprisingly, we found 22% of the 8,855 expressed genes enriched for H3K27me3. Asubset of genes that were enriched for H3K27me3 in WT gained H3K4me2/3 in kmt6 (1,780 genes), and an overlapping set of genes showed increasedexpression. Almost 95% of the remaining 2,720 annotated silent genes showed no enrichment for either H3K27me3 or H3K4me2/3. In these cases absenceof H3K27me3 is insufficient for expression, which suggests a requirement for additional factors for gene expression. Taken together, we show that absenceof H3K27me3 allows expression of 14% of all annotated genes, resulting in derepression of predominantly secondary metabolite pathways and otherspecies-specific functions, including potentially secreted pathogenicity factors. This study provides the framework for novel targeted strategies to controlthe “cryptic genome” and specifically secondary metabolite expression.88

CONCURRENT SESSION ABSTRACTSSecondary metabolism in Botrytis cinerea: the grey and pink sides of a pathogen. M. Viaud 1 , H. Sghyer 1 , J. Schumacher 2 , A. Simon 1 , B. Dalmais 1 , J.M.Pradier 1 . 1) INRA, BIOGER, Av. L. Brétignières, 78850 Grignon, France; 2) Department of Botany and Plant Pathology, Purdue University, West Lafayette, IN47906, USA.Sequencing the genome of the grey mould fungus Botrytis cinerea revealed 40 secondary metabolism (SM) gene clusters corresponding to thebiosynthesis of mostly unknown compounds including polyketides (21), terpenes (11) and non-ribosomal peptides (8). The two identified phytotoxicmetabolites are the polyketide botcinic acid (BOA) and the sesquiterpene botrydial (BOT). Transcriptomic studies previously identified the correspondingBOA and BOT gene clusters. Deletion of the key enzyme BcBOA6 (PolyKetide Synthase) or BcBOT2 (SesquiTerpene Cyclase) encoding genes did not causeany defects, while bcboa6/bcbot2 double mutants were significantly impaired in necrotrophic development on several hosts suggesting that the twocompounds have a redundant function. We are currently investigating how the BOT and BOA gene clusters are regulated: The BOA cluster contains a gene(bcboa13) encoding a C6H6 zinc finger transcription factor (TF). Surprisingly, while bcboa6 mutant has no virulence defect, bcboa13 mutant shows adrastic reduced virulence which is even more pronounced than that of the bcboa6/bcbot2 double mutant lacking both known toxins. A wholetranscriptome analysis of bcboa13 mutant is underway to determine whether other genes apart from those of the BOA cluster are regulated by this TF. Inopposite to the BOA cluster, the BOT cluster does not contain any TF-encoding gene. We therefore set up a Yeast-One-Hybrid library that contains themajority of B. cinerea TFs (393 out of 406) and screened it with the promoter of bcbot2. This led to the identification of a C2H2 TF called BcYOH1.Inactivation of bcyoh1 gene and expression analysis revealed that this TF acts as a global regulator of SM, regulating the expression of genes of the BOT,BOA, and 20 other SM clusters. As in other fungi, the Velvet complex takes part in regulation of light-dependent development and SM in B. cinerea. Geneinactivation of bcvel1, however, does not significantly modify the production of BOA and BOT. Instead, BcVEL1 plays a significant role in the regulation ofoxalic acid (OA) formation and pigmentation: it regulates negatively the synthesis of melanin and positively the synthesis of OA and bikaverin (BIK) anotherpolyketidic pigment that is only produced in rare pink strains of B. cinerea.Is fungal secondary metabolism regulated by competing insects? Annika Regulin 1 , Nancy Keller 2 , Frank Kempken 1 . 1) Department of Botany, Christian-Albrechts University, Kiel, Germany; 2) Department Medical Microbiology and Immunology, Dept of Bacteriology, UW-Madison, USA.Fungi synthesize an astonishing variety of secondary metabolites, some of which belong to the most toxic compounds in the living world. Even thoughlittle is known about the benefit of these metabolites, the ability to regulate the secondary metabolism might be seen as an evolutionary adaptation.Presumably fungi regulate secondary metabolites (e.g. mycotoxin) in response to confrontation with natural competitors like insects to guarantee efficientexploitation of environmental resources (1-3). Admittedly it should be mentioned that secondary metabolites are not the only defence mechanisms offungi (4). In order to enlighten the biological function of these secondary metabolites with reference to chemical defence reactions of insect-fungalinteractions, we utilized complementary approaches of experimental ecology and functional genomic techniques. The vinegar fly Drosophila melanogasterand its natural antagonist Aspergillus nidulans are used as an ecology model system. To analyse fungal up- or down regulated target genes in theinteraction of A. nidulans with Drosophila larvae microarray analysis was performed. This led to the identification of secondary metabolite genes up- ordown-regulated under these conditions. Quantitative RT-PCR was employed to analyze secondary metabolite gene expression at different time points.Fungal single, double and triple mutations of identified up-regulated genes are currently analyzed in confrontation assays to identify potentialmodifications in gene expression and the survival rate of larvae concerning to chemical defense reaction of fungus-insect interaction compared to wildtype. This could reveal insights about the biological function of secondary metabolite genes and clusters such as stc and mdp.(1.) Rohlfs, M., Albert, M., Keller, N. P., and Kempken, F. (2007) Biol Lett 3, 523-25. (2.) Kempken, F., and Rohlfs, M. (2010) Fungal Ecol 3, 107-14. (3.)Rohlfs, M., Trienens, M., Fohgrub, U., and Kempken, F. (2009) in "The Mycota XV. (Anke, T., Ed.), Springer Heidelberg, New York, Tokyo, pp. 131-51 (4.)Kempken, F. (2011) Mol Ecol 20, 2876-77.27th Fungal Genetics Conference | 89

CONCURRENT SESSION ABSTRACTSSaturday, March 16 2:00 PM–5:00 PMHeatherLight Sensing and Circadian RhythmsCo-chairs: Luis Larrondo and Reinhard FischerCircadian rhythms in gene expression in Aspergillus nidulans. Maria Olmedo 1,2 , Julio Rodriguez-Romero 3,4 , Julian Röhrig 3 , Martha W. Merrow 1,2 , ReinhardFischer 3 . 1) Institute for Medical Psychology, Ludwig-Maximilians-Universität-München, München, Bavaria, Germany; 2) Department of MolecularChronobiology, University of Groningen, The Netherlands; 3) Institute for Applied Biosciences, Karlsruhe Institute of Technology, Germany; 4) Centre forPlant Biotechnology and Genomics Universidad Politécnica de Madrid(UPM) Campus de Montegancedo Autopista M-40 (Km 38) 28223-Pozuelo de Alarcón(Madrid).The circadian clock is an endogenous timekeeper that allows organisms to predict cyclic changes in their environment derived from the rotationalmovement of the Earth. The fungus Neurospora crassa has substantially contributed to the understanding of the mechanism of the circadian clock, mainlydue to the presence of an easily assayable rhythm in conidiation. Our understanding of circadian rhythms tells us that they are present in most organisms(from bacteria to humans), yet they have been difficult to detect in other fungal species, likely due to the absence of a clear readout. To circumvent thisproblem, we have adopted a tool developed for the study of the clock in Neurospora and other model organism, namely bioluminescent reporter proteins,to study circadian rhythms in Aspergillus nidulans, where a previous report has suggested a circadian clock (Greene et al. 2003). The characterization of theAspergillus clock represents a further step in the understanding of the biology of this fungal genus, which is of great importance for medical, industrial andagricultural reasons. We have produced strains with the promoters of Neurospora clock controlled gene homologs conJ, ccgA and gpdA fused to the (A.nidulans optimized) firefly luciferase ORF. We observe an oscillation in the bioluminescence signal in these strains in constant conditions, implying thepresence of an endogenous oscillator. This free running rhythm is detectable in both constant light and darkness and is entrained by environmentalsignals. Using these reporters we are studying the contribution of the wc (white collar) homologs and other photoreceptors to the Aspergillus clock. Theprotein WC-1 is a central molecule of the best-studied Neurospora oscillator, which also comprises the protein FRQ (FREQUENCY). The absence of a FRQhomolog in Aspergillus implies that the characterization of its clockwork may unveil new components of additional Neurospora oscillators.Circadian clock-gated cell division cycles in Neurospora crassa. C. Hong 1 , J. Zamborszky 1 , M. Baek 1 , K. Ju 1 , H. Lee 1 , L. Larrondo 2 , A. Goity 2 , A. Csikasz-Nagy 3 .1) Molecular and Cellular Physiology, University of Cincinnati, Cincinnati, OH; 2) Departamento de Genética Molecular y Microbiología, Facultad deCiencias Biológicas, Pontificia Universidad Católica de Chile, Casilla 114-D, Santiago, Chile; 3) Randall Division of Cell and Molecular Biophysics, andInstitute for Mathematical and Molecular Biomedicine, King’s College London, London, SE1 UL, UK.Asynchronous nuclear divisions are readily observed in filamentous fungi such as Ashbya gossypii and Neurospora crassa. Our computational simulations,however, predict synchronous circadian clock-gated mitotic divisions if the division cycles of such multinucleated organisms are coupled with circadianrhythms. Based on this hypothesis, we investigate the coupling between the cell cycle and the circadian clock in Neurospora crassa. First, we show WC-1-dependent light-induced expression of stk-29 mRNA (homolog of wee1), which suggests that there exists a conserved coupling between the clock and thecell cycle via STK-29 in Neurospora as in mammals. Second, we demonstrate that G1 and G2 cyclins, CLN-1 and CLB-1, respectively, show circadianoscillations with luciferase bioluminescence reporters. Moreover, clb-1 and stk-29 gene expressions show circadian clock-dependent light-induced phaseshifts, which may alter the timing of divisions. Third, we show circadian clock-dependent synchronized nuclear divisions by tracking nuclear morphologywith histone hH1-GFP reporter. Synchronized divisions occur late in the evening, and they are abolished in the absence of circadian rhythms (frq KO ). Ourfindings demonstrate the importance of circadian rhythms for synchronized mitotic cycles and establish Neurospora crassa as an ideal model system toinvestigate mechanisms that couple the cell cycle and the circadian clock.Light regulates growth, stress resistance and metabolism in the fungal pathogen Aspergillus fumigatus. Kevin K. Fuller, Carol S. Ringleberg, Jennifer J.Loros, Jay C. Dunlap. Genetics, Geisel School of Medicine at Darmouth, Hanover, NH.Light serves as an important environmental cue that influences developmental and metabolic pathways in a variety of fungi. Interestingly, orthologs of aconserved blue light receptor, WC-1, promote virulence in two divergently related pathogenic species, Cryptococcus neoformans and Fusarium oxysporum,suggesting that photosensory systems may be conservatively linked to fungal pathogenesis. Aspergillus fumigatus is the predominant mold pathogen ofimmunocompromised patients, but if and how the organism responds to light has not been described. In this report, we demonstrate that the fungus canindeed sense and distinctly respond to both blue and red portions of the visible spectrum. Included in the A. fumigatus photoresponse is a reduction inconidial germination kinetics, increased hyphal pigmentation, enhanced resistance to acute ultra-violet and oxidative stresses, and an increasedsusceptibility to cell wall perturbation. Through gene deletion analysis we have found that the WC-1 ortholog, LreA, is a bone fide blue light receptor in A.fumigatus that is required for the photopigmentation response. However, the DlreA mutant retains several blue light mediated responses, including thegermination and stress resistance phenotypes , suggesting other blue light receptors are operative in this fungus. We also show that the putative red lightsensing phytochrome, FphA, is involved with some, but not all, blue light specific phenotypes, indicating a complex interaction between red and blue lightphotosystems in A. fumigatus. Finally, whole genome microarray analysis has revealed that A. fumigatus displays broad patterns of gene induction andrepression upon exposure to light. Affected genes are largely metabolic and include those involved in lipid and sterol synthesis, respiration, carbohydratecatabolism, amino acid metabolism and metal ion homeostasis. Taken together, these data demonstrate the importance of the photic environment on thephysiology of A. fumigatus and provide a foundation for future studies into an unexplored area of this important pathogen.90

CONCURRENT SESSION ABSTRACTSShedding light on Botrytis biology: characterization of the WC1 photoreceptor and FRQ homologues in the necrotrophic plant pathogen Botrytiscinerea. Paulo Canessa 1 , Montserrat Hevia 1 , Julia Schumacher 2 , Paul Tudzynski 3 , Luis F Larrondo 1 . 1) Depto. Genética Molecular y Microbiologia, PontificiaUniversidad Catolica de Chile, Santiago, Chile; 2) Purdue University, USA; 3) West F Wilhelms Univ Muenster Germany.The non-pathogenic fungus Neurospora crassa has been a premier model for the study of circadian clocks, but it is the only fungus where clocks havebeen molecularly characterized. In Neurospora the circadian oscillator is based on the transcriptional control of frq expression by the White Collar complex(WCC) composed of WC-1, a blue-light photoreceptor and WC-2, its functional partner. The WCC activates frq expression until FRQ promotes thephosphorylation of the WCC, shutting down its transcriptional activity. Then FRQ is progressively phosphorylated and degraded setting the bases ofcircadian rhythmicity. While circadian clocks modulate defense in plants, little is known about its participation in fungal pathogenicity. In an effort tocharacterize light responses and the circadian clock in B. cinerea, we have generated KO of homologues of wc-1 and frq (bcwcl1 and bcfrql). Our resultsindicate that light can modulate B. cinerea growth and activates transcriptional responses. Most -but not all of themare mediated by BcWCL1. Moreover,the obtained bcwcl1 KO mutant strains exhibit enhanced conidiation patterns and susceptibility to hydrogen peroxide. Virulence assays show significantlyreduced lesion formation under constant light conditions (LL) and light:dark cycles (LD), but not under constant darkness (DD). Further analysis of thebcwcl1 KO strains indicates non-altered ROS patterns in planta but delayed penetration on onion epidermis. RT-qPCR experiments have confirmed dailyoscillations in bcfrql levels under LD and DD conditions. Using a BcFRQL-luciferase translational reporter, we have further observed oscillatory levels of theBcFRQL protein under both temperature cycles and constant culture conditions. Both BcFRQL and bcfrql respond to light pulses, while under oscillatoryculture conditions, BcFRQL anticipates cyclical-environmental changes, a key characteristic of circadian behavior. Finally, bcfrql KO mutants also exhibitimpaired pathogenicity, with a drastic reduction in spore production. To our knowledge, these results provide the first evidence of functionallight/circadian machineries in B. cinerea, showing that in this necrotroph, homologues of circadian clock components modulate the plant pathogeninteraction from a fungal perspective. Fondecyt 1090513, postdoc 3110127, AT24121100, ICGEB CRPCHI0902.The transcription factor FL is phosphorylated and interacts with a trehalose related protein in Neurospora crassa. Carmen Ruger-Herreros 1 , GencerSancar 2 , Michael Brunner 2 , Luis M. Corrochano 1 . 1) Departamento de Genetica, Universidad de Sevilla, Spain; 2) BZH, Universität Heidelberg, Germany.Several environmental cues, including light, promote a developmental transition in Neurospora crassa that leads to the formation of conidia. Conidiationis controlled by FLUFFY (FL), a zinc finger transcription factor. Light activates the transcription of fl through the transient binding of the WC complex to thefl promoter. Light also activates the transcription of several conidiation genes in Aspergillus nidulans, and their Neurospora homologs have been identifiedin the Neurospora genome. We have assayed the activation by light of the Neurospora homologs of A. nidulans conidiation genes (flbA, flbC, flbD, medAand stuA), and the Neurospora conidiation gene con-10 as a control. Unlike con-10, none of the Neurospora homologs of the A. nidulans conidiation geneswere induced by light in vegetative mycelia. However, we found that deletion fl resulted in light-dependent mRNA accumulation for all the conidiationgenes. This result indicated that the absence of FL allows the binding of the WC complex to the promoter of these genes to activate transcription in a lightdependentmanner. We have assayed the amount of WC proteins in the Dfl and wild type strains but we did not find any difference between the twostrains. We expect to identify additional genes deregulated by the absence of FL after massive sequencing of total RNA (RNAseq) using a Dfl strain andwild-type strain in dark and light conditions. We have investigated the role of FL during conidiation in Neurospora using a tagged version of FL. FL ispresent in vegetative mycelia but the amount increses after light exposure. We observed several forms of FL due to phosphorylation, and and we havedetermined by mass spectrometry that FL is phosphorylated in several residues. We have immunoprecipitated FL to identify proteins that may interactwith FL. We have found a protein that interacts with FL in different growth conditions. This protein has been described in other organisms and plays a rolein the ability to grow in the presence of trehalose. Since FL is a transcription factor, we have use FL::3XFLAG strain to do ChIPseq in order to identify theputative binding sites of FL to the DNA. We expect that the results from these experiments will help us to understand in more detail the role of FL in theactivation of gene transcription during development.Regulation of gene expression in response to light in Trichoderma atroviride. Jose Cetz 1 , Nohemi Carreras-Villaseñor 1 , Monica Garcia-Esquivel 1 , UlisesEsquivel-Naranjo 2 , Jose M. Villalobos-Escobedo 1 , Cei Abreu-Goodger 1 , Alfredo H. Herrera-Estrella 1 . 1) National Laboratory of Genomics for Biodiversity,CINVESTAV-IPN, Irapuato, Irapuato, Mexico; 2) Facultad de Ciencias Naturales. Universidad Autónoma de Queretaro. Queretaro, Mexico.Trichoderma is used as a photomorphogenetic model due to its ability to conidiate upon exposure to light. In total darkness, T. atroviride growsindefinitely as a mycelium provided that nutrients are not limiting. However, a brief pulse of blue light given to a radially growing colony inducessynchronous sporulation. Photoconidiation in Trichoderma is controlled by the blr1 and blr2, orthologs of the N. crassa white-collar genes. We appliedhigh-throughput sequencing technology to RNA samples from the wild type strain grown in the dark or after exposure to a pulse of white or blue-light, aswell as from a photoreceptor mutant (Dblr-1) exposed to white light. We identified 331 transcripts regulated by white light and 204 responsive specificallyto blue light, both induced and repressed, the majority of them blr1 dependent. Among the genes identified there is a set of transcription factors. We haveobtained gene disruption mutants of several of the transcription factors, and all potential light receptors. Using this strategy we have obtained mutantsthat do not conidiate in response to light, as well as mutants that do not require this stimulus to conidiate, and a conection between light and carbonmetabolism. Further, transcriptional analysis in RNAi machinery mutants indicated that light induced conidiation is defective in the Ddcr2, Ddcr1Ddcr2,Drdr3 and Dago2 mutants, and significant differences were found in the set of light responsive genes between the dicer mutants and the wild type. Thesedata indicate that in T. atroviride, the RNAi machinery plays an important role in controlling development.27th Fungal Genetics Conference | 91

CONCURRENT SESSION ABSTRACTSGenome-wide analysis of light responses in Mucor circinelloides. Victoriano Garre, Sergio López-García, Eusebio Navarro, Santiago Torres-Martínez.Departamento de Genética y Microbiología, Universidad de Murcia, 30100 Murcia, Spain.Light regulates developmental and physiological processes in a wide range of fungi. Particularly, Zygomycete fungi have developed complex mechanismsto control the responses to light that await detailed characterization at molecular level. The zygomycete Mucor circinelloides is a good model for thispurpose because its genome has been sequenced and several molecular tools are available. Mucor, like other Zygomycetes, has three white collar-1 genes(mcwc-1a, mcwc-1b and mcwc-1c) that code for proteins which present characteristics of photoreceptors. Each mcwc-1 gene controls a specific responseto light. Thus, mcwc-1a and mcwc-1c control phototropism and photocarotenogenesis, respectively, whereas the mcwc-1b function in regulation by lighthas not been proved. In order to deepen in the regulation by light in Mucor, a systematic approach using microarrays was followed to characterize whitelight-inducible transcriptional changes in wild-type and knockout mutants for each mcwc-1 gene. Analysis of microarray data revealed that light is mainly apositive signal for transcription in Mucor, as in other fungi, since 123 genes were up-regulated in the wild-type strain in response to light, whereas only 26were down-regulated, considering a threshold of threefold change. Genes strongly induced by light included genes known to be up-regulated by light, likethe carotenogenic gene carB (74-fold), cryptochrome (45-fold) and mcwc-1c (22-fold), supporting reliability of the microarray data. Although many of upregulatedgenes code for proteins implicated in protection against light-induced damage, several of them code for protein involved in signal transductionthat could be involved in light responses like phototropism. Transcriptomic analysis of mcwc-1 mutants showed that induction of around 60% of the genesis mediated by mcwc-1a, whereas only 1% is mediated by mcwc-1c and none is mediated by mcwc-1b, suggesting that mcwc-1a is the main photoreceptor.Searching for cis-acting regulatory motifs upstream of genes regulated by mcwc-1a identified consensus sequences similar to those found in lightregulated genes of Neurospora crassa. Moreover, the identification of a small group of genes regulated by the three mcwc-1 genes points out that thethree proteins form complexes to regulate gene expression. Funded by MINECO (BFU2012-32246), Spain.Shedding light on secondary metabolite cluster gene expression, sporulation, UV-damage repair and carotenogenesis in the rice pathogen Fusariumfujikuroi. Phillipp Wiemann, Bettina Tudzynski. Institut für Biologie und Biotechnologie der Pflanzen Westfälische Wilhelms-Universität MünsterSchlossplatz 8 48143 Münster Germany.The rice pathogen Fusarium fujikuroi produces economically important secondary metabolites like gibberellic acids and carotenoids as well asmycotoxins like bikaverin and fusarin C. Their production is activated in response to environmental stimuli such as light, pH or nutrient availability. In thisstudy, we evaluate the effects of light and different putative light receptors on growth and differentiation as well as secondary metabolism. Bimolecularfluorescence complementation proved that homologs of the Neurospora crassa White Collar proteins in F. fujikuroi (WcoA and WcoB) form a nuclearlocalized complex (WCC) that is needed for full functionality. Deletion and complementation of both genes revealed that the WCC represses bikaverin geneexpression in constant light conditions and induces immediate light-dependent carotenoid gene expression as shown by northern blot analyses.Additionally the WCC represses conidiogenesis in response to light. The effects observed regarding bikaverin and carotenoid gene expressions as well asconidiogenesis are antagonistically to the ones observed in the velvet mutant, making a connection between the WCC and the velvet complex feasible,similarly to the situation in Aspergillus nidulans. Since carotenoid production was maintained in both wcoA and wcoB single as well as in wcoA/B doublemutants in constant light conditions, we focused on characterization of additional putative light receptors in F. fujikuroi. Deletion of the phytochrome-likeencodinggene fph1 did not show any significant phenotype. Deletion of phl1, coding for a cryptochrome/photolyase demonstrated impaired carotenoidbiosynthesis gene expression upon exposure to light. Additionally, gene expression and HPLC analyses of these mutants demonstrated loss of fusarin Cgene expression and concomitant production formation compared to the wild type, suggesting a distinct transcriptional activity for this barelycharacterized class of enzymes. Finally UV mutagenesis experiments and qRT-PCR demonstrate that WcoA, WcoB and Phl1 are involved in UV-damagerepair most likely by transcriptionally activating phr1, encoding a CPD-photolyase. The data presented here allow us to draw a first model of how lightreceptors function in a signaling network in the rice pathogen F. fujikuroi.92

CONCURRENT SESSION ABSTRACTSSaturday, March 16 2:00 PM–5:00 PMFred Farr ForumFungal Evo-DevoCo-chairs: Steve Harris and Brian ShawThe Molecular Foundations of the Fungal Lifestyle. Antonis Rokas. Department of Biological Sciences, Vanderbilt University, Nashville, TN.A defining characteristic of Fungi is that, in contrast to plants and animals, they are typically embedded in their food and digest it externally in thepresence of competitors (think of the blue lines of mold in blue cheese). Thus, different fungi specialize in “eating” different foods (hence their diverseprimary metabolism), and because digestion happens externally, fungi have also evolved potent food defense mechanisms (hence their diverse secondaryor specialized metabolism). In many ways, the genes involved in fungal primary metabolism are their “teeth”, whereas the genes involved in secondarymetabolism are their “horns”, “spines” and “claws”. I will use examples from our recent studies on fungal human pathogens, domesticated fungi andfungal genome evolution to argue that metabolism and more generally physiology is fundamental to the fungal lifestyle. In contrast to plants and animals,in which most phenotypic evolution proceeds largely through developmental changes affecting growth and morphological form, phenotypic evolution inthe fungal kingdom occurs largely through changes in metabolism.Gene expression and regulation during conidial morphogenesis in Neurospora crassa. Daniel J. Ebbole, Shengli Ding, Dawoon Chung, HeatherWilkinson, Shaw Brian. Plant Pathology & Microbiol, Texas A&M Univ, College Station, TX.The regulatory pathway controlling conidiation in N. crassa consists of five genes, acon-4 (aconidiate), fl (fluffy), acon-3, csp-1 (conidial separation) andcsp-2. The first three genes are required for the transition of aerial hyphae from filamentous growth to the budding pattern resulting in proconidial chainformation. Maturation of the proconidial chain to individual conidia relies on the activation of genes for two additional transcription factors, csp-1 and csp-2 that are required for proper interconidial septum formation and spore release. High throughput mRNA sequencing defined a set of genes induced duringconidiation and allowed us to identify genes for cell wall degrading enzymes required for conidial separation. Based on expression patterns and epistasisamong the regulators, we defined the order of gene action required for conidial morphogenesis from the initiation of budding to the release of matureconidia.Comparative developmental morphology in lentinoid mushrooms: toward a new fungal evo-devo? David S. Hibbett. Biology, Clark University, Worcester,MA.Fungi produce a mind-boggling diversity of complex forms, including mushrooms, puffballs, coral fungi, and others. Reconstructing patterns ofmorphological transformations has been a major focus of fungal molecular systematics, and developmental morphology has been a traditional source ofcharacters for fungal taxonomy. However, few studies have explicitly compared developmental programs in a phylogenetic context, and research into thegenetic bases of morphological evolution in fungi has lagged far behind that in plants and animals. In this talk, I will review three cases of naturallyoccurring developmental variation in the polyphyletic “lentinoid” fungi, which may suggest profitable avenues for studies in fungal evo-devo: 1) Normallyagaricoid species of Lentinus, Neolentinus and Lentinellus may be induced to produce coralloid forms under conditions of light deprivation or lowtemperatures. These phenomena provide examples of phenotypic plasticity that may reflect evolvability of developmental programs. 2) Panus rudis has ashort lateral stipe and fruits directly on wood, while the closely related P. fulvus has a dramatically elongated stipe and often fruits from sclerotiaimmersed in leaf litter. The developmental transformation between forms suggests a case of hypermorphosis, with a delayed onset of pileus initiation in P.fulvus. Light-induced formation of pilei in P. rudis may provide clues to the mechanisms of offset in stipe elongation. 3) Lentinus tigrinus is a gilledmushroom that has a naturally occurring “secotoid” mutant that has an enclosed, puffball-like hymenophore. The secotioid morphology appears to beconferred by a recessive allele at a single locus. Resolving the gene(s) responsible for the secotioid phenotype may provide clues to the evolution ofgasteromycetes. Complete genome sequences have been (or are being) produced for species of Lentinus, Panus, Lentinellus and Neolentinus, which willprovide opportunities to study the mechanisms underlying inter- and intraspecific developmental variants in lentinoid mushrooms.The Cdc42 GTPase module and the evolution of conidiophore architecture in Aspergillus. Steven D. Harris. Center Plant Sci Innovation, Univ Nebraska,Lincoln, NE.Conidiophores are reproductive structures that enable filamentous fungi to produce and disseminate large numbers of asexual spores. The diversity inconidiophore morphology is sufficiently large to serve as a basis for fungal systematics. Aspergillus and Penicillium species are members of the familyTrichocomaceae that form conidiophores with characteristic architecture. Whereas the Penicillium conidiophore appears to be a modified branchedhyphal structure, the Aspergillus conidiophore is seemingly more complex and includes additional cell types. Here, it is proposed that the “aspergillioid”conidiophore may have evolved from a “penicillioid” ancestor via changes in expression of key regulators of the GTPases Cdc42 and RacA. In particular,mutations that affect these regulators in A. nidulans dramatically alter conidiophore morphology by eliminating terminal vesicles and permitting formationof branched stalks. Because the transcriptional regulatory network that controls conidiophore development in Aspergillus is well characterized, furtherstudy of how this network links to regulators of Cdc42 should provide fundamental insight into the evolution of developmental morphogenesis in fungi(i.e., fungal evo-devo).Cdc14 association with basal bodies in the oomycete Phytophthora infestans indicates potential new role for this protein phosphatase. Audrey M.V. Ah-Fong, Howard S. Judelson. Plant Pathology & Microbiology, University of California, Riverside, CA.The dual-specificity phosphatase Cdc14 is best known as a regulator of cell cycle events such as mitosis and cytokinesis in yeast and animal cells.However, the diversity of processes affected by Cdc14 in different eukaryotes raises the question of whether its cell cycle functions are truly conservedbetween species. Analyzing Cdc14 in Phytophthora infestans should provide further insight into the role of Cdc14 since this organism does not exhibit aclassical cell cycle. Prior study in this organism already revealed novel features of its Cdc14. For example, instead of being post-translationally regulatedlike its fungal and metazoan relatives, PiCdc14 appears to be mainly under transcriptional control. It is absent in vegetative hyphae where mitosis occurs27th Fungal Genetics Conference | 93

CONCURRENT SESSION ABSTRACTSand expressed only during the spore stages of the life cycle which are mitotically quiescent, in contrast to other systems where it is expressedconstitutively. Since transformants overexpressing PiCdc14 exhibit normal nuclear behavior, the protein likely does not play a critical role in mitoticprogression although PiCdc14 is known to complement a yeast Cdc14 mutation that normally arrests mitosis. Further investigation into the role of PiCdc14uncovered a novel role. Subcellular localization studies based on fusions with fluorescent tags showed that PiCdc14 first appeared in nuclei during earlysporulation. During the development of biflagellated zoospores from sporangia, PiCdc14 transits to basal bodies, which are the sites from which flagelladevelop. A connection between Cdc14 and flagella is also supported by their phylogenetic distribution, suggesting an ancestral role of Cdc14 in basalbodies and/or flagellated cells. To help unravel the link between PiCdc14 and the flagella apparatus, searches for its interacting partners using both yeasttwo hybrid and affinity purification are underway. Together with colocalization studies involving known basal body/centrosome markers such as centrinand gamma-tubulin, the location and hence the likely roles of PiCdc14 will be revealed.Molecular Determinants of Sporulation in Ashbya gossypii. Jurgen W. Wendland, Lisa Wasserstroem, Klaus Lengeler, Andrea Walther. Yeast Genetics,Carlsberg Laboratory, Copenhagen V, Kopenhagen V, Denmark.Previously we have analysed the pheromone response MAPK signal transduction cascade in A. gossypii. The major findings were (i) deletion of bothpheromone receptor genes STE2 and STE3 did not inhibit sporulation whereas (ii) deletion of the transcription factor STE12 resulted in hypersporulation(Wendland et al. 2011). Here we present our analysis of key A. gossypii homologs of Saccharomyces cerevisiae sporulation specific genes. We show thatmutants in IME1, IME2, KAR4, and NDT80 are blocked in sporulation. Mutants in IME4, KAR4, and UME6 also confer a vegetative growth defect. IME4expression was found during vegetative growth while IME2 was not detected under these conditions. We performed transcriptional profiling of nonsporulatingstrains and determined a core set of about 50 down-regulated sporulation specific genes in these mutants. Interestingly, this set of downregulatedgenes is upregulated in the A. gossypii ste12 mutant providing regulatory evidence of the hypersporulation phenotype of this mutant. Othergenes identified in the RNAseq data indicated that during development of sporangia metabolic genes for nutrient uptake are active. Therefore weperformed Return-To-Growth assays with mutants inhibited in the sporulation pathway. These strains were kept under conditions in which the wild typeinitiates sporulation. This lead to induction of sporangium formation, a stage at which these strains remained. Supply of new nutrients resulted in hyphaloutgrowth in all mutants indicating that after initiation of the sporulation program A. gossypii can reverted to vegetative growth at different stages. Inaddition we identified differential regulation of two endoglucanases encoded by ENG1 and ENG2. While ENG1 was not differentially regulated, ENG2 wasdown-regulated in e.g. ime1 but strongly up-regulated in ste12. Deletion analysis of ENG2 showed that Eng2 is required for hyphal fragmentation intoindividual sporangia. We can thus provide a detailed overview of the genetic regulation of sporulation in A. gossypii. A comparison with S. cerevisiaehighlights the role of KAR4 in sporulation upstream of IME1. Finally, our study provides further evidence that the pheromone signaling response MAPKcascadein A. gossypii has a regulatory control function over sporulation alongside regulation of sporulation by nutritional cues.THE velvet regulators in Aspergilli. Heesoo Park, JJae-Hyuk Yu. Bacteriology, University of Wisconsin Madison, Madison, WI.The velvet regulators are the key players coordinating fungal growth, differentiation and secondary metabolism in response to various internal andexternal cues. All velvet family proteins contain the conserved velvet homology motif (~190 a.a.), and define a novel class of fungal specific transcriptionfactors with the DNA binding ability. Some velvet regulators form time and/or cell type specific complexes with other velvet regulators or non-velvetproteins. These complexes play differential roles in regulating growth, development, sporogenesis and toxigenesis. Among the velvet complexes, the VelB-VosA hetero-complex acts as a functional unit conferring the completion of sporogenesis (focal trehalose biogenesis and spores wall completion), and thelong-term viability of spore, and the attenuation of conidial germination in the model filamentous fungus Aspergillus nidulans. Both velB and vosA areactivated by AbaA in developing cells, and the VelB-VosA complex plays a dual role in activating genes associated with spore maturation and in exertingnegative feedback regulation of developmental genes. Interestingly, the VelB-VosA complex plays similar yet somewhat distinct roles in spore maturation,dormancy and germination in Aspergillus fumigatus and Aspergillus flavus. A comprehensive model depicting the roles of the velvet regulators in aspergilliis presented.A network of HMG-box transcription factors regulates sexual cycle in the fungus Podospora anserina. J. Aït-Benkhali 1,2 , E. Coppin 1,2 , S. Brun 1,2,3 , T.Martin 4 , C. Dixelius 4 , R. Debuchy 1,2 . 1) Univ Paris-Sud, Institut de Génétique et Microbiologie, Orsay, France; 2) CNRS, Institut de Génétique etMicrobiologie, Orsay, France; 3) UFR des Sciences du Vivant, Université Paris-7 Diderot, Paris, France; 4) Department of Plant Biology and Forest Genetics,Uppsala BioCenter, Swedish University of Agricultural Sciences and Linnean Center for Plant Biology, Uppsala, Sweden.High-mobility group B proteins are eukaryotic DNA-binding proteins characterized by the HMG-box functional motif. These transcription factors play apivotal role in global genomic functions and in the control of genes involved in specific developmental or metabolic pathways. The filamentous ascomycetePodospora anserina contains 12 HMG-box genes. Of these, four have been previously characterized; three are mating-type genes that control fertilizationand development of the fruiting-body, whereas the last one encodes a factor involved in mitochondrial DNA stability. Systematic deletion analysis of theeight remaining uncharacterized HMG-box genes indicated that none were essential for viability, but that seven were involved in the sexual cycle. TwoHMG-box transcription factors display striking features. Pa_1_13940, an ortholog of SpSte11 from Schizosaccharomyces pombe, is a pivotal activator ofmating-type genes in P. anserina, whereas Pa_7_7190 is a repressor of several phenomena specific to the stationary phase, most notably hyphalanastomoses. Constitutive expression of mating-type genes in a DPa_1_13940 strain did not restore fertility, indicating that Pa_1_13940 has additionalfunctions related to sexual reproduction besides activating mating-type genes. RT-qPCR analyses of HMG-box genes in different HMG-box deletion strainsindicated that Pa_1_13940 is at the hub of a network of several HMG-box factors that regulate the sexual cycle. Complementation experiments with astrain deleted for mating-type genes revealed that this network control fertility genes in addition to mating-type target genes. This study points to thecritical role of the HMG-box members in sexual reproduction in fungi, as 11 out of 12 members were involved in the sexual cycle in P. anserina.Pa_1_13940 and SpSte11 are conserved transcriptional regulators of mating-type genes, although P. anserina and S. pombe have diverged 1.1 billion yearsago. Two HMG-box genes, SOX9 and its upstream regulator SRY, play also an important role in sex determination in mammals. The mating-type genes andtheir upstream regulatory factor form a module of HMG-box genes similar to the SRY/SOX9 module, suggesting it may be ancestral in Opisthokonta.94

CONCURRENT SESSION ABSTRACTSSaturday, March 16 2:00 PM–5:00 PMKilnEnvironmental MetagenomicsCo-chairs: Chris Schadt and Betsy ArnoldMicrobial Responses to a Changing Climate: Implications for the Future Functioning of Terrestrial Ecosystems. Donald R. Zak. University of Michigan, AnnArbor, MI.Soil harbors a phylogenetically diverse community of microorganisms whose physiological activity mediates the biogeochemical cycling of carbon andnitrogen at local, regional, and global scales. These microbial communities are structured by the physical environment as well as the availability of growthlimitingresources (i.e., organic compounds in plant detritus). Presently, human activity is manipulating both the physical conditions and the availability oflimiting resources to soil microbial communities at a global scale, but the implications of doing so for the future functioning of ecosystems is presentlyunclear. In this presentation, I will discuss the ways in which humans are manipulating the ecological constraints on microbial communities in soil, thecompositional and functional responses that may result, and identify gaps in our knowledge that limit our ability to anticipate the response of microbialcommunities and ecosystem processes in a changing environment. Using an array of metagenomic approaches, I will provide evidence that rates ofatmospheric N deposition expected in the near future can down regulate the transcription of fungal genes with lignocellulolytic function, thereby alteringmicrobial community composition, slowing plant litter decay, and increasing soil C storage. This mechanism is not portrayed by any biogeochemical modelsimulating ecosystem response to atmospheric N deposition, and it demonstrates that microbial communites in soil may respond to a changingenvironment in ways that have unanticipated consequences for the future functioning of terrestrial ecosystems.The Interaction of Mycoplasma-related Endobacteria with their Arbuscular Mycorrhizal Fungal Host. Mizue Naito 1 , Teresa Pawlowska 2 . 1) Dept. ofMicrobiology, Cornell University, Ithaca, NY; 2) Dept. of Plant Pathology & Plant-Microbe Biology, Cornell University, Ithaca, NY.Arbuscular mycorrhizal fungi (AMF), comprising the monophyletic phylum Glomeromycota, are obligate biotrophs, and form symbiotic associations with80% of terrestrial plants. AMF associate symbiotically with the roots of plants, and are specialized in the transfer of nutrients from the soil to the planthost. In return for increased nutrient uptake, the plants supply AMF with up to 20% of their photosynthetically derived carbohydrates. Thus, AMFsymbiosis contributes significantly to global nutrient cycling and terrestrial ecosystems. AMF have been known to harbour two types of bacteria in theircytoplasm: (i) the Burkholderia-related Candidatus Glomeribacter gigasporarum and (ii) a Mycoplasma-related bacteria, which we refer to as Mycoplasmarelatedendobacteria (MRE). MRE live freely in the AMF cytoplasm, and have been found associated with all lineages of AMF worldwide. Virtually nothingis known about the MRE, such as their evolution, biological capabilities, and whether they are mutualists or parasites of their AMF hosts. In order tounderstand the nature of this symbiosis, and determine the role that the MRE play in arbuscular mycorrhizae, next generation sequencing (Roche 454 andIllumina) was performed on MRE isolated from 3 distinct AMF hosts, Claroideoglomus etunicatum, Funneliformis mosseae, and Racocetra verrucosa.Phylogenetic reconstruction and divergence dating using 22 conserved genes have revealed that MRE form a novel monophyletic subclade of theMycoplasmas and have diverged from their Mycoplasma relatives at least 400 million years ago, which may indicate the establishment of the MRE-AMFassociation to be quite ancient. Analysis of annotated genes have revealed novel proteins that are likely to play a role in interacting directly with the fungalhost. Preliminary data suggest that MRE are important in enabling the completion of the life cycle of their AMF hosts.Metagenomic analysis reveals hidden fungal diversity in grass rhizosphere and tree foliage. Ning Zhang 1 , Stephen Miller 1 , Shuang Zhao 1 , Hayato Masuya 2 .1) Plant Biology and Pathology, Rutgers Univ, New Brunswick, NJ; 2) Dept Forest Microbiology, Forestry and Forest Products Research Institute,Matsunosato 1, Tsukuba, Ibaraki 305-8687, JAPAN.The diversity of microorganisms on earth remains poorly understood. Unculturable fungi inhabiting rhizosphere, phyllosphere, and other less studiedniches are thought to represent a large fraction of the unknown diversity. In this study, we used both culture-dependent method and Illuminametagenomic sequencing approach to explore fungal diversity in two environments: grass (Poa pratensis, Kentucky bluegrass) rhizosphere and tree(Cornus spp., dogwood) foliage. For the grass rhizosphere sample, Illumina metagenomic analysis identified 1,192 fungal genera from 20.8 million reads,while the culture-based method identified 21 genera. For the Cornus sample, metagenomic analysis identified 73 fungal genera from 6.6 million reads,while 22 genera were isolated from culture. From both cases, we found that metagenomic sequencing analysis revealed significantly higher fungaldiversity than culture-based method, which will help us better understand the diversity and role of fungi in the ecosystem.Host-to-pathogen gene transfer facilitated infection of insects by a pathogenic fungus. Weiguo Fang, Xiaoxuan Chen. College of Life Sciences, ZhejiangUniversity, Hangzhou, Zhejiang, China.Inspite being of great concern to human health and the management of plants and animals, the mechanisms facilitating host switching of eukaryoticpathogens remain largely unknown. The endophytic insect-pathogenic fungus Metarhizium robertsii evolved directly from endophytes and itsentomopathogenicity is an evolutionarily acquired characteristic. We found that M.robertsii acquired a sterol carrier (Mr-NPC2a) from an insect byhorizontal gene transfer (HGT). Mr-NPC2a increased the amount of ergosterol in hyphal bodies by capturing sterol from insect hemolymph, and thusmaintained cell membrane integrity and improved fungal survival rate. On the other hand, the reduction in sterol (substrate for molting hormonesynthesis) in insect hemolymph elongated larval stage, which allows the fungus to fully exploit host tissues and produce more conidia. This is first report ofHGT from host to a eukaryotic pathogen, and the host gene ultimately improved the infectivity of the pathogen.Structure and function of soil fungal communities across North American pine forests. Kabir Peay 1 , Jennifer Talbot 1 , Dylan Smith 1 , Rytas Vilgalys 2 , JohnTaylor 3 , Thomas Bruns 3 . 1) Dept. of Biology, Stanford University, Stanford, CA; 2) Dept. of Biology, Duke University, Durham, NC; 3) Plant & MicrobialBiology, UC Berkeley, Berkeley, CA.Fungi are a critical component of the diversity and function of terrestrial ecosystems. They regulate decomposition rates, facilitate plant nutrient uptakeand have a profound impact on agriculture and economics. Understanding the forces that structure fungal communities thus has important theoretical and27th Fungal Genetics Conference | 95

CONCURRENT SESSION ABSTRACTSpractical implications. While ecologists have long recognized the importance of scale on ecological processes, fungal communities have primarily beenstudied at small-scales, focusing on deterministic processes. To rectify this knowledge gap we are using next generation sequencing techniques to surveysoil fungi in North American pine forests with a sampling design that allows us to examine community structure from roots-to-biomes. Our results showthat soil fungal communities in these ecosystems are highly diverse and are structured primarily by large-scale, macroecological processes, rather thansmall-scale deterministic processes. Our results also show that there is high functional redundancy in soil fungal communities. This work demonstratesthat increasing the scale of observation is critical to a complete understanding of the ecological dynamics of soil fungi and the ecosystem processes theymediate.Genomic analysis of Mortierella elongata and its endosymbiotic bacterium. Gregory Bonito 1 , Andrii Gryganskyi 1 , Christopher Schadt 2 , Dale Pelletier 2 , AmySchaefer 3 , Gerald Tuskan 2 , Jessy Labbé 2 , Sofia Robb 4 , Rebecca Ortega 1 , Francis Martin 5 , Mitchel Doktycz 2 , Kurt LaButti 6 , Matt Nolan 6 , Robin Ohm 6 , IgorGrigoriev 6 , Rytas Vilgalys 1 . 1) Duke University, Durham NC; 2) Oak Ridge National Laboratory, Oak Ridge TN; 3) University of Washington, Seattle WA; 4)University of California, Riverside CA; 5) Institut National de la Recherche Agronomique, Nancy France; 6) Joint Genome Institute, Walnut Creek CA.Mortierella belong to a group of basal fungi (Mortierellomycotina) common to soils and the rhizosphere and endosphere of many plant species.Mortierella species are known for rapid growth and abundant lipid production. Mortierella elongata is one species commonly isolated from forest soils andhealthy plant roots where it grows asymptomatically as an endosymbiont. Mortierella elongata is a heterothallic species but can also reproduce asexuallythrough chlamydospores and sporangiospores. Recent reports indicate that some isolates of M. elongata host endosymbiotic bacteria, which may betransmitted vertically via spores. However, it is still unclear whether all Mortierella species host endosymbionts or whether these are lineage-specificassociations. Given the geographically widespread distribution of Mortierella elongata and its ubiquitous presence in forest soils and plants we chose tosequence its genome through the JGI Forest Metatranscriptome CSP. We also sought to assemble the genome of the bacterial endosymbiont to addresswhether there are genomic signatures of co-adaptation or co-evolution in the genomes of Mortierella and its endosymbiotic bacterium, which may impactthe function and growth of Mortierella elongata. The 50 Mb genome of M. elongata was sequenced to 112x coverage. Of the 220,113 putative proteinsidentified in M. elongata, 109,093 appear to be unique (e.g. only ~50% have orthologs in other fungal species having sequenced genomes). The M.elongata genome appears to be enriched in genes related to tryptophan metabolism, siderophore group nonribosomal peptides, glucan 1,4-alphaglucosidases, and in lipid metabolism (e.g. sphingolipids, etherlipids, and glycerophopholids) compared to genome sequences of other basal fungi. Theendosymbiotic bacterium sequenced along with the M. elongata isolate is related to Glomeribacter (endosymbiont of Gigospora, Scutellospora, and otherGlomeromycota) within the Burkholdariales. The ~2.6 MB endosymbiont genome is larger than that of Glomeribacter but quite reduced compared to freelivingisolates of Burkholdaria. The reduced genome size of this bacterium, and the fact that it has thus far evaded pure culture isolation, supports the viewthat this is an ancient and obligate symbiosis.Integrative genomics of poplar-fungal pathogen interactions. Richard C. Hamelin. Forest Sciences, University of British Columbia, Vancouver, BC, Canada.Poplar is an important tree, both from an ecosystemic point of view as riparian species, and as a commercial agro-forestry crop for the production ofwood and paper products and increasingly as a source of bioenergy. Fungi in the Urediniales and the Dothideomycetes are responsible for the mostimportant diseases of poplars. In most tree-fungal pathogen interactions, a few founder species are key determinants of the outcome. To betterunderstand these interactions at the landscape level and predict their outcome, we are using a variety of genome-based approaches. We have appliedDNA barcoding and multigene phylogeny to poplar pathogens to assess species diversity and host specificity both from environmental and herbariumderivedsamples. We have found a high level of fidelity in host tracking of pathogens vis-à-vis their host. However, this fidelity tends to break down whenpoplar is grown in intensive plantations or when interspecific hybrids are planted. To further investigate and understand this pattern, we sequenced thegenomes of 12 poplar pathogens with different host specificity. By comparing these genomes we identified core gene sets as well as genes that are uniqueto each species and are candidate determinants of the interaction outcomes. Annotation of these genome sequences with customized pipelines allowedus to assign putative functions to the candidate genes and detect effector-like sequence profiles. RNAseq profiling of the interaction of poplar rust ondifferent hosts and their hybrids confirms the uniqueness of expression patterns in host-specific infections. In addition, we are using population genomere-sequencing approaches to detect selection patterns at the genome level. Several of the candidate genes identified in the upstream analyses encodesecreted proteins and possess a signature of positive selection. Integration of this data can be used as predictive resources to determine the outcome ofpoplar-pathogen interactions in the environment.Fungal pathogen and endophyte genetics within the context of forest community dynamics. M.-S. Benitez 1 , M. H. Hersh 2 , L. Becker 1 , R. Vilgalys 3 , J. S.Clark 1,3 . 1) Nicholas School of the Environment, Duke University, Durham, NC; 2) Department of Biology, Eastern Michigan University, Ypsilanti, MI; 3)Department of Biology, Duke University, Durham, NC.Fungal pathogens play important roles in forest community dynamics, particularly through negative-density dependent regulation. Negative-densitydependence regulation is hypothesized to be regulated by the presence of host-specific pathogens. Studies on forest pathogens, however, indicate thepredominance of generalist seedling pathogens, capable of infecting more than one host species. To understand the mechanisms through which“generalist” pathogens contribute to forest-community dynamics we conducted extensive surveys of seedling pathogens in temperate hardwood forestsof the eastern U.S.A. Species in the genera Colletotrichum and Ilyonectria were among the most commonly isolated and recovered amplicon sequencefrom seedlings of multiple host species showing disease symptoms. Further, co-infection by both Colletotrichum and Ilyonectria species decreases hostsurvival, as quantified by posterior model probabilities. To investigate molecular mechanisms associated with multi-host generalism and co-infection, andto determine whether these “generalist” pathogens are distinct species or species-complexes, the genomes of three common species in our dataset (e.g.C. fioriniae, C. gloesporoides and Ilyonectria europea) were sequenced. The largest genome of the three belonged to Ilyonectria at 63.66 Mb, which alsocontained the highest number (22,250) of genes. The smallest genome belonged to C. fioriniae with 50.04 Mb and 15,777 genes. Genome size and numberof predicted genes appears expanded, confirming their role as seedling pathogens. For instance, three out of four polysaccharide lyase (PL) enzymedomains found in fungal genomes, are enriched in these three species. PL enzymes are relevant in plant pathogenicity since they may contribute to initialstages of host penetration. The genome sequence of these fungal groups will serve as a reference set for population level studies to address hostspecificityand local adaptation within our isolate database.96

CONCURRENT SESSION ABSTRACTSSaturday, March 16 2:00 PM–5:00 PMNautilusDimorphic TransitionsCo-chairs: Anne Dranginis and Alex AndrianopoulosEpigenetic Switching Regulates the Yeast-Hyphal Transition in Candida albicans. Haoyu Si 1 , Allison Porman 1 , Matthew Hirakawa 1 , Stephen Jones 1 , AaronHernday 2 , Alexander Johnson 2 , Richard Bennett 1 . 1) Mol Microbiol & Immunology, Brown University, Providence, RI; 2) Mol Microbiol & Immunology,UCSF, San Francisco, CA.Candida albicans is a dimorphic yeast that is normally found as a commensal organism in the mammalian gastrointestinal tract. It is also a prevalentopportunistic pathogen able to infect multiple mucosal and internal sites in the human body. A principle feature of C. albicans biology is its ability to growin multiple phenotypic states, including both yeast and filamentous forms. Phenotypic plasticity is also exemplified by the “white-opaque switch”, in whichcells can reversibly transition between the white and opaque states. White and opaque forms differ in multiple aspects including their shape, theirinteraction with host immune cells, their mating competency, and their pathogenesis. Furthermore, white cells are induced to form hyphal filaments whengrown at 37°C, neutral pH, or in the presence of serum, whereas opaque cells do not form filaments in response to these conditions, and this differencecould explain the decreased virulence of opaque cells in models of systemic infection. In this study, we show that opaque cells can undergo the yeastfilamenttransition in response to environmental cues, but that these cues are distinct from those that induce the transition in white cells. For example,growth on low phosphate medium or medium containing the sugar sorbitol induced efficient filamentous growth in opaque cells, while these conditionsdid not induce filamentous growth in white cells. Genetic dissection of the regulation of opaque cell filamentation showed extensive overlap with theregulation of filamentation in white cells, including roles for the established transcriptional regulators Ume6, Efg1, and Tup1. However, genes induced byfilamentous growth in opaque cells showed only limited overlap with those induced during white cell filamentation. Together, these studies indicate thatC. albicans white and opaque cells are both capable of undergoing filamentation but do so in response to different environmental signals and generatedistinct transcriptional profiles, reflecting intrinsic differences in the programming of the two phenotypic states.Extracellular and intracellular signaling orchestrates morphotype-transition and virulence in human pathogen Cryptococcus neoformans. Linqi Wang,Xiuyun Tian, Rachana Gyawali, Xiaorong Lin. Biology, College Station, TX.Interactions with the environment and divergent species drive the evolution of microbes. To sense and rapidly respond to these dynamic interactions,“simple” microbes developed bet-hedging social behaviors, including the construction of heterogeneous biofilm communities and transition betweendifferent morphotypes. The human fungal pathogen Cryptococcus neoformans can undergo morphotype transition between the yeast and the filamentousform. Most recently, we demonstrated that the zinc-finger regulator Znf2 bridges the bi-direction yeast-hypha transition and virulence in this pathogen.One of Znf2 downstream targets is extracellular protein Cfl1. Cfl1 is a cell-wall bound adhesin and a signaling molecule when it is released. This matrixprotein Cfl1 plays a similar but less prominent role than Znf2 in orchestrating morphogenesis and virulence in C. neoformans. Through transcriptomeanalyses and screening Znf2 downstream targets by overexpression, we identified an additional player in the control of morphogenesis and biofilmformation. This factor is an intracellular RNA-binding protein Pum1. As expected, Pum1 affects filamentation in a Znf2 dependent manner. However, theeffect of Pum1 on morphogenesis is independent of Cfl1. The pum1D cfl1D double mutant shows a more severe defect in filamentation than either of thesingle mutant, indicating that Pum1 and Cfl1 act in two parallel pathways. Two of Pum1’s targets, Fad1 and Fad2, form a Cryptococcus-specific adhesinfamily. Like Cfl1, these two extracellular adhesins show regulatory roles in conducting morphogenesis and virulence in C. neoformans and thus may beinvolved in extracellular signaling transduction. Our results indicate that complex regulatory cascades composed of extracellular and intracellular circuitsmay be responsible for mediating morphological transition in response to the cues in the environments and the host.Histoplasma strain variations and differences in pathogenic-phase transcriptomes. Jessica A. Edwards 1 , Chenxi Chen 2 , Megan M. Kemski 1 , Thomas K.Mitchell 2 , Chad A. Rappleye 1 . 1) Microbiology, Ohio State University, Columbus, OH; 2) Plant Pathology, Ohio State University, Columbus, OH.The morphological dimorphism of Histoplasma capsulatum reflects an underlying change in gene expression that is essential for pathogenesis. In theyeast-phase, Histoplasma infects the mammalian lung and proliferates within phagocytic cells. Geographically distinct strains of Histoplasma exhibitdifferences in their relative virulence and in their pathogenic mechanisms. The close similarity in the genome sequences of these diverse strains suggeststhat phenotypic variations result from gene expression differences rather than gene content. To better understand how the transcriptional programtranslates into morphological and pathogenic differences between strains, we profiled the yeast-phase transcriptomes of two Histoplasma strains byRNAseq methodology. For both strains, about 50% of sequence reads align to the genome providing evidence for approximately 9000 genes. Quantitativecomparisons reveal about 200 genes are at least 10-fold differentially expressed between strains, and these include genes related to Histoplasmapathogenesis (SOD3, YPS3, AGS1). The genes encoding the secreted calcium-binding protein (CBP1), histone proteins (H2B, H3, and H4) and an ammoniumtransporter are among the most highly expressed genes overall. Using GFP-transcriptional fusions and their introduction into both strain backgrounds, wedemonstrate that dissimilarity in the transcriptional activity of individual genes reflects variations in the trans-acting factors between strains rather thanthe sequence of the promoters, themselves. These studies lay an essential foundation to facilitate discovery of the factors that contribute to strain-specificvirulence differences of Histoplasma.27th Fungal Genetics Conference | 97

CONCURRENT SESSION ABSTRACTSThe C 2H 2 transcription factor HgrA promotes hyphal growth in the dimorphic pathogen Penicillium marneffei. Hayley E. Bugeja, Michael J. Hynes, AlexAndrianopoulos. Department of Genetics, University of Melbourne, Parkville, VIC, Australia.Penicillium marneffei (recently renamed Talaromyces marneffei) is well placed as a model experimental system for investigating fungal growth processesand their contribution to pathogenicity. An opportunistic pathogen of humans, P. marneffei is a dimorphic fungus that displays multicellular hyphal growthand asexual development (conidiation) in the environment at 25°C and unicellular fission yeast growth in macrophages at 37°C. We have characterised thetranscription factor hgrA (hyphal growth regulator), which contains a C 2H 2 DNA binding domain closely related to that of the stress-response regulatorsMsn2/4 of Saccharomyces cerevisiae. HgrA is not required for controlling yeast growth in response to the host environment, nor does it appear to have akey role in response to stress agents, but is both necessary and sufficient to drive the hyphal growth program. hgrA expression is specific to hyphal growthand its deletion affects multiple aspects of hyphal morphogenesis and the dimorphic transition from yeast cells to hyphae. Loss of HgrA also causes cellwall defects, reduced expression of cell wall biosynthetic enzymes and increased sensitivity to cell wall, oxidative, but not osmotic stress agents. As well ascausing apical hyperbranching during hyphal growth, overexpression of hgrA prevents conidiation and yeast growth, even in the presence of inductivecues. HgrA is a strong inducer of hyphal growth and its activity must be appropriately regulated to allow alternative developmental programs to occur inthis dimorphic pathogen.A conserved splicing factor is required for vesicle transport in Ustilago maydis. Nikola Kellner 1 , Kai Heimel 1,2 , Florian Finkernagel 3 , Theresa Obhof 1 , JoergT. Kaemper 1 . 1) Dept. of Genetics, Karlsruhe Institute of Technology, Karlsruhe, Germany; 2) Dept. of Molecular Microbiology and Genetics, Georg-August-University Göttingen, Göttingen, Germany; 3) Institute for Molecular Biology and Tumor Research, Marburg, Germany.In the corn smut fungus Ustilago maydis, sexual development is initiated by the fusion of two yeast-like haploid sporidia, resulting in a filamentousdikaryon that is capable to infect the plant. Growth as a dikaryon requires an elaborate coordination of the cell cycle, the migration and distribution of thenuclei and polar hyphal growth. We have identified the U. maydis Num1 protein with a pivotal function during these processes. Num1 is homologous toSPF27, a core component of the evolutionary conserved Prp19/CDC5 complex (NTC). The NTC contributes to splicing efficiency and fidelity, but is alsoinvolved in cell cycle checkpoint control, response to DNA damage or formation and export of mRNP-particles. Deletion of num1 in U. maydis has noobvious phenotype in sporidia, however, hyphae exhibit polarity defects; in addition, the num1 mutation affects the cell cycle and cell division. Weidentified Cdc5 and Prp19, two conserved components of the Prp19/CDC5 complex, as Num1 interactors. In line with the function of the NTC, wedemonstrated by means of a genome-wide mRNA-Seq analysis that splicing in num1 deletion strains is impaired on a global level. In addition to the NTCcomponents, several proteins with putative functions during vesicle-mediated transport processes were identified as Num1 interactors; in particular theconventional kinesin 1 motor protein Kin1 was shown to physically interact with Num1. Both num1- and kin1-deletion strains exhibit identical phenotypeswith respect to filamentous and polar apical growth, the morphology of vacuoles, the subcellular distribution of the Dynein motor protein as well as themotility of early endosomes, strongly corroborating a genetic interaction between Num1 and Kin1. Our data implicate a previously unidentified connectionbetween a component of the splicing machinery and cytoplasmic transport processes. As the num1 mutation also affects cytoplasmic mRNA transport, theprotein might constitute a novel functional interconnection between the two disparate processes of splicing and trafficking.N-acetylglucosamine (GlcNAc) Triggers a Morphogenetic Program in Systemic Dimorphic Fungi. Sarah A. Gilmore 1 , Shamoon Naseem 2 , James B.Konopka 2 , Anita Sil 1 . 1) Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA; 2) Department ofMolecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY.Cellular differentiation is an essential process for the development and growth of multicellular eukaryotic organisms. Similarly, many unicellularorganisms undergo a program of cellular differentiation to produce a new cell type specialized for survival in a distinct environmental niche. Systemicdimorphic fungal pathogens, such as Histoplasma capsulatum (Hc) and Blastomyces dermatitidis (Bd), can switch between a unicellular parasitic yeast formadapted for growth within mammals and an infectious soil-growing filamentous form as part of their natural life cycles. Temperature is thought to be thepredominant environmental cue that promotes cellular differentiation of systemic dimorphic fungi; however, work with other fungi indicates thatadditional environmental cues including CO2, light, and nutrient availability can influence how an organism responds to its environment. Recent worksuggests that the ubiquitous monosaccharide N-acetylglucosamine (GlcNAc) can play a role in cell signaling in fungi. We identified GlcNAc as a potentinducer of the yeast-to-filament transition in Hc and Bd. Micromolar concentrations of exogenous GlcNAc were sufficient to induce a robust morphologicaltransition of Hc yeast cells to filamentous cells at room temperature, indicating that dimorphic fungal cells may be sensing GlcNAc, or one of its catabolicbyproducts, to promote filamentation. Using GlcNAc as a tool to induce a robust and more synchronous phase transition of Hc yeast cells to filaments, weexamined the temporal regulation of the Hc transcriptome during morphogenesis to reveal candidate genes involved in establishing the filamentousgrowth program. Two genes we identified during transcriptome analysis included NGT1 and NGT2, which encode GlcNAc major facilitator superfamilytransporters. RNAi depletion of NGT1 or NGT2 rendered Hc cells unable to respond to exogenous GlcNAc. Furthermore, wild type levels of NGT1 and NGT2transcripts were important for efficient Hc yeast-to-filament conversion even in the absence of exogenously added GlcNAc. These data suggest that Ngt1and Ngt2 may monitor endogenous GlcNAc as part of an autoregulatory system that allows Hc to regulate its filamentous growth.A GATA transcription factor encoded by SREB functions as a global regulator of transcription in Blastomyces dermatitidi. Amber Marty, Aimee T.Broman, Christina Kendziorski Gregory M. Gauthier. University of Wisconsin - Madison, 1550 Linden Drive, Microbial Sciences Building, Madison, WI,53706.The thermally dimorphic fungi infect several million people each year including those with normal immune defenses. These fungi grow as mold in the soil(22 o C) where they produce infectious conidia. Upon soil disruption, aerosolized conidia are inhaled into the lungs (37 o C) where they convert into yeast.This reversible, temperature-dependent phase transition defines the lifestyle of the dimorphic fungi. In Blastomyces dermatitidis, we discovered SREB(siderophore biosynthesis repressor in Blastomyces), which encodes a GATA transcription factor that promotes the conversion to mold at 22 o C and alsoregulates iron homeostasis. To begin to dissect how SREB affects the transcriptional response to temperature, we used gene expression microarrays andchromatin immunoprecipitation with quantitative real-time PCR (ChIP-qPCR). For microarrays, RNA was isolated from SREB and wild type (WT) isolates atbaseline (yeast) and 6, 24, and 48-hours after a drop in temperature to 22 o C. LIMMA and EBarrays were used to identify differentially expressed (DE)genes. For ChIP, we engineered SREB to contain an in-frame, C-terminal 3x-HA tag. SREB-3xHA was cross-linked to its DNA binding targets in vivo using 1%formaldehyde at 37 o C and 48-hours at 22 o C. Following chromatin shearing and reversal of cross-links, enrichment for SREB-3xHA binding (vs. isogeniccontrol) at GATA motifs was assessed by qPCR. Gene expression microarray analyses indicated that SREB was a global regulator of transcription at 37 o Cand 22 o C. Gene Ontology enrichment demonstrated SREB was involved with diverse processes including iron ion binding, amino acid transport, metabolic98

CONCURRENT SESSION ABSTRACTSprocess, and fatty acid biosynthesis. Complementary analysis using weighted gene co-expression network analysis identified modules with DE genesenriched for transmembrane transport, metabolic process, transcription factor activity, translation, and fatty acid biosynthesis. To identify candidate genesfor ChIP-qPCR, we integrated gene expression microarray data with genome-wide in silico analysis of GATA transcription factor binding motifs. Using thisapproach, we identified a subset of genes bound and regulated by SREB including SIDA, MIRB, WD, HAPX, and PDH at 37 o C and 22 o C. In contrast,enrichment for SREB-3xHA binding of ECI1 occurred at 22 o C, but not 37 o C. In conclusion, integration of microarray analysis, in silico GATA motifs, GOenrichment, and ChIP-qPCR indicates that SREB affects pleiotropic events in B. dermatitidis.Functional Analysis of Genes in Regions of Introgression in Coccidioides. Bridget M. Barker. Immunology & Infectious Diseases, Montana State Univ,Bozeman, MT.Coccidioides immitis and C. posadasii are dimorphic fungi endemic to the Americas. Genomic analysis of sequenced strains of C. posadasii and C. immitisreveals insights into the population biology of these organisms. There is strong evidence for hybridization and introgression, such that for many of the C.immitis strains, there are several regions that have a closer match to C. posadasii, but few regions within C. posadasii matching C. immitis. Multiplehybridization regions were located in several genomes analyzed, and at least one region containing ten genes exhibits a pattern consistent withintrogression in C. immitis. This conserved region was further evaluated in a larger collection of isolates. Approximately half of the C. immitis isolatescontain the C. posadasii fragment, and the majority of those are from the southern California and Mexico populations. The region of introgressionrepresents a unique opportunity to functionally assess genes that are likely to be relevant for species-specific virulence and adaptation to mammalianhosts or the environment. This region has a shared recombination point flanking a metalloproteinase, Mep4; genes that are highly expressed in theparasitic phase; and genes of unknown function. Importantly, evolutionary selection has preserved this region in multiple strains of C. immitis furtheremphasizing the possible role in virulence of these genes. Variation among strains for virulence in murine models of coccidioidomycosis has beenobserved, but has not been tested in the context of the newly discovered species or with a targeted underlying genetic mechanism hypothesis to test.Gene deletion mutants are being generated for three genes in the conserved introgression region to determine effects on in vitro growth andmorphological change under host relevant conditions.27th Fungal Genetics Conference | 99

CONCURRENT SESSION ABSTRACTSSaturday, March 16 2:00 PM–5:00 PMScrippsTropic Growth and FusionCo-chairs: Andre Fleissner and Nick ReadRole of the cell fusion gene idcA in fungal mutualism. Carla J. Eaton 1,2 , Cornelia Staerkel 1 , Barry Scott 1,2 . 1) Institute of Molecular BioSciences, MasseyUniversity, Palmerston North, New Zealand; 2) Bio-Protection Research Centre, Massey University, Palmerston North, New Zealand.Maintenance of the mutualistic association between the fungal endophyte Epichloë festucae and perennial ryegrass relies on a number of importantsignalling pathways including ROS signalling by the NADPH oxidase (Nox) complex, and the cell integrity- and stress-activated MAP kinase pathways.Perturbation of these signalling pathways leads to a dramatic switch from mutualistic to pathogenic-like association with perennial ryegrass. Interestingly,all E. festucae ‘symbiotic regulator’ genes identified to date are involved in vegetative cell fusion in other fungi, suggesting hyphal fusion may play animportant role in maintenance of the mutualistic association. To investigate this putative link, the role of the cell fusion gene IDC1 (ham-5) was examined.IDC1 is of particular interest as in addition to its role in cell fusion it is also linked to Nox signalling in Podospora anserina. Disruption of E. festucae idcAleads to a dramatic symbiotic switch from mutualistic to pathogenic-like association with perennial ryegrass. Infected plants are severely stunted anddisplay precocious senescence. Biomass of the DidcA mutant in planta is significantly increased relative to the wild-type strain, and hyphae extensivelycolonise host vascular tissues. Formation of intra-hyphal hyphae by the DidcA mutant in planta is also abundant, possibly due to defective hyphal fusion ordefects in septation. The importance of idcA for maintenance of mutualistic association with perennial ryegrass supports the hypothesis that hyphal fusionis required for establishment of an interconnected hyphal network essential for mutualism.Role of extracellular calcium in budding yeast cell fusion. Pablo S. Aguilar. Cell Membranes Laboratory, Institut Pasteur de Montevideo, Montevideo,Uruguay.The molecular details of membrane fusion during yeast mating are poorly understood. The tetraspanner protein Prm1 is one of the few knowncomponents that acts at the step of bilayer fusion. In its absence, mutant mating pairs lyse or arrest in the mating reaction with tightly apposed plasmamembranes. The absence of another tetraspanner, Fig1p, which controls pheromone-induced Ca 2+ influx, yields similar cell fusion defects. Althoughextracellular Ca 2+ is not required for efficient cell fusion of wild-type cells, cell fusion in prm1 mutant mating pairs is dramatically reduced when Ca 2+ isremoved. A genetic screen was conducted to uncover genes that promote mating-dependent lysis in the absence of extracelular Ca 2+ . The role of differentcandidates in relation to Prm1p will be reviewed here.The role of calcium and calmodulin during cell fusion and colony initiation in Neurospora crassa. Chia-Chen Chang, Nick Read. Fungal Cell Biology Group,Institute of Cell Biology, University of Edinburgh, Edinburgh EH9 3JH.Calcium is an ubiquitous signalling molecule which regulates many important processes in filamentous fungi including spore germination, hyphal growth,mechanosensing, stress responses, circadian rhythms, and virulence. Transient increases in cytosolic free calcium ([Ca 2+ ] c) act as intracellular signals. As theprimary intracellular Ca 2+ receptor, calmodulin (CaM) converts these Ca 2+ signals into responses by regulating the activity of numerous target proteins. Wehave found that both Ca 2+ -free medium and two CaM antagonists (calmidazolium and trifluoperazine) selectively inhibit a form of cell fusion called conidialanastomosis tube (CAT) fusion that occurs during colony initiation in the fungal model Neurospora crassa. GFP labelled CaM localized as dynamic particlesassociated with the plasma membrane and moved around within the cytoplasm in both germ tubes and CATs. In particular, CaM showed a dynamicaccumulation at two growing tips of CATs that exhibit chemoattraction towards each other. CaM also localized at developing septa in germ tubes. The b-tubulin inhibitor, benomyl, reduced the movement of CaM in the cytoplasm. Moreover, the absence of extracellular Ca 2+ inhibited the recruitment of CaMto CAT tips as well as inhibiting CAT chemoattraction. The deletion of the myosin-5 (myo-5) gene caused the mis-localization of CaM in tips of growinggerm tube and CATs. This suggests that the movement of cytoplasmic CaM involves transport along microtubules, and the recruitment of CaM to tipsinvolves myosin-5 along F-actin and is dependent on extracellular Ca 2+ .LFD-1 is a component of the membrane merger machinery during cell-cell fusion in Neurospora crassa. Javier Palma-Guerrero, N. Louise Glass. Plant andMicrobial Biology Department, UC Berkeley, Berkeley, CA.Cell-cell fusion is an essential part of the development of most eukaryotic organisms, playing an important role both during sexual development andvegetative growth. In the filamentous fungus Neurospora crassa, cell fusion events occur during all stages of the life cycle. This, together with thesequenced and annotated genome and the genetic tools available, makes this fungus a good model system for dissecting cell fusion. Plasma membranemerger is the last step in the cell-cell fusion process, occurring after cell wall remodeling, and it is the definitive event that allows for cytoplasm mixingbetween the fusing cells. Although molecular mechanisms associated with intracellular membrane fusion are well characterized, the molecularmechanisms of plasma membrane merger between cells are poorly understood. Only one gene encoding a protein involved in this last step of cell fusionhad been previously identified in N. crassa: Prm-1, a deletion of which results in strains that show a »50% reduction in vegetative and sexual cell fusion.We have identified a second gene, lfd-1, which is also involved in plasma membrane merger in N. crassa. LFD-1 is a plasma membrane protein only presentin ascomycete filamentous fungi. N. crassa strains carrying a deletion of lfd-1 results in a reduction in both vegetative and sexual cell fusion, and having asimilar, but less severe, phenotype than a Prm-1 deletion strain. Strains carrying both Prm-1 and lfd-1 deletions indicate that LFD-1 acts independently ofPRM-1. Strains carrying Prm-1 or lfd-1 mutations result in increased cell lysis during cell-cell fusion, a phenotype that was enhanced by reducingextracellular calcium concentration. These results suggest that the lysis phenotype associated with cell fusion events is due to membrane damage causedby defects during membrane merger, and which may be repaired in a calcium dependent process. Our results indicate that both PRM-1 and LFD-1 areimportant, but non-essential components of the cell fusion membrane merger machinery.100

CONCURRENT SESSION ABSTRACTSSpecific Structural Features of Sterols Affect Cell-Cell Signaling and Fusion in Neurospora crassa. Martin Weichert 1 , Ewald Priegnitz 1 , Raphael Brandt 1 ,Thorben Nawrath 2 , Stefan Schulz 2 , André Fleissner 1 . 1) Institut für Genetik, Technische Universität Braunschweig, Spielmannstrasse 7, 38106Braunschweig, Germany; 2) Institut für Organische Chemie, Technische Universität Braunschweig, Hagenring 30, 38106 Braunschweig, Germany.Sterols are major constituents in the plasma membrane of eukaryotic cells. They modulate the physical properties of the lipid bilayer, e.g. fluidity. Byinteracting with certain lipids and proteins in the plasma membrane, sterols cluster into microdomains which might act as platforms for many biologicalfunctions, such as signal transduction. In the early stages of colony formation in Neurospora crassa, germinating spores direct their growth towards eachother, establish physical contact, and fuse. Cell-to-cell signaling requires the coordinated dynamic recruitment of the MAP kinase MAK-2 and thecytoplasmic protein SO to the tips of interacting cells. Subsequent plasma membrane fusion is facilitated by the transmembrane protein PRM1. Here, wereport that mutants affected in the biosynthesis of ergosterol, the major sterol in most fungal species, show distinct defects during germling fusion.Deletion of erg-2, which encodes an enzyme mediating the last step in the pathway, strongly impairs both directed growth and cell fusion. Interestingly,both MAK-2 and SO mislocalize at the tips of interacting Derg-2 germlings. In contrast, the absence of ERG-10a and ERG-10b, two enzymes with redundantfunction that act upstream of ERG-2, does not affect cell-to-cell communication. However, Derg-10a Derg-10b germling pairs show DPrm1-like deficienciesin plasma membrane merger. By relating the sterol composition and fusion competence of several erg mutants, we find that not the absence of ergosterolbut the accumulation of sterol intermediates specifically impairs distinct steps of germling fusion. While the presence of two double bonds in the sterolside chain provokes Derg-2-like deficiencies, an altered double bond arrangement in the sterol ring system causes DPrm1-like defects. During sexualdevelopment, cell fusion precedes the fertilization of fruiting bodies. Unlike the defects during germling fusion, female and male mating partners of Derg-2and Derg-10a Derg-10b efficiently fuse, suggesting that alterations in the sterol composition specifically impair signaling mechanisms mediating vegetativecell fusion. These data suggest that specific structural features of sterols differentially affect membrane properties and functions, such as the membranerecruitment of proteins, the assembly of signaling complexes, and plasma membrane fusion.Co-option of a sex pheromone receptor and MAPK signalling pathway for chemotropism of Fusarium oxysporum towards plant host compounds. DavidTurra, Federico Rossi, Antonio Di Pietro. Departamento de Genética, Universidad de Córdoba, 14071 Córdoba, Spain.Fungal hyphae explore the surrounding environment in search of nutrient sources, mating partners or host organisms by sensing gradients of tropicallyactive cues. Chemotropism is crucial for fungal development and virulence, but the underlying mechanisms are poorly understood. Here we followed agenetic approach to dissect chemotropism in the soilborne plant pathogen Fusarium oxysporum. A plate assay was used to measure directed growth ofgerm tubes towards different classes of compounds, including carbon and nitrogen sources, sex pheromones, plant secondary metabolites and tomatoroot exudate. F. oxysporum mutants lacking the mitogen activated protein kinase (MAPK) Fmk1 or the transcription factor Ste12, two components of theconserved Pathogenicity MAPK cascade, were impaired in chemotropism towards nutrients, but fully responsive to a-pheromone and root exudate. Bycontrast, Rho1 and Mpk1, two components of the cell integrity MAPK cascade, were specifically required for directed growth towards a-pheromone androot exudate. Deletion of the seven transmembrane G protein coupled receptor Ste2 abolished the chemotropic response to a-pheromone and,unexpectedly, also to tomato root exudate. Our results provide evidence for co-option of a cognate sex pheromone receptor and a conserved MAPKsignalling pathway for chemotropism of F. oxysporum towards plant host compounds.Characterization of new STRIPAK complex interaction partners in the filamentous ascomycete Sordaria macrospora. Britta Herzog, Yasmine Bernhards,Berit Habing, Eva Reschka, Sabine Riedel, Stefanie Pöggeler. Institute of Microbiology and Genetics, Department of Genetics of EurkaryoticMicroorganisms, Georg-August-University Göttingen, Germany.Using Sordaria macrospora as model organism we investigate the complex process of fruiting-body development and involved proteins in thisfilamentous ascomycete. This differentiation process is regulated by more than 100 developmental genes. Recently, we have shown that a homologue ofthe human STRIPAK (striatin-interacting phosphatase and kinase) complex engages a crucial role in sexual development in fungi. The S. macrospora striatinhomologue PRO11 and its interaction partner SmMOB3 are key components of this complex (Bloemendal et al., 2012). PRO11 contains a conserved WD40repeat domain and is supposed to function as scaffolding protein linking signaling and eukaryotic endocytosis (Pöggeler and Kück, 2004). SmMOB3(phocein) is a member of the MOB family (Bernhards and Pöggeler, 2011). Beside their important role in multicellular development and hyphal fusion bothproteins seem to be involved in vesicular trafficking and endocytosis.By means of yeast two-hybrid screens and GFP-Trap analysis we identified several new interaction partners of PRO11 and SmMOB3. Similar to PRO11and SmMOB3, a multitude of them are predicted to be involved in vesicular trafficking and are localized to the ER or to the Golgi. Here, we show theresults of a detailed analysis of the new STRIPAK complex interaction partners. Initially, we isolated the cDNA of the genes and confirmed the interactionby yeast two-hybrid. For further characterization and to get knowledge about their cellular functions we created knock-out strains and analyzed theirmorphological phenotypes. For localization and expression studies we constructed EGFP-tagged fusion proteins and expressed them in S. macrospora.Bernhards and Pöggeler, 2011; Curr Genet 57 (2): 133-49.Bloemendal et al., 2012; Mol Microbiol 84 (2): 310-23.Pöggeler and Kück, 2004; Eukaryot Cell 3 (1): 232-40.27th Fungal Genetics Conference | 101

CONCURRENT SESSION ABSTRACTSCharacterisation of contact-dependant tip re-orientation in Candida albicans hyphae. Darren Thomson, Silvia Wehmeier, Alex Brand. Aberdeen FungalGroup, Aberdeen University, Aberdeen, United Kingdom.Candida albicans is a pleiomorphic fungus that lives as a commensal yeast in the human body but can become pathogenic in susceptible patient groups.Virulence is strongly linked with the production of penetrative hyphae that can adhere to and invade a wide range of substrates, including blood vessels,organ tissue, keratinised finger-nails and even soft medical plastics. Using live-cell imaging and nanofabricated surfaces, we are characterising the spatiotemporaldynamics of contact-induced hyphal tip behaviour (thigmotropism). To test whether tip re-orientation responses positively correlate with levelsof hyphal adhesion, we generated substrates with increasing adhesive force. Hyphal tip re-orientation was absent in poorly-immobilised hyphae and athreshold adhesive force was required sub-apically to generate the hyphal tip pressure required for re-orientation. Interestingly, sub-threshold adhesionresulted in sub-apical hyphal bending. Localization of fluorescent protein markers for the Spitzenkörper and the Polarisome (Mlc1-YFP and Spa2-YFP,respectively) showed that C. albicans hyphal tips grow in an asymmetric, ‘nose-down’ manner on a surface. Additionally, hyphal tips can detect surfacestiffness and show a distinct preference for nose-down growth on the softer of two substrates. Localisation of fluorescent cell-cycle reporter proteins overtime revealed that hyphal tip contact slowed the cell-cycle, suggesting that tip-contact perturbs cell-cycle mechanics. Finally, we examined the role ofcytoskeleton regulators in thigmotropism and determined the force that can be generated by the hyphal tip. Our results suggest that C. albicans hyphaecan exert sufficient force to penetrate human epithelial tissue without the need for secreted enzyme activity. This is consistent with the observed hyphalpenetration of medical-grade silicone, which has a similar Young’s modulus to human cartilage.102

LISTING OF ALL POSTER ABSTRACTSBiochemistry and Metabolism1. Heme regulation in Aspergillus fumigatus., Nicola Beckmann2. Key Steps in the Biosynthesis of the Fungal Virulence FactorGliotoxin., Pranatchareeya Chankhamjon3. Identification of a gene cluster mediating the biosynthesis of theAspergillus fumigatus cell wall and secreted polysaccharide,galactosaminogalactan., Fabrice N. Gravelat4. A fasciclin-like protein in Aspergillus fumigatus., Thomas Hartmann5. Characterization of fumiquinazoline biosynthesis in Aspergillusfumigatus., Fang Yun Lim6. Identification of local and cross-chromosomal biosynthetic geneclusters in filamentous fungi using gene expression data. Mikael R.Andersen7. N-glycan profiling of Aspergillus nidulans using solid-phase glycanextraction and mass spectrometry. Diana Anyaogu8. Targeting of AcvA to peroxisomes increases penicillin production inAspergillus nidulans. Andreas Herr9. Characterization of the 3-methyl orsellinic acid gene cluster inAspergillus nidulans. Jakob B. Nielsen10. Induction of sclerotia and Aspergillus section Nigri. Jens Frisvad11. Analyzing the impact of compartmentalization on organic acidproduction in Aspergillus niger. Matthias G. Steiger12. Subcellular localization of aphidicolin biosynthesis enzymes fromPhoma betae expressed heterologously in Aspergillus oryzae. K. Gomi13. Increased production of fatty acids and triglycerides in Aspergillusoryzae by modifying fatty acid metabolism. Koichi Tamano14. Improved Properties of Thermostable Cellobiohydrolase in aTreatment of Cellulosic Material. Taija Leinonen15. The phytopathogenic fungus Botrytis pseudocinerea is resistant tothe fungicide fenhexamid due to detoxification by a cytochrome P450monooxygenase Cyp684. Danièle Debieu16. Evolutionary rewiring of ubiquitination targets in Candida albicanspromotes efficient carbon assimilation in host niches. Alistair J. P.Brown17. Can-Hsp31 is important for Candida albicans growth and survival.S. Hasim18. Influence of N-glycans on α-/β-(1,3)-glucanase and α-(1,4)-amylasefrom Paracoccidioides brasiliensis yeast cells. Fausto Bruno Dos ReisAlmeida19. Cell wall structure and biosynthesis in oomycetes and true fungi: acomparative analysis. Vincent Bulone20. Investigating the function of a putative chitin synthase fromPhytophthora infestans. Stefan Klinter21. Deciphering cell wall structure and biosynthesis in oomycetesusing carbohydrate analyses and plasma membrane proteomics. HugoMelida22. Identification and characterization of the chitin synthase genes inthe fish pathogen Saprolegnia parasitica. Elzbieta Rzeszutek23. Role of Ccr4-mediated mRNA turnover innucleotide/deoxynucleotide homeostasis and Amphotericin Bsusceptibility in Cryptococcus neoformans. D. Banerjee24. WITHDRAWN25. Blue light induce Cordyceps militaris fruiting body formation andcordycepin production. Chun-Hsiang Yang26. Insight into alkaloid diversity of the epichloae, protectivesymbionts of grasses. Carolyn A. Young27. Extracellular polysaccharide degrading capabilities of variousAgaricus bisporus strains during compost cultivation. A.Patyshakuliyeva28. Reconstruction of the rubrofusarin biosynthetic pathway inSaccharomyces cerevisiae. Rasmus J. N. Frandsen29. Expression and purification of hydrophobin fusion proteinstargeted to intracellular protein bodies in T. reesei . Nina K. Aro30. Metabolic adaptations in Phytophthora infestans and the role of aphosphagen kinase system in energy metabolism. Meenakshi Kagda31. Platforms for secondary metabolite analysis in filamentous fungi.Uffe H. Mortensen32. Transcriptional analysis of oxalate degradation in the white rotbasidiomycete Dichomitus squalens. Miia R. Mäkelä33. Creation of temperature-influenced hyphal growth mutants in abasidiomycete fungus through the use of UV mutagenesis. Stephen J.Horton34. Functional Analysis of a Novel Diaminopimelate Decarboxylasefrom the Oomycete Saprolegnia parasitica. Paul Morris35. Living on Air?: Ustilago maydis cells grow without being providednitrogen in their growth media. Michael H. Perlin36. Saprotrophic metabolism of the White-Nose Syndrome fungusGeomyces destructans in bat hibernacula. Hannah Reynolds37. Cellulose acting enzymes of the white-rot fungus Dichomitussqualens: expression of the genes and characterization of theenzymes. Johanna Rytioja38. Metabolomics of growth and type B trichothecenes production inFusarium graminearum. N. Ponts39. Diversity of telomeric sequences and telomerase RNA structureswithin Ascomycetes. Xiaodong Qi40. Characterizing a putative three-step formaldehyde oxidationpathway in Neurospora crassa. Ethan Addicott41. Nitrate assimilation in Neurospora crassa. Oleg Agafonov27th Fungal Genetics Conference | 103

LISTING OF ALL POSTER ABSTRACTS42. Protein kinases affecting glycogen accumulation and likelyregulating the glycogen synthase phosphorylation status inNeurospora crassa. T. Candido43. Endogenous ergothioneine is required for wild type levels ofNeurospora crassa conidiogenesis and conidial survival, but does notprotect against uv-induced kill or mutagenesis. Lynn Epstein44. Thiolutin inhibits protein turnover in Neurospora and yeast. LindaLauinger45. Characterization of a Phanerochaete chrysosporium glutathionetransferase reveals a novel structural and functional class withligandin properties for wood extractive molecules. Melanie Morel46. Secretome analysis of Trichoderma harzianum cultivated in thepresence of Fusarium solani cell wall or glucose. Marcelo HS Ramada47. Analysis of carbon catabolite repression (CCR) during cellulaseformation by Trichoderma reesei (Hypocrea jecorina) using twodimensionaldifferential gel electrophoresis (2D-DIGE). Roberto N.Silva48. Transposon-associated evolution of a fungal NRPS. Daniel Berry49. Velvet family control of penicillin production in Penicilliumchrysogenum: PcVelB binding to isopenicillin N synthase suggests anovel regulatory mechanism. Sandra Bloemendal50. Genome mining reveals the evolutionary origin and biosyntheticpotential of basidiomycete polyketide synthases. Gerald Lackner51. Engineering Cyclic Peptide Biosynthesis in Poisonous Mushrooms.Hong Luo52. Spatial assessment of oxidative and enzymatic reactions in brownrotted wood. Jon R. Menke53. Molecular biological basis for statin resistance in naturally statinproducingorganisms. Ana Rems54. Molecular genetic characterization of secondary metabolismpathways in Asperillus species. Clay Wang55. A branched biosynthetic pathway is involved in production ofroquefortine and related compounds in Penicillium chrysogenum.Arnold Driessen56. A biosynthetic gene cluster for the antifungal metabolitephomenoic acid in the plant pathogenic fungus, Leptosphaeriamaculans. Candace Elliott57. Exploring and Manipulating Pleuromutilin Production. Patrick M.Hayes58. Improvement of Monascus pilosus for the production of functionalfoods by overexpression of the laeA gene. In H. Lee59. Molecular genetics studies on secondary metabolism inChaetomium globosum reveal involvement of aureonitol andchaetoglobosins in gene regulation and sexual reproduction. TakehitoNakazawa60. Identification of T. asperellum CAZyme genes. Lasse Bech61. Identification of a lactose permease of Trichoderma reesei that isrequired for cellulase gene expression. Christa Ivanova62. The diversity of the Mannosylerythritol lipids depends on theperoxisomal targeting of the Mannosylerythritol acyl transferasesMac1 and Mac2 in Ustilago maydis. Björn Sandrock63. Metabolic adaptation of the oomycete Phytophthora infestansduring colonization of plants and tubers. Carol E. Davis64. Multi-copper oxidase genes of Heterobasidion irregulare. BastianDoernte65. The two novel class II hydrophobins of Trichoderma stimulateenzymatic hydrolysis of polyethylene terephthalate (PET). Irina S.Druzhinina66. Analysis of polyketide synthase gene clusters in Cladoniametacorallifera genome. J.-S. Hur67. Inhibition of benzoate 4-monooxygenase (CYP53A15) fromCohliobolus lunatus by cinnamic acid derivatives. Branka Korosec68. Higher yields of cyclodepsipetides from Scopulariopsis brevicaulisby random mutagenesis. Linda PaunCell Biology and Development69. Generation of pathogenic diploids from heterogeneous conidialpopulations of Aspergillus flavus. Farhana Runa70. Inhibition of appressorium formation of Magnaporthe oryzae byroxithromycin and its possible molecular target. Akira Ishii71. Identification of novel genes involved in induction ofappressorium development triggered by plant-derived signals inColletotrichum orbiculare. Sayo Kodama72. Unique protein domains regulate Aspergillus fumigatus RasAlocalization and signaling during invasive growth. Jarrod R. Fortwendel73. Light regulates growth, stress resistance and metabolism in thefungal pathogen Aspergillus fumigatus. Kevin K. Fuller74. Analysis of critical domains in calcineurin A required for septaltargeting and function in Aspergillus fumigatus. Praveen R. Juvvadi75. Calcium imaging and measurement during growth and response tostresses in Aspergillus fumigatus. Alberto Muñoz76. WITHDRAWN77. The copper transporter ctpA in Aspergillus fumigatus is critical forconidial melanization and virulence in an invertebrate infectionmodel. Srijana Upadhyay78. Aspergillus nidulans SNXA HRB1 is an SR/RRM family protein thatrescues defects in the CDC2/CYCLINB pathway. Sarah Lea Anglin79. The Aspergillus nidulans MAPK module AnSte11-Ste50-Ste7-Fus3controls development and secondary metabolism. Oezguer Bayram80. Functional analysis of sterol transporter in filamentous fungusAspergillus nidulans. Nicole Bühler104

LISTING OF ALL POSTER ABSTRACTS81. Mechanisms of cellular resistance to copper and arsenic inAspergillus nidualans. Steven H. Denison82. Functional characterization of Aspergillus nidulans ANID_05595.1:a possible homologue of the polarisome component Pea2. Nathan W.Gross83. Aspergillus nidulans septin interactions and post-translationalmodifications. Yainitza Hernandez-Rodriguez84. A highly conserved sequence motif is required for PkcAlocalization to septation sites and protein function in Aspergillusnidulans. Terry Hill85. The MpkB MAP kinase plays a role in autolysis and conidiation ofAspergillus nidulans. Kwang-Yeop Jahng86. Beyond green mining: analysis of fungal cytochemistry using goldnanoparticles. Susan GW Kaminskyj87. Aspergillus nidulans as an experimental system to identify novelcell wall growth and maintenance genes through identification of antifungaldrug resistance mutations. Susan Kaminskyj88. Aspergillus nidulans cell walls lacking galactofuranose are moresusceptible to glucan degrading enzymes. Susan Kaminskyj89. The GATA-type transcription factor NsdD is a key regulator ofconidiation and secondary metabolism in Aspergillus. Mi-Kyung Lee90. THE velvet regulators in Aspergilli. Heesoo Park91. Coordinated regulation of asexual development, cell death andautolysis by the C2H2 zinc finger transcription factor BrlA inAspergillus nidulan. István Pócsi92. Whole-genome sequencing identifies novel alleles of genesrequired for organelle distribution and motility in Aspergillusnidulans. Samara Reck-Peterson93. Two methyltransferase protein complexes control fungaldevelopment and secondary metabolite production. Oezlem SarikayaBayram94. Control of Multicellular Development by the Physically InteractingDeneddylases DEN1/DenA and COP9 Signalosome. Josua Schinke95. Visualization of apical membrane domains in Aspergillus nidulansby Photoactivated Localization Microscopy (PALM). Norio Takeshita96. Cellular morphogenesis of Aspergillus nidulans conidiophores: asystematic survey of protein kinase and phosphatase function.Lakshmi Preethi Yerra97. The Putative Guanine Nucleotide Exchange Factor RicA MediatesUpstream Signaling for Growth and Development in Aspergillus. Jae-Hyuk Yu98. Evidence for a role of peroxisomes in microtubule organization.Ying Zhang99. Autophagy promotes survival in aging submerged cultures of thefilamentous fungus Aspergillus niger. Maria A. Burggraaf100. Inactivation of flbA results in increased secretome complexityand reduced secretion heterogeneity in colonies of Aspergillus niger.Pauline Krijgsheld101. Functional characterization of A. niger class III and class V chitinsynthases and their role in cell wall integrity. Jean-Paul Ouedraogo102. Exploiting transcriptomic signatures of Aspergillus niger touncover key genes important for high protein traffic through itssecretory pathway. Joohae Park103. Identification of two Golgi-localized putative UDPgalactofuranosetransporters with overlapping function in Aspergillusniger. Joohae Park104. Maltose permease-encoding mRNA is cleaved under inductioncondition of amylolytic gene expression in Aspergillus oryzae. MizukiTanaka105. Functional characterisation of Rac GTPase in Botrytis cinereareveals impact on polarity, cell cycle and pathogenicity. Anna Minz-Dub106. Light matters: The transcription factor LTF1 regulates virulenceand light responses in the necrotrophic plant pathogen Botrytiscinerea. Julia Schumacher107. Functional analysis of genes in the mating type locus of Botrytiscinerea. Jan van Kan08. The role of hydrophobins in sexual development of Botrytiscinerea. Jan van Kan109. The pescadillo homolog, controlled by Tor, coordinatesproliferation and growth and response in Candida albicans yeast.Tahmeena Chowdhury110. Uncovering the mechanisms of thermal adaptation in Candidaalbicans. Michelle Leach111. Characterisation of contact-dependant tip re-orientation inCandida albicans hyphae. Darren Thomson112. Cdc14 association with basal bodies in the oomycetePhytophthora infestans indicates potential new role for this proteinphosphatase. Audrey M. V. Ah-Fong113. Colletotrichum orbiculare Bub2-Bfa1 complex, a spindle positioncheckpoint (SPOC) component in Saccharomyces cerevisiae, isinvolved in proper progression of cell cycle. Fumi Fukada114. Metazoan-like mitotic events in the basidiomycetous buddingyeast Cryptococcus neoformans - a human fungal pathogen. L.Kozubowski115. Distinctive Mitotic Localization of a Novel Suppressor of nimA1Provides New Insight into NIMA Function. Jennifer R. Larson116. Investigating Cell Cycle-Regulated Control of AppressoriumMorphogenesis in the Rice Blast Fungus Magnaporthe oryzae. WasinSakulkoo117. THE ROLE AND TRAFFIC OF CHITIN SYNTHASES IN Neurosporacrassa. R. Fajardo118. DFG5 and DCW1 cross-link Cell Wall Proteins into the Cell WallMatrix. Stephen J. Free119. Cell wall biology to illuminate mechanisms of pathogenicity in27th Fungal Genetics Conference | 105

LISTING OF ALL POSTER ABSTRACTSPhytophthora infestans. Laura Grenville-Briggs120. Analysis of the cell wall integrity (CWI) pathway in Ashbyagossypii. Klaus B. Lengeler121. Dynamics of exocytic markers and cell wall alterations in anendocytosis mutant of Neurospora crassa. Rosa R. Mouriño-Pérez122. Comprehensive genome-based analysis of cell wall biosynthesisin the filamentous phytopathogen Ashbya gossypii. P. Philippsen123. cAMP regulation in Neurospora crassa conidiation. WilhelmHansberg124. Ste12 is a negative regulator of conidiation and cell wall lyticenzymes production in response to nitrogen deprivation and light inTrichoderma atroviride. Maria Fernanda Nieto-Jacobo125. Black holes in fungal virulence: loss of RNAi in C. gattii outbreakstrains reveals a novel RNAi factor. Marianna Feretzaki126. The Crz1/Sp1 transcription factor of Cryptococcus neoformans isactivated by calcineurin and regulates cell wall integrity. Sophie Lev127. A Fungal Adhesin Guides Community Behaviors by Autoinductionand Paracrinal Signaling. Linqi Wang128. The PacC Signal Transduction Pathway regulates SexualDevelopment in Neurospora crassa. Stephen J. Free129. Aspergillus flavus MAP kinase AflMpkB positively regulatesdevelopmental process but not aflatoxin production. Kap-Hoon Han130. Subcellular localization and kinase activity of GK4, aPhytophthora infestans GPCR-PIPK involved in actin cytoskeletonorganisation. Chenlei Hua131. External calcium ions and deletion of per-1 gene suppressed theabnormal morphology of och-1 and frost mutants in Neurosporacrassa. Masayuki Kamei132. Functional analysis of carbonic anhydrases from the filamentousascomycete Sordaria macrospora. Ronny Lehneck133. The Coprinopsis cinerea cag1 (cap-growthless1) gene, whosemutation affects cap growth in fruiting body morphogenesis, encodesthe budding yeast Tup1 homolog. H. Muraguchi134. Adaptation of the microtubule cytoskeleton to multinuclearityand chromosome number in hyphae of Ashbya gossypii as revealed byelectron tomography. P. Philippsen135. High resolution proteomics of spores, germlings and hyphae ofthe phytopathogenic fungus Ashbya gossypii. P. Philippsen136. Indoor Fungal Growth and Humidity Dynamics. Frank J. J. Segers137. Essentiality of Ku70/80 in Ustilago maydis is related to its abilityto suppress DNA damage signalling at telomeres. Jose Perez-Martin138. Magnaporthe oryzae effectors with putative roles in cell-to-cellmovement during biotrophic invasion of rice. Mihwa Yi139. Functional characterization of autophagy genes Smatg8 andSmatg4 in the homothallic ascomycete Sordaria macrospora. Stefanie106Poeggeler140. Laser microdissection and transcriptomics of infection cushionsformed by Fusarium graminearum. Marike Boenisch141. Biochemical and biophysical analysis of the CarO rhodopsin ofFusarium fujikuroi. Jorge García-Martínez142. Roles of membrane and organellar calcium channels andtransporters in controlling pulsatile [Ca 2+ ] c signatures. Hye-Seon Kim143. Characterization of positive regulator for asexual and sexualreproduction in the cereal head blight pathogen Gibberella zeae.Jungkwan Lee144. Functional analysis of Elongator complex protein 3 in Gibberellazeae. Y. J. Lee145. Functional analyses of regulators of G protein signaling (FgRGS)and GzGPA proteins in Gibberella zeae. A. R. Park146. A novel gene, GEA1, is required for ascus cell wall developmentin the ascomycete fungus, Gibberella zeae. H. SON147. A systems-biology approach to build gene-regulatory networkmodels connecting osmotic stress responses and asexualdevelopment in Fusarium graminearum. A. Thompkins148. Starvation enhances heterokaryon formation betweenincompatible strains of Fusarium oxysporum. Shermineh Shahi149. Requirements for horizontal chromosome transfer in the plantpathogenic fungus Fusarium oxysporum. Ido Vlaardingerbroek150. Characterization of the endocytotic proteins Yel1-Arf3-Gts1 inAshbya gossypii and the role of Gts1 in endocytosis, actin localizationand filamentous growth. Therése Oskarsson151. A Late Embryogenesis Abundant (LEA) protein in Neosartoryafischeri confers protection against desiccation. Martin Richard vanLeeuwen152. Coordination of polarized secretion by the exocyst complex iscritical for filamentous growth and cytokinesis in Ustilago maydis.Michaela Wehr153. Localization of Neurospora crassa Cell Fusion Proteins. Ci Fu154. Identification of novel Neurospora crassa genes involved inhyphal fusion by tanscriptomic analysis. Wilfried Jonkers155. N-acetylglucosamine (GlcNAc) Triggers a Morphogenetic Programin Systemic Dimorphic Fungi. Sarah A. Gilmore156. How water influences fungal growth on "real" materials. H. P.Huinink157. Identification and characterization of two genes required in thecontrol of a cell degeneration in the filamentous fungi Podosporaanserina. Herve Lalucque158. Dynein drives oscillatory nuclear movements in thephytopathogenic fungus Ashbya gossypii and prevents nuclearclustering. P. Philippsen159. Quantification of the thigmotropic response of Neurosporacrassa to microfabricated slides with ridges of defined height andtopography. Karen Stephenson

LISTING OF ALL POSTER ABSTRACTS160. Specificity determinants of GTPase recognition by RhoGEFs inUstilago maydis. Britta A. M. Tillmann161. Moisture dependencies of P. Rubens on a porous substrate. K. A.van Laarhoven162. Localization of Gα proteins during germination in the filamentousfungus, Neurospora crassa. Ilva Esther Cabrera163. Deciphering the roles of the secretory pathway key regulatorsYPT-1 and SEC-4 in the filamentous fungus Neurospora crassa. E.Sanchez164. Functional characterization of CBM18 proteins, an expandedfamily of chitin binding genes in the Batrachochytrium dendrobatidisgenome. Jason Stajich165. The exocyst complex is necessary for secretion of effectorproteins during plant infection by Magnaporthe oryzae. Yogesh K.Gupta166. Functional analysis of protein ubiquitination in the rice blastfungus Magnaporthe oryzae. Yeonyee Oh167. The role of autophagy in Cryphonectria hypovirus 1 (CHV1)infection in Cryphonectria parasitica. M. Rossi168. Neurospora crassa protein arginine methyl transferases areinvolved in growth and development and interact with the NDRkinase COT1. D. Feldman169. Role of tea1 and tea4 homologs in cell morphogenesis in Ustilagomaydis. Flora Banuett170. Sex determination directs uniparental mitochondrial inheritancein Phycomyces blakesleeanus. Viplendra P. S. Shakya171. Exploring the role of a highly expressed, secreted tyrosinase inHistoplasma capsulatum mycelia. Christopher F. Villalta172. Hypobranching induced by both anti-oxidants and ROS controlgene knockouts in Neurospora crassa. Michael K. Watters173. Septum formation starts with the establishment of a septal actintangle (SAT) at future septation sites. Diego Delgado-Álvarez174. Characterization of the Neurospora crassa STRIPAK complex.Anne Dettmann175. Does the CENP-T-W-S-X tetramer link centromeres tokinetochores? Jonathan Galazka176. Proper actin ring formation and septum constriction requirescoordination of SIN and MOR pathways through the germinal centrekinase MST1. Yvonne Heilig177. Regulatation of the BUD3-BUD4 landmark complex by the NDRkinases DBF2 and COT1 during septum formation in Neurosporacrassa. Yvonne Heilig178. Development of a Protein-Protein Interaction Platform inNeurospora Crassa. Shouqiang Ouyang179. Specific Structural Features of Sterols Affect Cell-Cell Signalingand Fusion in Neurospora crassa. Martin Weichert180. The role of NADPH oxidases in Neurospora crassa cell fusion.Nallely Cano-Dominguez181. DYNAMICS OF THE PROTEINS BUD-2 AND BUD-5 DURING CELLPOLARIZATION IN NEUROSPORA CRASSA. E. Castro-Longoria182. The role of calcium and calmodulin during cell fusion and colonyinitiation in Neurospora crassa. Chia-Chen Chang183. Deletion of cAMP phosphodiesterase pde-2/acon-2 gene causesthe enhanced osmotic sensitivity in os-1 and os-2 mutants of N. M.Fujimura crassa.184. Genetic analysis of GNB-1 and CPC-2 with the G alpha subunits inHeterotrimeric G protein signaling in Neurospora crassa. AMrutaGarud185. Communication Interference during Cell Fusion in Neurosporacrassa is controlled by a Region under Balancing Selection in theHeterokaryon Incompatibility Locus het-c. Jens Heller186. The N. crassa Bem46 protein: alternative splicing and eisosomalassociation. Frank Kempken187. The alternative oxidase induction pathway is involved insenescence associated with over-replication of a mitochondrialplasmid in Neurospora crassa. John Kennell188. Relationship among mutagen sensitivity, senescence andmitochondrial morphology in the ultraviolet sensitive-5 mutant ofNeurospora crassa. Kiminori Kurashima189. Localization of EGL-1 and EGL-2, two GPI anchored cell wall β (1-3) endoglucanases, at hyphal apices and septa, and in interconidialsepta in Neurospora crassa. Leonora Martinez190. Stability of a G protein alpha subunit in genetic backgroundslacking the G beta subunit or a cytosolic guanine nucleotide exchangefactor. Alexander V. Michkov191. Functional analysis the Saccharomyces cerevisiae Ste20, Cla4homologue in Neurospora crassa. Yuhei Nogami192. Dissecting the Pathway of Cellulase Secretion in Neurosporacrassa. Trevor Starr193. Towards understanding the endoplasmic reticulum associateddegradation process of misfolded glycoproteins in Neurospora crassa.Georgios Tzelepis194. Withdrawn195. Genetic analysis of the role of peroxisomes in the virulence andsurvival in Fusarium graminearum. K. Min196. roGFP and anti-oxidant defences in the rice blast fungusMagnaporthe oryzae. Marketa Samalova197. Dimorphism and virulence in pathogenic zygomycetes. Soo ChanLee198. Genetic analysis of the components of the ime-2 mediatedsignaling events during nonself recognition and programmed celldeath (PCD) in Neurospora crassa. Joanna A. Bueche199. PRO45 is a component of the conserved STRIPAK complex inSordaria macrospora. Steffen Nordzieke27th Fungal Genetics Conference | 107

LISTING OF ALL POSTER ABSTRACTS200. Molecular Determinants of Sporulation in Ashbya gossypii.Jurgen W. Wendland201. VELVET is regulated by ENV1 and impacts development ofTrichoderma reesei. Hoda Bazafkan202. Sexual reproduction and mating type function in the penicillinproducing fungus Penicillium chrysogenum. Julia Böhm203. Exponentiate complexity: non-mating GPCRs in thebasidiomycete Schizophyllum commune. Daniela Freihorst204. Characterization of new STRIPAK complex interaction partners inthe filamentous ascomycete Sordaria macrospora. Britta Herzog205. Hypocrea jecorina meiosis generates segmentally aneuploidprogeny to enhance production of xylan-degrading hemicellulases. T.-F. Wang206. Deletion of MAT 1-2-1 gene results in mating type switching inCeratocystis fimbriata. Brenda D. Wingfield207. Mannitol is essential for the development of stress resistantascospores in Neosartorya fischeri. Timon T. Wyatt208. A small lipopeptide pheromone with limited proline substitutionscan still be active. Thomas J. Fowler209. Function of Ras proteins in fungal morphogenesis ofSchizophyllum commune. E.-M. Jung210. The developmental PRO40/SOFT protein participates in signalingvia the MIK1/MEK1/MAK1 module in Sordaria macrospora. InesTeichert211. Map-based identification of the mad photosensing genes ofPhycomyces blakesleeanus. Silvia Polaino Orts212. The C 2H 2 transcription factor HgrA promotes hyphal growth inthe dimorphic pathogen Penicillium marneffei. Hayley E. Bugeja213. Involvement of a specific ubiquitin ligase in the assembly of thedynein motor. Michael Plamann214. Identification and characterization of new alleles required formicrotubule-based transport of nuclei, endosomes, and peroxisomes.K. Tan215. Pheromone-induced G2 cell cycle arrest in Ustilago maydisrequires inhibitory phosphorylation of Cdk1. Sónia M. Castanheira216. Microtubule-dependent mRNA transport and mitochondrialprotein import in Ustilago maydis. T. Langner217. “The vacuole” of Neurospora crassa may be composed ofmultiple compartments with different structures and functions. BarryJ. Bowman218. Comparisons of two wild type A mating type loci and derivedself-compatible mutants in the basidiomycete Coprinopsis cinerea.Ursula Kües219. Transformation of an NACHT-NTPase gene NWD2 suppresses thepkn1 defect in fruiting body initiation of the Coprinopsis cinereamutant Proto159. Ursula Kües108220. Dynamics of the actin cytoskeleton in Phytophthora infestans.Harold MeijerComparative and Functional Genomics221. A novel approach for functional analysis of genes in the rice blastfungus. Sook-Young Park222. Distribution and evolution of transposable elements in theMagnaporthe oryzae/grisea clade. Joelle Amselem223. Alternative structural annotation of Aspergillus oryzae andAspergillus nidulans based on RNA-Seq evidence. Gustavo C. Cerqueira224. Improved Gene Ontology annotation for biofilm formation,filamentous growth and phenotypic switching in Candida albicans.Diane O. Inglis225. Genome sequencing of Verticillium albo-atrum pathotypes inorder to understand wilt disease in hop production. J. Jakse226. Aegerolysin proteins from Aspergillus species. Nada Krasevec227. Assembly, Annotation, and Analysis of Multiple MycorrhizalFungal Genomes. Alan Kuo228. Comparative reannotation of 21 Aspergillus genomes. Asaf A.Salamov229. Using the phenotypic information in the PHI-base database toexplore pathogen genomes, transcriptomes and proteomes. MartinUrban230. RNA-Seq analysis reveals new gene models and alternativesplicing in Fusarium graminearum. Theo vanderLee231. Comparison of transcriptome technologies in the MpkA deletionmutant of Aspergillus fumigatus. Clara Baldin232. Spliceosome twintrons ( “stwintrons”) revealed by fungal nucleargenomes. Erzsébet Fekete233. NGS data revealed that the NSDA sterile mutant contains amutation in the SCF ubiquitin ligase subunit gene, culC, in Aspergillusnidulans. Kap-Hoon Han234. Whole genome sequencing of two Aspergillus oryzae strainsisolated from Meju, a traditional brick of dried fermented soybean, inKorea. Kap-Hoon Han235. Systematic analysis of the uncharacterized genes, which widelyconserved among filamentous fungi, in Aspergillus oryzae.K. Iwashita236. Penicillium purpurogenum degrades lignocellulose. What can welearn of this process by analyzing the genome, transcriptome andsecretome of the fungus? Jaime Eyzaguirre237. Functional genomics of lignocellulose degradation in theBasidiomycete white rot Schizophyllum commune. Robin A. Ohm238. Functional characterization of genes expressed in early infectionstages by the phytopathogenic fungus Botrytis cinerea. J. Espino

LISTING OF ALL POSTER ABSTRACTS239. Regulation of biofilm formation in Candida parapsilosis. LindaHolland240. Functional analysis of the Mps1 MAP kinase pathway in the riceblast fungus Magnaporthe oryzae. E. Grund241. A RNA-Seq directed functional genomics screen to identify novelcell wall genes in the hyphal tip of Neurospora crassa. Divya Sain242. Identification of centromeres in the plant pathogenZymoseptoria tritici (synonym Mycosphaerella graminicola). KlaasSchotanus243. Loss of the RNAi pathway in VGII Cryptococcus gattii sheds lighton the intact system in Cryptococcus neoformans. R. Blake Billmyre244. A chemical-genetic map of a human fungal meningitis pathogen.Jessica C. S. Brown245. Whole genome sequencing of high-mortality and low-mortalitystrains of Cryptococcus neoformans var. grubii to discover geneticdeterminants of virulence. Tami R. McDonald246. Identification of high temperature-regulated genes controlled bySch9 through comparative transcriptome analysis in Cryptococcusneoformans. Dong-Hoon Yang247. Genome-wide analysis of eleven white- and brown-rotPolyporales provides insight into mechanisms of wood decay. ChiakiHori248. Genomic context and distribution of effector genes in Fusariumoxysporum. Sarah Maria Schmidt249. Whole genome sequencing reveals new links between diverseplant pathogens; an expanded AvrLm6-like gene family in Venturiaspecies. Jason Shiller250. Oömycetes Protein Array Project. Samantha Taylor251. Extensive chromosomal reshuffling drives evolution of virulencein an asexual pathogen. Ronnie de Jonge252. Genomic census of transmembrane proteins of the marinefungus, Corollospora maritima. Derek Johnson253. Fungal Calcium Signaling Database (FCSD). Venkatesh Moktali254. Evolutionary genomic analysis of cytochrome P450 proteins inthe subphyla Pezizomycotina. Venkatesh Moktali255. Uncovering the evolutionary pressures shaping theGlomeromycota-Glomeribacter endosymbiosis. Stephen J. Mondo256. The Unique family of Telomere-Linked Helicases in Fungi. OlgaNovikova257. Evolution of proteins containing intein- and Hedgehog-like Vintdomains in Fungi. Olga Novikova258. The genome and development-dependent transcriptome ofPyronema confluens: a window into fungal evolution. MinouNowrousian259. Genome and transcriptome analysis of the mycoparasiteClonostachys rosea. Kristiina Nygren260. The mitochondrial genomes of Fusarium circinatum, F.verticillioides and F. fujikuroi are unexpectedly similar. G. Fourie261. Comparative pathogenomics: next generation dissection ofmechanisms of pathogenesis on plants. Donald M. Gardiner262. Characterisation of stuA homologue in Fusarium culmorum.Matias Pasquali263. Comparative analysis of noncoding sequences in the Gibberellafujikuroi species complex. Christian Sieber264. Virulence of Fusarium circinatum on Pinus species. S. L. Slinski265. Understanding the remodeling of the wheat grain genomeexpression during infection, a gate to get new insights on themolecular cross-talk controlling the development of the interactionbetween the wheat and Fusarium graminearum.. Chetouhi Cherif266. Genetic and epigenetic changes in Fusarium graminearumfollowing serial subculture. Heather E. Hallen-Adams267. Evolutionary and functional analysis of mitosis-related kinasegenes in Fusarium graminearum. Huiquan Liu268. Functional analysis of A MADS-box transcription Mcm1 inFusarium graminearum . Chen fang Wang269. Genome sequencing of the Fusarium graminearum speciescomplex in Korea. Sung-Hwan Yun270. Identification and functional analysis of virulence genes indifferent host-pathogenic forms of Fusarium oxysporum. P. van Dam271. Protocol for generating gene knock-out transformants of thefungal pathogen Verticillium albo-atrum. M. Flajsman272. Functional analysis of catalase-peroxidase encoding genes in thefungal wheat pathogen Zymoseptoria tritici. A. Mirzadi Gohari273. Efficient recycling of selective marker genes with the Cre-loxPrecombination system via anastomosis in Cryphonectria parasitica.Dong-Xiu Zhang274. RNA-seq reveals the pleiotropic regulating functions of thetranscription factor XYR1 in Trichoderma reesei. Liang Ma275. Tyrosinase an important enzyme for melanin production in theoomycete Saprolegnia parasitica. Marcia Saraiva276. Comparative transcriptomics of Cordyceps bassiana tounderstand expression levels of its NRPS related genes. B. Shrestha277. Global analysis of the Colletotrichum gloeosporioides genomeand transcriptome reveals a conserved role for pacC pH regulation infungi. Noam Alkan278. WITHDRAWN279. Anisogramma anomala: a unique fungus with a huge genome.Guohong Cai280. Leptosphaeria maculans 'brassicae': "Transposable Elementschanged my life, I feel different now". Jonathan Grandaubert281. Genomics of fungal interactions for bioenergy crops. IgorGrigoriev27th Fungal Genetics Conference | 109

LISTING OF ALL POSTER ABSTRACTS282. Genomic analysis of oleaginous fungi. H. Koike283. Genome sequencing, assembly and annotation of a marine fungalisolate of Scopulariopsis brevicaulis using three different nextgeneration sequencing technologies. Abhishek Kumar284. Genome sequencing, assembly and annotation of a marine fungalisolate of Pestalotiopsis using next generation sequencingtechnologies. Abhishek Kumar285. Genome sequencing, assembly and annotation of a marine fungalisolate of Calcasporium using different next generation sequencingtechnologies. Abhishek Kumar286. Genome sequencing, assembly and annotation of three marinefungal isolates using different next generation DNA sequencingmethods for pharmaceutically important secondary metabolites.Abhishek Kumar287. From Shiitake genomics to comparative mushroom genomics:Mushroomics. Hoi Shan Kwan288. Comparative genomics of Ceratocystis polonica and Ophiostomabicolor, two bark beetle-associated pathogenic fungi. Ljerka Lah289. The largest fungal mitochondrial genome of a basidiomycetecontains signs of genetic flexibility and recombination events. Taina K.Lundell290. De Novo Assembly of Fungal Genomes and Detection ofStructural Variation using Extremely Long Single-Molecule Imaging.Nicholas R. Rhind291. Discovering host specificity candidate genes of Sporisoriumreilianum by genotyping mixed-variety offspring. J. Schirawski292. Comparing comparative “omics” in Coccidioides spp. Emily A.Whiston293. The transcriptional response during cell-fusion incompatibility inPodospora anserina. Sven J. Saupe294. Comparative Analysis of Putative Rhodopsins in Early DivergingFungal Lineages. Steven Ahrendt295. Molecular Tools to Silence and Confirm Genes in PhytophthoraSojae. Felipe R. Arredondo296. Functional Characterization of Transcription Factor Genes,MoNIT4 and MoLEU3, in Magnaporthe oryzae. Jaehyuk Choi297. Comparative genomic analysis of world-wide Magnaportheoryzae isolate collection. Jaeyoung Choi298. Functional characterization of two genes encoding putativeZn(II) 2Cys 6 transcription factors, MoCOD1 and MoCOD2 inMagnaporthe oryzae. Hyunjung Chung299. Comparative proteomics of monoclonal-antibody enrichedhaustoria from three races of Puccinia triticina. Christof Rampitsch300. RNA-seq analyses of gene expression in the microsclerotia ofVerticillium dahliae. Steven J. Klosterman301. Exploring the Genome Diversity of Mycorrhizal Fungi toUnderstand the Evolution and Functioning of Symbiosis. Francis M.110Martin302. Known unknown genes: evolution of eukaryotic BEM46.Abhishek Kumar303. Sugar ‘cubed’ - A Comparative Systems Analysis of Plant Cell WallPolysaccharide Recognition and Degradation Using the ModelFilamentous Fungus Neurospora crassa. J. P. Benz304. Building upon whole genome resequencing in Neurospora. KevinMcCluskey305. Genome based phylogeny of early diverging fungal lineages. A. P.Gryganskyi306. Comparative analysis of 35 basidiomycete genomes revealsdiversity and uniqueness of the phylum. Robert Riley307. Functional analysis of roles of expanded genes in fruiting bodydevelopment in Coprinopsis cinerea. Jinhui Chang308. Comparative analysis of fungal kinomes. Abhishek Kumar309. VeA, VelB and FluG affect conidiation and aflatoxin production ofAspergillus flavus. L. Scharfenstein310. The Environmental Molecular Sciences Laboratory molecularanalysis capabilities for fungal biology. S. E. Baker311. Comparative Genomics and Transcriptomics of InsectPathogenesis. Kathryn E. Bushley312. Integrated transcriptional profiling and analysis for identificationof Cryptococcus neoformans genes regulated during humancryptococcal meningitis. Y. Chen313. Structural and functional characterization of microRNA-like RNAsin the penicillin producing fungus Penicillium chrysogenum. TimDahlmann314. Metatranscriptomic analysis of ectomycorrhizal root clusters inPinus taeda: new methodologies for assessing functional geneexpression in situ. H.-L. Liao315. Transcriptomic response of Neurospora crassa germinatingconidia to chitosan in sub-lethal dose. Federico Lopez-Moya316. Transcriptomic analysis of the interaction between Trichodermaharzianum and the phytopathogen Sclerotinia sclerotiorum using theRNA-seq approach. Andrei S. Steindorff317. Genome evolution of the original Saccharomyces carlsbergensislager yeast strain, Unterhefe No1, as revealed by whole genomesequencing. Andrea Walther318. Retention of genes in a secondary metabolite gene cluster thathas degenerated in multiple lineages of the Ascomycota. Daren W.Brown319. Functional characterization of unique non-ribosomal peptidesynthetase genes in the cereal fungal pathogen Cochliobolus sativus.Yueqiang Leng320. Phylogenomics unveils secondary metabolites specific tomycoparasitic lineages in Hypocreales. C. Alisha Owensby321. Genome and transcriptome sequence of the apomictic fungusArnium arizonense (Podospora arizonensis). R. Debuchy

LISTING OF ALL POSTER ABSTRACTS322. Role of MAP kinase pathways in the pathogenicity of the wheatpathogen Mycosphaerella graminicola . Elisabetta Marchegiani323. WITHDRAWN324. Ancient and abundant MITEs in epichloae genomes. DamienFleetwood325. Exploring the biomass modifying enzymes of new filamentousfungal isolates from Vietnam, using secretome and transcriptomeanalyses. George E. Anasontzis326. Fusarium Comparative Transcriptomics and TranscriptionalRegulatory Network Reconstruction. L. Guo327. The mycorrhizal genome initiative (MGI): Identification ofsymbiosis-regulated genes by using RNA-Seq. A. Kohler328. Transcriptome, secreted enzymes and systematics of the whiterot basidiomycete Phlebia radiata. Jaana Kuuskeri329. Characterization of molecular mechanisms underlying the multidrug-resistantphenotypes of Mycosphaerella graminicola fieldisolates. Sabine Fillinger330. Candidate pathogenesis gene identification via Ustilago maydis`first gen` genomic analyses. M. E. Donaldson331. A biocontrol agent among pathogens : How Pseudozymaflocculosa genome relates to singular lifestyle. F. Lefebvre332. A tale of two poplar pathogens - Moving from sequence tofunction. B. Dhillon333. Defining Open Chromatin Regions in Coprinopsis cinerea Oidia byFAIRE. Virginia K. Hench334. Ensembl Fungi - genome-scale data portal from fungal species.Uma Maheswari335. A draft genome of the ectomycorrhizal fungus Rhizopogonvesiculosus: Characterization of mating system and heterozygositywithin the dikaryon. Alija Mujic336. Diverse Lifestyles and Strategies of Plant Pathogenesis Encodedin the Genomes of Eighteen Dothideomycetes Fungi. Robin A. Ohm337. Domains of meiotic DNA recombination and gene conversion inCoprinopsis cinerea (Coprinus cinereus). Patricia J. Pukkila338. FungiDB: An integrated functional genomics database for fungi.Raghuraman Ramamurthy339. Letters from the front: The Microbotryum violaceum genome andtranscriptome project. Su San Toh340. The Aspergillus and Candida Genome Databases: RecentDevelopments and Future Plans. Martha B. Arnaud341. The Trichoderma reesei polyketide synthase gene pks1 isnecessary for yellow-green pigmentation of conidia and is involved inthe establishment of environmental fitness. Lea Atanasova342. Functional Analysis of Genes in Regions of Introgression inCoccidioides. Bridget M. Barker343. Classification and accurate functional prediction of carbohydrateactiveenzymes by recognition of short, conserved peptide motifs.Peter K. Busk344. The mechanism of introner-like element multiplication in fungi.Jérôme Collemare345. Fungi use prion folds for signal transduction processes involvingSTAND proteins. Asen Daskalov346. RNA silencing in poplar anthracnose fungus Colletotrichumgloeosporioides. Simeng Li347. Comparative Genomics of L and S Morphotypes of Aspergillusflavus. Mana Ohkura348. Searching for Functional Mobile Elements in Coprinopsis cinerea.Marilee A. Ramesh349. Genome-wide analysis of small RNA machineries in fungalkingdom. Jiayao Wu350. Comparative Analysis of Malassezia Mating Loci. Jun XuEducation and Professional Development351. The internet effectiveness for gaining students enrolled at collegeof education The scientific facts and concepts about Biofuel issueaccording to the Responsibility Spiral model. Khlood S. AlSheikh352. ComGen Authentic Research Experiences (C-ARE): Fungal geneticanalysis. Andrea Gargas353. Facilitating an Interdisciplinary Learning Community AmongstUndergraduate Research Fellows By Emphasizing Scientific Inquiry asthe Unifying Thread. Virginia K. HenchGene Regulation354. YAB- An Agrobacterium-based vector system for direct cloning ofeukaryotic gene constructs via yeast recombination. M. AlejandraMandel355. Removal of C4-methyl Sterol Accumulation in a SREBP-nullMutant of Aspergillus fumigatus Restores Hypoxia Growth. Sara J.Blosser356. VeA Regulates Conidiation, Gliotoxin Production and ProteaseActivity in the Opportunistic Human Pathogen Aspergillus fumigatus.Sourabh Dhingra357. HapXcess and C-terminal truncation impairs Aspergillusfumigatus' iron homeostasis. Fabio Gsaller358. The CCAAT-Binding-Complex mediates Iron Regulation inAspergillus fumigatus. Hubertus Haas359. Protein kinase A signaling in Aspergillus fumigatus: Identificationof downstream targets. Juliane Macheleidt360. Aspergillus nidulans galactofuranose biosynthesis affectsantifungal drug sensitivity. Md. Kausar Alam27th Fungal Genetics Conference | 111

LISTING OF ALL POSTER ABSTRACTS361. WITHDRAWN362. Sulfur metabolism regulatory mutations induce environmentalstress response. J. Brzywczy363. RNA 3' tagging - signalling transcript degradation andtranslational repression. Mark X. Caddick364. A novel C2H2 type finger transcription factor, MtfA, regulatesmycotoxin biosynthesis and development in Aspergillus nidulan. AnaM. Calvo-Byrd365. Histidine 704 of the Aspergillus nidulans GATA factor AreA isrequired for nuclear export. Damien Downes366. Redundant Nuclear Localization Signals Mediate Nuclear Importof the Aspergillus nidulans Transcription Activator of NitrogenMetabolic Genes AreA. Cameron C. Hunter367. Conditional expression of the phospho-transmitter gene ypdAand the signaling interaction of YpdA with response regulators; SskAand SrrA in Aspergillus nidulans. Mayumi Nakayama368. WITHDRAWN369. FigA, a putative member of low-affinity calcium system, isinvolved in both asexual and sexual differentiation in Aspergillusnidulans. Shizhu Zhang370. The Saccharomyces cerevisiae FUS3 homologue MAKB inAspergillus niger is a central regulator connecting differentiation andsecondary metabolite production with nutrient availability and light.Bert-Ewald Priegnitz371. Engineering and characterizing protein secretion in Aspergillusniger. Y. Zheng372. Design of culture conditions for secondary metabolite productionof fungi based on large-scale transcriptome data. M. Umemura373. The effect of the clbR overexpression on cellulose degradingenzyme production in Aspergillus aculeatus. S. Tani374. Regulation of the α-tubulin B-encoding gene during earlydevelopment of the phytopathogenic fungus Botrytis cinerea. Y. FaivreTalmey375. The regulation of D-galacturonic acid utilization in Botrytiscinerea. Lisha Zhang376. Epigenetic Regulation of Subtelomeric Gene Noise in Candidaalbicans. Matthew Z. Anderson377. Signaling and Cell Behavior: Pheromone Response in Candidaalbicans. Ching-Hsuan Lin378. Transcriptional regulatory networks controlling the early hypoxicresponse in Candida albicans. A. Nantel379. Gene expression and function during invasive Candida infection.Wenjie Xu380. Effects of histone H3 point mutations on centromeremaintenance. Steven Friedman381. Analysis of the transcriptional regulation of genes involved in thesynthesis and organization of the cell wall of Ustilago maydis during112infection of an alternatiive host. Angélica Mariana Robledo Briones382. Expression of the Trichophyton rubrum ace2 and pacC genesduring degradation of keratinized substrates. Larissa Silva383. Control and Function of Two Fatty Acid Regulators in Neurosporacrassa. Erin L. Bredeweg384. Characterization of genomic targets for the Neurospora crassahypothetical transcription factor NCU04390 by ChIP-sequencing. R.Gonçalves385. The KMT6 Histone H3 K27 Methyltransferase RegulatesExpression of Secondary Metabolites and Development in Fusariumgraminearum. Kristina M. Smith386. Circadian clock-gated cell division cycles in Neurospora crassa. C.Hong387. Protein Binding Microarrays and high-throughput real-timereporters studies: Building a four-dimensional understanding oftranscriptional networks in Neurospora crassa. L. F. Larrondo388. Glycogen metabolism is regulated by the circadian clock inNeurospora crassa. S. Virgilio389. Genetic and Molecular Dissection of the Neurospora CircadianOscillatory System. Qijun Xiang390. Non-optimal codon usage determines the expression level,structure and function of the circadian clock protein FREQUENCY.Mian Zhou391. Two putative long non-coding RNAs upstream of transcriptionfactor Znf2 may regulate morphogenesis (or dimorphic transition) inCryptococcus neoformans. Nadia Chacko392. Introns in Cryptococcus neoformans. Guilhem Janbon393. Unravelling of sexual differentiation mediated by Ire1 via Hxl1-independent manners in Cryptococcus neoformans. Kwang-Woo Jung394. Functional analysis of PUF mediated post-transcriptionalregulation in Cryptococcus neoformans. Jan Naseer Kaur395. Determining the direct targets of two master regulators of sexualdevelopment in Cryptococcus neoformans. Matthew E. Mead396. Post-transcriptional gene regulation contributes to hosttemperature adaptation and virulence in Cryptococcus neoformans.Amanda L. Misener Bloom397. Protein arginine methylation in post-transcriptional generegulation and stress adaptation of Cryptococcus neoformans. J. T.Graham Solomons398. UVE1 is a Photo-regulated Gene Required for the Protection ofMitochondrial DNA in Cryptococcus neoformans from UV InducedDNA Damage. Surbhi Verma399. Uve1 endonuclease protects Cryptococcus neoformans from UVdamage through regulation by the White collar complex. SurbhiVerma400. Multiple laccase genes in Schizophyllum commune. S. Madhavan

LISTING OF ALL POSTER ABSTRACTS401. Regulation of DNA repair genes expression by UV stress inNeurospora crassa. Tsukasa Takahashi402. Diverse classes of small RNAs originating from genomic hotspots,tRNA and the mitochondrial genome in Phytophthora infestans.Sultana N. Jahan403. Epigenetic control of effector gene expression in the plantpathogen fungus Leptosphaeria maculans. Jessica Soyer404. Discovering the link: The NOX-GSA network for sexualdevelopment and ascospore germination in Sordaria macrospora.Daniela Dirschnabel405. The bZIP transcription factor Atf1 acts as a global regulator forsecondary metabolite production in Fusarium fujikuroi. Sabine E.Albermann406. Role of the Vivid ortholog of Fusarium fujikuroi VvdA incarotenoid biosynthesis and development. Marta Castrillo Jimenez407. Gene expression of secondary metabolism gene clusters bydifferent Fusarium species during in planta infection. J. Espino408. A cis-acting factor modulating the transcription of FUM1 inFusarium verticillioides. Matias Pasquali409. Shedding light on secondary metabolite cluster gene expression,sporulation, UV-damage repair and carotenogenesis in the ricepathogen Fusarium fujikuroi. Phillipp Wiemann410. Fgap1-mediated response to oxidative stress in trichotheceneproducingFusarium graminearum. N. Ponts411. Functional analyses of FgLaeA in Fusarium graminearum. Sung-Hwan Yun412. Molecular cloning and differential expression of two novel Family1 β-glucosidases genes from the rare fungus Stachybotrys microspora.Salma Abdeljalil413. The transcriptional factors XYR1 and CRE1 regulate theexpression of Cellulolytic and Xylanolytic genes at carbon sourcedependent-manner in Hypocrea jecorina (Trichoderma reesei).Amanda C. C. Antoniêto414. Characterization of tannic acid-inducible and hypoviral-regulatedCpsHsp1 expression level of the chestnut blight fungus Cryphonectriaparasitica. J.-H. Baek415. Artificial miRNA constructs for Phytophthora sojaetransformation. Stephanie R. Bollmann416. RNAi-dependent epimutations evolve antifungal drug resistancein the zygomycete fungal pathogen Mucor. Silvia Calo Varela417. Heterochromatic marks are involved in the repression of plantregulatedsecondary metabolism in Epichloë festucae and forsymbiotic interaction with the host perennial ryegrass. Tetsuya Chujo418. Cellulose Degradation Regulator 2 Induces Expression of aConserved Core of Genes for Plant Cell Wall Saccharification inNeurospora crassa and Aspergillus nidulans. Samuel T. Coradetti419. The transcriptional repressor CRE-1 regulates glycogenmetabolism in Neurospora crassa. Fernanda B. CupertinoJuwen C. DuBois420. Transcriptional Response to Hypoxia in the Dimorphic FungusHistoplasma capsulatum . Juwen C. DuBois421. The Neurospora crassa SEB1 transcription factor binds to STREmotif and modulates stress responses through different pathways. F.Freitas422. WITHDRAWN423. Regulation of glycolysis and gluconeogenesis by antisensetranscription in Aspergillus nidulans? Michael Hynes424. ClbR and its paralog, ClbR2, regulate gene expression of cellulasegenes in response to cellobiose in Aspergillus aculeatus. E. Kunitake425. Expression of a bacterial xylanase in Trichoderma reesei underthe egl2 and cbh2 glycosyl hydrolase gene promoters. HelenaNevalainen426. A single argonaute gene participates in exogenous andendogenous RNAi and controls different cellular functions in the basalfungus Mucor circinelloides. F. E. Nicolas-Molina427. The transcription factor, AtrR, regulates the expression of ABCtransporter genes and ergosterol biosynthesis genes in aspergilli.Ayumi Ohba428. Effects of intron deletions in production of a heterologousprotease in Trichoderma reesei. Marja Paloheimo429. Novel core promoter elements in the oomycete Phytophthorainfestans and their influence on expression pattern detected bygenome-wide analysis. Laetitia Poidevin430. New insights into the phosphate-sensing network in Neurosporacrassa. Antonio Rossi431. Carbon source and light dependent regulation of gene clusters inTrichoderma reesei (Hypocrea jecorina). Monika Schmoll432. Identification of regulators for enzyme production inTrichoderma reesei using genome-wide approaches. M. Valkonen433. The central core of the response to light and injury and theirregulation by RNAi machinery in the filamentous fungus Trichodermaatriviride. J. M. Villalobos-Escobedo434. RNA-mediated Gene Silencing in Candida albicans: Reduction ofFungal Pathogenesis by Use of RNAi Technology. M. Moazeni435. RNAi machinery controls the asexual reproduction induced bylight of the filamentous fungus Trichoderma atroviride. N. Carreras-Villaseñor436. Genome-wide analysis of light responses in Mucor circinelloides.Victoriano Garre437. Gene expression profiling of the basidiomycetous fungusLentinula edodes after light stimulation. H. Sano438. Further Characterization of Surface Recognition Mechanisms inMagnaporthe oryzae. Guanghui Wang439. Evidence of Microbial Epigenetics; Loss-of-function mutant of the27th Fungal Genetics Conference | 113

LISTING OF ALL POSTER ABSTRACTSBck1 Homolog, CpBCK1, from the chestnut blight fungusCryphonectria parasitica resulted in the sectoring accompanied withthe changes in DNA methylation. J.-M. Kim440. NUP-6 (Importin α) is required for DNA methylation inNeurospora crassa. Andrew D. Klocko441. Identification and characterization of a Blastomyces dermatitidismutant with a bidirectional defect in the phase transition. Gregory M.Gauthier442. Transcriptional regulation of peptidases and nitrogentransporters during the assimilation of organic nitrogen by theectomycorrhizal fungi Paxillus involutus. Firoz ShahEvelina Y. Basenko443. Characterization of genome maintenance components inNeurospora crassa using whole-genome high-throughput approach.444. Circadian regulation and carbon catabolite repression inNeurospora crassa: Two integrated regulatory systems? Rodrigo Díaz-Choque445. Opposing activities of the HCHC and DMM complexes maintainproper DNA methylation in Neurospora crassa. Shinji Honda446. The transcription factor FL is phosphorylated and interacts with atrehalose related protein in Neurospora crassa. Carmen Ruger-Herreros447. Transcriptomic profiling of fumonisin B biosynthesis by Fusariumverticillioides. N. Ponts448. Differential transcriptome analysis of Zymoseptoria triticiinfecting wheat reveals novel effectors. Stefano F. F. Torriani449. Role of the xprG gene in autolysis, secondary metabolism andasexual development in Aspergillus nidulans. Margaret E. Katz450. Fugal-specific sirtuin HstD coordinates the secondary metabolismand development via the LaeA. M. Kawauchi451. Improved flavor production by manipulation of the Ehrlichpathway in ascomycetes. D. Ravasio452. Suppressor mutagenesis of a ΔlaeA mutant reveals novelregulators of secondary metabolism in Aspergillus nidulans. AlexandraSoukup453. A network of HMG-box transcription factors regulates sexualcycle in the fungus Podospora anserina. R. Debuchy454. Sclerotinia sclerotiorum MAT genes function in fertility andapothecial morphogenesis. Benjamin Doughan455. The Sclerotinia sclerotiorum mating type locus (MAT) contains a3.6-kb region that is inverted in every generation. Patrik Inderbitzin456. Repression of the phosphor-transmitter gene ypdA resulting ingrowth defect in Aspergillus fumigatus. Daisuke Hagiwara457. Unravelling the GTPase polarity complex in Claviceps purpurea.Andrea Herrmann458. Atypical Gβ and RACK homolog Gib2 is a signal transducingadaptor protein affecting growth and virulence of Cryptococcusneoformans. Ping Wang459. Consequences of the loss of transcription factors SreA and HapXon siderophore biosynthesis and iron homeostasis in the perennialryegrass endophyte, Epichloë festucae. Natasha T. Forester460. Who is to blame: defining the host responses that lead to ToxAinducedsusceptibility. Iovanna Pandelova461. RNA silencing of pacC increases aflR transcript levels underalkaline pH conditions in Aspergillus flavus. Benesh M. Somai462. High-throughput prediction and functional validation ofpromoter motifs regulating gene expression in spore and infectionstages of Phytophthora infestans. H. Judelson463. Cooperative regulation of Aspergillus nidulans cellulase genes bytranscription factors McmA and ManR/ClrB. Tetsuo Kobayashi464. Transcription factor shuttling during cellulase induction inTrichoderma reesei. Alex Lichius465. Trichophyton rubrum ap-1 gene expression in response toenvironmental challenges. Nalu TA Peres466. Vib-1 is required for cellulose utilization in Neurospora crassa. YiXiong467. The stringency of start codon selection in the filamentous fungusNeurospora crassa. Matthew S. Sachs468. A temperature-dependent complex transcriptional networkcontrols cell shape and virulence in Histoplasma capsulatum. SinemBeyhan469. Evolutionary analysis of Dicer proteins: a preliminary analysis tostudy of microRNAs in the mushroom, Coprinopsis cinerea. XuanjinCheng470. Effect of the trp1 gene on transformation frequencies inCoprinopsis cinerea. Bastian Doernte471. The first promoter for conditional gene expression inAcremonium chrysogenum: iron starvation-inducible mir1 P . FabioGsaller472. Mutagenic effect of high-LET ion beam irradiation in Neurosporacrassa. Liqiu Ma473. The Mad complex binds to light-regulated promoters inPhycomyces blakesleeanus. Alejandro Miralles-Duran474. Down Regulation of sidB Gene by Use of RNA interferenceTechnology in the Filamentous Fungi Aspergillus nidulans. S, Rezaie475. SmallRNA mediated meiotic silencing of a transposable elementin Neurospora crassa. Yizhou WangPathogenic and Mutalistic Interactions476. Functional Characterization of Small, Cysteine-Rich SecretedEffectors from the Filamentous Fungus Magnaporthe oryzae. WilliamC. Sharpee477. Penetration-specific effectors from Phytophthora parasiticafavour plant infection. Edouard Evangelisti478. Transcriptional regulatory circuits necessary for appressorium-114

LISTING OF ALL POSTER ABSTRACTSmediated plant infection by Magnaporthe oryzae. Miriam Oses- Ruiz479. Differential activation of ammonium transporters during theaccumulation of ammonia by Colletotrichum gloeosporioides and itseffect on appressoria formation and pathogenicity. Dov B. Prusky480. Functional analysis of Nbs1 of Magnaporthe oryzae. K. Sasaki481. Influence of hypoxia on antifungal susceptibility, sterole patternand biomarker release of Aspergillus spp. Ulrike Binder482. Sit and wait: Special features of Aspergillus terreus inmacrophage interactions and virulence. M. Brock483. Identification and characterization of an RXLR-like effector familyfrom medically relevant fungi. Shiv D. Kale484. A role for PalH-mediated signal transduction in A. fumigatusvirulence and cell wall integrity: An exploitable target for combinationtherapy? M. Bertuzzi485. Aspergillus fumigatus trehalose-6-phosphate regulates innateimmune responses and virulence through modulation of fungal cellwall composition. Robert A. Cramer486. Fungal lipoxygenases: a novel instigator of asthma? Gregory J.Fischer487. F-box protein 15 (Fbx15) links virulence of Aspergillus fumigatusto protein degradation and stress response. Bastian Jöhnk488. The sfp-type phosphopantetheinyl transferase, PPTA, is criticalfor the virulence of Aspergillus fumigatus. A. E. Johns489. Characterization of effectors of the barley pathogenRhynchosporium commune. Daniel Penselin490. Molecular and genetic basis guiding the establishment of amutualistic relationship between Epichloë festucae and perennialryegrass. Sladana Bec491. Puccinia graminis and Brachypodium distachyon: contrastingprofiles of host-pathogen incompatibility. Melania Figueroa492. Magnaporthe oryzae has evolved two distinct mechanisms ofeffector secretion for biotrophic invasion of rice. Martha C. Giraldo493. Trichoderma rhizosphere’s competency, endophytism and plantcommunication: A molecular approach. Artemio Mendoza494. Ustilago bromivora - Brachypodium distachyon: a novelpathosystem. Franziska Rabe495. T-DNA mediated insertional mutagenesis: evidence of a newgene implied in the early phase of pathogenic development of Botrytiscinerea. Nathalie Poussereau496. The NADPH Oxidase Complexes in Botrytis cinerea. UlrikeSiegmund497. A putative function of small RNAs in the plant pathogen Botrytiscinerea. Arne Weiberg498. The Role of Quorum-sensing Molecules in Interactions betweenCandida albicans and its Host. Jessica C. Hargarten499. The Role of ISW2 for in vitro and in vivo ChlamydosporeProduction in Candida albicans. Ruvini U. Pathirana500. Nutrient immunity and systemic readjustment of metalhomeostasis modulate fungal iron availability during the developmentof renal infections. Joanna Potrykus501. Identification of the gut fungi in humans with nonconventionaldiets. Mallory Suhr502. The mutational landscape of gradual acquisition of drugresistance in clinical isolates of Candida albicans. Dawn Thompson503. Yeast-Hypha transition and immune recognition of Candidaalbicans influenced by defects in cell signal transduction pathways.Pankaj Mehrotra504. GPI PbPga1 of Paracoccidioides brasiliensis is a surface antigenthat activates macrophages and mast cells through the NFκB signalingpathway. C. X. R. Valim505. Cladosporium fulvum effector Ecp6 outcompetes host immunereceptor for chitin binding through intrachain LysM dimerization.Andrea Sánchez-Vallet506. Genotypic and phenotypic characterization of Setosphaeriaturcica reveals population diversity and a candidate virulence genelocation. Santiago MiderosSantiago Mideros507. The secretome is linked to virulence in the yeast pathogenCryptococcus. Santiago Mideros508. Post-Transcriptional Regulation of the ER Stress Response inCryptococcus neoformans. Virginia E. Havel509. A morphogenesis regulator controls cryptococcal neurotropism.Xiaorong Lin510. Extracellular and intracellular signaling orchestratesmorphotype-transition and virulence in human pathogenCryptococcus neoformans. Linqi Wang511. Evidence for alkaloid diversity and independent hybridizationevents of Elymus endophytes. Nikki D. Charlton512. The functional characterization of candidate genes involved inhost specialization of Zymoseptoria grass pathogens. Stephan Poppe513. Diversity and Phylogeny of genus Suillus (Suillaceae, Boletales)from Pakistan (Asia). Samina Sarwar514. Saprolegnia species can switch hosts to cause infection: a newinsight into host pathogen interaction. Mohammad N. Sarowar515. The Plant-Microbe Interfaces project: defining and understandingrelationships between Populus and the rhizosphere microbiome.Christopher Schadt516. Do the fungal homologs of Verticillium dahliae effector Ave1 actas virulence factors? Jordi C. Boshoven517. The candidate effector repertoire of closely related Venturiapathogens of the Maloideae revealed by whole genome sequence andRNA sequencing analyses. Joanna Bowen27th Fungal Genetics Conference | 115

LISTING OF ALL POSTER ABSTRACTS518. Thermodynamic characterization of RXLR and RXLR-like effectorsbinding to phosphatidylinositiol-phosphates. Kelly Drews519. Investigating virulence effectors in the poplar-poplar rustpathosystem. Sebastien Duplessis520. Functional analysis of Aphanomyces euteiches effectors, alegume root pathogen. E. Gaulin521. Participation of effector proteins from Trichoderma spp. ininteraction with Arabidopsis thaliana. P. Guzman-Guzman522. The role of LysM effectors in fungal fitness. Anja Kombrink523. WITHDRAWN524. Functional analysis and localization of SnTox1, a necrotrophiceffector produced by the wheat pathogen Stagonospora nodorum.Zhaohui Liu525. Host-targeting protein 3 (SpHtp3) from the oomyceteSaprolegnia parasitica translocates specifically into fish cells in a pHand tyrosine O-sulfate-dependent manner. Lars Löbach526. Ave1-like orthologs in Venturia: another expanded effectorfamily emerges. Kim M. Plummer527. Identification of targets of mycorrhizal effector proteins inplanta. Natalia Requena528. Structural basis for interactions of the Phytophthora sojae RXLReffector Avh5 with phosphatidylinositol 3-phosphate and for host cellentry. Brett M. Tyler529. Dose-dependent induction of plant immunity by application ofthe Fusarium mycotoxin deoxynivalenol. Christian A. Voigt530. How Oomycete Pathogens Exploit PI3P to Target Secreted RxLREffectors into Host Cells. Q. Wang531. Identification and functional assay of Phytophthora sojaeavirulence effectors. Yuanchao Wang532. Evaluating the translocation- and phospholipid binding abilitiesof the Phytophthora infestans AVR3a and Phytophthora sojae Avr1bRxLR-leaders. Stephan Wawra533. Identifying essential effectors from the soybean pathogenPhytophthora sojae. Hua Z. Wise534. The LysM effector, Ecp6, is a virulence factor in the interaction ofthe hemibiotroph, Setosphaeria turcica, but not the necrotroph,Cochliobolus heterostrophus, with their common host, maize.Dongliang Wu535. Nematode-trapping fungi eavesdrop on nematode pheromones.Yen-Ping Hsueh536. Molecular diagnosis to discriminate pathogen and apathogenspecies of the hybrid Verticillium longisporum on the oilseed cropBrassica napus. Van Tuan Tran537. Investigating the Pathogenicity of Armillaria. Kathryn Ford538. Detoxification of nitric oxide by flavohemoglobin and thedenitrification pathway in the maize pathogen Fusariumverticillioides. Thomas Baldwin539. Family disintegration: One Fusarium verticillioides betalactamasegene at a time. Scott E. Gold540. A Fungal Metallo-Beta-Lactamase Necessary forBiotransformation of Maize Phytoprotectant Compounds. Scott E.Gold541. Nectria haematococca DNase: role in dynamics and localizationof pea root infection. D. Huskey542. FvSNF1, a protein kinase of Fusarium virguliforme that affectsSDS development in Soybean. K. T. Islam543. Functional and molecular analysis of AstA sulfate transporter inpathogenic Fusarium sambucinum with respect to its virulence andability to infect potato. Sebastian Pilsyk544. WITHDRAWN545. The adenylate cyclase of the cereal pathogen Fusariumgraminearum controls infection structure development, mycotoxinproduction and virulence to wheat. Jörg Bormann546. The ATF/CREB transcription factor Atf1 is essential for fullvirulence, deoxynivalenol production and stress tolerance in the plantpathogen Fusarium graminearum. Jörg Bormann547. The stress-activated protein kinase FgOS-2 is a key regulator inthe life cycle of the cereal pathogen Fusarium graminearum. JörgBormann548. Innate Immunity in Fusarium graminearum. Vong shian Simon IpCho549. Balanced posttranslational activation of eukaryotic translationinitiation factor 5A is required for pathogenesis in Fusariumgraminearum. Ana Lilia Martinez-Rocha550. The Con7 transcription factor, essential for pathogenicity,regulates the expression of genes involved in glycolysis and virulencein Fusarium oxysporum. M. Isabel G. Roncero551. Fusarium oxysporum produces volatile organic compounds thatenhance the growth and stress resistance of Arabidopsis thaliana.Vasileios Bitas552. Lipolytic system of the tomato pathogen Fusarium oxysporumf.sp. lycopersici. G. A. Bravo Ruiz553. Role of glycogen metabolism in the pathotypic behavior ofFusarium oxysporum f.sp. lycopersici on tomato plants. C. CorralRamos554. Identification of chemoattractant compounds from tomato rootexudate that trigger chemotropism in Fusarium oxysporum. El GhalidMennat555. TOR-mediated control of virulence functions in the transkingdompathogen Fusarium oxysporum. Gesabel Y. Navarro Velasco556. Components of the urease complex govern virulence of Fusariumoxysporum on plant and animal hosts. Katja Schaefer557. Knock-out of the Fusarium oxysporum f.sp. lycopersici homologsof the DNA-methylation genes DIM2 and HP1 does not affect effectorgene expression. Charlotte van der Does116

LISTING OF ALL POSTER ABSTRACTS558. Mechanistic investigation of Trichoderma cf. harzianum SQR-T037 mycoparasitism against Fusarium oxysporum f. sp. cubense 4,(banana wilt disease). Jian Zhang559. Epichloënin A, a unique siderophore of Epichloae endophytesand its role in restricting fungal growth in planta. Linda J. Johnson560. Interaction between the saprotrophic fungus Serpula lacrymansand living pine roots. Nils OS Högberg561. You turn me on: Pyrenophora tritici-repentis genes differentiallyregulated early during infection of wheat. V. A. Manning562. Characterisation of genes in Quantitative Trait Loci affectingvirulence in the basidiomycete Heterobasidion annosum s.l. Ake Olson563. Elevation of FPP synthase activity in Trichoderma atrovirideresults in higher biocontrol abilities. Sebastian Pilsyk564. The life history of Ramularia collo-cygni. Maciej Kaczmarek565. Mechanical stress sensing in Epichloë fungal symbionts duringcolonization of grasses. Kahandawa G. S. U. Ariyawansa566. Aspergillus flavus hypertrophy and hyphal entry by Ralstoniasolanacearum is mediated by bacterial type three secretion systemfunction. Joe E. Spraker567. Vegetative hyphal fusion in epichloae endophytes. Jun-ya Shoji568. Oxygen and the stomatal cue: Dissecting stomatal tropism inCercospora zeae-maydis. R. Hirsch569. Host colonisation processes by symbiotic epichloid fungi areregulated through cAMP. Christine R. Voisey570. Role of VCP1 and SCP1 proteases in the mutitrophic behaviour ofthe nematophagous fungus Pochonia chlamydosporia. Nuria Escudero571. Cellular development integrating primary and induced secondarymetabolism in the filamentous fungus Fusarium graminearum. H.Corby Kistler572. DNA double-strand breaks generated by yeast endonuclease I-Sce I induce ectopic homologous recombination and targeted genereplacement in Magnaporthe oryzae. T. Arazoe573. Investigation of the Magnaporthe oryzae proteome andphosphoproteome during appressorium formation. William L. Franck574. Characterization of the binding site and downstream targets ofthe MST12 transcription factor in Magnaporthe oryzae. Guotian Li575. CorA Magnesium transporters are key regulators of growth andpathogenicity in the rice blast fungus M. oryzae. Md Hashim Reza576. Magnaporthe oryzae AVR-Pia protein: induction of resistancereaction in Pia rice by the recombinant AVR-Pia and preparation ofanti-AVR-Pia antibody. Y. Satoh577. Homologous recombination causes the spontaneous deletion ofAVR-Pia in Magnaporthe oryzae. T. Sone578. The interactome of pathogenicity factors in the rice blast fungusMagnaporthe oryzae. Xiaoying Zhou579. Interaction between phenolic and oxidant signaling inCochliobolus heterostrophus. Benjamin A. Horwitz580. Mode of Action of Chitosan: Antifungal and Gene Modulatorfrom Natural Origin. Luis V. Lopez-Llorca581. Unraveling the metabolome: how zombie ant fungiheterogeneously control ant brains. Charissa de Bekker582. Gene expression of fungal aldehyde dehydrogenases inectomycorrhiza. Catarina Henke583. Interaction of ectomycorrhizal fungi with environment. KatrinKrause584. Genetic exchange in an arbuscular mycorrhizal fungus;Rhisophagus irregularis. Pawel Rosikiewicz585. A surface hydrophobin in ectomycvorrhiza interaction. DominikSenftleben586. Response of Alternaria brassicicola to the antifungal activity ofisothiocyanates. Benoit Calmes587. Redox regulation of an AP-1-like transcription factor, YapA, in thefungal symbiont Epichloë festucae. Gemma M. Cartwright588. Genomic approaches to understand pathogenesis in thebasidiomycete pathogen of food and energy crops, Rhizoctonia solani.Jonathan P. Anderson589. The Cpc1 (CpcA/Gcn4) regulator of the cross-pathway control ofamino acid biosynthesis is required for plant infection of the vascularpathogen Verticillium longisporum. Susanna A. Braus-Stromeyer590. Fungal-Specific Transcription Factor AbPf2 ActivatesPathogenicity in Alternaria brassicicola. Yangrae Cho591. WITHDRAWN592. Host-to-pathogen gene transfer facilitated infection of insects bya pathogenic fungus. Weiguo Fang593. Characterization of the CoPRF1 mutant of Colletotrichumorbiculare defective in evasion of host defense responses. YasuyukiKubo594. CPS1 mutants in Coccidioides are avirulent and act as anattenuated vaccine in the valley fever mouse model. Marc J. Orbach595. Elucidating the response of wheat to the exposure ofStagonospora nodorum effectors. Peter S. Solomon596. Nep1-like proteins of the downy mildew Hyaloperonosporaarabidopsidis trigger immunity, but not necrosis, in the Arabidopsishost. Guido Van den Ackerveken597. Genes important for in vivo survival of the human pathogenPenicillium marneffei. Harshini C. Weerasinghe598. Oxalate-minus mutants of Sclerotinia sclerotiorum via T-DNAinsertion accumulate fumarate in culture and retain pathogenicity onplants. Liangsheng Xu599. Molecular characterization of fungi associated with superficialblemishes of potato tubers in Al-Qasim region, Saudi Arabia. RukaiaM. Gashgari27th Fungal Genetics Conference | 117

LISTING OF ALL POSTER ABSTRACTS600. Patterns of Distribution of Bacterial Endosymbionts in LowerFungi. Olga Lastovetsky601. Phylogenetic and genomic analysis of a novel, nematophagousspecies of Brachyphoris. S. Sharma Khatiwada602. The proteome of the traps of the nematode-trapping fungusMonacrosporium haptotylum . K.-M. Andersson603. Sequencing the in planta transcriptomes of Colletotrichumspecies provides new insights into hemibiotrophy. Richard J. O'Connell604. Biological activities of natural products synthesized by themammalian fungal pathogen, Histoplasma capsulatum. A. Henderson605. From antagonism to synergism: roles of natural phenazines inbacterial-fungal interactions between Pseudomonas aeruginosa andAspergillus fumigatus. Yun Wang606. Genomic analysis of Mortierella elongata and its endosymbioticbacterium. Gregory Bonito607. Diversity and Content of Maize Leaf Endophytes are CorrelatedWith Maize Genotype. Alice C. L. Churchill608. Characterisation of epichloae endophytes from the Triticeae andtheir potential use in modern cereals. Richard D. Johnson609. The Interaction of Mycoplasma-related Endobacteria with theirArbuscular Mycorrhizal Fungal Host. Mizue Naito610. The Velvet gene is required for mutualism between Epichloëfestucae and perennial ryegrass. Mostafa Rahnama611. An examination of phosphate solubilization and hormoneproduction by two Penicillium species growing in the rhizoplane. TimRepas612. The role of Epichloe festucae RacA interacting proteins, PakA,PakB and RhoGDI, on cell polarity in culture and synchronized growthin Lolium perenne. Barry Scott613. A Host-Induced Gene Silencing Approach to Control MycotoxinContamination in Corn. B. H. Bluhm614. CDiT1, a novel type of proteinaceous toxin secreted by thenecrotrophic pathogen of the roots of tomato Pyrenochaetalycopersici. Pierre-Henri Clergeot615. Nonhost-specific phytotoxicity of the polyketide-derived toxinsolanapyrone A produced by Ascochyta rabiei and Alternaria solani.W. Kim616. Crosstalk of the unfolded protein response and regulatorypathways controlling pathogenic development in Ustilago maydis. KaiHeimel617. Transcriptional profiling of the APSES genes in Trichophytonrubrum during growth on human nail. Elza A. S. Lang618. Carbohydrate binding proteins of two Leptosphaeria pathogensof Brassica napus. Rohan G. T. Lowe619. Domains for plant uptake of Ustilago maydis secreted effectors.Anupama Ghosh620. Lipid metabolism influences virulence in Ustilago maydis. Scott118Lambie621. Functional characterization of the putative cell surface receptorfor hydrophobicity, Msb2, in Ustilago maydis.Marino Moretti622. The U. maydis effector Pit2 inhibits maize cysteine proteases tosuppress host defense. Andre Mueller623. The Ustilago maydis MAP Kinase signaling pathway:Identification of direct MAP kinase targets by phospho-peptideenrichment. Vikram Naik624. See1 : A novel organ specific effector in the Ustilago maydis -maize interaction. Amey Redkar625. Investigation of unconventionally secreted proteins in Ustilagomaydis. Stefanie Reissmann626. Identification of a key regulator for the developmental switchleading to sporogenesis in Ustilago maydis. Marie Tollot627. Genetic characterization of virulence in the Pyrenophora teres f.teres - barley pathosystem. Timothy L. Friesen628. Illumina-based genetic linkage map for wheat leaf rust. David L.Joly629. The deletion of the Histoplasma capsulatum RYP1 homolog inCoccidioides posadasii is avirulent. M. Alejandra Mandel630. Cpkk2, a MEK from Cryphonectria parasitica is necessary formaintenance of CHV1 virus infection. M. Turina631. Deep RNAseq of wheat leaf infection by M. graminicola identifiesphase-specific in planta expressed genes and varying transcriptionalcontributions of fungal chromosomes. Mikaël S. Courbot632. A genomic analysis of the infection strategies employed byPhoma medicaginis a necrotrophic fungal pathogen of alfalfa and themodel legume Medicago truncatula. Angela H. Williams633. Two G protein-coupled receptors, GprC and GprD, regulatedensity-dependent development in Aspergillus flavus. Katharyn J.Affeldt634. Characterization of genes encoding putative secreted proteinsduring pathogenesis in Magnaporthe oryzae. Seongbeom Kim635. The biosynthesis of oxalate is entirely dependent onoxaloacetate acetylhydrolase in Sclerotinia sclerotiorum . X. Liang636. In vivo efficacy of antifungal treatment of Aspergillus terreusinfections and the influence on host immune response in Galleriamellonella. Elisabeth Maurer637. Recognition and response to non self in Podospora anserina: amodel of the fungal immune system. Mathieu Paoletti638. Increased late blight resistance in HIGS potato lines targeting a P.infestans gene. N. Temme639. WITHDRAWNPopulation and Evolutionary Genetics

LISTING OF ALL POSTER ABSTRACTS640. Fertility in Aspergillus fumigatus and the identification of anadditional ‘supermater’ pair. Céline M. O'Gorman641. Understanding the dynamic plant pathogen Ramularia collo-cygniat both the sequence and field level. James Fountaine642. Comparing germination dynamics of co Kolea C. K. Zimmermannidia and ascospores from natural isolates of Neurospora crassa.643. Cryptococcus gattii and the origins of outbreaks. Dee Carter644. Evolution of the mating type locus in the species within andclosely related to the pathogenic Cryptococcus species complex. ShengSun645. Evolutionary history and genetic diversity of Exobasidium sp., thecause of an emerging disease of blueberry. Marin Brewer646. Microsatellite markers reveal population structure and geneticdiversity in the blueberry pathogen Monilinia vaccinii-corymbosi.Kathleen M. Burchhardt647. Genetic diversity of Australian Pyrenophora tritici-repentisisolates using microsatellites. Caroline Moffat648. WITHDRAWN649. Fungal community composition analysis by Internal TranscribedSpacer (ITS) sequencing using Illumina MiSeq. Robin A. Ohm650. Estimation of genetic diversity of Ramularia collo-cygnipopulations using nuclear SSR markers to infer its potential to adaptto environmental changes. Marta Piotrowska651. Alkaloid genotype profiling of tall fescue endophytes todetermine influence of ancestral progenitors. J. E. Takach652. Evolution of the pan-secretome among lineages of Magnaportheoryzae attacking different host-plants. E. Fournier653. Exploiting the high evolutionary potential of Leptosphaeriamaculans minimises severity of blackleg disease of canola. Barbara J.Howlett654. Experimental demonstration of Crozier's paradox in fungi. DuurK. Aanen655. A completely unknown lifecycle in mushrooms: cyclicalinbreeding and haplo-diploidy. Duur K. Aanen656. Diversity and evolution of ABC proteins in basidiomycetes. FredO. Asiegbu657. Co-evolution and life cycle specialization of plant cell walldegrading enzymes in a hemibiotrophic pathogen. Patrick C. Brunner658. Recombination landscape of the plant pathogenic fungusZymoseptoria tritici (syn. Mycosphaerella graminicola). D. Croll659. The evolution of Sfp1 mediated, cell size control in Ascomycetefungi. Toni M. Delorey660. Cryptic population subdivision, sympatric coexistence and thegenetic basis of local adaptation in Neurospora discreta. PierreGladieux661. WITHDRAWN662. Evolutionary genomics of NRPS gene clusters in Beauveria and itsallies. J.-G. Han663. Neurospora presents a model for the evolution of matingsystems. Christopher Hann-Soden664. Profiling conditionally dispensable chromosomes of the plantpathogenicfungus Zymoseptoria tritici (syn. Mycosphaerellagraminicola). Ronny Kellner665. Take a walk on the wild side: evolutionary consequences ofresistance to apple scab introgressed from a wild host. C. Lemaire666. Genomic footprint of adaptive divergence in Ophiostomamontium, a fungal symbiont associated with the mountain pinebeetle. J. F. Mao667. Ecological context in symbioses: when is your enemy also yourfriend? Georgiana May668. Population shifts and mating-type heterokaryosis in Aspergillusflavus. Rodrigo A. Olarte669. Structural variation of trichothecene mycotoxins has resultedfrom multiple evolutionary processes in the fungal order Hypocreales.R. H. Proctor670. Evidence for birth-and-death evolution and horizontal transfer ofthe fumonisin mycotoxin biosynthetic gene cluster in Fusarium. R. H.Proctor671. Chemotype predominance in Fusarium graminearum is notdirectly affected by the use of the fungicides trifloxystrobin andisopyrazam. Matias Pasquali672. Evolution of races within f.sp lycopersici of Fusarium oxysporum.BV. Chellappan673. Detection of Mitochondrial DNA Heteroplasmy in the progeny ofcrossed genetically divergent isolates of Arbuscular Mycorrhizal Fungi.Maryam Nadimi674. Evolution of mode of infection in the rice blast fungus and alliedspecies. Ning Zhang675. WITHDRAWN676. Population genomic analysis reveals a complex evolutionaryhistory of Neurospora tetrasperma.Padraic Corcoran677. Rapid evolution of female-biased genes: a novel example fromthe eukaryotic model organism Neurospora crassa. HannaJohannesson678. Fungal Community Dynamics During Biomass Degradation in theCow Rumen Determined by ITS Sequencing. Matthias Hess679. A Systematic and Genomic Description of Undulatus ophiodiicola,Formerly Referred to as Chrysosporium ophiodiicola, an EmergingFungal Pathogen of Snakes. Mana Ohkura680. Pair-wise linkage disequilibrium decay among linked loci suggests27th Fungal Genetics Conference | 119

LISTING OF ALL POSTER ABSTRACTSmeiotic recombination in natural populations of Sclerotiniasclerotiorum. Renuka Nilmini Attanayake681. Population genomics of Suillus brevipes. Sara Branco682. Ugly, understudied and undertreated: population genomics ofthe most common human fungal pathogen - the dermatophyteTrichophyton rubrum. M. Gajdeczka683. Poppr: an R package for genetic analysis of populations withmixed reproduction. Zhian N. Kamvar684. The heterothallic fungus Cercospora beticola contains fragmentsof both mating type genes. Melvin D. Bolton685. Population structure from mountain to coast of twoLophodermium endophytes; a case study comparing rare andcommon fungal species in pine needles. Ryoko Oono686. Fungal pathogen and endophyte genetics within the context offorest community dynamics. M.-S. Benitez687. Discovery of Sexual Reproduction in the Black Aspergilli. HeatherL. Darbyshir688. Culture-based survey of soil fungi from bat hibernacula. AndreaGargasOther Topics689. Understanding the cellular basis of Azole resistance in Aspergillusfumigatus. Michael J. Bromley690. Chemically Induced Haploinsufficiency Screens to Identify DrugMechanism of Action in Aspergillus Fumigatus. D. A. Macdonald691. Antifungal Pisum sativum defensin 1 Induces a non-ApoptoticDeath in Aspergillus nidulans. Caroline M. Fernandes692. Is fungal secondary metabolism regulated by competing insects?Frank Kempken693. Eisosome distribution and localization in the meiotic progeny ofAspergillus nidulans. V. Sophianopoulou694. Fungal-bacterial interactions: Bacillus subtilis forms biofilm onAspergillus niger hyphae. Isabelle Benoit695. Co-cultivations of fungi: microscopic analysis and influence onprotein production. Isabelle Benoit696. Improving heterologous protein production in Aspergillusvadensis . Ourdia Bouzid697. Production and characterization of esterases from Chaetomiumthermophilum and their applicability in biomass conversion. XiaoxueTong698. Antioxidant adaptation by Eugenol and its derivatives and theiraffect on the expression of virulence in candida species. Aijaz Ahmad699. Elevation of chitin is linked with multiparallel mechanisms inresponse to C. albicans cell wall stress. F. Nogueira700. Prezygotic and postzygotic control of uniparental mitochondrialDNA inheritance in Cryptococcus neoformans. Rachana Gyawali701. SIS, a sex genome defense mechanism operating in Cryptococcusneoformans. Xuying Wang702. Effects of the use of biocontrol agent (Phlebiopsis gigantea) onfungal communities of Picea abies stumps. E. Terhonen703. Diversity of yeast and mold species in different cheeses. NabarajBanjara704. Heterologous expression and characterization of soil organicmatter-specific proteases secreted by the ectomycorrhizal fungusPaxillus involutus. Morten N. Grell705. Effector proteins in fungal defense against fungivorousnematodes: Targets and functional significance. Markus Künzler706. Microfluidic platforms for monitoring interactions between fungiand bacteria. Martina A. Stöckli707. WITHDRAWN708. The French Fusarium Collection: a living resource for mycotoxinresearch. N. Ponts709. Chemical genetics: Discovery of novel fungicides and their targetsin the phytopathogen Fusarium graminearum. G. Subramaniam710. Functional characterization of an Aspergillus flavus polyketidesynthase gene necessary for the synthesis of a sclerotium-specificpigment. J. W. Cary711. Functional Analysis of the Pleurotus ostreatus Manganese-Peroxidase Gene Family. Yitzhak Hadar712. Temperature- and pH characteristics of endo-cellulases inRhizophlyctis rosea. Bo Pilgaard713. Applying unconventional secretion of the endochitinase Cts1 toexport heterologous proteins in Ustilago maydis. K. Schipper714. A new method for fungal genetics: flow cytometry ofmicroencapsulated filamentous microcolonies. D. Canovas715. DNA methylation dynamics during development in the rice blastfungus. Junhyun Jeon716. pH-enotype array: a novel phenomics platform for filamentousfungi. Jaejin Park717. GFP analysis of meiotic recombination in Neurospora Δmsh-2homozygotes. David E. A. Catcheside718. Residual recombination in Neurospora crassa spo11 mutanthomozygotes occurs during meiosis. David E. A. Catcheside719. Controlled synthesis of gold nanoparticles by Neurospora crassaextract and their SERS properties. Katrin Quester720. Cellulase production in Neurospora crassa. M. Reilly721. Identification and Functional analysis of New Neurospora crassaNonself Recognition Loci. Jiuhai Zhao722. Combinatorial cationic and oxidative stresses promote the killingof Candida albicans cells by human neutrophils. Alistair J. P. Brown120

LISTING OF ALL POSTER ABSTRACTS723. Phosphoproteomic analysis of the aquatic fungus Blastocladiellaemersonii during germination. J. Crestani724. Towards an accurate genome: high-throughput proteogenomicvalidation of Stagonospora nodorum genes via sub-cellularproteomics. Kar-Chun Tan725. Evolutionary Imprint of Fungal PKS-NRPS Catalytic Domains.Daniela Boettger726. Secondary metabolism and development is mediated by LlmFcontrol of VeA subcellular localization in Aspergillus nidulans.Jonathan M. Palmer727. Overproduction of phleichrome by synthetic inducers and cloningof polyketide synthase genes in phytopathogenic fungusCladosporium phlei. D.-H. Kim728. Symbiotic fungal endophytes that confer tolerance for plantgrowth in saline soil. Susan GW Kaminskyj729. Stable cesium and radiocesium response of Schizophyllumcommune. Matthias Gube730. The completion of meiosis in Ustilago maydis requires an Ndt80ortholog. B. J. Saville731. Impact of changes in the target P450 CYP51 enzyme associatedwith altered triazole-sensitivity in the Wheat pathogenMycosphaerella graminicola. Michael Csukai732. Molecular Evolutionary Analysis and Synteny of Fungal GALGenes. Julien S. Gradnigo733. Meiotic Drive: A Single Gene Conferring Killing and Resistance inFungal Spore Killer. Pierre Grognet734. Alkaliphilic fungi from soda lakes and soda soils. Alexey A. Grum-Grzhimaylo735. A new method for gene mining and enzyme discovery. Y. Huang736. Occurrence of dsRNA mycovirus (LeV-FMRI 2427) in ediblemushroom Lentinula edodes and its meiotic stability. J.-M. Kim737. Analysis of fungal communities associated with grapevine wooddiseases, based on fungal ITS pyrosequencing. Nicolas Lapalu738. The Antidepressant Sertraline Provides a Promising TherapeuticOption for Neurotropic Cryptococcal Infections. Xiaorong Lin739. Analysis of gene expression of proteases of Trichoderma sppduring confrontation with plant pathogens in vivo. Valdirene Monteiro740. Analysis of differential protein profile during antagonism ofTrichoderma harzianum and Sclerotinia sclerotiorum. ValdireneMonteiro741. Effector proteins in fungal defense against fungivorousnematodes: Diversity and regulation of expression. D. F. Plaza27th Fungal Genetics Conference | 121

FULL POSTER SESSION ABSTRACTSBiochemistry and Metabolism1. Heme regulation in Aspergillus fumigatus. Nicola Beckmann 1 , Ernst R. Werner 2 , Hubertus Haas 1 . 1) Division of Molecular Biology, Biocenter, InnsbruckMedical University, Austria; 2) Division of Biological Chemistry, Biocenter, Innsbruck Medical University, Austria.Sufficient iron supply is indispensable for survival of almost all organisms. However, an excess of iron is potentially toxic. In the opportunistic humanpathogenicfungus Aspergillus fumigatus the ability to adapt to iron limitation represents a crucial virulence factor. Iron regulation is tightly interconnectedwith heme metabolism, as iron-containing heme is an essential cofactor of a variety of cellular processes, e.g. respiration, sterol biosynthesis, oxidativestress detoxification and also reductive iron assimilation. Most knowledge on fungal heme regulation derives from studies in Saccharomyces cerevisiae. A.fumigatus, as well as most other fungal species, lack homologs of key heme regulators found in S. cerevisiae. The goal of this study is to elucidate hemedependentregulation in A. fumigatus (wt). As a first step, we generated a mutant strain (DhemA) lacking the gene encoding aminolevulinic acid synthase(HemA), which catalyzes the committed step in heme biosynthesis. This mutation offers the possibility to control the cellular heme content bysupplementation with aminolevulinic acid (ALA). Growth of DhemA was blocked at ALA concentrations below 20 mM, but fully restored by addition of 200mM on solid as well as in liquid media. Supplementation with protoporphyrin IX (PpIX), the iron free heme precursor, and hemin (chloroprotoporphyrin IXiron(III)) supported growth of DhemA, proving that A. fumigatus is able to utilize exogenous porphyrins. Nevertheless, A. fumigatus in contrast to severalother fungal species is not able to utilize hemin as iron source. Under iron starvation, ALA supplementation led to a tremendous accumulation of PpIX inboth DhemA and wt, which indicates that HemA represents the major rate limiting step in heme biosynthesis when iron is scarce. In DhemA, ALArestriction transcriptionally increased the heme-biosynthetic coproporphyrinogen(III)oxidase and the putative heme receptor CFEM3. Additionally, ALAlimitation decreased the resistance of DhemA to oxidative stress and the triazole antifungal drug posaconazole, which underlines the crucial role of hemein detoxification and sterol biosynthesis. This work was supported by the Austrian Science Foundation grant FWF P21643-B11 to HH.2. Key Steps in the Biosynthesis of the Fungal Virulence Factor Gliotoxin. Pranatchareeya Chankhamjon 1 , Daniel H. Scharf 2 , Kirstin Scherlach 1 , NicoleRemme 1 , Andreas Habel 1 , Thorsten Heinek 2 , Martin Roth 3 , Axel A. Brakhage 2 , Christian Hertweck 1 . 1) Biomolecular Chemistry, HKI, Jena, Jena, Germany; 2)Molecular and Applied Microbiology,HKI, Jena, Jena, Germany; 3) Bio Pilot Plant,HKI, Jena, Jena, Germany.The prototype of epipolythiodioxopiperazine (ETP) family, gliotoxin, is an infamous virulence factor of the human pathogen Aspergillus fumigatus,notably the leading cause of invasive aspergillosis in the immunocompromised patients. Its toxicity has been attributed to the unusual intramoleculardisulfide bridge, which is the functional motif of all ETPs. A number of studies showed that the diketopiperazine core of gliotoxin is assembled by a nonribosomalpeptide synthetase. However, downstream pathway steps have remained elusive, mainly because of the scarcity and instability of pathway inthe mediates produced. Here we present the critical role of a specialized glutathione S-transferase (GST), GliG, in the enzymatic sulfurisation and the keystep of epidithiol formation by an unprecedented twin carbon-sulfur lyase, GliI. Our studies not only unveil the understanding of key steps in thebiosynthesis pathway of an important virulence factor, but also outline a new function of microbial GSTs and gain insights into the formation oforganosulfur compounds.3. Identification of a gene cluster mediating the biosynthesis of the Aspergillus fumigatus cell wall and secreted polysaccharide, galactosaminogalactan.Fabrice N. Gravelat 1 , Mark J. Lee 1 , Alexander Geller 1 , Dan Chen 2 , Anne Beauvais 3 , Hong Liu 4 , William C. Nierman 2 , Jean-Paul Latge 3 , Thierry Fontaine 3 , ScottG. Filler 4 , Donald C. Sheppard 1 . 1) Microbiology & Immunology Department, McGill University, Montréal, Qc, Canada; 2) J. Craig Venter Institute, Rockville,Maryland, USA; 3) Aspergillus Unit, Institut Pasteur, Paris, France; 4) Division of Infectious Diseases, Los Angeles Biomedical Research Institute at Harbor-UCLA Medical Center, Torrance, California, USA.Aspergillus fumigatus is the most common cause of invasive mold disease in humans. Although adherence of fungal hyphae to host constituents is acritical early step in the pathogenesis of invasive aspergillosis, the molecular mechanisms underlying this process have not been elucidated. Using aforward genetic approach, we identified a glucose epimerase, Uge3, which is required for adherence of hyphae to a wide variety of substrates.Biochemical analyses confirmed that Uge3 is required for the synthesis of the secreted glycan galactosaminogalactan (GAG), which in turn functions as thedominant adhesin of A. fumigatus hyphae and is required for virulence. However, the biochemical and regulatory pathways governing GAG synthesisremain unknown. Using comparative transcriptome analysis, we found that uge3 is found within a cluster of 5 co-regulated genes on chromosome 3.Interestingly, 3 of the 5 proteins (Uge3, Gtb3 and Ega3) encoded by these genes are predicted to contain conserved domains involved in polysaccharidemetabolism. The 2 other proteins (Sph3 and Esr3) have no homologs in other organisms. We hypothesized that this cluster of genes may be required forGAG biosynthesis. To test this hypothesis, we constructed deletion mutants of two of the cluster genes: sph3, encoding a cell surface spherulin 4-likeprotein; and esr3, encoding an extracellular serin-rich protein. Phenotypic analysis of both the Desr3 and Dsph3 mutant strains confirmed that deletion ofthese genes resulted in both impaired GAG production and impaired adherence, similar to the phenotype of the Duge3 mutant strain. Gene deletion forthe 2 remaining genes is ongoing. Collectively, these data suggest that the 5 gene cluster identified on chromosome three is likely a carbohydratebiosynthetic cluster required for the synthesis of GAG. Importantly, this is the first description of a gene cluster for the biosynthesis of a cell wallpolysaccharide in A. fumigatus, and suggests the possibility that other similar gene clusters may govern the synthesis of glycans in this fungus. Thediscovery of this cluster, and the subsequent characterization of the role of each of the component elements, may provide insight into the synthesis andfunction of GAG.4. A fasciclin-like protein in Aspergillus fumigatus. Thomas Hartmann 1 , Lei Sun 2 , Cheng Jin 1 . 1) Institute of Microbiology, Chinese Academy of Sciences,Beijing, China; 2) Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.The saprophytic filamentous fungus Aspergillus fumigatus has been gaining importance as an opportunistic human pathogen over the past decades, asadvances in modern medicine have created a growing group of patients susceptible to potentially deadly invasive aspergillosis. The role of fungal adhesionduring infection progression is still poorly understood and this work aims to focus on this neglected aspect of the infection process. Fasciclin-like proteinscan be found in a wide variety of organisms, where they often perform functions related to surface adhesion. Here we describe a fasciclin-like protein in A.fumigatus. The Fas protein was first discovered as differentially expressed in an O-mannosyltransferase Dpmt1 deletion strain, which indicates that it maybe substantially glycosylated. We generated Dfas deletion strains in two different A. fumigatus strain backgrounds. The Dfas deletion strains grewnormally on plates and in solution and neither bright field microscopy, nor TEM revealed phenotypical differences to the wildtype strains. Interestingly, theDfas deletion strains showed reduced adhesion to hydrophobic plastic surfaces, but adhered normally to glass slides. Whether these altered adhesiveproperties have an effect on the strains’ virulence in mammalian hosts such as mice remains to be investigated.122

FULL POSTER SESSION ABSTRACTS5. Characterization of fumiquinazoline biosynthesis in Aspergillus fumigatus. Fang Yun Lim 1 , Brian Ames 2 , Christopher Walsh 2 , Nancy Keller 1 . 1) Medicalmicrobiology and Immunology, University of Wisconsin-Madison, Madison, WI; 2) Biological chemistry and molecular pharmacology, Harvard MedicalSchool, Boston, MA.The fumiquinazolines (FQs) comprise a related, sequentially generated family of bioactive peptidyl alkaloids that are signature metabolites of Aspergillusfumigatus. The FQ framework is built by nonribosomal peptide synthetase (NRPS) machinery with anthranilate as a key non-proteinogenic amino acidbuilding block. Despite being prevalent across the species, its gene cluster has not been characterized. Prior bioinformatic analysis coupled withheterologous expression of the putative A. fumigatus proteins termed here FmqA -FmqD led to the identification of a four-enzymatic process that buildsincreasingly complex FQ scaffolds. Briefly, FmqA, a trimodular NRPS condenses alanine, tryptophan, and anthranilic acid to form fumiquinazoline F (FQF).The tandem action of a flavoprotein (FmqB) and a monomodular NRPS (FmqC) converts FQF to fumiquinazoline A (FQA). Finally, FmqD, a FAD-dependentoxidoreductase converts FQA to the heptacyclic fumiquinazoline C (FQC). Interestingly, FmqD contains an N-terminus signal peptide predicted forextracellular transport. This study is aimed at providing in vivo validation to the FQ biosynthetic framework and characterizing how cellular localization ofFmqD affects production of FQC in A. fumigatus. We found that the conidial metabolite, FQC, is the predominant FQ moiety in two wild type isolates and isselectively accumulated in the conidia. Targeted single gene deletions of FmqA through FmqD coupled with metabolomic profiling of the singlebiosynthetic gene mutants supported previous biochemical prediction of FQ biosynthesis. Fluorescent microscopy of mutants bearing a C-terminal FmqD-GFP fusion showed that FmqD is localized to the cell wall of the fungus and this localization is abolished when the signal peptide is removed. Future studieswill elucidate if cell wall localization of FmqD is crucial for FQC production.6. Identification of local and cross-chromosomal biosynthetic gene clusters in filamentous fungi using gene expression data. Mikael R. Andersen 1 , JakobB. Nielsen 1 , Andreas Klitgaard 1 , Lene M. Petersen 1 , Tilde J. Hansen 1 , Lene H. Blicher 1 , Charlotte H. Gottfredsen 2 , Thomas O. Larsen 1 , Kristian F. Nielsen 1 ,Uffe H. Mortensen 1 . 1) Department of Systems Biology, Technical University of Denmark, Kgs Lyngby, Denmark; 2) Department of Chemistry, TechnicalUniversity of Denmark, Kgs Lyngby, Denmark.Biosynthetic pathways of secondary metabolites from fungi are currently subject to an intense effort to elucidate the genetic basis for these compoundsdue to their large potential within pharmaceutics and synthetic biochemistry. The preferred method is methodological gene deletions to identifysupporting enzymes for key synthases one cluster at a time.In earlier work we presented a method for using a gene expression compendium to accurately predict co-regulated gene clusters in general, and inparticular the members of gene clusters for secondary metabolism. A benchmarking of the method in Aspergillus nidulans by comparison to previous genedeletion studies showed the method to be accurate in 13 out of 16 known clusters and nearly accurate for the remaining three.In this work, we have expanded the algorithm to identify cross-chemistry between physically separate gene clusters (super clusters), and validate thisboth with legacy data and experimentally by prediction and verification of a new supercluster consisting of the non-ribosomal peptide synthetase (NRPS)AN1242 (on chr VIII) and the prenyltransferase AN11080 (on chromosome V) as well as identication of the shared product compound nidulanin A.We also propose further implications of the gene clustering, as our analysis shows that approximately 10 % of the genes seem to be non-randomly(p

FULL POSTER SESSION ABSTRACTS9. Characterization of the 3-methyl orsellinic acid gene cluster in Aspergillus nidulans. Jakob B. Nielsen 1 , Marie L. Klejnstrup 1 , Paiman K. Jamal 1 , Dorte K.Holm 1 , Michael L. Nielsen 1 , Anna M. Kabat 2 , Charlotte H. Gotfredsen 3 , Thomas O. Larsen 1 , Uffe H. Mortensen 1 . 1) Center for Microbial Biotechnology,Department of Systems Biology, Technical University of Denmark; 2) Center for Systems Microbiology, Department of Systems Biology, TechnicalUniversity of Denmark; 3) Department of Chemistry, Technical University of Denmark.With the aim of mapping the polyketome of Aspergillus nidulans we have made a library of strains, which individually overexpress PKS genes from anectopic locus. A screen of this collection on different media demonstrated that overexpression of AN6448 (pkbA) leads to increased production of 3-methyl orsellinic acid. An inspection of the DNA sequence surrounding this gene uncovered a putative gene cluster including a gene, AN6446 (pkbR), withhomology to transcription factors. Based on this observation, we decided to overexpress pkbR. A qRT-PCR analysis of this strain was used to delineate theborders of the gene cluster as well as to stimulate formation of cichorine, cichorinic acid, nidulol and a novel cichonidulol dimer, just to name a few of theproducts that we have linked to this gene cluster. Subsequent deletion of all genes in the cluster has allowed us to propose a comprehensive model for thebiosynthetic pathway of this cluster.10. Induction of sclerotia and Aspergillus section Nigri. Jens Frisvad, Lene Petersen, Ellen Lyhne, Thomas Larsen. CMB, Dept Systems Biol, Kgs. Lyngby,Denmark.The purpose of this study was to induce sclerotium production in Aspergillus niger and other black Aspergilli. Some species in Aspergillus section Nigri areknown for their production of sclerotia, especially A. carbonarius, A. tubingensis (few isolates), A. sclerotioniger, A. sclerotiicarbonarius, A. costaricaensis,A. piperis, A. japonicus, and A. aculeatus. A. heteromorphus was reported in 1955 to produce sclerotia, but this could not be confirmed in later studies.There are also un-confirmed data on sclerotium production in Aspergillus niger, but often isolates reported to produce sclerotia were not A. niger anyway.Induction of sclerotium production in Aspergillus niger is important, since this may help in inducing the perfect state in this important industrial fungus. Byscreening several media, we were able to develop some media and use some growth conditions that induced sclerotium production in Aspergillus nigerand other species hitherto not reported to produce sclerotia. Earlier French beans were suggested as inducers of sclerotium production, but we could notrepeat this with any isolate of A. niger. However by using media such as white rice and brown rice or adding different fruits to CYA (Czapek yeastautolysate agar) and incubate at 25 C we were able to induce sclerotium production in certain strains of A. niger. Old strains used for citric acid production,or full genome sequenced strains, were not induced to produce sclerotia, but several fresh strains from different foods did produce abundant sclerotia onthe different media, at 25 C, but not 37 C. One older classical citric acid producer from NRRL produced many sclerotia, however. Sclerotium producingisolates also contained aflavinines, confirmed by HPLC-DAD-MS-MS, secondary metabolites only produced in the sclerotia, and detected in A. niger for thefirst time. Other species, such as A. ibericus, A. neoniger, A. heteromorphus, A. fijiensis, A. luchuensis (formerly A. acidus), A. aculeatinus and A.saccharolyticus could also produce sclerotia on fruit media. The sclerotia contained many sclerotium-specific secondary metabolites.11. Analyzing the impact of compartmentalization on organic acid production in Aspergillus niger. Matthias G. Steiger 1,2* , Marzena L. Blumhoff 1,2,3 ,Diethard Mattanovich 1,2 , Michael Sauer 1,2 . 1) Austrian Centre of Industrial Biotechnology (ACIB GmbH), Muthgasse 11, 1190 Vienna, Austria; 2) Universityof Natural Resources and Life Sciences, Department of Biotechnology, Muthgasse 18, 1190 Vienna, Austria; 3) University of Applied Sciences FH-CampusVienna, School of Bioengineering, Muthgasse 86, 1190 Vienna, Austria.Aspergillus niger is a well-established host organism for the production of carboxylic acids. Acids like citric, gluconic and oxalic acids can already beproduced by A. niger and high titers are obtained. The formation of carboxylic acids involves the shuttling of intermediate metabolites between differentintracellular compartments and utilizes different enzymatic capabilities of the respective compartment. The knowledge about the involved shuttlingmechanisms and the localization of the necessary enzymes is still fragmentary. Using fluorescence microscopy, it is possible to characterize theintracellular localization of GFP tagged proteins and hence mitochondrial leader sequences can be functionally tested. In order to analyze the influence ofthe compartmentalization on the organic acid production, we have chosen itaconic acid as a target substance. Itaconic acid, which was selected by the USDepartment of Energy as one of the 12 building block chemicals for the industrial biotechnology, is currently produced by A. terreus. Heterologousexpression of the A. terreus cadA gene also enables the formation of itaconic acid in A. niger although only low titers are obtained. We set out tocharacterize the influence of the compartmentalization on the productivity and re-engineered the enzymatic cascade by flipping the enzymatic activities ofthe cis-aconitic acid decarboxylase and aconitase between the mitochondrion and the cytosol. We will present new leader sequences for mitochondrialtargeting in A. niger alongside with results about the positive impact of the enzymatic re-localization on the itaconic acid production.12. Subcellular localization of aphidicolin biosynthesis enzymes from Phoma betae expressed heterologously in Aspergillus oryzae. A. Ban 1 , M. Tanaka 1 ,R. Fujii 2 , A. Minami 2 , H. Oikawa 2 , T. Shintani 1 , K. Gomi 1 . 1) Graduate Sch Agriculture Sci, Tohoku Univ, Sendai, Japan; 2) Graduate Sch Sci, Hokkaido Univ,Sapporo, Japan.In recent years, a lot of biosynthesis gene clusters involving in secondary metabolite biosynthesis from filamentous fungi have been revealed, and thusthe attempts to produce these valuable metabolites at high yield have been actively made. To this end, Aspergillus oryzae is an attractive host forheterologous secondary metabolites production because of its less productivity for own secondary metabolites, which leads to the production for themetabolite of interest at a highly pure grade. Actually, the number of reports has been increasing recently, in which biosynthetic genes involved in fungalsecondary metabolite biosynthesis were heterologously overexpressed in A. oryzae. On the other hand, it would be necessary to consider the cellularcompartments where the target secondary metabolite is synthesized in filamentous fungi to produce it efficiently in the heterologous host, A. oryzae.However, currently there is very little knowledge about the spatial distribution of the biosynthesis enzymes of the secondary metabolite in fungi.Therefore, in this study, we examined the subcellular localization of the enzyme proteins encoded by a gene cluster involved in aphidicolin biosynthesisfrom Phoma betae, which were expressed as GFP-fusion proteins in A. oryzae. The gene cluster of aphidicolin in P. betae contains 4 genes encodingbiosynthesis enzymes (geranylgeranyl diphosphate synthase [GGS], aphidicolan-16b-ol synthase [ACS], cytochrome P450 monooxygense 1 [P450-1], andP450-2), a gene for transporter (TP), and a gene for transcription factor. We constructed 4 biosynthesis enzymes and the transporter that each was fusedto GFP under the a-amylase gene promoter, and introduced into A. oryzae. Similarly, the organelle marker proteins fused to RFP were also constructed andexpressed simultaneously with GFP-fusion proteins to identify the organelle where the biosynthesis enzyme was localized. Fluorescent microscopyrevealed that GGS and ACS were distributed in the cytoplasm and P450-1 was located in endoplasmic reticulum (ER). Interestingly, all of GFP-fused P450-2was not observed in ER when only P450-2 was expressed, but mostly localized in ER when coexpressed with P450-1. In addition, TP fused to GFP waslocalized mainly on the plasma membrane and also rarely observed on other organelles such as vacuole.124

FULL POSTER SESSION ABSTRACTS13. Increased production of fatty acids and triglycerides in Aspergillus oryzae by modifying fatty acid metabolism. Koichi Tamano 1 , Kenneth Bruno 2 ,Tomoko Ishii 1 , Sue Karagiosis 2 , David Culley 2 , Shuang Deng 2 , James Collet 2 , Myco Umemura 1 , Hideaki Koike 1 , Scott Baker 2 , Masayuki Machida 1 . 1) NationalInstitute of Advanced Industrial Science and Technology (AIST); 2) Pacific Northwest National Laboratory (PNNL).Biofuels are attractive substitutes for petroleum based fuels. Biofuels are considered they do not contribute to global warming in the sense they arecarbon-neutral and do not increase carbons on the globe. Hydrocarbons that are synthesized by microorganisms have potential of being used as biofuelsor the source compounds. In the hydrocarbon compounds synthesized by A. oryzae, fatty acids and triglycerides are the source compounds of biodieselthat is fatty acid methyl ester. We have increased the production by modifying fatty acid metabolism with genetic engineering in A. oryzae. Firstly,enhanced-expression strategy was used for the increase. For four enzyme genes related to the synthesis of palmitic acid [C16:0-fatty acid], the individualenhanced-expression mutants were made. And the fatty acids and triglycerides in cytosol were assayed by enzyme assay kits, respectively. As a result,both fatty acids and triglycerides were most synthesized in the enhanced-expression mutant of fatty acid synthase gene at 2.1-fold and 2.2-fold more thanthe wild-type strain, respectively. Secondly, gene disruption strategy was used for the increase. Disruptants of several enzyme genes related to long-chainfatty acid synthesis were made individually. And one of them showed drastic increase in fatty acid synthesis. In the future, further increase in the synthesisis expected by utilizing genetic engineering in A. oryzae.14. Improved Properties of Thermostable Cellobiohydrolase in a Treatment of Cellulosic Material. Taija Leinonen 1 , Susanna Mäkinen 1 , Kari Juntunen 1 ,Merja Niemi 2 , Juha Rouvinen 2 , Jari Vehmaanperä 1 , Terhi Puranen 1 . 1) Roal Oy, Rajamäki, Finland; 2) University of Eastern Finland, Department ofChemistry, Joensuu, Finland.Production of biofuels i.e. bioethanol from lignocellulosic material is a promising alternative technology for using biomass as a renewable and cleansource of energy instead of consuming limited natural resources e.g. fossil fuels, and releasing increasing amounts of CO 2. Enzymatic hydrolysis isconsidered to be the most promising technology for converting cellulosic biomass into fermentable sugars. Enzymatic total hydrolysis of (ligno)cellulosicsubstrates requires at least cellobiohydrolases, endoglucanases and beta-glucosidases. Previously cloned thermostable glycoside hydrolase family 7cellobiohydrolase (CBHI) from Acremonium thermophilum was expressed in Trichoderma reesei. The purified A. thermophilum CBHI was crystallized, andthe structure of the catalytic domain of the protein was determined at a resolution of 1.8 Å revealing the overall structure of the catalytic core of theenzyme to be similar with the previously determined structures of glycoside hydrolase family 7 cellobiohydrolases. In the biomass hydrolysis experimentsthe A. thermophilum CBHI, as part of the enzyme mixture, was shown to have enhancing effect on hydrolysis yield as compared to the Trichoderma reeseienzyme. To achieve even further improvements in thermal stability and hydrolysis performance, several single and combined amino acid mutations weredesigned based on the resolved 3D-structure. The data obtained from the mutants demonstrates that thermal stability and hydrolysis performance of theA. thermophilum CBHI protein can be increased by introducing single mutations as well as mutation combinations to the molecule.15. The phytopathogenic fungus Botrytis pseudocinerea is resistant to the fungicide fenhexamid due to detoxification by a cytochrome P450monooxygenase Cyp684. Saad Azeddine, Alexis Billard, Jocelyne Bach, Catherine Lanen, Anne-Sophie Walker, Sabine Fillinger, Danièle Debieu. INRAUR1290 BIOGER CCP, avenue Lucien Brétignières F78850 Thiverval-Grignon, France.The Botrytis species complex responsible for the grey mould disease found on grapevines is composed of two species: Botrytis cinerea, to a large extent(roughly 90%), and Botrytis pseudocinera. Despite their genetic polymorphism, these species cannot be morphologically distinguished. However, they dodiffer in their response to several fungicides, especially to the sterol biosynthesis inhibitor fenhexamid. While B. cinerea is sensitive to this hydroxyanilidefungicide, B. pseudocinerea is naturally resistant. Because a strong synergism was found on B. pseudocinerea between fenhexamid and sterol 14ademethylationinhibitors (DMIs) known to inhibit Cyp51, a cytochrome P450 monooxygenase, it was hypothetized that the detoxification of fenhexamid bya cytochrome P450 monooxygenase similar to Cyp51 is involved in the resistance B. pseudocinerea displays. To test this, we sought the geneoverexpressed in the presence of fenhexamid with the highest similarity to cyp51. Taking into account the Cyp P450 classification based on homology andphylogenetic criteria, this gene, whose function remains unknown, belongs to the Cyp684 family. It was then deleted in a B. pseudocinerea strain. Cyp684knock out mutants exhibit a loss of fenhexamid resistance and synergism between DMIs and fenhexamid, showing that the Cyp684, cytochrome P450protein is responsible for B. pseudocinerea’s natural resistance to fenhexamid and is involved in fenhexamid detoxification. Although cyp684 is alsopresent in B. cinerea, which is sensitive to fenhexamid, a polymorphism was observed between the two species: in B. pseudocinerea the cyp684 promotershows a deletion of 25 bp. We are currently establishing the cyp684 expression profiles in both species in order to analyze the impact of the promoterdeletion on its expression. Metabolization studies are also being conducted to identify metabolites that would help in understanding the enzymaticfunctions of Cyp684 and to determine to what extent Botrytis sp. is sensitive to these metabolites.16. Evolutionary rewiring of ubiquitination targets in Candida albicans promotes efficient carbon assimilation in host niches. Alistair J P Brown, DoblinSandai, Zhikang Yin, Laura Selway, David Stead, Janet Walker, Michelle D Leach, Iryna Bohovych, Iuliana V Ene, Stavroula Kastora, Susan Budge, Carol AMunro, Frank C Odds, Neil A R Gow. School of Medical Sciences, University of Aberdeen, Aberdeen, United Kingdom.Pathogens must assimilate carbon to grow and infect their host. Interesting questions remain about the regulation of carbon assimilation in Candidaalbicans despite the wealth of knowledge about this major fungal pathogen of humans. C. albicans is classified as a Crabtree-negative yeast because itcontinues to respire in the presence glucose [J Med Vet Mycol 26, 195]. How then can C. albicans be exquisitely sensitive to sugars, down-regulatingtranscripts involved in the utilization of alternative carbon sources following exposure to 0.01% glucose [Molec Biol Cell 20, 4845]? We have now shownthat there is a significant dislocation between the transcriptome and proteome in C. albicans: glucose triggers the decay of key transcripts but the enzymesthey encode are retained. This allows the simultaneous assimilation of alternative carbon sources such as fatty acids, carboxylic acids and sugars by C.albicans. This contrasts with the situation in Saccharomyces cerevisiae where simultaneous carbon assimilation is prevented by catabolite inactivation[Arch Micro 134, 187; Arch Micro 147, 231]. We show that C. albicans has retained the molecular apparatus that mediates ubiquitin-mediated, glucoseacceleratedprotein degradation. For example, S. cerevisiae isocitrate lyase (ScIcl1) is degraded rapidly when expressed in C. albicans. However, C. albicansisocitrate lyase (CaIcl1) lacks critical ubiquitination sites that mediate this catabolite inactivation. Furthermore, other C. albicans enzymes involved ingluconeogenesis and the glyoxylate cycle appear to lack such sites, whereas glycolytic enzymes are ubiquitinated (e.g. Fba1, Pgk1, Eno1). Therefore therehas been significant rewiring of ubiquitination targets in C. albicans compared to S. cerevisiae. This metabolic flexibility probably enhances efficientcolonisation of host niches that contain complex mixtures of nutrients.27th Fungal Genetics Conference | 125

FULL POSTER SESSION ABSTRACTS17. Can-Hsp31 is important for Candida albicans growth and survival. S. Hasim, N. Ahmad hussin, K. Nickerson. Biological Science, Univesity of NebraskaLincoln, Lincoln, NE.Candida albicans is an opportunistic pathogen that is able to grow as budding yeast, pseudohyphae, and hyphae. A key feature of C. albicans is its abilityto grow in diverse microenvironments and develop complex and highly efficient responses in order to survive within the host environment. The C. albicansHsp31 (ORF19.251) gene encodes a protein that belongs to the DJ1/PfpI family with close homology to other fungal Hsp31-like proteins. Despite intensivestudy, the function of these fungal Hsp31 proteins is unknown. The crystal structure of Can-Hsp31 was solved to 1.6 Å resolution. Its structure is similar tothose of the E.coli and S. cerevisiae Hsp31 proteins except that Can-Hsp31 is a monomer in the crystal while all other known homologues are dimers. Inthis report, we show that the C. albicans’ Hsp31 is important for growth and survival under various stress conditions.18. Influence of N-glycans on a-/b-(1,3)-glucanase and a-(1,4)-amylase from Paracoccidioides brasiliensis yeast cells. Fausto Bruno Dos Reis Almeida 1 ,Valdirene Neves Monteiro 2 , Roberto Nascimento Silva 3 , Maria Cristina Roque-Barreira 1 . 1) Cellular and Molecular Biology, University of Sao Paulo, RibeiraoPreto, Brazil; 2) University of Goias, Anapolis, Brazil; 3) Biochemistry and Immunology, University of Sao Paulo, Ribeirao Preto, Brazil.Paracoccidioides brasiliensis (Pb) is a temperature-dependent dimorphic fungus and the causative agent of paracoccidioidomycosis, the most prevalentsystemic mycosis in Latin America. The cell wall (CW) of Pb is a network of glycoproteins and polysaccharides, such as chitin, glucan and N-glycosylatedproteins, that may perform several functions. N-glycans are involved in glycoprotein folding, intracellular transport, secretion, and protection fromproteolytic degradation. Our group has been describing the role of N-acetylglucosaminidase (NAGase) in fungal growth, exerted through participation inchitin metabolism and CW remodeling. In addition, by assessing yeast cells cultured with tunicamycin (TM), we determined that N-glycans play importantroles in growth and morphogenesis of Pb yeasts and are required for the fungal NAGase function. In this study, we verify the influence of TM-mediatedinhibition of N-linked glycosylation on a- and b-(1,3)-glucanase, as well as the a-(1,4)-amylase, produced by Pb yeast cells. The treatment of Pb with 15 mgTM/ml did not interfere with a- and b-(1,3)-glucanase production, secretion or on enzyme structure. The absence of N-glycans did not affect pH optimum(5.5) or temperature optimum (45 °C). Moreover, the fully- and under-glycosylated forms of the enzymes had similar Km and Vmax values. On the otherhand, a-(1,4)-amylase demonstrated lower enzymatic activity when underglycosylated, although no difference was detected between the pH andtemperature optimums of the two forms. Our results corroborates with the recent observation that a-(1,4)-amylase from Pb plays important roles on thefungal CW a-(1,3)-glucan biosynthesis. However, interestingly the Pbaglucan gene, that encode to a-(1,3)-glucanase, had its expression increased by 2.5-fold in Pb cells treated with TM when evaluated by qRT-PCR, suggesting an indirect influence of TM on CW glucan synthesis. Genes encoding to UPR(Unfolding Protein Response) and CW synthesis showed their expression increased, corroborating with our data. Analyses investigating the effect of N-glycans in mycelium cells are under way in our laboratory. Our results suggest that N-glycans do not play direct effect on a- and b-(1,3)-glucanase activityproduced by yeasts cells but indirect effect by affecting a-(1,4)-amylase.19. Cell wall structure and biosynthesis in oomycetes and true fungi: a comparative analysis. Vincent Bulone. Sch Biotech, Royal Inst Biotech (KTH),Stockholm, Sweden.Cell wall polysaccharides play a central role in vital processes like the morphogenesis and growth of eukaryotic micro-organisms. Thus, the enzymesresponsible for their biosynthesis represent potential targets of drugs that can be used to control diseases provoked by pathogenic species. One of themost important features that distinguish oomycetes from true fungi is their specific cell wall composition. The cell wall of oomycetes essentially consists of(1®3)-b-glucans, (1®6)-b-glucans and cellulose whereas chitin, a key cell wall component of fungi, occurs in minute amounts in the walls of some oomycetespecies only. Thus, the cell walls of oomycetes share structural features with both plants [cellulose; (1®3)-b-glucans] and true fungi [(1®3)-b-glucans, (1®6)-b-glucans and chitin in some cases]. However, as opposed to the fungal and plant carbohydrate synthases, the oomycete enzymes exhibit specific domaincompositions that may reflect polyfunctionality. In addition to summarizing the major structural differences between oomycete and fungal cell walls, thispresentation will compare the specific properties of the oomycete carbohydrate synthases with the properties of their fungal and plant counterparts, withparticular emphasis on chitin, cellulose and (1®3)-b-glucan synthases. The significance of the association of these carbohydrate synthases with membranemicrodomains similar to lipid rafts in animal cells will be discussed. In addition, distinguishing structural features within the oomycete class will behighlighted with the description of our recent classification of oomycete cell walls in three different major types. Genomic and proteomic analyses ofselected oomycete and fungal species will be correlated with their cell wall structural features and the corresponding biosynthetic pathways.20. Investigating the function of a putative chitin synthase from Phytophthora infestans. Stefan Klinter, Laura Grenville-Briggs, Hugo Mélida, VincentBulone. School of Biotechnology, Division of Glycoscience, Royal Institute of Technology (KTH), Stockholm, Sweden.The oomycete Phytophthora infestans is a plant pathogen that causes potato late blight, a devastating disease associated with tremendous economiclosses. In contrast to true fungi, oomycetes are traditionally described as cellulosic micro-organisms. Indeed, in addition to other b-glucans, cellulose is amajor polysaccharide in the mycelial cell wall of P. infestans while chitin and other N-acetylglucosamine (GlcNAc)-based carbohydrates are absent fromhyphal walls. However, a putative chitin synthase gene (chs) is present in the genome. Bioinformatic analysis identified the C-terminal region of thepredicted protein to be highly similar to glycosyltransferase family 2 proteins, such as fungal chitin synthases, while the N-terminal domain is moredivergent. Orthologous putative chs genes are present in all sequenced oomycete genomes and phylogenetic analysis shows the oomycete gene productsform a new clade separate from the fungal lineage. The P. infestans chs transcript is highly abundant in older mycelium. However, no chitin synthaseactivity was detectable in microsomal fractions assayed with radioactively-labeled UDP-GlcNAc, the natural substrate of chitin synthase. Suprisingly, hyphalgrowth was severely retarded in the presence of low micromolar concentrations of the chitin synthase inhibitor nikkomycin Z, a structural analogue ofUDP-GlcNAc. Microscopic analysis of nikkomycin Z-treated hyphae revealed frequent tip swelling and bursting. Similarly, transient RNA-mediated silencingof the chs gene resulted in severely reduced growth, and hyphae showed a hyper-branched morphology with swollen tips. As a first step to determine theprecise function of the P. infestans chs gene, we have cloned and expressed it in Saccharomyces cerevisiae.21. Deciphering cell wall structure and biosynthesis in oomycetes using carbohydrate analyses and plasma membrane proteomics. Hugo Melida 1 ,Vaibhav Srivastava 1 , Erik Malm 1 , J. Vladimir Sandoval-Sierra 2 , Javier Dieguez-Uribeondo 2 , Vincent Bulone 1 . 1) Division of Glycoscience, Royal Institute ofTechnology (KTH), Stockholm, Sweden; 2) Mycology Department, Royal Botanical Garden (CSIC), Madrid, Spain.Some oomycete species are severe pathogens of economically important animals or plants. Proteins involved in cell wall metabolism represent excellenttargets for disease control. The objective of our work was to determine the fine cell wall polysaccharide composition of selected species and identify thecorresponding membrane-bound biosynthetic enzymes and other proteins involved in cell wall remodeling. In the first instance, we performed a detailedcarbohydrate analysis of the mycelial cell walls of 11 oomycete species from 2 major orders, the Saprolegniales and Peronosporales. We then selected thefish pathogen Saprolegnia parasitica for in-depth proteomics analysis. Our results indicate the existence of 3 clearly different cell wall types. This126

FULL POSTER SESSION ABSTRACTSbiochemical distinction is in agreement with the taxonomic grouping based on molecular markers of the species studied. The 3 cell wall types aredistinguishable by their cellulose content and the fine structure of their 1,3-b-glucans. Furthermore, unique features were found in each case. Type I cellwalls (e.g. Phytophthora) are devoid of N-acetylglucosamine (GlcNAc) but contain glucuronic acid and mannose; type II (e.g. Achlya, Dictyuchus,Leptolegnia and Saprolegnia) contain up to 5% GlcNAc and residues indicative of cross-links between cellulose and 1,3-b-glucans; type III (e.g.Aphanomyces) are characterized by the highest GlcNAc content (> 5%) and the occurrence of unusual carbohydrates that consist of 1,6-linked GlcNAcresidues. Analysis of the recently sequenced genome of S. parasitica was combined with quantitative mass spectrometry-based proteomics (label-free andiTRAQ) to characterize the plasma membrane proteome of hyphal cells. This strategy allowed us to experimentally identify a total of 677 plasmamembrane proteins, including several key cell wall polysaccharide synthases, e.g. cellulose, 1,3-b-glucan and chitin synthases, some of which arespecifically enriched in plasma membrane microdomains similar to lipid rafts in animal cells.22. Identification and characterization of the chitin synthase genes in the fish pathogen Saprolegnia parasitica. Elzbieta Rzeszutek, Sara Diaz, VincentBulone. School of Biotechnology, Division of Glycoscience, Royal Institute of Technology (KTH), Stockholm, Sweden.The oomycete Saprolegnia parasitica is a fungus-like microorganism responsible for fish diseases and huge losses in aquaculture. The analysis of the cellwall composition of the microorganism and the characterization of key enzymes involved in cell wall biosynthesis may facilitate the identification of newtarget proteins for disease control. The cell wall of hyphal cells of S. parasitica consists mainly of cellulose, b-(1®3)- and b-(1®6) glucans, whereas chitin ispresent in minute amounts only. The main objective of this work was to test the effect of nikkomycin Z, a competitive inhibitor of chitin synthase (CHS), onthe growth of S. parasitica. Genome mining allowed the identification of six different putative chs genes whose actual occurrence in the genomic DNA ofthe microorganism was confirmed by Southern blot analysis. The expression of the chs genes in the mycelium was analyzed using Real-Time PCR. Theresults revealed a higher expression level of four of the six genes while the two others exhibited undetectable levels of expression in the mycelium. Thissuggests that the latter genes are most likely primarily involved in chitin formation at a different developmental stage. The presence of nikkomycin Zincreased the expression level of one of the genes, chs3, suggesting that the corresponding product is involved in forming the abnormal branchingstructures in the hyphae exposed to the inhibitor. The capacity of the mycelium to synthesize chitin was demonstrated by performing in vitro synthesisreactions using cell-free extracts. CHS activity was measured in intact cell membranes as well as in detergent-extract of membranes. The polysaccharidesynthesized in vitro was characterized by enzymatic hydrolysis with a specific chitinase. Our data demonstrate that CHS represent promising targets ofanti-oomycete drugs, even though the amount of chitin in the cell wall of S. parasitica does not exceed a few percent.23. Role of Ccr4-mediated mRNA turnover in nucleotide/deoxynucleotide homeostasis and Amphotericin B susceptibility in Cryptococcus neoformans.D. Banerjee, J. Panepinto. Department of Microbiology and Immunology.University at Buffalo, SUNY, Buffalo, NY.Ccr4 mediated deadenylation is the first and rate limiting step in eukaryotic mRNA decay. The end products of mRNA degradation are nucleosidemonophosphates (NMPs) which are then converted to nucleotides (NDPs and NTPs) and deoxynucleotides (dNTPs) in downstream reactions. A C.neoformans degradation deficient ccr4D mutant exhibits replication stress sensitivity and stabilizes ribosomal protein (RP) transcripts during carbonstarvation, suggesting that ccr4D mutant is deficient in intracellular nucleotide stores. Analysis of gene expression showed an up-regulation of thenucleotide synthesis machinery in ccr4D mutant even under unstressed conditions consistent with our hypothesis. Time-kill assays in the presence ofmycophenolic acid (MPA), an inhibitor of guanine nucleotide de novo synthesis, showed a reduction in the viability of ccr4D mutant that was rescued bythe addition of exogenous guanine, suggesting that the salvage pathway is indeed functional. These results suggest that the degradation of mRNAtranscripts lead to the production of NMPs that replenish NTP/dNTP pools in C. neoformans during starvation stress. The fungicidal efficacy ofAmphotericin B (AmpB) is enhanced by the use of Flucytosine, a pyrimidine analog, suggesting a synergy between AmpB and nucleotide deficiency forcryptococcosis treatment. We compared the sensitivity of wild type (H99), ccr4D mutant and H99-FOA strain (de novo mutant of pyrimidine synthesis) to acombination of AmpB and NTP/dNTP inhibitors. Both mutants exhibited higher sensitivity to AmpB which was unaltered by additional stressors. H99exhibited an increased sensitivity to the combination of AmpB with both NTP and dNTP inhibitors, compared to AmpB alone. Taken together, these datasuggest that nucleotide depletion, either by a pharmacologic agent or a mutation predisposes the cells to enhanced AmpB mediated cell death.Thus ouroverall hypothesis is that Ccr4 mediated mRNA turnover results in the maintenance of intracellular NTP/dNTP pools to promote growth, virulence, stresstolerance and also modulates Amp B susceptibility in C. neoformans. Results from these studies will identify a novel role of the mRNA degradationmachinery in C. neoformans pathogenesis and stress tolerance and also aid in the identification of new anti-cryptococcal drug targets.24. WITHDRAWN25. Blue light induce Cordyceps militaris fruiting body formation and cordycepin production. Chun-Hsiang Yang 1 , Shun-Kuo Sun 2 , Su-Der Chen 1 . 1)Biotechnology and Animal science, National Ilan University, Ilan, Taiwan; 2) Bionin Biotechnology, INC.Cordyceps militaris is a very important fungal medicine in Chinese. The fruiting body of Cordyceps militaris has been described by many researcherscontaining biological activies, such as being able to inhibit cell proliferation, provide anti-ageing activity, inhibit protein synthesis and lowing cardiovascularrisk. Cordyceps militaris has been grown and harvested by many Chinese people, and were able to obtain its fruiting body with orange collar and barshape. Solid cultivation as carried out 3 to 4 days after mycelium seeding from liquid culture, then fruiting body formation can been induced by light( 12hours per day). In this research, the light-inducing mechanism of fruiting body formation was studied. The results showed the fruiting body was induced byblue light but not red light. Cordycepin, the most important compound with medical potential of Cordyceps militaris, is mainly stored in fruiting body,rather than in mycelium from liquid culture. Cordycepin production depends on various factors, including: wave length of light and culture in solid orliquid. The results also showed the relationship between cordycepin production and blue light sensor in Cordyceps militaris, which might contain LOVdomain.26. Insight into alkaloid diversity of the epichloae, protective symbionts of grasses. Carolyn A. Young 1 , Nikki D. Charlton 1 , Johanna E. Takach 1 , Ginger A.Swoboda 1 , Bradley A. Hall 1 , Kelly D. Craven 2 , Christopher L. Schardl 3 . 1) Forage Improvement Division, The Samuel Roberts Noble Foundation, Ardmore,OK; 2) Plant Biology Division, The Samuel Roberts Noble Foundation, Ardmore, OK; 3) Plant Pathology, University of Kentucky, Lexington, KY.Cool season grasses from the subfamily Pooideae often form symbiotic associations with fungal endophytes known collectively as the epichloae (Epichloëand Neotyphodium species). The epichloae consist of both sexual (nonhybrid) and asexual (hybrid and nonhybrid) species that can produce the bioactiveanti-herbivore compounds, ergot alkaloids, indole-diterpenes, lolines and peramine. Epichloae can exhibit considerable chemotypic diversity within thepathways of these four alkaloid classes as well as the combination of alkaloids that can be produced by an individual, and as such, may equate to fitnessbenefits for the host. The current genome sequencing efforts, whereby at least 10 epichloae have been sequenced, now allows us to develop simple27th Fungal Genetics Conference | 127

FULL POSTER SESSION ABSTRACTSapproaches to rapidly screen large cool-season grass populations to identify endophyte diversity. Molecular analyses of endophyte genetic traits fromamong and between host populations allow us to explore resident endophyte incidence and diversity present in single host species. PCR with genomicDNA extracted from individual plants (seeds or tillers) is used to determine endophyte incidence within a line and to predict alkaloid chemotypes at theEAS (ergot alkaloids), LOL (lolines), IDT/LTM (indole-diterpenes) and PER (peramine) loci. The presence or absence of genes at each locus can be used topredict the likely pathway end product for a given endophyte-infected plant line. Phylogenetic analyses of housekeeping and mating-type genes are usedto infer hybrid versus nonhybrid origins as well as hybrid ancestral progenitors. Sequence analyses of alkaloid genes encoding key pathway steps provideallele copy number and can be used to determine progenitor origins to further support the phylogenetic relationships. Grass collections across multiplehost tribes have recently been evaluated and considerable endophyte chemotypic diversity was identified. Multiple endophyte species were able toindependently associate with some grass host species and often both hybrid and nonhybrid endophytes could be found within a population. In manycases, chemotypic diversity of the hybrids may have arisen from independent hybridization events and as such, this alkaloid diversity likely translates intodifferences in fitness and persistence of the host.27. Extracellular polysaccharide degrading capabilities of various Agaricus bisporus strains during compost cultivation. A. Patyshakuliyeva, J. Yuzon, R. P.de Vries. CBS-KNAW Fungal Biodiversity Centre, Utrecht, The Netherlands.In the temperate forests of North America and Europe, basidiomycetes such as Agaricus bisporus are renowned for their ecological significance in thecycling of carbon from dead plant matter. A. bisporus is also the most widely produced mushroom in the world and has been cultivated for centuries.However, little is known about the interaction between A. bisporus and its most preferred substrate, composted plant matter. In this study, a wide array ofextracellular polysaccharide degrading enzymes was studied under semi-commercial conditions to understand the carbon nutritive needs of the fungi.Various time points were sampled from filling of the beds, vegetative growth and development and maturation of fruiting bodies. Clear correlations in theenzymatic activities were observed from different stages of development of A. bisporus between compost, casing layer and fruiting bodies. This couldsuggest that vegetative mycelia and the fruiting body divide their metabolic roles as vegetative mycelium of A. bisporus provides nutrients for the growthof fruiting bodies, while fruiting bodies aims on reproduction. This was also confirmed by identification of the expression of genes encoding plant andfungal polysaccharide modifying enzymes in compost, casing layer and fruiting bodies.28. Reconstruction of the rubrofusarin biosynthetic pathway in Saccharomyces cerevisiae. Rasmus J N Frandsen 1 , Peter Rugbjerg 1 , Michael Naesby 2 , UffeH Mortensen 1 . 1) Systems Biology - CMB, Technical University of Denmark, Kgs. Lyngby, Denmark; 2) Evolva SA, Duggingerstrasse 23. CH-4153 Reinach,Switzerland.The aromatic heptaketide rubrofusarin is a common core substructure of several fungal pigments, including rubrofusarin B, aurofusarin, nigerone,nigerasperone A, chaetochromin, ustilaginoidin and parasperone A. Compounds that are produced by a wide variety of different filamentous fungi such asFusarium graminearum, Aspergillus niger, Aspergillus parasiticus, Chaetomium gracile and Ustilaginoidea virens. Previous reverse genetics analysis of theaurofusarin biosynthetic pathway, by targeted gene replacement in F. graminearum (Fg), has resulted in the formulation of a six step biosynthetic pathwaythat includes rubrofusarin as an intermediate. In the current study we have used heterologous expression in Saccharomyces cerevisiae to test whether allthe enzymes required for biosynthesis of rubrofusarin have been identified. Successful reconstruction of the rubrofusarin pathway is dependent on theheterologous co-expression of four genes: the Fg polyketide synthase PKS12, the Fg dehydratase aurZ, the Fg O-methyltransferase aurJ and the Aspergillusfumigatus phosphopantetheine transferase npgA. To eliminate potential problems with intron splicing of the fungal genes in S. cerevisiae the requiredcoding sequences were de novo synthetized in codon optimized versions. The four genes were expressed individually from four different single copyplasmids, each with a unique auxotrophic marker. Co-expression of the codon optimized version of PKS12 with npgA did not result in production of anynew metabolites. However, surprisingly co-expression of a cDNA version of PKS12, assembled from gDNA by USER-fusion, resulted in production of theexpected product YWA1. Additional co-expression of the codon optimized dehydratase encoding aurZ gene lead to production of nor-rubrofusarin, andsubsequent introduction of the O-methyltransefase gene aurJ yielded rubrofusarin. These results support the previously proposed biosynthetic route forthe formation of rubrofusarin in F. graminearum. The utilized bottom-up approach shows that formation of rubrofusarin is dependent only on thecombined action of PKS12, AurZ and AurJ in F. graminearum, and likely also in other fungal species that produce compounds with a rubrofusarin core. Thelatter is further supported by sequence base homology searches in the available relevant fungal genome sequences.29. Expression and purification of hydrophobin fusion proteins targeted to intracellular protein bodies in T. reesei . Nina K. Aro, Marika Vitikainen, JussiJoensuu, Eero Mustalahti, Markku Saloheimo. Biotechnology, VTT Technical Research Centre, 02044 VTT, VTT, Finland.Recombinant protein production is a fast growing market area. The need for novel production platforms is growing together with the number of newapplications for recombinant proteins. The ascomycete T. reesei is an excellent producer of hydrolytic enzymes. However, heterologous protein productionin T. reesei is often suffering from low product yields due to protease degradation and inefficiency in heterologous protein secretion. We have previouslydemonstrated a novel recombinant protein production system for T. reesei using GFP as a model protein. This system uses hydrophobin, a small andamphipathic fungal protein, as a fusion tag for purification and ER retention signal for targeting the produced protein to intracellular protein bodies. TheGFP-HFBI fusion protein can be extracted from total protein lysate by aqueous two-phase separation system. We have now further optimised theexpression for GFP-HFBI fusion and demonstrated the applicability of this production concept for two additional proteins, glucose oxidase (GOX) and tissueplasminogen activator (tPA). Effect of C- and N-terminal hydrophobin fusion on productivity and extraction in two-phase separation system will bediscussed. The new production concept is aiming at widening the spectrum of recombinant proteins that can be produced efficiently in T. reesei.128

FULL POSTER SESSION ABSTRACTS30. Metabolic adaptations in Phytophthora infestans and the role of a phosphagen kinase system in energy metabolism. Meenakshi Kagda, HowardJudelson. Plant Pathology and Microbiology, University of California, Riverside, CA 92521.Nutrient acquisition and metabolic adaptation to host-derived nutrients is an important aspect of pathogen biology. An understanding of the metabolicadaptations made by Phytophthora infestans, an important pathogen of potato and tomato, to optimize nutrient uptake from diverse host tissues andwithin the microenvironments of the host will lead to a better understanding of host-pathogen relationships. In order to study metabolic adaptations of P.infestans, transcriptional profiling and live cell imaging using promoter-fluorescent protein fusions will be used. Preliminary results demonstrated thedifferential gene expression of many metabolic genes of P. infestans grown on different natural hosts and that grown on rich media. The next step involvesanswering the question: Are some metabolic genes expressed in a stage-specific or time-dependent manner? In addition, the role of enzymes involved inenergy homeostasis and metabolite channeling are being studied. The roles of two such genes encoding putative creatine kinases are being elucidatedusing subcellular localization, substrate utilization and loss of function studies.31. Platforms for secondary metabolite analysis in filamentous fungi. Uffe H. Mortensen 1 , Jakob B. Nielsen 1 , Diana C. Anyaogu 1 , Dorte K. Holm 1 , Lene M.Petersen 1 , Morten T. Nielsen 1 , Mikael S. Joergensen 1,2 , Kristian F. Nielsen 1 , Pia F. Johannesen 2 , Dominique A. Skovlund 2 , Thomas O. Larsen 1 . 1) DTU SystemsBiology, Technical University of Denmark, Kgs. Lyngby, Denmark; 2) Department 463, Fungal Gene Technology, Novozymes, Denmark.We are developing versatile methods that allows for rapid and simple genetic manipulation of filamentous fungi. Currently, we use our methods forelucidation of pathways for secondary metabolite production in a number of different species. The platform includes simple systems for gene targetingand defined expression platforms for pathway reconstitution. Alternatively, if few or no genetic tools are available for the fungus, we use AMA1 basedplasmids for transformation. All DNA handling prior to fungal transformation is based on assembly by efficient USER cloning that allows for many DNAfragments to be merged in a single cloning step. Examples of pathway reconstitution will be presented including functional transfer of the entire geodinproducing gene cluster from Aspergillus terreus into A. nidulans. In an attempt to map the first intermediates of polyketide pathways in a fungal species,we have individually expressed all PKS genes from A. niger as a starting point for pathway elucidation. Using this approach we identified a PKS generesponsible for production of 6-MSA. Next, we individually deleted all genes in the corresponding gene cluster in A. niger to further map the pathway.Theses analyses suggest that 6-MSA is a precursor of Yanuthone D/E. In a similar study, we have identified a related PKS gene in A. aculeatus that alsoproduces 6-MSA when expressed in A. nidulans. The corresponding gene cluster in A. aculeatus contains a gene encoding a transcription factor. Using ourAMA1 based expression system, this gene has been overexpressed in A.aculeatus. As a result a new 6-MSA based compound has been identified. Lastly,using a knock-in/knock-out platform in Trichoderma reesei we use the same principles to uncover a gene cluster that is responsible for a very complexfamily of sorbicillinoids.32. Transcriptional analysis of oxalate degradation in the white rot basidiomycete Dichomitus squalens. Miia R. Mäkelä, Johanna Rytioja, Outi-MaariaSietiö, Sari Timonen, Annele Hatakka, Kristiina Hildén. Department of Food and Environmental Sciences, University of Helsinki, Helsinki, Finland.Basidiomycetous white rot fungi are the most efficient degraders of lignocellulose with a unique ability to mineralize the recalcitrant lignin polymer.Lignocellulose decay involves a complex enzymatic system, but is also suggested to be promoted by the fungal secretion of oxalic acid. White rot fungisynthesize oxalate as a metabolic waste compound and typically secrete it to their environment in millimolar quantities. As oxalate is a toxic compound,regulation of its intra- and extracellular concentration is extremely crucial for fungi and also for lignocellulose degradation since high oxalate levels areshown to inhibit the decomposition reactions. Therefore, specific oxalate-converting enzymes, namely oxalate decarboxylases (ODCs) that work inconjunction with formate-degrading formate dehydrogenases (FDHs), are recognized as key fungal enzymes in lignocellulose decay. Dichomitus squalens isa white rot fungus that degrades effectively all the wood polymers, i.e. cellulose, hemicelluloses and lignin, and secretes oxalic acid during its growth onwood. The genome of D. squalens harbours 5 putative ODC and 3 putative FDH encoding genes, while these numbers differ in other fungi based oncomparative genomics. In order to enlighten the roles of the multiple oxalic-acid catabolising enzymes of D. squalens, the expression of the odc and fdhgenes was followed with quantitative real-time RT-PCR when the fungus was grown on its natural substrate, i.e. Norway spruce (Picea abies) wood. Inaddition, the effect of organic acid (oxalic acid) and inorganic acid (HCl) supplementation on the relative transcript levels of the oxalate-catabolizing geneswas examined in the submerged liquid cultures of D. squalens. The results show for the first time the sequential action of ODC and FDH at the transcriptlevel in a white rot fungal species. The constitutive expression of odc1 suggests the pivotal role of the corresponding enzyme during the growth of D.squalens on wood. In addition, the strong upregulation of the transcription of odc2 in oxalic-acid amended cultures indicates the distinct roles of individualODC isoenzymes.33. Creation of temperature-influenced hyphal growth mutants in a basidiomycete fungus through the use of UV mutagenesis. Stephen J. Horton, CarlyWender, Suhasini Padhi. Dept Biological Sci, Union Col, Schenectady, NY.Filamentous fungi have been used extensively in industry for decades, most prominently for the purposes of protein expression. An emerging technologyis the use of fungi in the production of ecologically-friendly materials used in packaging and insulation, materials presently manufactured using nonrenewablepetroleum-based technologies. The growth characteristics of the mycelia used in these manufacturing processes play a pivotal role in theproperties of the final product. We decided to utilize the classical approach of UV mutagenesis to create new strains of a basidiomycete fungus that wouldpotentially have growth characteristics more suited than the wild type to particular industrial applications. One example of this would be the production offungal strains with a wider temperature spectrum for growth, a factor applicable to the industrial reality of the fluctuating ambient temperature found innon-laboratory conditions. Protoplasts from our wild-type strain were subjected to 70,000 microjoules/cm2 of UV irradiation at a wavelength of 254 nm.Survivors were allowed to recover overnight at 28°C, and then plated at a selective temperature of 40°C. Out of an estimated 6400 protoplasts irradiated,we observed 48 colonies of various sizes after 11 days growth at the selective temperature of 40°C. This temperature was chosen because it approachesthe maximum permissible growth temperature for this fungus. We further characterized 22 of these presumed mutants and were able to sort them intobroad categories based upon their growth rates over the range of 28°C to 37°C. The categories were: (1) strains that grew well at the normal laboratorytemperature (28°C), but less well at elevated temperatures (33.5°C and 37°C), (2) strains that grew best at elevated temperature (33.5°C), and (3) strainsthat grew at relatively similar rates at all three temperatures. The morphology of the hyphae (density, branching pattern) was also found to differ betweenthe growth mutants. Selected mutants will be analyzed by both RNA seq and genomic sequencing approaches in an effort to identify any common genesthat may have been altered as a result of the mutagenesis regime.27th Fungal Genetics Conference | 129

FULL POSTER SESSION ABSTRACTS34. Functional Analysis of a Novel Diaminopimelate Decarboxylase from the Oomycete Saprolegnia parasitica. Lingxiao ge 1 , Josie Hug 1 , Stan Oome 2 , PaulMorris 1 . 1) Biological Sci, Bowling Green State Univ, Bowling Green, OH; 2) Plant-Microbe Interactions, Utrecht University, The Netherlands.In bacteria and plants, the lysine precursor L, L-diaminopimelate (DAP) is first converted to meso-diaminopimelate by an epimerase. Then meso-DAP isconverted to lysine by a DAP decarboxylase. Comparative analysis of seven sequenced oomycete genomes, revealed that only Saprolegnia parasiticacontains a predicted epimerase. Sequence homology in all of the predicted DAP decarboxylases in oomycetes is strongly conserved, suggesting that theseproteins have similar biochemical activity. The oomycete DAP gene appears to have been acquired by horizontal transfer from Archaea sp. Notably, theseparticular Archaea sp. have all the genes needed to synthesize lysine, except for epimerase. Thus we postulated that the oomycete DAP might be a novelenzyme capable of converting L, L-DAP directly to lysine. To test this hypothesis, we codon-optimized the DAP gene from S. parasitica and expressed it inan E. coli DAP mutant. Complementation assays of the mutant expressing the S parasitica gene in lysine-minus media indicate that the gene functions as aDAP decarboxylase. To determine the substrate specificities of the S. parasitica DAP gene, we have developed an HPLC method to separate the D, L, andmeso isomers of chemically synthesized DAP. Authentic L, L-DAP has also been purified from the culture filtrates of an E coli epimerase mutant. Functionalassays of the affinity-purified protein will enable us to characterize the substrate specificities of the oomycete enzyme. If the S. parasitica DAP enzyme canutilize L, L-DAP as a substrate, then the retention of epimerase in this genome may indicate that meso-DAP is incorporated into the cell wall of this groupof organisms.35. Living on Air?: Ustilago maydis cells grow without being provided nitrogen in their growth media. Michael H Perlin, Michael Cooper. Dept Biol andProgram on Disease Evolution, Univ Louisville, Louisville, KY, USA.Nitrogen is an essential nutrient for all living creatures. Ammonium is one of the most efficiently used and thus preferred, sources of nitrogen. As withother dimorphic fungi, yeast-like cells of Ustilago maydis, the fungal pathogen of maize, switch to filamentous growth when starved fornitrogen/ammonium. U. maydis carries two genes, ump1 and ump2, encoding ammonium transporters that facilitate both uptake of ammonium and thefilamentous response to its absence. While no obvious phenotype is observed when ump1 is deleted, cells without ump2 are unable to filament inresponse to low ammonium, although they can still grow. Surprisingly, ump1ump2 double mutants can also grow on low ammonium. More amazing still,both wild type and mutant cells continue to grow, even after strenuous efforts were made to remove all nitrogen sources from their growth media. Toinvestigate these unusual observations further, we grew wild type and mutant cells in the absence or presence of added nitrogen, as ammonium orsupplied as 15 N gas. Septum bottles with rich, low ammonium and no ammonium media were inoculated with rinsed overnight wild type and mutant cells,injected with +0.1% 15 N 2 and were then incubated for seven days. The resulting biomass was sampled for microscopic examination, collected by filtration,dried and loaded into tin sample capsules for d 15 N analysis by the Stable Isotope Research Unit at Oregon State University. The wild type cells under rich,minimal and no ammonium conditions had mean d 15 N ratios of 0.7, 10.8 and 45.2, respectively, while the mutant cells had mean d 15 N ratios of 3.29, 49.5and 134.8, respectively, for these growth conditions. This indicated significant incorporation of the 15 N tracer from the injected gas into the cellularbiomass. We are currently investigating additional candidate genes that may play a role in this novel capability by a fungus.36. Saprotrophic metabolism of the White-Nose Syndrome fungus Geomyces destructans in bat hibernacula. Hannah Reynolds 1 , Tom Ingersoll 2 , HazelBarton 1 . 1) Department of Biology University of Akron Akron, OH 44325; 2) National Institute for Mathematical and Biological Synthesis (NIMBioS)University of Tennessee Knoxville, TN 37996.Geomyces destructans (Myxotrichaceae, Leotiomycetes), an emerging epizootic disease of hibernating bats in North America has arisen from apredominately saprotrophic genus. We have isolated multiple, non-infectious Geomyces species from cave surfaces and healthy bats for physiological andgenetic comparison with G. destructans to better understand its disease ecology. In particular, we are interested in 1) whether G. destructans retainssaprotrophic ability, acting as a facultative rather than an obligate pathogen and 2) identifying the microhabitats that support Geomyces and presumablyG. destructans growth. Identifying an environmental niche for G. destructans would aid in understanding future disease ecology. Comparative genomicsindicates the presence of multiple enzymes involved in saprotrophic metabolism, including endoglucanases, b-glucosidases and chitinases, while in vitrosaprotrophic assays demonstrate similar cellulase and lipase functions in both pathogenic and non-pathogenic Geomyces. To understand the nativemicrobial habitats that might inhibit or promote G. destructans growth we used molecular phylogenetic analyses of environmental fungal ITS sequences toexamine both the overall fungal diversity and the diversity of Geomyces in multiple cave microhabitats. Knowledge of the specific habitat of G. destructanswill allow us to determine the likelihood for saprophytic growth within caves and estimate the role that subsidies can play in disease ecology. Indeed,disease modeling indicates that an environmental subsidy for the growth of G. destructans increases the likelihood of bat host extinction events.37. Cellulose acting enzymes of the white-rot fungus Dichomitus squalens: expression of the genes and characterization of the enzymes. JohannaRytioja, Aila Mettälä, Kristiina Hildén, Annele Hatakka, Miia Mäkelä. Food and Environmental Sciences, University of Helsinki, Helsinki, Finland.Plant biomass is a diverse raw material that has great potential to be exploited e.g. in second generation biorefinery applications. In order to overcomethe economic and technological thresholds in biomass utilization, novel cellulose attacking enzymes and optimal enzyme mixtures are needed. Thesynergistic effect of cellulose hydrolyzing enzymes, namely endoglucanases (E.C. 3.2.1.4), cellobiohydrolases (CBH, E.C. 3.2.1.91) and b-glucosidases (E.C.3.2.1.21), during cellulose degradation is a well-defined phenomenon, which has also been reported for cellulose oxidizing enzymes. The fungal producedoxidative enzymes related to cellulose degradation include cellobiose dehydrogenases (CDH, E.C. 1.1.99.18) and the proteins of glycoside hydrolase (GH)family 61 (www.cazy.org).Basidiomycetous white-rot fungi are able to efficiently degrade all the wood polymers, i.e. cellulose, hemicelluloses and lignin. Their lignin-modifyingoxidoreductases (peroxidases and laccases) are rather well characterized, whereas their cellulose acting enzymes (CAZymes) have so far gained lessattention. In the large screening of hydrolytic enzymes of basidiomycetous fungi from the Fungal Biotechnology Culture Collection (FBCC, University ofHelsinki), the white-rot fungus Dichomitus squalens was found to produce high cellulolytic activity and appeared as a promising source of novel CAZymes.In this work, the expression of selected hydrolytic and oxidative CAZyme encoding genes (cdh, four cbhs, five putative gh61s) was followed withquantitative real-time RT-PCR during the growth of D. squalens on solid Norway spruce (Picea abies) wood and in semi-solid microcrystalline cellulose(Avicel) -peptone liquid medium. The enzymatic activities of cellulases and xylanase as well as lignin-modifying oxidoreductases were measured from thesemi-solid cultures. In addition, CBHI and CDH enzymes of D. squalens were purified and characterized.130

FULL POSTER SESSION ABSTRACTS38. Metabolomics of growth and type B trichothecenes production in Fusarium graminearum. L. Legoahec 1 , V. Atanasova-Penichon 1 , N. Ponts 1 , C.Deborde 2,3 , M. Maucourt 3,4 , S. Bernillon 2,3 , A. Moing 2,3 , F. Richard-Forget 1 . 1) 1INRA, UR1264 MycSA, 71 avenue Edouard Bourlaux, BP81, F-33140 Villenaved’Ornon, France; 2) INRA, UMR1332 Fruit Biology and Pathology, 71 avenue Edouard Bourlaux, BP81, F-33140 Villenave d’Ornon, France; 3) MetabolomeFacility of Bordeaux Functional Genomics Center, IBVM Centre INRA de Bordeaux, F-33140 Villenave d'Ornon, France; 4) Univ. Bordeaux, UMR1332 FruitBiology and Pathology, Centre INRA de Bordeaux, F-33140 Villenave d'Ornon, France.The plant fungal pathogen Fusarium graminearum can produce type B trichothecenes, a family of sesquiterpene molecules with toxic properties uponhuman or animal ingestion. Deoxynivalenol, or DON, and its acetylated forms belong to this family of secondary metabolites and are frequentcontaminants of cereals worldwide. The biosynthesis of trichothecenes initiates with the condensation of two molecules of farnesyl pyrophosphate, at theend of the mevalonate pathway in Fusarium, and is under the control of various factors such as the redox parameters of the environment or the carbonsource. For example, supplementing liquid submerged cultures of F. graminearum with caffeic acid, a phenolic acid with known antioxidant properties,reduces the accumulation of DON and its acetylated forms in the medium. Such a result, however, gives a partial glimpse of the effect of phenolic acids,from the trichothecene production point of view only. The present study analyzes F. graminearum metabolome in conditions when DON and its acetylatedforms are produced. Liquid chromatography coupled with mass spectrometry and proton nuclear magnetic resonance were used to characterize themetabolites produced by the fungus, secreted in the culture medium or not, over the course of 14 days. Fifty-two polar and semi-polar metabolites wereidentified in the culture medium, i.e., the exo-metabolites, and/or in the mycelium, i.e., the endo-metabolites, comprising amino acids and derivatives,sugars, polyketides, and terpenes including trichothecenes and DON precursors. Sample composition varied over time in terms of primary metabolites aswell as secondary metabolites. Data analysis further revealed correlations, positive or negative, between metabolic pathways. In the presence of caffeicacid, metabolomic profiles were modified, counting those resulting from primary metabolism even though fungal biomass production was not affected bythe treatment. Several metabolites affected by the treatment were identified for both the exo- and endo-metabolome, in particular DON and itsprecursors. For the first time, these results expose a unique outlook of a hidden aspect of Fusarium’s response to antioxidant treatment.39. Diversity of telomeric sequences and telomerase RNA structures within Ascomycetes. Xiaodong Qi, Yang Li, Dustin P. Rand, Julian J-L Chen.Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ, 85287-1604.Telomeres are specialized DNA-protein complexes that cap chromosome ends. Telomeric DNA is composed of repetitive short sequences synthesized bytelomerase, an RNA-containing DNA polymerase. The integral telomerase RNA (TER) contains telomerase provides a short template for telomeric DNAsynthesis and two highly conserved structural elements essential for enzymatic function. Fungal telomerase from budding and fission yeasts has beenstudied extensively. We have recently developed Neurospora crassa as a new fungal model organism for telomere and telomerase studies, and haveidentified TER structural domains highly conserved in vertebrate and Pezizomycotina, but not in budding yeasts (Qi et al. 2012). N. crassa telomeraseprocessively synthesizes the (TTAGGG)n telomere repeats, an attribute conserved in vertebrate but not yeast telomerases. In contrast, both budding andfission yeast telomerases synthesize irregular telomere repeats non-processively. Two structural elements of TER, the template-pseudoknot and the threewayjunction (TWJ) domain, are conserved in vertebrates, Pezizomycotina as well as Taphrinomycotina. Both of these elements are necessary fortelomerase activity in vitro for Pezizomycotina and Taphrinomycotina telomerases, while the TWJ is dispensable for budding yeast telomerase.Furthermore, spliceosome-mediated TER 3’-end processing is conserved in Pezizomycotina and Taphrinomycotina, but not in budding yeasts. Incomparison, the budding yeast (e.g. S. cerevisiae) TER employs a nuclease-mediated mechanism for the 3’ end processing. Our results indicate thatPezizomycotina telomerase preserved ancestral features that budding and fission yeast species lost during evolution and supports N. crassa as an excellentmodel for the study of telomere and telomerase. (Reference: Qi, X., Y. Li, S. Honda, S. Hoffmann, M. Marz. A. Mosig, J.D. Podlevsky, P.F. Stadler, E. Selkerand J.J.-L. Chen (2012) The common ancestral core of vertebrate and fungal telomerase RNAs. Nucleic Acids Research 40: doi:10.1093/nar/gks980.).40. Characterizing a putative three-step formaldehyde oxidation pathway in Neurospora crassa. Ethan Addicott 1 , Kolea Zimmerman 2 , Anne Pringle 2 . 1)Faculty of Arts and Sciences, Harvard College, Harvard University, Cambridge, MA; 2) Organismic and Evolutionary Biology, Harvard University, Cambridge,MA.Using bioinformatic analyses, we identified 13 Neurospora genes that code for putative secreted-proteins. One of these proteins, NCU01056 - a proposedS-(hydroxymethyl)glutathione synthase, is implicated in a highly conserved formaldehyde oxidation pathway involving two other genes, NCU06652 - anNAD and GSH dependent formaldehyde dehydrogenase and NCU0173- an S-formylglutathione hydrolase. Knockout strains for the three genes in thispathway were obtained from the FGSC and confirmed by PCR. We conducted standard phenotypic assays on the three knock-outs and WT controls,including growth morphology, growth rate, and mating ability. Additionally, growth in the presence of methanol, the compound just upstream offormaldehyde in the pathway, was tested by biomass and flow cytometry. Two key observations were made: (1) NCU06652 knockouts showed significantgrowth defects compared to the WT (2) Knockouts for NCU01056 (hypothesized to be upstream of the critical enzyme) showed increased pigmentation onSC media (3) NCU6652 knockouts germinated significantly slower than other strains in the presence of methanol compared to a control treatment. Thedata suggest NCU06652 is involved in the critical oxidation step of the pathway and that the absence of NCU01056 may induce stress, which points to itsrole in the formation of a formaldehyde-glutathione complex, immediately upstream of NCU06652. The fact that NCU01056 codes a secreted protein maysuggest that N. crassa may detoxify formaldehyde extracellularly or in membrane-bound vesicles. Further exploration will involve determining a doseresponsecurve for formaldehyde, confirming the localization of the proteins, and investigating the GSH balance in each of the strains.41. Nitrate assimilation in Neurospora crassa. Oleg Agafonov, Tina Marie Monge Are, Peter Ruoff. Centre for Organelle Research, University of Stavanger,Stavanger, Norway.Nitrogen is one of the essential components for a variety of cellular elements. Regulation of nitrogen assimilation can be critical for the evolutionaryadvantage of an organism and it has been extensively studied in filamentous fungi Neurospora crassa. Nitrate is an important source of inorganic nitrogenfor N. crassa, but it is not utilized unless favored nitrogen sources such as ammonium, glutamine or glutamate are absent in the environment. It wasshown that nitrate is transported into the cell by high affinity transporter, NIT10, where it is stepwise reduced, first, by nitrate reductase, NIT3 to nitrite,and then by nitrite reductase, NIT6 to ammonia, which is then converted to organic nitrogen in a form of glutamate, making it available for furtherutilization by the cell.Although biochemical pathways of nitrate assimilation have been extensively studied, there is a certain disagreement in literature about therequirement of functional nitrate reductase activity for nitrate uptake. In the paper by Schloemer and Garrett, 1974, it was shown that nitrate transport isnot dependent upon nitrate reduction. However, later Unkles et. al., 2004, concluded that functional nitrate reductase is required for the nitrateaccumulation in Neurospora crassa.The goal of this work was to investigate nitrate assimilation and involvement of nitrate reductase in this process in N. crassa. Nitrate disappearance from27th Fungal Genetics Conference | 131

FULL POSTER SESSION ABSTRACTSthe media and weight of the mycelium were measured in nit-10 (KO) and nit-3 (KO) strains in comparison to a wild type strain. The nit-10 (KO) mutant wasnot able to grow on nitrate as a sole nitrogen source, and no nitrate depletion from the media was observed. Therefore, it was concluded that NIT10 is theonly active transporter. The accumulation of nitrate in the mycelium was measured and it was found that in the nit-3 (KO) mutant it was 9 times higherand in the nit-10 (KO) mutant 3 times higher than in the wild type strain. Therefore, we concluded that nitrate reductase is not required for the nitratetransport.42. Protein kinases affecting glycogen accumulation and likely regulating the glycogen synthase phosphorylation status in Neurospora crassa. T.Candido 1 , A. P. Felício 2 , R. Gonçalves 1 , F. Cupertino 1 , F. Freitas 1 , M. C. Bertolini 1 . 1) UNESP - IQ - Araraquara, Araraquara, SP., Brazil; 2) Departamento deGenética e Evolução, UFSCar, São Carlos, SP, Brazil.The ability to sense and respond appropriately to environmental changes is required for all living organisms and reversible phosphorylation of proteinsmediated by protein kinases plays a key role in this aspect. In this work we describe the results of a screen aimed to identify protein kinases regulatingglycogen metabolism in Neurospora crassa. The glycogen synthase (GS) and glycogen phosphorylase (GP), the regulatory enzymes in glycogen synthesisand degradation processes, respectively, are highly regulated by phosphorylation, however the protein quinases that phosphorylate them in N. crassahave not been identified. In this work, a set of mutant strains individually knocked-out in genes encoding proteins kinases was used. The glycogen levelswere quantified under normal growth temperature (30°C) and under heat stress (45°C). From 84 mutant strains, 37 strains presented glycogenaccumulation profile different from the wild-type strain suggesting that the missing protein kinase is implicated in glycogen metabolism control. Amongthe protein kinases selected most are Ser/Thr protein kinases, and it is important to mention proteins already characterized as regulators of glycogenmetabolism, such as the Saccharomyces cerevisiae Pho85 and Snif1 proteins. The GSN activity was quantified in the selected strains grown under normaltemperature (30°C) and under heat stress (45°C) in the presence and absence of the allosteric activator glucose-6-phosphate (G6P). The ratio -/+ G6P isconsidered as an index of phosphorylation, lower levels correlating with higher phosphorylation. Some protein kinases were implicated in glycogenmetabolism control by likely influencing the GSN phosphorylation status. The GSN phosphorylation profile in the mutant strains were analyzed in 2D-PAGEfollowed by Western blot using polyclonal GSN antibody. Some mutant strains showed phosphorylation profile different from the wild-type strain and theresults revealed putative proteins kinases not yet described as able to phosphorylate GSN. The expression of glycogen synthase (gsn) and glycogenphosphorylase (gpn) genes was analyzed by qRT-PCR in the mutant strains and the results showed that some protein kinases regulate the expression ofboth genes. Supported by FAPESP and CNPq.43. Endogenous ergothioneine is required for wild type levels of Neurospora crassa conidiogenesis and conidial survival, but does not protect againstuv-induced kill or mutagenesis. Lynn Epstein, Marco Bello, John Mogannam. Plant Pathology, University of California, Davis, CA. 95616-8680.Ergothioneine (EGT) is a histidine derivative that apparently is only synthesized by fungi (except in the Saccharomycotina), and by some bacteria in theCyanophyta and Actinomycetales. Although plants and animals do not synthesize EGT, they acquire it from the environment; EGT is concentrated in animalcells with an EGT-specific transporter. Bello et al. (2012, Fungal Genet Biol 49:160) showed that the concentration of EGT is 5x greater in Neurosporacrassa conidia than in mycelia, and that growth of strain NcDEgt-1 with a knockout in gene NCU04343 is indistinguishable from the wild type. Toinvestigate the function of EGT, wild type (Egt+) and NcDEgt-1 were crossed and six Egt+ and six Egt- sib strains were analyzed. Compared to the Egt+ sibs,Egt- sibs had a highly significant reduction (59 + 6%, + SE) in the number of conidia produced on Vogel’s agar; the detransformed mean of the Egt- sibs was1.5 x 10 5 conidia/cm 2 with a detransformed 95% confidence interval (CI 95) from 1.2 x 10 5 to 1.8 x 10 5 conidia/cm 2 whereas the Egt+ sibs had a mean of 3.6 x10 5 conidia/cm 2 and a CI 95 from 2.9 x 10 5 to 4.6 x 10 5 conidia/cm 2 . The concentration of EGT in wild type conidia did not increase with increasing exposureto light during conidiogenesis. Seven-day-old conidia were stored at 30 °C at 97% and 51% relative humidity (RH) for a time course to either 17 or 98 days,respectively. Life expectancies (LE) were calculated from logistic curves fitted to percentage germination as a function of days in storage in two trials. At97% RH, Egt+ sibs had a LE = 11.0 + 0.2 days whereas Egt- sibs had a highly significantly lower LE = 8.4 + 0.2 days, a 23 + 8% reduction. At 51% RH, Egt+ sibshad a LE = 71 + 1 days whereas Egt- sibs had a highly significantly lower LE = 58 + 1 days, an 18 + 3% reduction. We tested the hypothesis that EGT protectsagainst uv-induced kill or mutagenesis. There were no significant differences between the germinability of Egt+ and Egt- sibs after exposure to 0 to 400Joules/m 2 of 254 nm light. There also were no significant differences between the Egt+ and Egt- sibs in the mtr mutation rate to fluorophenylalanineresistance after exposure of conidia to 0 to 400 Joules/m 2 of 254 nm light. Consequently, our in vivo analysis indicates that EGT does not protect againstuv-induced kill or mutagenesis.44. Thiolutin inhibits protein turnover in Neurospora and yeast. Linda Lauinger, Michael Brunner, Axel Diernfellner. BZH, Heidelberg, Germany.Proteasome inhibitors are a powerful tool for the characterization of proteins in vivo. In yeast as well as in filamentous fungi, however, the availableproteasome inhibitors, like e.g. MG132 do not function due to the barrier posed by the cell wall of the organisms and an efficient evacuation of themolecules out of the cells. The dithiole thiolutin has been shown to be a potent inhibitor of RNA polymerases in prokaryotes and fungi. In the filamentousfungus Neurospora crassa, thiolutin efficiently suppresses transcription, indicating that the drug is cell permeable and not subject to a significant efflux bythe multidrug resistance system. Our data indicate that thiolutin also significantly inhibits protein turnover. Concomitant with the increase in proteinstability after treatment with thiolutin, we observe an accumulation of ubiquitinated protein species. Thus, our findings suggest that thiolutin may be apleiotropic inhibitor suppressing both, RNA polymerase as well as the proteasomal activity.45. Characterization of a Phanerochaete chrysosporium glutathione transferase reveals a novel structural and functional class with ligandin propertiesfor wood extractive molecules. Yann Mathieu 1,2,6 , Pascalita Prosper 3,4 , Marc Buée 2 , Stéphane Dumarçay 5 , Frédérique Favier 3,4 , Eric Gelhaye 1,2 , PhilippeGérardin 5 , Luc Harvengt 6 , Jean-Pierre Jacquot 1,2 , Tiphaine Lamant 1,2 , Edgar Meux 1,2 , Sandrine Mathiot 3,4 , Claude Didierjean 3,4 , Melanie Morel 1,2 . 1)Université de Lorraine, IAM, UMR 1136, IFR 110 EFABA, Vandoeuvre-les-Nancy, F-54506, France; 2) INRA, IAM, UMR 1136, Vandoeuvre-les-Nancy, F-54506, France; 3) Université de Lorraine, CRM2, UMR 7036, Vandoeuvre-les-Nancy, F-54506, France; 4) CNRS, CRM2, UMR 7036, Vandoeuvre-les-Nancy, F-54506, France; 5) Université de Lorraine, LERMAB, EA 1093, Vandoeuvre-les-Nancy, F-54506, France; 6) Laboratoire de biotechnologie, Pole Biotechnologieet Sylviculture Avancée, FCBA, Campus Foret-Bois de Pierroton, 33610 Cestas, France.Glutathione transferases (GSTs) form a superfamily of multifunctional proteins with essential roles in cellular detoxification processes. A new fungalspecific class of GST has been highlighted by genomic approaches. The biochemical and structural characterization of one isoform of this class inPhanerochaete chrysosporium revealed original properties. The three-dimensional structure showed a new dimerization mode and specific features bycomparison with the canonical GST structure. An additional b-hairpin motif in the N-terminal domain prevents the formation of the regular GST dimer andacts as a lid, which closes upon glutathione binding. Moreover, this isoform is the first described GST that contains all secondary structural elements,including helix a4’ in the C-terminal domain, of the presumed common ancestor of cytosolic GSTs, i.e. glutaredoxin 2. A sulfate binding site has been132

FULL POSTER SESSION ABSTRACTSidentified close to the glutathione binding site and allows the binding of 8-anilino-1-naphtalene sulfonic acid (ANS). Competition experiments betweenANS, which has fluorescent properties, and various molecules, showed that this GST binds glutathionylated and sulfated compounds, but also woodextractive molecules such as vanillin, chloro nitrobenzoic acid, hydroxyacetophenone, catechins and aldehydes in the glutathione pocket. This enzymecould thus function as a classical GST through the addition of glutathione mainly to phenethyl isothiocyanate but alternatively and in a competitive way, itcould also act as a ligandin of wood extractive compounds. These new structural and functional properties, lead us to propose that this GST belongs to anew class that we name GSTFuA for Fungal specific GST class A.46. Secretome analysis of Trichoderma harzianum cultivated in the presence of Fusarium solani cell wall or glucose. Marcelo HS Ramada 1,3 , Andrei SSteindorff 1 , Carlos Bloch Jr. 3 , Cirano J Ulhoa 2 . 1) Brasilia University, Cell Biology Department, Brasilia, DF, Brazil; 2) Federal University of Goias, BiochemistryDepartment, Goiania, GO, Brazil; 3) EMBRAPA CENARGEN, Mass Spectrometry Laboratory, Brasilia, DF, Brazil.Trichoderma harzianum is a fungus well known for its potential as a biocontrol agent of many fungal phytopathogens. The aim of this study was toevaluate the potential of T. harzianum ALL42 to control Fusarium solani, a phytopathogen fungus that causes several losses in common bean and soy cropsin Brazil and to evaluate the secreted proteins of T. harzianum ALL42 when its spores were inoculated and incubated in culture media supplemented (TLE)or not (MM) with nitrogen sources and in the presence or not of F. solani cell walls (FsCW). In the absence of FsCW, the media were supplemented withglucose (GLU). T. harzianum was able to control the phytopathogen growth and started to sporulate in its area after 7 days in a dual culture assay,indicating that it had successfully parasitized the host. T. harzianum was able to grow in TLE+FsCW, MM+FsCW, TLE+GLU, but unable to grow in MM+GLU.Protein quantification showed that TLE+FsCW and MM+FsCW had 45 and 30 fold, respectively, more proteins than TLE+GLU, and this difference wasobserved in the bidimensional gels, as the two supernatants from media supplemented with FsCW had around 200 spots and the one supplemented withglucose only had 18. TLE+FsCW and MM+FsCW had above 80% of spot similarity. A total of 100 proteins were excised from all three conditions andsubmitted to mass spectrometry analysis. 85 out of 100 proteins were identified. The only protein observed in all three conditions is a small protein, calledepl1, involved in eliciting plant-response against phytopathogens. An aspartic protease, previously described as related to mycoparasitism, was only foundwhen T. harzianum was grown with glucose. Gene expression was evaluated and confirmed the gel results. In the media supplemented with FsCW,different hydrolases like chitinases, beta-1,3-glucanases, glucoamylases, alpha-1,3-glucanases, and proteases were identified. Some proteins like a smallcystein-rich, alpha-L-arabinofuranosidase and NPP1, with no known function in mycoparasitism were also identified. T. harzianum ALL42 is able to inhibitthe growth and parasitize F. solani and showed a complex and diverse arsenal of proteins that are secreted in response to the presence of the cell walls,with novel proteins not previously described in mycoparasitism studies.47. Analysis of carbon catabolite repression (CCR) during cellulase formation by Trichoderma reesei (Hypocrea jecorina) using two-dimensionaldifferential gel electrophoresis (2D-DIGE). Wellington Pedersoli, Lilian Castro, Amanda Antonieto, Vitor M. Faça, Roberto N. Silva. Biochemistry andImmunology, University of São Paulo, Ribeirao Preto, Sao Paulo, Brazil.The production of cellulases by Trichoderma reesei (Hypocrea jecorina) is fundamental for the production of second generation biofuels from cellulosicwastes. The complex of cellulases of fungi T. reesei is strongly induced by sophorose and cellulose and also antagonized by glucose. Thus, the objective ofthis study was to compare the differential secretome using 2D-DIGE of T. reesei at induction and repression conditions. The fungus T. reesei (QM9414)were grown in media containing 1% cellulose for 24, 48 and 72 hours and 1% glucose for 24 and 48 hours in a orbital shaker at 180 rpm at 28°C. Afterfiltration of each medium the supernatant was used for protein determination. Samples from each time in the different conditions were mixed inproportional amounts and submitted to isoelectric focusing in 18 cm strip of pH 4.0 - 7.0 and subsequently subjected to electrophoresis on twodimensionalgel - differential (2D-DIGE) for the separation of proteins. The comparative analysis PDQuest software (BioRad) 130 selected spotsdifferentially expressed, wherein the inhibitor glucose showed 41 distinct protein spots and inductor cellulose 89 spots. However, only the 34 spots and 63spots from glucose and cellulose respectively could be spotted. Identification of these spots was carried out using the mass spectrometer XEVO TQ-S(Waters). A total of 57 proteins were identified, 30 and 27 proteins from cellulose and glucose conditions respectively. The proteins identified from glucosecondition were amidases, proteases, isoamyl alcohol oxidases and protein belonging to Glycosyl Hydrolases 16, 17, 43 and 54. On the other hand, theproteins identified from cellulose condition were Glycosyl Hydrolases 3, 7, 54, 55, 72, two Cip 1 (envolved in the degradation of cellulose, but with nofunction described), two proteases and a protein responsible for the regulation of stress. Two other proteins, one of each condition were identified asunknown. Thus, the identification of these proteins will allow a better understanding of the mechanism formation of cellulases by T reesei and thuscontribute to improve the production of second generation ethanol. Supported by: FAPESP, CAPES and FAEPA-HC/USP-RP.48. Transposon-associated evolution of a fungal NRPS. Daniel Berry 1 , Carolyn Young 2 , Paul Dijkwel 1 , Barry Scott 1 . 1) Massey University, Palmerston North,New Zealand; 2) The Samuel Roberts Nobel Foundation, Ardmore, Oklahoma.Epichloë endophytes systemically colonise the aerial tissues of cool season grasses to form mutually beneficial symbiotic associations. A defining featureof these associations is fungal synthesis of a range of secondary metabolites that protect the host from biotic and abiotic stress. One key bioprotectivemetabolite is the insect feeding deterrent, peramine, which is synthesized by the two-module non-ribosomal peptide synthetase (NRPS) PerA. This NRPShas an A-T-C-A-M-T-R domain structure where the Adenylation-domains provide specificity for and activation of the two amino acid substrates, theThiolation-domains bind the reaction intermediates, the Condensation-domain catalyzes formation of the peptide bond, the Methylation-domainmethylates the substrate amino acid, and the Reductase-domain cyclizes and releases the finished product from the NRPS. The perA gene is foundexclusively within the Epichloë, where it is widespread among the different species, but peramine production is somewhat more discontinuous. We showthat transposon-mediated deletion of the R-domain present in some E. typhina and E. festucae peramine deficient isolates is associated with a change inthe predicted substrate specificity code of the first A-domain. Phylogenetic analysis of this domain groups the peramine negative isolates togetherwhereas the phylogeny based on the whole gene matches the species phylogeny. The recombination mechanism responsible for the evolution of thisnovel NRPS is still to be resolved.49. Velvet family control of penicillin production in Penicillium chrysogenum: PcVelB binding to isopenicillin N synthase suggests a novel regulatorymechanism. Sandra Bloemendal, Katarina Kopke, Birgit Hoff, Sarah Milbredt, Alexandra Katschorowski, Ulrich Kück. Christian Doppler Laboratory forFungal Biotechnology, Ruhr-University Bochum, Universitätsstr. 150, 44780 Bochum, Germany.The filamentous fungus Penicillium chrysogenum is the main industrial producer of the pharmaceutically relevant beta-lactam antibiotic penicillin. Allthree biosynthesis genes are found in a single cluster and the expression of these genes is known to be controlled by a complex network of globalregulators. It is supposed that subunits of the velvet complex, which were recently detected for P. chrysogenum, function as such global regulators,although the exact regulatory mechanism still have to be elucidated. Core components of this complex are PcVelA and PcLaeA, which regulate secondary27th Fungal Genetics Conference | 133

FULL POSTER SESSION ABSTRACTSmetabolite production, hyphal morphology, conidiation, and pellet formation [1]. Here we describe the characterization of PcVelB, PcVelC, and PcVosA asnovel subunits of this velvet complex. Using yeast two-hybrid analysis and bimolecular fluorescence complementation (BiFC), we demonstrate that allvelvet proteins are part of an interaction network. Functional analyses using single and double knockout strains generated by the FLP/FRT recombinationsystem [2] clearly indicate that velvet subunits have opposing roles in the regulation of penicillin biosynthesis and light-dependent conidiation. Moststrikingly, a direct interaction of PcVelB with an enzyme of the penicillin biosynthesis pathway, the isopenicillin N synthase was identified during yeast twohybridanalysis with PcVelB as bait. This surprising interaction was confirmed with BiFC in vivo, thereby localizing the interaction in dot-like structures inthe cytoplasm. Our discovery of a direct interaction of the isopenicillin N synthase with a subunit of the velvet complex implies a novel regulatorymechanism how enzymes of penicillin biosynthesis are regulated at the molecular level. The results provided here contribute to our fundamentalunderstanding of the function of velvet subunits as part of a regulatory network mediating signals responsible for morphology and secondary metabolism,and will be instrumental in generating mutants with newly derived properties that are relevant to strain improvement programs.[1] Hoff B, Kamerewerd J, Sigl C, Mitterbauer R, Zadra I, Kürnsteiner H, Kück U (2010) Eukaryot Cell: 9:1236-50[2] Kopke K, Hoff B, Kück U (2010) Appl Environ Microbiol 76:4664-4674.50. Genome mining reveals the evolutionary origin and biosynthetic potential of basidiomycete polyketide synthases. Gerald Lackner, Mathias Misiek,Jana Braesel, Dirk Hoffmeister. Pharmaceutical Biology, Friedrich-Schiller-University Jena, Germany.Polyketide biosynthesis is a rich source of pharmaceutically active secondary metabolites present in fungi. Besides lipid-lowering drug lovastatin, manyinfamous toxins are produced via this pathway. While abundant in Aspergillus, only few polyketides have been isolated from basidiomycetes. Highthroughput genome sequencing projects, however, now help estimate the genetic capacity of basidiomycetes to biosynthesize polyketide derivatives. Byinspection of 35 sequenced basidiomycete genomes we identified and annotated 111 iterative type I and three type III polyketide synthase (PKS) genes.Phylogenetic analyses of KS genes imply that all main families of fungal PKS had already evolved before the Ascomycota and Basidiomycota diverged. Acomparison of genomic data and metabolomic records shows that the number of polyketide genes surpasses the number of known polyketidesconsiderably. This work might serve as a guide for upcoming genomic mining projects to discover novel polyketide derivates from mushrooms.51. Engineering Cyclic Peptide Biosynthesis in Poisonous Mushrooms. Hong Luo, John S. Scott Craig, Robert M. Sgambelluri, Sung-Yong Hong, Jonathan D.Walton. Department of Energy Plant Research Laboratory, Michigan State University, E. Lansing, MI 48824, United States.Ninety percent of fatal mushroom poisonings are caused by alpha-amanitin and related bicyclic peptides found in some species of Amanita, Galerina,Lepiota, and Conocybe. We showed that the amatoxins (mainly amanitins) and related phallotoxins are synthesized on ribosomes in A. bisporigera and theunrelated mushroom G. marginata. The primary gene products are short (34-35 amino acid) proproteins that are initially processed by a dedicated prolyloligopeptidase. A genome survey sequence of A. bisporigera suggested that it has a repertoire of over 40 cyclic peptides, all produced on a singlebiosynthetic scaffold. Members of this extended gene family are characterized by conserved upstream and downstream amino acid sequences, includingtwo invariant proline residues, flanking a six to ten-amino acid “hypervariable” region that encodes the amino acids found in the mature toxins (orpredicted toxins). The evidence indicates that A. bisporigera has evolved a combinatorial strategy that could in principle biosynthesize billions of smallcyclic peptides. In order to study the other steps in amanitin biosynthesis, and to engineer novel cyclic peptides, we have developed a transformationstrategy for the amanitin-producing mushroom G. marginata. This first transformation method uses Agrobacterium-mediated transformation followed byhygromycin selection. Taking advantage of this platform, we are introducing artificial toxin genes that are deliberately designed to provide insights into thepathway. The synthetic genes include those that encode the cyclic octapeptide beta-amanitin, the heptapeptides phalloidin and phallacidin, examples ofthe toxin gene family known from A. bisporigera but not G. marginata, and randomly generated artificial sequences. Currently, thousands of transformantshave been generated through an efficient pipeline and the transformants are being analyzed for production of the expected products. If successful, thenovel peptides will be screened in a number of assays including RNA polymerase (the site of action of alpha-amanitin), membrane ion channels,pathogenic bacteria, and cancer cell lines.52. Spatial assessment of oxidative and enzymatic reactions in brown rotted wood. Jon R. Menke 1 , Jae San Ryu 2,5,6 , Gerald N. Presley 1 , Shona M. Duncan 1 ,Joel A. Jurgens 3 , Robert A. Blanchette 3 , Timothy R. Filley 4 , Kenneth E. Hammel 2,5 , Jonathan S. Schilling 1 . 1) Department of Bioproducts and BiosystemsEngineering, University of Minnesota, St. Paul, MN; 2) Department of Bacteriology, University of Wisconsin, Madison, WI; 3) Department of PlantPathology, University of Minnesota, St. Paul, MN; 4) Department of Earth and Atmospheric Sciences and the Purdue Climate Change Research Center,Purdue University, West Lafayette, IN; 5) Institute for Microbial and Biochemical Technology, U.S. Forest Products Laboratory, Madison, WI; 6) Eco-Friendliness Research Department, Gyeongsangnam-do Agricultural Research and Extension Services, Republic of Korea.Brown rot fungi are theorized to use coordinated free radical oxidations and enzymatic reactions to consume wood. Though likely incompatible in vitro, itis proposed these reactions occur concurrently during brown rot of wood. We mapped and then compared fungal growth, wood modifications related tonon-enzymatic mechanisms, and cellulase activity in thin spruce ‘wafers’ to investigate the degree of spatial coincidence of these reactions in wooddegraded by Postia placenta. Nearly coincident oxidative and enzymatic reaction fronts were observed behind the most advanced hyphal tips, suggesting afine-scale (likely sub-micron) spatial or biochemical extracellular mechanism may protect both hyphae and enzymes from oxidative stress. To furtherinvestigate a possible role of enzymatic reactions in the primary depolymerization of lignocellulose during brown rot, we have initiated a study totemporally assess the depth of penetration of an endoglucanase into wood cells during this process. Previous studies have used the marker proteinsinsulin (5.7 kDa), myoglobin (17.6 kDa), and ovalbumin (44.4 kDa); and immunocytochemical electron microscopy to demonstrate the ability of the twosmaller proteins to infiltrate the cell walls of rotted wood. The Postia placenta endoglucanase Cel5B (PpCel5B) has a theoretical molecular weight of 34.6kDa, which is considerably lower than the molecular weight of ovalbumin. Our approach involves using a polyclonal antibody raised against PpCel5Bheterologously expressed in Pichia pastorius. This antibody will be used to assess the extent to which a native brown rot endoglucanase is able topenetrate Pinus resinosa cells. Given the common supposition that pore size prevents brown rot fungal endoglucanases from accessing wood secondarywalls, even in late decay stages, this work will provide a direct assessment of enzyme ingress.53. Molecular biological basis for statin resistance in naturally statin-producing organisms. Ana Rems, Rasmus Frandsen. DTU Systems Biology, TechnicalUniversity of Denmark, Kongens Lyngby, Denmark.Secondary metabolites can be toxic to the organism producing them; therefore gene clusters for biosynthesis of secondary metabolites often includegenes responsible for the organism’s self-resistance to the toxic compounds. One such gene cluster is the compactin (ML-236B) cluster in Penicilliumsolitum. Compactin is an inhibitor of 3-hydroxy-3-methyl-glutaryl-CoA (HMG-CoA) reductase, and is used as a precursor for production of the cholesterolloweringdrug pravastatin. The compactin gene cluster includes two genes encoding proteins that may confer the self-resistance to compactin and its134

FULL POSTER SESSION ABSTRACTSsecretion [1]. The mlcD gene encodes a putative 'HMG-CoA reductase-like protein’, and mlcE encodes a putative efflux pump. However, the function ofthese two putative proteins has not yet been confirmed. We aim to elucidate the biological basis for compactin resistance in the compactin-producingorganism. A codon-optimized version of the mlcD gene was inserted into the Saccharomyces cerevisiae genome. The constructed yeast strain was testedfor sensitivity to lovastatin, a statin structurally similar to compactin, by growing the strain on media containing lovastatin. The strain showed an increasedresistance to lovastatin compared to the wild-type strain. Furthermore, we investigated if MlcD confers the resistance by functional complementation ofthe endogenous HMG-CoA reductases in S. cerevisiae. There are two isozymes of HMG-CoA reductase in yeast, HMG1 and HMG2, both involved in thesterol biosynthetic pathway, which leads to the synthesis of ergosterol. Following deletion of HMG1 and HMG2 genes in S. cerevisiae, we inserted the mlcDgene into the knockout mutants, and tested the resulted strains for sensitivity to lovastatin. The HMG1 and HMG2 knockout mutants were unable to growon minimal media and had an increased sensitivity to lovastatin on rich media. However, insertion of the mlcD gene restored the growth of the yeastmutants and increased their resistance to lovastatin. These results show that MlcD complements the activity of the deleted HMG-CoA reductases, enablingsynthesis of ergosterol in yeast. In addition MlcD confers statin resistance by being insensitive to the inhibiting effects of statins. Reference: [1] Abe Y.,Suzuki T., Ono C., Iwamoto K., Hosobuchi M., Yoshikawa H. Mol Genet Genomics 2002, 267, 5:636-46.54. Molecular genetic characterization of secondary metabolism pathways in Asperillus species. Clay Wang 1 , Yiming Chiang 1 , Nancy Keller 3 , KennethBruno 4 , Scott Baker 4 , Chun jun Guo 1 , James Sanchez 1 , Benjamin Knox 4 , Alexandra Soukup 3 , Jin Woo Bok 3 , Manmeet Ahuja 2 , Ruth Entwistle 2 , Liz Oakley 2 ,Shu-lin Chang 1 , Hsu-Hua Yeh 1 , Mike Praseuth 1 , Berl Oakley 2 . 1) Pharma Sci & Chemistry, Univ Southern California, Los Angeles, CA; 2) Department ofMolecular Biosciences, University of Kansas; 3) Department of Medical Microbiology and Immunology and Department of Bacteriology, University ofWisconsin Madison; 4) Pacific Northwest National Laboratory.Advances in next generation DNA sequencing have provided a large number of fungal genome sequences in public databases. Within these genomes arelarge numbers of cryptic secondary metabolism pathways. Data will be presented where we use a comparative genomics approaches to identify theproducts of these cryptic pathways. Next we use a gene knock out approach to create mutants followed by isolation and characterization of intermediatesand shunt products. Using this approach we have been able to identify the products of a meroterpenoid pathway in A. terreus.55. A branched biosynthetic pathway is involved in production of roquefortine and related compounds in Penicillium chrysogenum. Hazrat Ali 1,2 , MarcoRies 3 , Jeroen Nijland 1,2 , Peter Lankhorst 4 , Thomas Hankemeier 3,5 , Roel Bovenberg 4,6 , Rob Vreeken 3,5 , Arnold Driessen 1,2* . 1) Molecular Microbiology,Groningen Biomolecular Sciences and Biotechnology Institute, Zernike Institute for Advanced Materials, University of Groningen, The Netherlands; 2)Kluyver Centre for Genomics of Industrial Fermentations, Julianalaan 67, 2628BC Delft, The Netherlands; 3) 3Division of Analytical Biosciences,Leiden/Amsterdam Center for Drug Research, Leiden University, The Netherlands; 4) DSM Biotechnology Center, Alexander Fleminglaan 1, 2613 AX Delft,The Netherlands; 5) Netherlands Metabolomics Centre, Leiden University, Leiden, The Netherlands; 6) Synthetic Biology and Cell Engineering, GroningenBiomolecular Sciences and Biotechnology Institute, University of Groningen, The Netherlands.Profiling and structural elucidation of secondary metabolites produced by the filamentous fungus Penicillium chrysogenum and derived deletion strainswere used to identify the various metabolites and enzymatic steps belonging to the roquefortine/meleagrin pathway. Major abundant metabolites of thispathway were identified as histidyltryptophanyldiketopiperazine (HTD), dehydrohistidyltryptophanyldiketopiperazine (DHTD), roquefortine D,roquefortine C, glandicoline A, glandicoline B and meleagrin. Specific genes could be assigned to each enzymatic reaction step. The nonribosomal peptidesynthetase RoqA accepts histidine and tryptophan as substrates leading to the production of the diketopiperazine HTD. DHTD, previously suggested to bea degradation product of roquefortine C, was found to be derived from HTD involving the cytochrome P450 oxidoreductase RoqR. Thedimethylallyltryptophan synthetase RoqD prenylates both HTD and DHTD yielding the products roquefortine D and roquefortine C, respectively. This leadsto a branch in the otherwise linear pathway. Roquefortine C is subsequently converted into meleagrin with glandicoline A and B as intermediates, involvingtwo monooxygenases (RoqM and RoqO) and a methyltransferase (RoqN). It is concluded that roquefortine C and meleagrin are derived from a branchedbiosynthetic pathway rather than a linear pathway as suggested in literature.56. A biosynthetic gene cluster for the antifungal metabolite phomenoic acid in the plant pathogenic fungus, Leptosphaeria maculans. Candace Elliott 1 ,Damien Callahan 2 , Daniel Schwenk 3 , Markus Nett 4 , Dirk Hoffmeister 3 , Barbara Howlett 1 . 1) School of Botany, University of Melbourne, Melbourne,Australia; 2) Metabolomics Australia, School of Botany, The University of Melbourne, Victoria 3010, Australia; 3) Friedrich-Schiller-Universität, DepartmentPharmaceutical Biology at the Hans-Knöll-Institute, Beutenbergstrasse 11a, 07745 Jena, Germany; 4) Leibniz Institute for Natural Product Research andInfection Biology e.V., Hans-Knöll-Institute, Beutenbergstrasse 11a, 07745 Jena, Germany.Phomenoic acid, a long chain aliphatic carboxylic acid, is a major metabolite produced by Leptosphaeria maculans, a fungal pathogen of Brassica napus(canola). Early biosynthetic studies suggested that the methyl group derived from S-adenosylmethionine (SAM), whereas the incorporation pattern of[13C] acetate suggested a polyketidic origin of the linear portion of phomenoic acid (Devys et al., 1984). We have used domain modelling to predict acandidate polyketide synthase (PKS) for phomenoic acid biosynthesis. Of the 15 predicted polyketide synthases (PKS) in the L. maculans genome, sevenwere reducing with the appropriate domains (KS - keto-synthase; AT - acyltransferase; DH - dehydratase; MT- methyltransferase; ER - enoylreductase; KR -ketoreductase; ACP- acyl carrier protein) for the biosynthesis of phomenoic acid. The most highly expressed of these seven genes, PKS2, was silenced to10% of that of wild type levels and the resultant mutant produced 25 times less phomenoic acid than the wild type did, indicating that PKS2 is involved inphomenoic acid biosynthesis. This gene is part of a cluster and nearby genes are co-regulated. A two-fold reduction in the expression of the adjacenttranscriptional regulator C6TF, led to at least a 20-fold reduction in expression of PKS2, as well as of other genes in the cluster (P450, YogA, RTA1 andMFS), but not of the adjacent ChoK or a hypothetical gene (Hyp). This down-regulated mutant also showed a marked reduction in phomenoic acidproduction. Phomenoic acid is toxic towards another canola pathogen Leptosphaeria biglobosa ‘canadensis’, but L. maculans and to a lesser extent thewheat pathogen, Stagonospora nodorum are more tolerant. Phomenoic acid may play a role in allowing L. maculans to outcompete other fungi in itsenvironmental niche.57. Exploring and Manipulating Pleuromutilin Production. Patrick M Hayes 1 , Russell J Cox 2 , Andy M Bailey 1 , Gary D Foster 1 . 1) School of BiologicalSciences, University of Bristol, Woodland Road, Bristol BS8 1UG, UK; 2) School of Chemistry, University of Bristol, Bristol BS8 1TS, UK.Antibiotic resistance has arisen in a significant number of human pathogens, antibiotic discovery and development is, however, currently not keepingpace. This has lead to the reinvestigation of some naturally produced antibiotic compounds which act in a manner that avoids common resistancemechanisms. Pleuromutilin is one such compound with activity against bacteria such as Methicillin Resistant Staphylococcus aureus (MRSA). Pleuromutilinis generally synthesised at a low titre by its native producer Clitopilus passeckerianus and as such research into its biosynthesis may enable yield increases.This project has taken a multifaceted approach to manipulate the Pleuromutilin biosynthetic gene cluster in a variety of fungal organisms. Within C.27th Fungal Genetics Conference | 135

FULL POSTER SESSION ABSTRACTSpasseckerianus gene silencing is being used to interfere with specific stages of biosynthesis, silenced transformed lines have been analysed bothchemically, to determine the impact of silencing on the metabolome of the organism, and via QRT-PCR, to determine the impact of silencing on thetranscription of the gene cluster. Heterologous expression of the entire gene cluster is being evaluated and analysed in the model basidiomyceteCoprinopsis cinerea. Engineered expression of the biosynthetic genes is being performed in the industrially relevant ascomycete Aspergillus oryzae to buildthe pathway in a stepwise manner. Progress in each of these areas will be presented.58. Improvement of Monascus pilosus for the production of functional foods by overexpression of the laeA gene. In H. Lee, Sang S. Lee, Jin H. Lee. Dept.of Advanced Fermentation Fusion Science & Technology, Kookmin Univ, Seoul, South Korea.Filamentous fungi Monascus species have been used to ferment rice producing red mold rice (RMR). They produce several bioactive compounds duringfermentation, however, they should have a potential to produce other bioactive compounds considering that most of fungi has many silent secondarymetabolite (SM) gene clusters. Therefore, we thought that Monascus species could be improved for functional food production by activation of such silentSM gene clusters. We overexpressed the laeA gene that is known to encode a global positive regulator of secondary metabolism under the alcA promoterin Monascus pilosus. An OE::laeA transformant produced more secondary metabolites including ones not detected under an uninduced condition. RMRfermented with the Monascus pilosus OE::laeA contained 7 times more monacolin K, a cholesterol lowering agent, than non-transformants increasing from2.45 to 15.59 mg/kg. In addition, the production of pigments was remarkably increased and antioxidant activity was increased as well. This study suggeststhat Monascus species that are important industrial fermentative fungi in Asia could be improved for the production of functional foods by overexpressionof the laeA gene.59. Molecular genetics studies on secondary metabolism in Chaetomium globosum reveal involvement of aureonitol and chaetoglobosins in generegulation and sexual reproduction. Takehito Nakazawa, kan'ichiro Ishiuchi, Satoru Sugimoto, Yasutaka Gotanda, Michio Sato, Hiroshi Noguchi, KenjiWatanabe. Pharmaceutical Sciences, University of Shizuoka, Shizuoka, Japan.Chaetomium globosum has been reported to produce many natural products, secondary metabolites, with complex structures biosynthetic pathways ofwhich are very interesting to be elucidated such as aureonitol, chaetoglobosins, chaetocins and chaetoviridins. Recently, we developed molecular geneticssystems for understanding the secondary metabolism in this fungus: High frequency gene targeting by Cg.ligD disruption and the pyrG marker recycling.Then, we identified biosynthetic gene clusters for various natural products that had been isolated from C. globosum, and analyzed their biosyntheticmechanisms/pathways. We also obtained new natural products by changing an epigenetic regulation. During these studies, we found that some ofmutations in biosynthetic genes allowed us to obtain interesting phenotypes: drastic changes in secondary metabolism profiles and defects in productionof sexual spores. Here, we present that aureonitol and chaetoglobosins would play a critical role of controlling the productivity of secondary metabolitesand producting of sexual spores, respectively. A mutated gene of artH responsible for biosynthesizing aureonitol exhibits activating chaetoviridinsbiosynthesis and inactivating chaetoglocin A biosynthesis. A qRT-PCR analysis shows that transcriptional expressions of biosynthetic genes forchaetoviridins are activated, whereas those for chaetoglocins are inactivated by the mutated artH gene. Supplementation with aureonitol toDartH strain isobserved to inactivate chaetoviridins biosynthesis and transcriptional expression of their biosynthetic gene cluster. These results strongly suggest thataureonitol could be involved in transcriptional regulation of secondary metabolism in C. globosum. On the other hand, we also find chaetoglobosins isessential for production of sexual spores: Mutations in its biosynthetic genes clearly impair meiotic process. It is anticipated that chaetoglobosins areinvolved in meiotic process, because the mutants don’t affect formation of fruiting bodies (perithecia). Previously, chaetoglobosins were shown to inhibitactin polymerization in vitro. Therefore, chaetoglobosins would play a role of regulating actin or actin-related protein in C. globosum, which has beenreported to be associated with regulation of meiotic process as well as morphogenesis in fungi.60. Identification of T. asperellum CAZyme genes. Lasse Bech 1 , Morten Nedergaard Grell 1 , Peter Kamp Busk 1 , Hai Zhao 2 , Lene Lange 1 . 1) Section forSustainable Biotechnology, Department of Biotechnology, Chemistry and Environmental Engineering, Aalborg University Copenhagen, Denmark; 2)Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, PR China.To understand the complexity of enzymatic hydrolysis of plant cell walls and to discover new enzymes, one approach is to analyze cell wall degradingenzymes (CWDEs) in the transcriptome of plant degrading fungi like species of the genus Trichoderma. The Trichoderma spp. are ubiquitous colonizers ofcellulosic materials and are often be found where decaying plant material is available as well as in the rhizosphere of plants. Trichoderma spp. aresuccessful colonizers of their habitats, which is shown both by their efficient utilization of the biomass as well as their secretion capacity for antibioticmetabolites and enzymes. This study shows the transcriptome of Trichoderma asperellum isolated from duckweed. Duckweed is an aquatic plant that hasbeen shown to clean eutrophic water reservoirs for the production of biomass, which can be used as feed, fertilizer and fuel through the biorefineryplatform. The fungus was optimized for the production of CWDEs, where more than 200 glucoside hydrolases from 47 different families were identified.Several group members exhibited novel traits such as larger residue differences in active site and substrate binding site, hence making them interestingsubject for expression and further characterization. This data was compared with the hydrolysis of duckweed by enzyme blend from T. asperellum. Theenzyme blend showed a promising degree of hydrolysis of duckweed indicating that T. asperellum is a candidate for on-site enzyme production for theenzymatic hydrolysis of certain duckweed species. The transcriptome data can further be used to map the expression of CWDEs under different conditionsthereby coming closer to understanding the relationship between CWDEs and the plant cell wall.61. Identification of a lactose permease of Trichoderma reesei that is required for cellulase gene expression. Christa Ivanova 1 , Jenny Bååth 2 , BernhardSeiboth 1,3 , Christian Kubicek 1,3 . 1) Institute of Chemical Engineering, University of Technology of Vienna, 1060 Vienna, Austria; 2) Lund University, SE-22100 Lund, Sweden; 3) Austrian Institute of Industrial Biotechnology (ACIB) GmBH c/o Institute of Chemical Engineering, University of Technology of Vienna,1060 Vienna, Austria.The disaccharide lactose has been shown to be a potent inducer of cellulases in T. reesei, and it is virtually the only soluble waste product that can beused for commercial enzyme production. To understand the complex regulatory mechanisms underlying cellulase induction by lactose, we performedcomparative transcriptomic analysis using oligonuclotide arrays. Among the 410 genes that were upregulated over four-fold on lactose, were all cellulases,cellulase-enhancing proteins, major hemicellulases and also 63 MFS- (major facilitator superfamily) -permeases. The MFS permeases are characterized by12-transmembrane helices and a well conserved motif between TMS (transmembrane segment) 2 and 9. In order to investigate the function of thesetransporters, we generated deletion strains in T. reesei. For this, the 14 most-upregulated transporter encoding genes were chosen. One of thesedisruptant strains showed strongly impaired growth on lactose and was therefore chosen for further analysis. The strain showed impaired growth onlactose, whereas growth on glucose, glycerol and cellobiose remained unaltered, suggesting that the transporter is required for lactose uptake. The strainwas devoid of cellulase gene expression during cultivation on lactose, whereas it formed cellulases upon incubation with sophorose suggesting that it is136

FULL POSTER SESSION ABSTRACTSinvolved in cellulase induction by this disaccharide. The deletion of the other 13 transporter encoding genes led to no reduction of growth on anycarbohydrate (including glucose and cellobiose), probably due to redundancy of their function. Our data show that lactose leads not only to the formationof a complete cellulase and hemicellulase system in T. reesei, but also to the transcription of a plethora of transporters likely associated with the uptake ofthe hydrolysis products.62. The diversity of the Mannosylerythritol lipids depends on the peroxisomal targeting of the Mannosylerythritol acyl transferases Mac1 and Mac2 inUstilago maydis. Johannes Freitag 1 , Julia Ast 1 , Uwe Linne 2 , Elisa Leisge 1 , Michael Bölker 1 , Björn Sandrock 1 . 1) Biology - Genetics, Philipps-UniversityMarburg, Marburg, Germany; 2) Chemistry, Philipps-University Marburg, Marburg, Germany.Under nitrogen starvation the smut fungus Ustilago maydis produces a bunch of secondary metabolites. Among these are the glycolipids Ustilagic acid(UA) and Mannosylerythritol lipid (MEL), which consist of a sugar moiety esterified with fatty acid side chains of variable length (from C2 - C18). Thebiosynthesis of UA is encoded by the UA gene cluster (11 genes). MEL production depends on the MEL gene cluster composed of the genes mat1, mmf1,mac1, emt1 and mac2. Deletion of mac1, mac2 or emt1 in U. maydis resulted in the complete loss of MELs. Medium-length fatty acids (C4-C14) are derivedfrom longer fatty acids (C16-C18) by partial peroxisomal b-oxidation. After bioinformatic analysis we have identified bona fide peroxisomal targetingsequences 1 (PTS1) at the C-termini of the two Mannosylerythritol lipid acyltransferases Mac1 and Mac2 but not in any other protein involved in thebiosynthesis of the MELs or the UAs. Here we show that Mac1 and Mac2 localize in peroxisomes, and that this localization depends on the PTS1 motifs.The analysis of glycolipid production by thin layer chromatography and mass spectrometry from wild type strain MB215 revealed a mixture of MELs withdifferent length of the fatty acid side chains ranging from C12, C14 and C16. Strains expressing both cytoplasmic variants Mac1DPTS and Mac2DPTSshowed a reduction of diversity of MELs. In these mutants MELs with C16 and C2 side chains are significantly overrepresented. This indicates that MELproduction is coupled to peroxisomal b-oxidation resulting in a more variable distribution in the length of fatty acid side chains. Currently, we investigatethe MEL production is strains lacking peroxisomes and the importance of MEL diversity for the life of U. maydis.63. Metabolic adaptation of the oomycete Phytophthora infestans during colonization of plants and tubers. Carol E. Davis, Howard S. Judelson. PlantPathology and Microbiology, University of California, Riverside, CA 92521.Phytophthora infestans is the causative agent of late blight and was responsible for the Irish famine in the 1840’s. Today it still continues to be a globalproblem and in the USA it has been reported that the economic loss on potato crops alone exceeds $6 billion per year. A successful phytopathogenicrelationship depends on the ability of the organism to adapt its metabolism during infection on various nutritional substrates (e.g., plant versus tuber) andat different times throughout infection when nutrients may be limiting. Investigation of this metabolic adaptation is key to understanding how P. infestanssucceeds as a pathogen. To do this, tomato plants and potato tubers were infected with zoospores using a “dipping” method. RNA was extracted at 3 dpiand 6 dpi and subsequently used in library preparation. Following this, the libraries were quality checked by analysis on a Bioanalyzer using a highsensitivity DNA chip. Using Illumina technology (50 bp, paired-end reads) RNA Sequencing was performed. For each sample an average of 262 million readswas obtained. As a reference for the in planta data, RNASeq was also performed on defined and complex media. Mining of the data shows that theexpression profiles of some pathways change, such as glycolysis and gluconeogenesis. Learning how metabolic adaptation occurs will prove useful in thedevelopment of novel control strategies for this plant pathogen.64. Multi-copper oxidase genes of Heterobasidion irregulare. Ming-Chen Hsieh, Bastian Doernte, Ursula Kües. Molecular Wood Biotechnology andTechnical Mycology, University of Goettingen, Goettingen, Germany.The species complex Heterobasidion consists of well-known wood decomposers that infect mainly conifers. The fungi are white rots that decay lignin andcellulose. Laccases are enzymes that potentially attack the lignin. In the sequenced genome of the North American species H. irregulare 18 multi-copperoxidase genes (mco) are found (1). Phylogenetic sequence analysis divides the encoded proteins into five subclusters of mcos. In total, 14 proteinsclustered in two different subfamilies of classical laccases whereas two others are found amongst ferroxidases/laccases (enzymes with often dualactivities), one under fungal Fet3-type ferroxidases and one with fungal ascorbate oxidases. The potential three-dimensional structures of all mcos werepredicted by homology modelling for further grouping. Six of the potential laccase genes were first chosen for subcloning and expression in theheterologous basidiomycete Coprinopsis cinerea. (1) Olson et al. (2012). Insight into trade-off between wood decay and parasitism from the genome of afungal forest pathogen. New Phytologist 194: 1001-1013.65. The two novel class II hydrophobins of Trichoderma stimulate enzymatic hydrolysis of polyethylene terephthalate (PET). Liliana E. Tenorio-Rammer 1 ,Doris Ribitsch 1 , Annemarie Marold 1 , Katrin Greimel 1 , Enrique Herrero Acero 1 , Georg M. Guebitz 1,2 , Christian Kubicek 1,3 , Irina S. Druzhinina 1,3 . 1) ACIB -Austrian Centre of Industrial Biotechnology, Petersgasse 14, 8010 Graz, Austria; 2) c/o Institute for Environmental Biotechnology, University of NaturalResources and Life Sciences, Vienna, Konrad Lorenz Strasse 20, 3430 Tulln, Austria; 3) Microbiology Group, Institute of Chemical Engineering, ViennaUniversity of technology, Vienna, Austria.Polyethylene terephthalate (PET), a thermoplastic polyester with excellent industrial properties, can be functionalised and/or recycled via hydrolysis bymicrobial cutinases. Here we tested whether hydrophobins (HFBs), small secreted fungal proteins containing eight positionally conserved cysteineresidues, would be able to enhance the rate of enzymatic hydrolysis of PET. To this end, we selected the genus of the mycoparasitic filamentous fungusTrichoderma as it has been previously shown to have the most proliferated arsenal of HFBs among all fungi. Consequently we used the phylogeneticapproach to identify the two novel class II HFBs (HFB4 and HFB7) from Trichoderma as the first candidates for the test. HFB4 and HFB7, produced in E. colias N-terminal glutathione-S-transferase fusion proteins, exhibited subtle structural differences reflected in the hydropathy plots which were correlatedwith unequal hydrophobicity and hydrophily respectively determined by water contact angle measurements. However they exhibited a dosage-dependentstimulation of PET hydrolysis by cutinase from Humicola insolens with HFB4 displaying an adsorption isotherm, whereas HFB7 was active only at very lowconcentrations and behaved inhibitory beyond them. We conclude that class II HFBs can stimulate the activity of cutinases on PET, but individual HFBs candisplay different properties in this process thus warranting a broader screening of HFBs for such industrial applications.27th Fungal Genetics Conference | 137

FULL POSTER SESSION ABSTRACTS66. Analysis of polyketide synthase gene clusters in Cladonia metacorallifera genome. J.-S. Hur 1 , J. A. Kim 1 , Y. J. Koh 1 , S. Kim 2 . 1) Korean Lichen ResearchCenter, Sunchon National University, Sunchon, South Korea; 2) Wildlife Genetic Resources Center, National Institute of Biological Resources, Korea.Lichen-forming fungi produce highly diverse and unique secondary compounds such as depsides, depsidones, dibenzofurans and depsones. Thebiosynthesis of secondary metabolites is governed by polyketide synthase (PKS). However, the molecular mechanisms underlying the biosynthesis of thesemetabolites are poorly understood. Here we present analysis of the structure of the PKS gene clusters responsible for secondary metabolite production inthe recently sequenced genome of lichen-forming fungus Cladonia metacorallifera. We found 37 type I polyketide synthase genes which were composedof 19 reducing PKSs, one partial reducing PKS and 17 non-reducing PKSs. Lichen-forming fungal PKS domains shared common structure with filamentousfungal PKSs. Phylogenetic analysis shows that some lichen-forming fungal PKSs constructed an unique clade in other filamentous fungal PKS clades.67. Inhibition of benzoate 4-monooxygenase (CYP53A15) from Cohliobolus lunatus by cinnamic acid derivatives. Branka Korosec 1 , Barbara Podobnik 2 ,Sabina Berne 3 , Neja Zupanec 1 , Metka Novak 1 , Nada Krasevec 1 , Samo Turk 4 , Matej Sova 4 , Ljerka Lah 1 , Jure Stojan 3 , Stanislav Gobec 4 , Radovan Komel 1,3 . 1)National Institute of Chemistry, Ljubljana, Slovenia; 2) Lek Pharmaceuticals d.d., Verovskova 57, SI-1000 Ljubljana, Slovenia; 3) Institute of Biochemistry,Faculty of Medicine, University of Ljubljana, Vrazov trg 2, SI-1000 Ljubljana, Slovenia; 4) Chair of Pharmaceutical Chemistry, Faculty of Pharmacy, Universityof Ljubljana, Askerceva cesta 7, SI-1000 Ljubljana, Slovenia.Fungal infections cause huge economic losses in agriculture. Some of the major phytopathogens also cause serious, and very often lethal, infections inhuman and animals. Plants may be a good source of antifungals since they have to defend themselves by producing numerous secondary metabolites,such as sterols, terpens, polycosanols and phenolic compounds. Successful development of antifungal compounds, based on natural defense molecules,could prove useful in combating infectious and toxin-producing fungi in both agriculture and medicine. In recent years several promising antifungal targetshave been under exploration. One of such is also fungal CYP53, member of the family of highly conserved CYP proteins, involved in detoxification ofbenzoate, a key intermediate in metabolism of aromatic compounds in fungi. High specificity and absence of homologue in higher eukaryotes assignCYP53A15 from the filamentous fungus Cochliobolus lunatus as interesting drug target. In our latest research we explored chemical properties ofisoeugenol for ligand-based similarity searching, and the homology model of CYP53A15 of Cochliobolus lunatus, for structure-based virtual screening of acomposite chemical library. Two cinnamic acid derivatives were amongst the highest scoring compounds. In the past few years, several other reportsabout antifungal activity of cinnamic acid derivatives have been published. In order to investigate the potential inhibitory activity on benzoate 4-monooxygenase (CYP53A15) we analyzed antifungal activity of 9 commercially available, and 10 representative cinnamic acid derivatives from our library.Furthermore, to obtain more information about structure-activity relationship 26 additional cinnamic acid esters and amides were synthesized andincluded in our assays. Among 45 cinnamic acid derivatives tested, 7 compounds have shown antifungal activity against C. lunatus, A. niger and P.ostreatus in in vivo inhibition tests. Compounds with antifungal activity were further evaluated for inhibition of CYP53A15 activity with spectral bindingtitration assay and HPLC. The best two inhibitors of CYP53A15 activity showed 70% inhibition at 600 mM concentration and were selected for furtheroptimization of new lead structures.68. Higher yields of cyclodepsipetides from Scopulariopsis brevicaulis by random mutagenesis. Linda Paun 1 , ElKbir Hihlal 1 , Annemarie Kramer 2 , AntjeLabes 2 , Johannes Imhoff 2 , Frank Kempken 1 . 1) Botanical Institute, Christian-Albrechts-University, Kiel, Germany; 2) Kieler Wirkstoff-Zentrum KiWiZ atGEOMAR, Kiel, Germany.The ascomycete Scopulariopsis brevicaulis, which was isolated from the marine sponge Tethya aurantium, produces two cyclodepsipeptides,scopularides A and B [1]. Both peptides exhibit activity against several tumor cell lines. Within the EU-project MARINE FUNGI (EU FP7, 265926) one of ouraims is to enhance the production of these secondary metabolites. We are in the process to establish two ways of random mutagenesis by both UVradiation and transposon-mediated. To this end we created UV-mutants and a miniaturised screening method was developed. UV-radiation wasperformed at 312 nm and the survival rate was set to 1 %. With this method a mutant library was established. To screen these mutants for highersecondary metabolites production, we developed a screening method which includes decreased cultivation volume, fast extraction and an optimised LC-MS analysis format. Using the UV mutagenesis, we identified several mutants with a higher scopularide production in comparison to the wild type. One ofthese mutants, which produces three times more biomass and more than double the amount of scopularide A, has been used for another round ofmutation. Next generation sequencing is being employed to identify the molecular genetic basis of the observed mutations. In parallel we employtransposable elements to introduce mutants [2]. The impact of transposons on gene expression as well as their ability to cause major mutations within thegenome makes them an interesting tool for random mutagenesis [3, 4, 5]. We employ the Vader transposon in its homologous host and found that itmostly integrates within or very close to genes thus it appears to be a useful tool for transposon-mediated mutagenesis in A. niger (6). At current we try toenhance its usability by modifying the Vader element. [1] Yu, Z.; Lang, G.; Kajahn, I.; Schmaljohann, R.; Imhoff, J. J. Nat. Prod. 2008, 71, 1052-1054 [2]Braumann I, van den Berg M, & Kempken F (2007) Fungal Genet Biol 44(12):1399-1414. [3] Daboussi MJ & Capy P (2003) Annu Rev Microbiol 57:275-299.[4] Kempken F (2003) Applied Mycology and Biotechnology, Vol. 3 Fungal Genomics, eds Arora DK & Khachatourians GG (Elsevier Science Annual ReviewSeries), pp 83-99. [5] Kempken F & Kück U (1998) BioEssays 20:652-659. [6] Hihlal E, Braumann I, van den Berg M, Kempken F (2011) Appl EnvironmentMicrobiol, 77: 2332-2336.Cell Biology and Development69. Generation of pathogenic diploids from heterogeneous conidial populations of Aspergillus flavus. Farhana Runa 1 , Ignazio Carbone 1 , DeepakBhatnagar 2 , Gary Payne 1 . 1) Plant Pathology, North Carolina State University, Raleigh , NC; 2) Southern Regional Research Center, USDA, New Orleans, LA.Aspergillus flavus, a major producer of aflatoxin, has emerged as an opportunistic pathogen for a wide range of hosts. Understanding genetic variationwithin strains of A. flavus is important for controlling disease and reducing aflatoxin contamination. Because conidia of A. flavus are multinucleated buthaploid, we wanted to know if nuclear condition or ploidy of conidia could be potential sources of genetic variation. The objective of our study is to detectnuclear heterogeneity and ploidy in conidial populations of A. flavus and determine their impact on fungal ecology. In order to examine heterokaryosis,protoplast of two different auxotrophic strains in which nuclei were labeled with yellow (EYFP) and cyan (ECFP) fluorescent markers were fused. Fusantsbetween the two strains were obtained through polyethylene mediated cell fusion and selection on minimal medium, which favored the growth of thefusants over that of either parental strain. Fusants selected for further study showed heterogeneous conidial populations with nuclei predominantlyexpressing either EYFP or ECFP, or a very few expressing both EYFP+ECFP. Conidia containing nuclei expressing only EYFP+ECFP were separated byFluorescence-Activated Cell Sorting (FACS) and found to contain both yellow and cyan fluorescent proteins in the same nuclei. Further characterization ofconidia having only one nucleus, but expressing both EYFP+ECFP, showed them to be diploids. Pathogenicity assays using Galleria mellonella showed that138

FULL POSTER SESSION ABSTRACTSdiploids are more virulent than the parental haploids. Our results suggest that conidial populations of A. flavus are predominantly homokaryotic but asmall percentage of conidia are heterokaryotic. Within a heterokaryon, a diploid nucleus could be formed by fusion of two haploid nuclei, which may allowthe generation of a pathogenic strain.70. Inhibition of appressorium formation of Magnaporthe oryzae by roxithromycin and its possible molecular target. Akira Ishii, Mayu Kumasaka,Megumi Narukawa, Takashi Kamakura. Applied Biological Science, Tokyo Univ. of Science, Noda, Chiba, Japan.Roxithromycin (RXM), a 14-membered macrolide which was originally active against prokaryote, has beneficial side effects such as anti-inflammatoryactivities were reported and actually applied to human. However, the mechanisms underlying these side effects are still unclear. In this study, we foundthat RXM inhibited appressorium formation of rice blast fungus Magnaporthe oryzae (M. oryzae). These results suggest that there are alternative targetsin broad eukaryotic organisms and it is interesting to identify the molecular target of the secondary effect on human using M. oryzae. Magnaporthe oryzaeis the causal agent of rice-blast disease. M. oryzae enters its host plant using a specialized infection structure known as an appressorium. Thedevelopmental stage of appressorium is sensitive to various chemical inhibitors, because large numbers of genes are involved in cellular differentiation.Since appressorium formation by M. oryzae can be observed on artificial surfaces, it can be a useful tool to search new activity of various chemicals. Weperformed phage display to search novel molecular target(s) of the antibiotic. We found that one candidate gene 32-11 may play important roles inappressorium formation. Expression of 32-11 gene, in a 32-11 mutant, was lower than wild type during developing infection structure, and the mutant wasless affected by RXM. Although germinate and formation of appressoria were normal. Over expression of 32-11 gene caused no effect to RXM sensitivity,germination nor appressorium formation compared with the wild type. Over expression of 32-11 caused no effect to RXM activity, germination orappressorium formation compare to the wild type. To investigate whether lower expression of 32-11 causes the less sensitivity to RXM, we introduced 32-11 over expression vector into 32-11 reduced mutant. These mutants restored their wild type phenotype. These results possibly suggest that the complexof 32-11 product and RXM affects another molecule which plays an important role in appressorium formation at M. oryzae.71. Identification of novel genes involved in induction of appressorium development triggered by plant-derived signals in Colletotrichum orbiculare.Sayo Kodama 1 , Ayumu Sakaguchi 2 , Yasuyuki Kubo 1 . 1) Laboratory of Plant Pathology, Graduate School of Life and Environmental Science, Kyoto PrefecturalUniversity, Kyoto, Japan; 2) National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan.Many plant pathogenic fungi initiate infection of host leaves by the germination of conidia and differentiation of appressoria at the tip of germ tubes.These morphological changes are triggered by various external signals such as physical or chemical signals from the plant surface. In our previous study,cucumber anthracnose fungus Colletotrichum orbiculare CoKEL2, a Schizosaccharomyces pombe tea1 homologue, encoding a kelch repeat protein wasidentified. The cokel2 mutants formed abnormal appressoria on glass slides, and those appressoria were defective in penetration hyphae developmentinto cellulose membranes, an artificial model substrate for fungal infection. In contrast, the cokel2 mutants formed normal appressoria on the hostcucumber plant and retained penetration ability. Moreover, when conidia were incubated in the presence of exudates from cucumber cotyledon, normalappressorium formation on the artificial substrate by the cokel2 mutants was restored. These results suggest that CoKEL2 is essential for normalmorphogenesis of appressoria and that there is a bypass pathway that transduces plant-derived signals for appressorium formation independent ofCoKEL2. These plant-derived signaling pathways for appressorium formation have not been characterized in fungal pathogens including C. orbiculare. Todetermine specific components of the plant-derived signaling pathway that leads to appressorium formation, we screened six cokel2 double mutants thatformed abnormal appressoria not only on artificial substrates but also on the host plant surface. Furthermore, reintroduction of CoKEL2 into those cokel2double mutants restored normal appressorium formation on artificial substrates, suggesting that cokel2 double mutants have defects in CoKEL2-independent and plant-derived specific signaling pathway for appressorium formation. We identified and characterized candidate mutated genes by wholegenome sequencing of the six cokel2 double mutants. To define the involvement of those candidate mutated genes in appressorium formation, weobserved the phenotypes of candidate geneD single mutants, cokel2D candidate geneD double mutants, and complementation strains. As expected,candidate geneD mutants in cokel2D back ground showed same phenotypes as those of screened cokel2 double mutants.72. Unique protein domains regulate Aspergillus fumigatus RasA localization and signaling during invasive growth. Rachel V. Lovingood 1 , Praveen R.Juvvadi 2 , William J. Steinbach 2 , Jarrod R. Fortwendel 1 . 1) Microbiology and Immunology, University of South Alabama, Mobile AL, USA; 2) PediatricInfectious Diseases, Duke University, Durham NC, USA.Invasive pulmonary aspergillosis (IPA) is propagated by inhalation of A. fumigatus spores that germinate and invade the lung tissue in search of nutrients.We have shown that the A. fumigatus RasA GTPase protein is necessary for hyphal morphogenesis, cell wall integrity, and virulence during IPA. Ourprevious studies focused on conserved protein domains regulating RasA localization and signaling. These studies revealed the requirement for plasmamembrane (PM)-localized Ras for proper signaling and regulation of A. fumigatus growth and virulence. Therefore, mechanisms controlling Ras localizationare of interest in designing novel antifungal Ras inhibitors. Although Ras pathways may represent valid antifungal targets, the importance of fungal-specificRas protein domains to Ras function in fungal pathogenesis remains unexplored. To address this important knowledge gap, we identified fungal-specificRas protein domains by comparing fungal Ras sequences to their human counterpart, H-ras. We hypothesized that such domains could serve as targetableareas to selectively inhibit the fungal Ras protein. This analysis revealed two areas of significant divergence with H-ras: i) the Invariant Arginine Domain(IRD), a novel domain conserved in the RasA homologs of every available fungal genome but not present in H-ras and ii) an extended hypervariable region(HVR). Truncation analysis of the HVR identified a serine-rich region that is necessary for localization to the PM and for RasA signaling during hyphalmorphogenesis. Interestingly, mutational analysis of the IRD produced a properly localized yet non-functional RasA protein. However, activation of the IRDRasA mutant was not altered suggesting a role for the IRD during interactions of RasA with downstream effectors. Further characterization of the IRD andHVR, and the protein interactions to which they contribute, will reveal fungal-specific aspects of Ras function and may define a new paradigm for Rassignal transduction in fungal organisms.73. Light regulates growth, stress resistance and metabolism in the fungal pathogen Aspergillus fumigatus. Kevin K. Fuller, Carol S. Ringleberg, Jennifer J.Loros, Jay C. Dunlap. Genetics, Geisel School of Medicine at Darmouth, Hanover, NH.Light serves as an important environmental cue that influences developmental and metabolic pathways in a variety of fungi. Interestingly, orthologs of aconserved blue light receptor, WC-1, promote virulence in two divergently related pathogenic species, Cryptococcus neoformans and Fusarium oxysporum,suggesting that photosensory systems may be conservatively linked to fungal pathogenesis. Aspergillus fumigatus is the predominant mold pathogen ofimmunocompromised patients, but if and how the organism responds to light has not been described. In this report, we demonstrate that the fungus canindeed sense and distinctly respond to both blue and red portions of the visible spectrum. Included in the A. fumigatus photoresponse is a reduction inconidial germination kinetics, increased hyphal pigmentation, enhanced resistance to acute ultra-violet and oxidative stresses, and an increased27th Fungal Genetics Conference | 139

FULL POSTER SESSION ABSTRACTSsusceptibility to cell wall perturbation. Through gene deletion analysis we have found that the WC-1 ortholog, LreA, is a bone fide blue light receptor in A.fumigatus that is required for the photopigmentation response. However, the DlreA mutant retains several blue light mediated responses, including thegermination and stress resistance phenotypes , suggesting other blue light receptors are operative in this fungus. We also show that the putative red lightsensing phytochrome, FphA, is involved with some, but not all, blue light specific phenotypes, indicating a complex interaction between red and blue lightphotosystems in A. fumigatus. Finally, whole genome microarray analysis has revealed that A. fumigatus displays broad patterns of gene induction andrepression upon exposure to light. Affected genes are largely metabolic and include those involved in lipid and sterol synthesis, respiration, carbohydratecatabolism, amino acid metabolism and metal ion homeostasis. Taken together, these data demonstrate the importance of the photic environment on thephysiology of A. fumigatus and provide a foundation for future studies into an unexplored area of this important pathogen.74. Analysis of critical domains in calcineurin A required for septal targeting and function in Aspergillus fumigatus. Praveen R. Juvvadi 1 , Jarrod R.Fortwendel 2 , Christopher Gehrke 1 , Frédéric Lamoth 1 , William J. Steinbach 1 . 1) Department of Pediatrics, Duke University Medical Center, Durham, NC; 2)Department of Microbiology and Immunology, University of South Alabama, Mobile, AL.Calcineurin, a calmodulin (CaM)-dependent protein phosphatase, is known to play key roles in virulence, growth and stress responses of pathogenicfungi. Critical understanding of calcineurin regulation and identifying the residues indispensable for calcineurin activity in vivo will pave the way fordevising new drug targets for combating invasive aspergillosis. Previous studies from our laboratory showed that the calcineurin complex (CnaA and CnaB)in Aspergillus fumigatus selectively localizes at the hyphal tip and septum to direct proper hyphal growth and regular septum formation. However, thedomains responsible for targeting and function of CnaA at the hyphal septum remain unknown. Here we performed extensive truncational and mutationalanalyses of the functional domains of CnaA to investigate the relevance of these domains for localization and function of CnaA at the septum. Importantlywe found that (i) CaM, the key protein known to activate calcineurin is not required for septal localization of CnaA but is required for its function at thehyphal septum, (ii) the PxIxIT substrate binding motif in CnaA is required for its localization at the hyphal septum, indicating it localizes at the septum byinteracting with other as yet unknown protein/s(iii) binding of CnaB subunit is not necessary for septal localization of CnaA but the regulatory subunit isrequired for its activation at the septum, and (iv) triple mutations in the catalytic active site do not affect septal localization of CnaA but completely blockhyphal growth revealing that both septal localization and activity of CnaA are required for proper hyphal growth.75. Calcium imaging and measurement during growth and response to stresses in Aspergillus fumigatus. Alberto Muñoz 1 , Margherita Bertuzzi 2 , JanBettgenhaeuser 1 , Elaine Bignell 2 , Nick Read 1 . 1) Fungal Cell Biology Group, University of Edinburgh, Edinburgh, United Kingdom; 2) Microbiology Section,Imperial College London, London, United Kingdom.Calcium signalling and homeostasis are essential for the growth, differentiation and virulence of filamentous fungi. During infection, A. fumigatus mustbalance concomitant demands to: (1) withstand toxic levels of exogenous calcium (3-5 mM) in the host environment which can be >100,000x that of thefungal cytosolic free calcium ([Ca 2+ ] c) concentration; (2) appropriately integrate homeostatic and stress-responsive adaptations; and (3) undergo normalcalcium signalling. There is evidence for calcium signalling regulating numerous processes including spore germination and hyphal tip growth. The lowresting level of [Ca 2+ ] c (50-100 nM) is maintained by Ca 2+ -pumps and -antiporters, and cytoplasmic Ca 2+ -buffering. However, [Ca 2+ ] c becomes an intracellularsignal when its concentration is transiently increased. We have developed two methods for measuring and imaging [Ca 2+ ] c: (1) 96-well plate luminometryusing the genetically encoded, bioluminescent aequorin; and (2) fluorescence microscopy using the genetically encoded calcium-sensitive, fluorescentprotein G-CaMP5. Aequorin is ideally suited for quantitative measurements of [Ca 2+ ] c calcium signatures in cell populations whereas fluorescence imagingof the G-CaMP5 is good for single cell and subcellular measurements of [Ca 2+ ] c. Using the aequorin methodology we have found that transient increases in[Ca 2+ ] c with specific, reproducible calcium signatures in A. fumigatus arise from exposure to stresses such as high external calcium. In our analysis, [Ca 2+ ] cspikes in actively growing hyphal tips have been imaged using G-CaMP5. Exposure of conidial germlings to high external calcium induces dramatic and verydynamic changes in [Ca 2+ ] c with the generation of localized [Ca 2+ ] c transients and waves. Furthermore, there is considerable heterogeneity in the [Ca 2+ ] cresponses of different germlings within the cell population. Calcium imaging and measurement using genetically encoded probes, particularly whencombined with pharmacological and genetic analyses, will provide major new insights into calcium signalling in filamentous fungi.76. WITHDRAWN77. The copper transporter ctpA in Aspergillus fumigatus is critical for conidial melanization and virulence in an invertebrate infection model. SrijanaUpadhyay, Xiaorong Lin. Biology, Texas A&M University, College Station, TX.Aspergillus fumigatus is an opportunistic pathogen that causes life-threatening invasive diseases in immunocompromised hosts. This fungus producesabundant, easily aerosolized, and heavily melanized conidia that are the infectious particles. The melanin, or the bluish green pigment coated on theconidial surface, is associated with fungal virulence and resistance to environmental stresses. This melanin is synthesized through the DHN melaninpathway by a cluster composed of six structural biosynthetic genes. Although all Aspergillus species produce conidial melanin, this DHN melanin genecluster found in A. fumigatus is not conserved in all species of this genus. In other species, laccases are critical for melanization and copper has beenshown to be critical for their activity. In A. nidulans, defective ygA that encodes a copper transporter results in reduction in conidial laccase activity andpoor conidial pigmentation. Whether copper is important for conidial melanization or whether it affects the function of the DHN gene cluster in A.fumigatus are not clear. In this study we have identified ctpA in A. fumigatus as the homolog of ygA in A nidulans and demonstrated its importance forconidial melanization under the copper limiting and the copper replete conditions. The defect in melanization caused by the deletion of the ctpA gene canbe remediated by addition of copper in the media or by the overexpression of the ctpA gene. Lack of melanin is caused by growing the wild type in thecopper-limiting conidiation or by the deletion of the ctpA gene. This renders the A. fumigatus conidia more immune-dominant, since these conidia cancause exacerbated immune-responses from the invertebrate host, larvae of Galleria mellonella. Furthermore, we have identified and characterizeregulators that play important roles in maintaining copper homeostasis and melanization in A. fumigatus.78. Aspergillus nidulans SNXA HRB1 is an SR/RRM family protein that rescues defects in the CDC2/CYCLINB pathway. Steven James 1 , Travis Banta 2 , JamesBarra 1 , Clifford Coile 2 , Ryan Day 2 , Cheshil Dixit 2 , Steven Eastlack 2 , Anh Giang 2 , Yulon Huff 2 , Julie Kobie 1 , Faustin Mwambutsa 2 , Mimi Nguyen 2 , AmandaOrzechowski 1 , Kristin Shingler 1 , Sarah Lea Anglin 2 . 1) Dept. Biology, Gettysburg College, Gettysburg, PA; 2) Dept. Biology, Millsaps College, Jackson, MS.Control of the eukaryotic G2/M transition by CDC2/CYCLINB is tightly regulated. To further characterize this regulation in Aspergillus nidulans, weconducted a screen for extragenic suppressors of nimX2 cdc2 that resulted in the identification of the cold-sensitive, G1-arresting snxA1 mutation. Our datashow that snxA1 suppresses defects in regulators of the G2/M transition, including nimX2 cdc2 , nimE6 cyclinB , and nimT23 cdc25 , but does not suppress the G1/SarrestingnimE10 cyclinB mutation or any of four S phase mutations. Furthermore, the snxA1 mutation or deletion of snxA alter localization patterns ofNIME CYCLINB at the restrictive temperatures for snxA1 and nimX2, supporting a role for SNXA in cell cycle control. snxA encodes the A. nidulans ortholog of140

FULL POSTER SESSION ABSTRACTSSaccharomyces cerevisiae Hrb1/Gbp2, nonessential shuttling mRNA binding proteins belonging to the SR (Serine-Arginine Rich) and RRM (RNA RecognitionMotif) protein family. snxA hrb1 is nonessential, its deletion phenocopies the snxA1 mutation, and overexpression of gDNAs or of alternatively spliced snxAcDNAs rescues snxA1 mutant phenotypes. SNXA HRB1 is predominantly nuclear, but is not retained in the nucleus during the partially-closed mitosis of A.nidulans. We further demonstrate that the snxA1 mutation does not suppress nimX2 by altering NIMX2 CDC2 /NIME CYCLINB kinase activity, suggesting that theeffects of SNXA1 on NIMX2 CDC2 /NIME CYCLINB may be due to altered localization of NIME CYCLINB . These data suggest a novel role in G2/M regulation for thisSR/RRM family member. This work was supported by the Mississippi INBRE funded by grants from the National Center for Research Resources(5P20RR016476-11) and the National Institute of General Medical Sciences (8 P20 GM103476-11) from the National Institutes of Health.79. The Aspergillus nidulans MAPK module AnSte11-Ste50-Ste7-Fus3 controls development and secondary metabolism. Oezguer Bayram 1* , OezlemSarikaya Bayram 1 , Yasar Luqman Ahmed 2 , Jun-Ichi Maruyama 1,4 , Oliver Valerius 1 , Silvio Rizzoli 3 , Ralf Ficner 2 , Stefan Irniger 1 , Gerhard Braus 1 . 1) Institute ofMicrobiology & Genetics, Department of Molecular Microbiology and Genetics, Georg-August-Universität, Grisebachstr. 8, D 37077 Goettingen, Germany;2) Department of Molecular Structural Biology, Institute for Microbiology and Genetics, Georg-August-Universität, Goettingen; 3) European NeuroscienceInstitute, Deutsche Forschungsgemeinschaft Center for Molecular Physiology of the Brain/Excellence Cluster 171, 37077 Göttingen; 4) Department ofBiotechnology, The University of Tokyo, Tokyo, Japan.The sexual Fus3 MAP kinase module of yeast is highly conserved in eukaryotes and transmits external signals from the plasma membrane to the nucleus.We show here that the module of the filamentous fungus Aspergillus nidulans (An) consists of the AnFus3 MAP kinase, the upstream kinases AnSte7 andAnSte11, and the AnSte50 adaptor. The fungal MAPK module controls the coordination of fungal development and secondary metabolite production. Itlacks the membrane docking yeast Ste5 scaffold homolog but similar to yeast the entire MAPK module interacts with each other at the plasma membrane.AnFus3 is the only subunit with the potential to enter the nucleus from the nuclear envelope. AnFus3 interacts with the conserved nuclear transcriptionfactor AnSte12 to initiate sexual development and phosphorylates VeA which is a major regulatory protein required for sexual development andcoordinated secondary metabolite production. Our data suggest that not only Fus3 but even the entire MAPK module complex of four physicallyinteracting proteins can migrate from plasma membrane to nuclear envelope.80. Functional analysis of sterol transporter in filamentous fungus Aspergillus nidulans. Nicole Bühler, R. Fischer, N. Takeshita. Microbiology, KarlsruheInstitut of Technology, Karlsruhe, Germany.A continuous flow of secretion vesicles from the hyphal cell body to the growing hyphal tip provides the delivery of proteins and lipids to the tip and isessential for cell wall and cell membrane extension at the tip. Apical sterol-rich plasma membrane domains (SRDs), which can be viewed using the sterolbindingfluorescent dye filipin, are gaining attention for their important roles in polarized growth of filamentous fungi. Although the importance of SRDs isbecoming clear, their exact roles and formation mechanisms remain rather unclear. Transport of sterol to hyphal tips is thought to be important for theSRDs organization. Oxysterol binding proteins, which are conserved from yeast to human and involved in vesicular trafficking, signalling, lipid metabolismand non-vesicular sterol transport. Saccharomyces cerevisiae has seven oxysterol binding protein homologues (OSH1-7). Their subcellular distributions areregulated respectively. The OSH proteins are thought to function as a sterol transporter between closely located membranes independently of vesicletransport. In the filamentous fungus Aspergillus nidulans, we found five OSH genes. To investigate their functions for the polarized growth and SRDsorganization, their localization are analyzed by GFP tagging. The gene-deletion strains are constructed and analyzed. Their expression levels are analyzedvia qRT-PCR.81. Mechanisms of cellular resistance to copper and arsenic in Aspergillus nidualans. Steven H. Denison. Natural Sciences, Eckerd College, St Petersburg,FL.Copper is an essential element for cells that is toxic in high concentrations. Understanding cellular mechanisms for survival in high concentrations ofcopper is important for at least two reasons. Firstly, copper is an important environmental contaminant. In addition, two human genetic disorders, Wilsonand Menkes diseases, result from impaired copper transport. I am using the filamentous fungus, Aspergillus nidulans, as a model organism forunderstanding cellular mechanisms of resistance to high concentrations of copper. I identified a gene in A. nidulans homologous to the coppertransporting ATPase-encoding genes mutated in Wilson and Menkes diseases. To determine the location of this copper transporter in A. nidulans cells, Iused fusion PCR to construct a GFP-tagged version of the gene, which was then transformed into A. nidulans in a gene replacement. In terms of its locationin the cell, the A. nidulans GFP-tagged protein behaves in the same way as the Menkes disease protein: it is located to an intracellular compartment(possibly the Golgi, as in human cells) in low copper medium but appears in the plasma membrane upon addition of excess copper to the medium. Inaddition, disruption of the A. nidulans copper transporter gene results in increased sensitivity to copper in the growth medium relative to wild type cellsand cells expressing the GFP-tagged protein. Taken together, these data suggest that the transporter functions in the plasma membrane in high copperenvironments to remove excess copper from cells. Arsenic is also an important environmental contaminant. To begin to understand the mechanism ofarsenic resistance in A. nidulans cells, I have GFP-tagged and disrupted a putative arsenic transporter gene from A. nidulans. Disruptants are more sensitiveto arsenic than wild type cells and cells expressing the GFP-tagged protein. The GFP fusion protein localizes to the plasma membrane, consistent with afunction for the protein in transporting arsenic across the plasma membrane, removing arsenic from cells.82. Functional characterization of Aspergillus nidulans ANID_05595.1: a possible homologue of the polarisome component Pea2. Nathan W Gross,Bradley Downs, Steven D Harris. Center for Plant Science Innovation, University of Nebraska-Lincoln, Lincoln, NE 68588-0660.Cell polarity is a defining feature of filamentous fungal growth. However, the complete molecular pathway that regulates this morphogeneticcharacteristic has not yet been elucidated. In Aspergillus nidulans, a germ tube emerges from a discrete location along the conidium following a briefperiod of isotropic swelling. Plasma membrane and cell wall components are continuously added to the apex of the germ tube via microtubule and actinmediated trafficking of vesicles to this region. As growth progresses, germ tube cells undergo cytokinesis and are compartmentalized by septa.Additionally, the cell wall becomes increasingly cross-linked throughout subapical regions forming a hypha, which continues to grow in the same polarizedmanner. ANID_05595.1 is located on chromosome 5, contig 96, and encodes a 946 amino acid hypothetical involucrin repeat protein. To investigate thefunction of ANID_05595.1 in A. nidulans, deletion mutants were generated using pyrG from Aspergillus fumigatus as a selectable marker. This mutationresulted in restricted colony growth, increased hyphal diameter, and dichotomous hyphal branching patterns. These phenotypes suggest thatANID_05595.1 function is important to the maintenance of polarized cell growth in A. nidulans and other ascomycetes. The hypothetical ANID_05595.1protein shares characteristics with Saccharomyces cerevisiae Pea2, a polarisome component required for bipolar budding and mating. Along withstructural similarities, the phenotypes observed in S. cerevisiae DPea2 are similar to A. nidulans D5595. This suggests that ANID_05595.1 may perform asimilar mechanistic function to Pea2 in A. nidulans.27th Fungal Genetics Conference | 141

FULL POSTER SESSION ABSTRACTS83. Aspergillus nidulans septin interactions and post-translational modifications. Yainitza Hernandez-Rodriguez 1 , Shunsuke Masuo 2 , Darryl Johnson 3 , RonOrlando 3,4 , Michelle Momany 1 . 1) Plant Biology, University of Georgia, Athens, GA, US; 2) Laboratory of Advanced Research A515, Graduate School of Lifeand Environmental Sciences, University of Tsukuba, Tennodai, Tsukuba, Ibaraki, JP; 3) Department of Chemistry, University of Georgia, Athens, GA, US; 4)Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, US.Septins are cytoskeletal elements found in fungi, animals, and some algae, but absent in higher plants. These evolutionarily conserved GTP bindingproteins form heteroligomeric complexes that seem to be key for the diverse cellular functions and processes that septins carry out. Here we usedAspergillus nidulans, a model filamentous fungus with well defined vegetative growth stages to investigate septin-septin interactions. A. nidulans has fiveseptins: AspA/Cdc11, AspB/Cdc3, AspC/Cdc12 and AspD/Cdc10 are orthologs of the “core-filament forming-septins” in S. cerevisiae; while AspE is onlyfound in filamentous fungi. Using S-tag affinity purification assays and mass spectrometry we found that AspA, AspB, AspC and AspD strongly interact inearly unicellular and multicellular vegetative growth. In contrast, AspE appeared to have little or no interactions with core septins in unicellular stagesbefore septation. However, after septation AspE interacted with other septins, for which we postulate an accessory role. AspE localized to the cortex ofactively growing areas and to septa, and localizations are dependent on other septin partners. Interestingly, core septin localizations can also depend onaccessory septin AspE, particularly post-septation. In addition, LC-MS/MS showed acetylation of lysine residues in AspA before septation and AspC afterseptation. Western blot analysis using an anti-acetylated lysine antibody showed that AspC is highly acetylated in all stages examined, while other septinsshowed acetylation post-septation. Though LC-MS analysis failed to detect phosphorylation of septins, this modification has been widely reported in fungalseptins. Using phosphatase treatments and Western Bloting, we found phosphorylation of AspD, but no other septins. This is interesting because AspDbelongs to a special group of septins that lack a C-terminal coiled-coil found in other septins. However, septin localization is not affected by the absence ofAspD/Cdc10, but by the absence of filamentous fungi specific septin AspE. Our data suggests that septin interactions and modifications change duringdevelopment and growth in A. nidulans, and that some modifications are septin specific.84. A highly conserved sequence motif is required for PkcA localization to septation sites and protein function in Aspergillus nidulans. Loretta Jackson-Hayes 1 , Terry Hill 1 , Darlene Loprete 1 , Claire DelBove 1 , Omolola Dawodu 2 , Jordan Henley 3 , Ashley Poullard 3 , Justin Shapiro 1 . 1) Rhodes College, Memphis, TN38112; 2) Rust College, Holly Springs, MS 38635; 3) Tougaloo College, Tougaloo, MS 39174.Many proteins with diverse functions contribute to cell wall synthesis in polarized growth and septation. Some of these proteins play similar roles at tipsand septa, while others are exclusively involved in one process or the other. In Aspergillus nidulans, wild type protein kinase C (PkcA) localizes to growinghyphal tips and septation sites, and a role for PkcA in cell wall synthesis is supported by the inability of PkcA mutant strains to exhibit resistance to cell wallperturbing agents. PkcA localization to septation sites is dynamic. Upon initiation of septum formation PkcA is organized as a ring at periphery of theseptation site. The ring constricts in synchrony with the actin/myosin contractile ring and dissipates when septa are fully matured. To determine whichdomains are important for septum site localization, green fluorescent protein tagged, domain-deleted versions of PkcA were constructed. The domainsthat are vital to A. nidulans maintenance of cell wall integrity were separately identified by growing the domain deleted stains in the presence of the cellwall stressor calcofluor white. We have determined that the localization signal and the domain responsible for resistance to calcofluor white are distinct.The PkcA septation site localization signal is found within a region having homology with C2 domains of PKC proteins found in other organisms.Observations of both N- and C- terminal truncations support the conclusion that the PkcA septation site localization signal lies within the final 20 aminoacids of the C2 domain. Removal of these amino acids causes PkcA mislocalization to the cytoplasm. Furthermore, removal of the localization signalrenders the resulting truncated proteins less able to complement calcofluor white hypersensitivity in a strain carrying a mutation in its PkcA gene,highlighting the requirement of proper localization for this aspect of PkcA function.85. The MpkB MAP kinase plays a role in autolysis and conidiation of Aspergillus nidulans. Ji Young Kang 1 , Keon-Sang Chae 2 , Dong-Min Han 3 , Kwang-YeopJahng 1 . 1) Dept Biol, Chonbuk Natl Univ Col Nat Sci, Jeonju, Jeonbuk, South Korea; 2) Dept Mol Biol, Chonbuk Natl Univ Col Nat Sci, Jeonju, Jeonbuk, SouthKorea; 3) Div Life Sci, Wonkwang Univ, Iksan, Jeonbuk, South Korea.The mpkB gene of Aspergillus nidulans encodes a MAP kinase homologous to Fus3p of Saccharomyces cerevisiae which is involved in conjugationprocess. MpkB is required for accomplishing successfully the sexual development at the anastomosis and post-karyogamy stages. The mpkB deletion strainproduced conidia under the repression condition of conidiation such as sealing in the dark and even in the submerged culture concomitant with persistentbrlA expression, implying that MpkB might have a role in timely regulation of brlA expression. The deletion of mpkB caused hyphal fragmentation,disorganization of mycelial balls and dry cell mass reduction in the submerged culture as well as the chiB, mutA and pepJ genes which are encoding cellwall hydrolytic enzymes to be transcribed highly in the culture. These results suggest that MpkB might play a role in regulation of BrlA-involving autolysis.86. Beyond green mining: analysis of fungal cytochemistry using gold nanoparticles. Fatemeh Farazkhorasani 1 , Martin Prusinkiewicz 2 , Kathleen MGough 1 , Susan GW Kaminskyj 2 . 1) Chemistry, University of Manitoba, Winnipeg, Canada; 2) Biology, University of Saskatchewan, Saskatoon, Saskatchewan,Canada.Cells including fungal hyphae and other microorganisms, as well as fungal growth medium including both complex and defined composition, can reducesolutions of HAuCl 4 to elemental gold nanoparticles (AuNPs). As described in 2012 Analyst 137:4934-42, we have shown that AuNPs formed by growingfungal hyphae can be used as analytical substrates for surface-enhanced Raman scattering (SERS) spectroscopic analysis. These SERS spectra are in thesame energy range as our Fourier-transform infrared (FTIR) spectroscopic studies that provided information about cell composition. However, SERS isorders of magnitude more sensitive, and analysis is limited to cell components within a few nanometers of the AuNP. Our current interest is the fungal cellwall, which forms a porous interface between the cell and its environment. Cell wall chemistry is intrinsically related to cell-environment interactions,particularly for pathogenesis. The fungal wall is about 25 % of fungal dry weight, and its synthesis and maintenance is estimated to require ~25 % of thefungal genome. Fungal walls are ~ 80 % carbohydrate. Minor structural differences in carbohydrate bonding can cause profound changes in theirmetabolism, which complicates analysis. Preliminary studies described in the Analyst paper showed that SERS-active AuNPs can be generated by livinghyphae. Higher Au concentrations produced larger AuNPs within and on the hypha, but in addition were lethal within 30 min. Lower Au concentrationsproduced clusters of smaller AuNPs on the cell wall surface, and were not lethal. These were also SERS-active. We are using SERS to probe the wallcomposition of engineered mutants in the Aspergillus galactofuranose biosynthesis pathway, which plays key roles in fungal growth and drug resistance.We expect the combination of fungal genetic engineering and high sensitivity/high spatial-resolution chemical analysis will provide novel informationabout fungal growth and infectivity.87. Aspergillus nidulans as an experimental system to identify novel cell wall growth and maintenance genes through identification of anti-fungal drug142

FULL POSTER SESSION ABSTRACTSresistance mutations. Xiaoxiao Sean He, Shengnan Jill Li, Susan Kaminskyj. Biology, University of Saskatchewan, Saskatoon, Saskatchewan, Canada.Systemic fungal infections are estimated to contribute to ~10% of hospital deaths. Systemic fungal infections are most dangerous for the young, the old,and the already sick, since their immune systems are less vigorous. Most antifungal drugs in current clinical use target ergosterol (polyenes) or theergosterol biosynthetic pathway (azoles and allylamines). Drugs against beta-glucan synthesis (echinocandins) are effective against aspergillosis andcandidaisis. The use of compounds that target fungal enzymes inevitably leads to the development and natural selection of drug resistant fungal strains.Not only are the anti-fungal drugs in current clinical use losing efficacy in some situations, but in addition the high level of conservation between animaland fungal physiology leaves relatively few relevant targets to explore. However, it is likely that for any drug-enzyme combination there will be relativelyfew mutations that could increase drug resistance while still maintaining enzyme function. We are using Aspergillus nidulans as an experimental modelsystem to assess the number and identity of mutations that lead to drug resistance. As proof of concept, we grew wild type A. nidulans on replicate platescontaining a sub-lethal concentration of Calcofluor. These developed fast-growing sectors beginning at ~ 5 d (70 rounds of mitosis). Preliminary resultsshow that many of these sectors harboured heritable, single-gene mutations. To date, mutated genes that confer robust, heritable resistance to Calcofluorthat were identified by next generation sequencing have roles in cell wall synthesis, cell wall integrity regulation, or drug detoxification. We suggest thisstrategy will be useful for predicting genetically-mediated anti fungal resistance adaptation and help us to be ahead in the drug-resistance arms race.88. Aspergillus nidulans cell walls lacking galactofuranose are more susceptible to glucan degrading enzymes. Biplab Paul 1 , Tanya Dahms 1 , SusanKaminskyj 2 . 1) Dept Chemistry and Biochemistry, Univ Regina, Regina, SK, Canada; 2) Dept Biology, Univ Saskatchewan, Saskatoon, SK, Canada.The cell wall of Aspergillus is a dynamic organ, consisting of a semi-permeable network of mannoprotein, and alpha- and beta-glucans. Thesecomponents are remodeled as fungal cell grows and responds to its environments. By weight, fungal walls are estimated to be 35-45% alpha-(1,3)-glucan,20-35% beta-(1,3)-glucan, 20-25% galactomannan, 7-15% chitin (beta-1,4-glucan), and 4% beta-(1,6)-glucan. Evidence from literature sources suggest thatthe Aspergillus wall 'core' is chitin and galactomannan linked to beta-1,6- and beta-1,6-glucan. Galactofuranose (Gal-f) appears to play a central role inAspergillus cell wall maturation. Previously, we showed that Gal-f biosynthesis is important for wild type chemical, physical, structural properties of the A.nidulans cell wall. We propose that the lack of Gal-f disrupts the proper packing of cell wall components, giving rise to more disordered surface subunitsand so to greater deformability. Here, we show results from an investigation of the susceptibility of Aspergillus Gal-f biosynthesis deletion strains to glucandegrading enzyme using atomic force microscopy. Topographic images of glucanase- and laminarinase-treated wildtype strains suggest that glucan is atleast one component of the cell surface subunits. Strains that lacked Gal-f were more susceptible to beta-1,3-glucanase.89. The GATA-type transcription factor NsdD is a key regulator of conidiation and secondary metabolism in Aspergillus. Mi-Kyung Lee 1 , Nak-Jung Kwon 1 ,Im-Soon Lee 2 , Jae-Hyuk Yu 1 . 1) Bacteriology, University of Wisconsin Madison, MADISON, WI, USA; 2) Department of Biological Sciences, KonkukUniversity, Seoul, Republic of Korea.Asexual development (conidiation for higher fungi) is the most common reproductive mode of many fungi; yet, its regulatory mechanisms remain to beunderstood. In this study, we carried out a multi-copy based genetic screen in the absence of the repressor of conidiation sfgA, which is designed toidentify a new set of negative regulator(s) of conidiation. Among over 100,000 colonies, 45 transformants showing altered conidiation were isolated, ofwhich 10 defined the nsdD gene (AN3152), a key activator of sexual fruiting. The others have defined AN7507, AN2009, AN1652, AN5833 and AN9141. Aseries of verification, genetic and mycotoxin analyses revealed that only NsdD is a true negative regulator of brlA (an essential activator of conidiation) andconidiation, and that NsdD acts downstream of fluG and flbA~E, but upstream of brlA. The removal of NsdD was sufficient to cause hyper-activeconidiation even in liquid submerged culture, as well as early and prolonged activation of brlA, suggesting that NsdD is indeed a key repressor of brlA andconidiation. Moreover, the deletion of nsdD results in hyper-active conidiation and altered production of mycotoxins in the opportunistic human pathogenAspergillus fumigatus and the aflatoxin-producing human/plant pathogen Aspergillus flavus. Importantly, we have discovered that nsdD encodes twodifferentially expressed mRNAs and polypeptides (b and a). Finally, the subsequent transient promoter analysis using the brlA promoter::luciferase fusionconstructs have revealed that NsdD negatively regulates the brlAb promoter activity. In summary, NsdD is a key negative regulator of conidiation actingdirect upstream of brlA in A. nidulans, and couples conidiation and mycotoxin biosynthesis in Aspergilli.90. THE velvet regulators in Aspergilli. Heesoo Park, JJae-Hyuk Yu. Bacteriology, University of Wisconsin Madison, Madison, WI.The velvet regulators are the key players coordinating fungal growth, differentiation and secondary metabolism in response to various internal andexternal cues. All velvet family proteins contain the conserved velvet homology motif (~190 a.a.), and define a novel class of fungal specific transcriptionfactors with the DNA binding ability. Some velvet regulators form time and/or cell type specific complexes with other velvet regulators or non-velvetproteins. These complexes play differential roles in regulating growth, development, sporogenesis and toxigenesis. Among the velvet complexes, the VelB-VosA hetero-complex acts as a functional unit conferring the completion of sporogenesis (focal trehalose biogenesis and spores wall completion), and thelong-term viability of spore, and the attenuation of conidial germination in the model filamentous fungus Aspergillus nidulans. Both velB and vosA areactivated by AbaA in developing cells, and the VelB-VosA complex plays a dual role in activating genes associated with spore maturation and in exertingnegative feedback regulation of developmental genes. Interestingly, the VelB-VosA complex plays similar yet somewhat distinct roles in spore maturation,dormancy and germination in Aspergillus fumigatus and Aspergillus flavus. A comprehensive model depicting the roles of the velvet regulators in aspergilliis presented.91. Coordinated regulation of asexual development, cell death and autolysis by the C2H2 zinc finger transcription factor BrlA in Aspergillus nidulans .István Pócsi 1 , Jae-Hyuk Yu 2 , Tamás Emri 1 . 1) Department of Microbial Biotechnology and Cell Biology, University of Debrecen, Debrecen, Hungary; 2)Departments of Bacteriology and Genetics, University of Wisconsin, Madison, WI, USA.Carbon starvation elicited in submerged cultures of Aspergillus nidulans triggers all various physiological responses affecting cell wall composition, stresstolerance, protein synthesis and primary and secondary metabolisms. Particularly, function of vacuoles and endoplasmic reticulum is drastically affectedleading to the re-utilization of cellular biopolymers through macroautophagy and the removal of damaged cells by apoptosis. Autolytic cell walldegradation is also an integrant part of this highly complicated and delicate regulatory process. Importantly, although the development of conidiophores isinitiated in carbon-starving submerged cultures, these structures are underdeveloped and only simple conidia are observable. There is an increasing bodyof evidence supporting the idea that the transcription factor BrlA, a well-studied central regulator of conidiation in aspergilli, is one of the most importantmaster controllers orchestrating development and autolysis in submerged culture of aspergilli. Major processes subjected to BrlA-dependent regulationunder these conditions include the production of autolytic enzymes, rodlet proteins and melanins. In fungal biology, the concerted and well-balancedregulation of conidiogenesis, cell death and autolysis is of primary importance because any overproduction of cell wall hydrolases may affect cell vitalityand colony propagation rather disadvantageously. The age-dependent production of autolytic hydrolases coincides with the synthesis of antimicrobial27th Fungal Genetics Conference | 143

FULL POSTER SESSION ABSTRACTSmetabolites and proteins, and all these carbon-starvation-associated products will affect markedly the microbiome in the ecological niche the autolyzingfungus occupies. A deeper understanding of the BrlA-mediated spatial and temporal regulatory mechanisms for conidiogenesis, cell death and autolysismay lead to the development of new industrial strains for heterologous protein production and/or novel biocontrol technologies.92. Whole-genome sequencing identifies novel alleles of genes required for organelle distribution and motility in Aspergillus nidulans. Kaeling Tan,Anthony Roberts, Martin Egan, Mark Chonofsky, Samara Reck-Peterson. Cell Biology, Harvard Med Sch, Boston, MA.Many organelles are transported long distances along microtubules in eukaryotic organisms by dynein and kinesin motors. To identify novel alleles andgenes required for microtubule-based transport, we performed a genetic screen in the filamentous fungus, Aspergillus nidulans. We fluorescently-labeledthree different organelle populations known to be cargo of dynein and kinesin in Aspergillus: nuclei, endosomes, and peroxisomes. We then used afluorescence microscopy-based screen to identify mutants with defects in the distribution or motility of these organelles. Using whole-genomesequencing, we found a number of single nucleotide polymorphisms (SNPs) that resulted in misdistribution of peroxisomes, endosomes, or nuclei. Some ofthese SNPs were novel alleles of cytoplasmic dynein/ nudA, Arp1/ nudK (dynactin), Lis1/ nudF, and kinesin-1/ kinA. Here, we characterize the in vivotransport defects in these novel mutants and analyze the single molecule in vitro motility properties of purified mutant motor proteins. We also describeour methods for using whole genome sequencing as a tool in mutagenesis studies in A. nidulans.93. Two methyltransferase protein complexes control fungal development and secondary metabolite production. Oezlem Sarikaya Bayram 1 , OezguerBayram 1 , Jong-Hwa Kim 2 , Keon-Sang Chae 3 , Dong-Min Han 4 , Kap-Hoon Han 2 , Gerhard Braus 1 . 1) Institute of Microbiology & Genetics, Dept. of MolecularMicrobiology and Genetics, Georg August University, Grisebachstr. 8, D 37077 Goettingen, Germany; 2) Department of Pharmaceutical Engineering,Woosuk University, Wanju, 565-701, Korea; 3) Division of Biological Sciences, Chonbuk National University, Jeonju, 561-756, Korea; 4) Division of LifeSciences, Wonkwang University, Iksan, 570-749, Korea.Coordination of development and secondary metabolism of the filamentous fungus Aspergillus nidulans requires the trimeric velvet complex consistingof VelB-VeA and the putative methyltransferase LaeA. We discovered a second trimeric protein complex for the same control mechanism consisting of anunusual zinc finger domain protein and even two subunits containing canonical methyltransferase domains. In contrast to velvet, which is assembled in thenucleus, the novel trimeric protein complex is formed at the plasma membrane. Functional green fluorescent protein fusions revealed that bothmethyltransferases are released from the membrane-bound zinc finger domain and migrate to the nucleus. The dimeric nuclear methyltransferasecomplex physically interacts with chromatin factors as heterochromatin protein and has an impact on the expression of asexual or sexual developmentalgenes as well as secondary metabolite gene clusters. Consistently, deletions of the corresponding genes result in defects in light response. Our resultssupport that a trimeric membrane complex initiates a signalling pathway which is mediated by two methyltransferases which transduce the signal tonuclear chromatin and affect gene expression. The interplay between the novel methyltransferase complex and the velvet complex remains to beelucidated.94. Control of Multicellular Development by the Physically Interacting Deneddylases DEN1/DenA and COP9 Signalosome. Josua Schinke 1 , MartinChristmann 1 , Tilo Schmaler 2 , Colin Gordon 3 , Xiaohua Huang 2 , Özgür Bayram 1 , Sina Stumpf 1 , Wolfgang Dubiel 2 , Gerhard Braus 1 . 1) Microbiology andGenetics, Georg-August-University, Göttingen, Niedersachsen, Germany; 2) Department of General, Visceral, Vascular and Thoracic Surgery, Division ofMolecular Biology, Charité - Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany; 3) MRC Human Genetics Unit, Western General Hospital,Crewe Road, Edinburgh EH4 2XU, UK.Deneddylases remove the ubiquitin-like protein Nedd8 from modified proteins. An increased deneddylase activity has been associated to various humancancers. In contrast, we show here that a mutant strain of the model fungus Aspergillus nidulans which is deficient in two deneddylases is viable but canonly grow as a filament and has lost most of the potential for multicellular development. The DEN1/DenA and the COP9 signalosome (CSN) deneddylasesphysically interact in A. nidulans as well as in human cells, and CSN targets DEN1/DenA for protein degradation. Fungal development responds to light andrequires both deneddylases for an appropriate light reaction. In contrast to CSN which is necessary for sexual development, DEN1/DenA is required forasexual development. The CSN-DEN1/DenA interaction which affects DEN1/DenA protein levels presumably balances cellular deneddylase activity. Adeneddylase disequilibrium impairs multicellular development and suggests that control of deneddylase activity is important for multicellulardevelopment.95. Visualization of apical membrane domains in Aspergillus nidulans by Photoactivated Localization Microscopy (PALM). Norio Takeshita 1 , YujiIshitsuka 2 , Yiming Li 2 , Ulrich Nienhaus 2 , Reinhard Fischer 1 . 1) Dept. of Microbiology, Karlsruhe Institute of Technology, Karlsruhe, Germany; 2) Institute forApplied Physics, Karlsruhe Institute of Technology.Apical sterol-rich plasma membrane domains (SRDs), which can be viewed using the sterol-binding fluorescent dye filipin, are gaining attention for theirimportant roles in polarized growth of filamentous fungi. The size of SRDs is around a few mm, whereas the size of lipid rafts ranges in general between10-200 nm. In recent years, super-resolution microscope techniques have been improving and breaking the diffraction limit of conventional lightmicroscopy whose resolution limit is 250 nm. In this method, a lateral image resolution as high as 20 nm will be a powerful tool to investigate membranemicrodomains. To investigate deeply the relation of lipid membrane domains and protein localization, the distribution of microdomains in SRDs wereanalyzed by super-resolution microscope technique, Photoactivated Localization Microscopy (PALM). Membrane domains were visualized by each markerprotein tagged with photoconvertible fluorescent protein mEosFP for PALM. Size, number, distribution and dynamics of membrane domains, anddynamics of single molecules were investigated. Time-laps analysis revealed the dynamic behavior of exocytosis.144

FULL POSTER SESSION ABSTRACTS96. Cellular morphogenesis of Aspergillus nidulans conidiophores: a systematic survey of protein kinase and phosphatase function. Lakshmi PreethiYerra, Steven Harris. University of Nebraska-Lincoln, Lincoln, NE.In the filamentous fungus Aspergillus nidulans, the transition from hyphal growth to asexual development is associated with dramatic changes inpatterns of cellular morphogenesis and division. These changes enable the formation of airborne conidiophores that culminate in chains of sporesgenerated by repeated budding of phialides. Our objective is to characterize the regulatory modules that mediate these changes and to determine howthey are integrated with the well-characterized network of transcription factors that regulate conidiation in A. nidulans. Because protein phosphorylationis likely to be a key component of these regulatory modules, we have exploited the availability of A. nidulans post-genomic resources to investigate theroles of protein kinases and phosphatases in developmental morphogenesis. We have used the protein kinase and phosphatase deletion mutant librariesmade available by the Fungal Genetics Stock Center to systematically screen for defects in conidiophore morphology and division patterns. Our initialresults implicate ANID_11101.1 (=yeast Hsl1/Gin4) in phialide morphogenesis, and also reveal the importance of ANID_07104.1 (=yeast Yak1) in themaintenance of cell integrity during asexual development. Additional deletion mutants with reproducible defects have been identified and will bedescribed in detail. We will also summarize initial results from double mutant analyses that attempt to place specific protein kinase deletions within theregulatory network that controls conidiation.97. The Putative Guanine Nucleotide Exchange Factor RicA Mediates Upstream Signaling for Growth and Development in Aspergillus. Nak-Jung Kwon 1 ,Hee Soo Park 2 , Seunho Jung 3 , Sun Chang Kim 4 , Jae-Hyuk Yu 1,2 . 1) Dept Bacteriology, University of Wisconsin, Madison, WI. USA; 2) Molecular andEnvironmental Toxicology Center, University of Wisconsin, Madison, WI, USA,; 3) Department of Bioscience and Biotechnology, and Center forBiotechnology Research in UBITA, Konkuk University, Seoul, Republic of Korea; 4) Department of Biological Sciences, Korea Advanced Institute of Scienceand Technology, Dae-Jon, Republic of Korea.Heterotrimeric G proteins (G proteins) govern growth, development, and secondary metabolism in various fungi. Here, we characterized ricA, whichencodes a putative GDP/GTP exchange factor for G proteins in the model fungus Aspergillus nidulans and the opportunistic human pathogen Aspergillusfumigatus. In both species, ricA mRNA accumulates during vegetative growth and early developmental phases, but it is not present in spores. The deletionof ricA results in severely impaired colony growth and the total (for A. nidulans) or near (for A. fumigatus) absence of asexual sporulation (conidiation). Theoverexpression (OE) of the A. fumigatus ricA gene (AfricA) restores growth and conidiation in the DAnricA mutant to some extent, indicating partialconservation of RicA function in Aspergillus. A series of double mutant analyses revealed that the removal of RgsA (an RGS protein of the GanB Gasubunit), but not sfgA, flbA, rgsB, or rgsC, restored vegetative growth and conidiation in AnricA. Furthermore, we found that RicA can physically interactwith GanB in yeast and in vitro. Moreover, the presence of two copies or OE of pkaA suppresses the profound defects caused by DAnricA, indicating thatRicA-mediated growth and developmental signaling is primarily through GanB and PkaA in A. nidulans. Despite the lack of conidiation, brlA and vosAmRNAs accumulated to normal levels in the ricA mutant. In addition, mutants overexpressing fluG or brlA (OEfluG or OEbrlA) failed to restore developmentin the AnricA mutant. These findings suggest that the commencement of asexual development requires unknown RicA-mediated signaling input in A.nidulans.98. Evidence for a role of peroxisomes in microtubule organization. Ying Zhang, Andreas Herr, Reinhard Fischer. Karlsruhe Institute of Technology,Karlsruhe, Germany.In Aspergillus nidulans spindle pole bodies (SPBs) and septum-associated microtubule-organizing centres (sMTOCs) polymerize cytoplasmic microtubules.Previously, we identified a novel MTOC-associated protein, ApsB (Schizosaccharomyces pombe mto1), whose absence affected MT formation fromsMTOCs more than from SPBs, suggesting that the two protein complexes are organized differently (Suelmann et al., 1998; Veith et al., 2005). In addition,we discovered that ApsB localizes to a subclass of peroxisomes apparently without a peroxisomal targeting motif. However, we found that ApsB interactswith the Woronin body protein HexA, which has a PTS1 motif at the C-terminus (Zekert er al., 2010). Our hypothesis is that ApsB is imported toperoxisomes by a piggyback import mechanism along with HexA. To further investigate the role of peroxisomes in microtubule orgnization, We created adeletion mutant of pexC. PexC is an essential protein for peroxisomal biogenesis (Heiland & Erdmann, 2005). The pexC mutant partially phenocopied theapsB mutant, which shows reduced sporulation and nuclear migration defects in comparison to wild type. The number of astral and cytoplasmicmicrotubules and the activities of sMTOCs and SPBs was reduced in the pexC mutant in comparison to wild type. sMTOC activity was more affected thanthe SPB activity, which again resembles the phenotype of the apsB mutant. In conclusion, peroxisomes play a role in microtubule organization throughApsB.99. Autophagy promotes survival in aging submerged cultures of the filamentous fungus Aspergillus niger. Maria A. Burggraaf 1,2 , Benjamin M. Nitsche 1,2 ,Gerda Lamers 1 , Vera Meyer 2,3 , Arthur F.J. Ram 1,2 . 1) Institute of Biology Leiden, Molecular Microbiology and Biotechnology, Leiden, The Netherlands; 2)Kluyver Centre for Genomics of Industrial Fermentation, Delft, The Netherlands; 3) Institute of Biotechnology, Applied and Molecular Microbiology, BerlinUniversity of Technology, Berlin, Germany.The filamentous fungus Aspergillus niger is an important and versatile cell factory commonly exploited for the industrial-scale production of a wide rangeof enzymes and organic acids. Although numerous studies have been conducted aiming at improving our knowledge of degradative cellular activities thatdetermine product yields in A. niger including secretion of proteases and the unfolded protein response, there is a catabolic pathway that has yet not beenstudied in this industrially exploited fungus, namely Autophagy. Autophagy is a well conserved catabolic process constitutively active in eukaryotes that isinvolved in cellular homeostasis by targeting of cytoplasmic content and organelles to vacuoles. Autophagy is strongly induced by limitation of nutrientsincluding carbon, nitrogen and oxygen and is clearly associated with cell death. We previously demonstrated that the accumulation of empty hyphalcompartments and secondary regrowth in carbon starved submerged batch cultures of A. niger were accompanied by a joint transcriptional induction ofautophagy genes. In this study we examined the role of autophagy by deleting the atg1, atg8 and atg17 orthologues in A. niger and phenotypicallyanalyzing the deletion strains in surface and submerged cultures. Our results indicate that atg1 and atg8 are essential for efficient autophagy whereasdeletion of atg17 has little to no effect on autophagy. Depending on the stressor, autophagy deficiency renders A. niger both more resistant and moresensitive to oxidative stress. Fluorescence microscopy showed that mitochondrial turnover upon carbon depletion in submerged cultures is severelyblocked in autophagy impaired mutants. Furthermore, automated image analysis demonstrated that autophagy promotes survival in maintained carbonstarved cultures of A. niger. Taken together, our results suggest that besides its function in nutrient recycling, autophagy plays important roles inphysiological adaptation by organelle turnover and protection against cell death upon carbon depletion in submerged cultures.27th Fungal Genetics Conference | 145

FULL POSTER SESSION ABSTRACTS100. Inactivation of flbA results in increased secretome complexity and reduced secretion heterogeneity in colonies of Aspergillus niger. PaulineKrijgsheld 1 , Benjamin M. Nitsche 2 , Harm Post 3 , Ana M. Levin 1 , Wally H. Müller 4 , Albert J.R. Heck 3 , Arthur F.J. Ram 2 , A.F. Maarten Altelaar 3 , Han A.B.Wösten 1 . 1) Microbiology and Kluyver Centre for Genomics of Industrial Fermentation, Utrecht University, Utrecht,The Netherlands; 2) Department ofMolecular Microbiology and Biotechnology, Institute of Biology Leiden and Kluyver Centre for Genomics of Industrial Fermentation, Leiden University,Leiden, The Netherlands; 3) Biomolecular Mass Spectrometry and Proteomics, Netherlands Proteomics Center, Bijvoet Center for Biomolecular Researchand Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands; 4) Biomolecular Imaging, Utrecht University, Utrecht, TheNetherlands.Aspergilli are among the most common fungi. They colonize substrates by secreting enzymes that degrade organic polymers into small products that canbe taken up by the fungus to serve as nutrient. Hyphae at the periphery of the colony are exposed to unexplored organic material, whereas the substrateis (partly) utilized in the colony center. Aspergillus niger is known for its capacity to secrete high amounts of proteins. Interestingly, the fungus secretesproteins in the central part and at the periphery of the colony but not in the sub-peripheral zone. The sporulating zone of the colony overlaps with thenon-secreting zone, indicating that sporulation inhibits protein secretion. Indeed, strain DflbA that is affected early in the sporulation program secretedproteins throughout the colony. In contrast, the DbrlA strain that still initiates but not completes sporulation did not show an altered spatial secretion. Thesecretome of 5 concentric zones of 7-dayold xylose-grown DflbA mutant colonies of A. niger was assessed by quantitative proteomics using stable isotopedimethyl labeling. In total 171 proteins were identified in the medium of the DflbA colonies, of which 33 proteins did not have a signal sequence forsecretion. Out of the 138 secreted proteins, 101 had previously not been identified in the secretome of the 5 concentric zones of xylose-grown wild-typecolonies. Moreover, 18 proteins had never been reported to be part of the secretome of A. niger. Taken together, inactivation of flbA, but not brlA resultsin spatial changes in secretion and in a more complex secretome. The latter may be explained by the fact that strain DflbA has a thinner cell wall comparedto the wild type, enabling efficient release of proteins. These results can implemented in the industry to improve A. niger as a cell factory.This research was financed by the Kluyver Centre for Genomics of Industrial Fermentation and by the Netherlands Proteomics Centre, which are part ofthe Netherlands Genomics Initiative/ Netherlands Organisation for Scientific Research.101. Functional characterization of A. niger class III and class V chitin synthases and their role in cell wall integrity. Jean-Paul Ouedraogo 1 , Arthur Ram 1 ,Vera Meyer 2 . 1) Molecular Microbiology and Biotechnology, Institut of Biology, Leiden, Netherlands; 2) Molecular and Applied Microbiology, Institut ofBiotechnology, Berlin University of Technology, Berlin, Germany.Class III and V chitin synthases play an important role in morphogenesis and cell wall integrity in many filamentous fungi. However, their function in thefilamentous fungus, A. niger has not yet been elucidated. To address this, deletion mutants of class III and V chitin synthase-encoding genes of A.niger,chsB and csmB, and their role in cell wall integrity have been studied. Deficiency in conidiation and abnormal swollen conidiophores have been observed inchsB and csmB deletion mutants. Using cell wall inhibitor reagents, it was shown that the mutants are hypersensitive towards cell wall stress. However,there are differences between them as regards susceptibility to the antifungal protein AFP. These results suggest that ChsB and CsmB play an importantrole during asexual development and in ensuring cell wall integrity of A. niger. Interestingly, the data indicate that only chitin synthase csmB is importantto counteract AFP inhibitory effects.102. Exploiting transcriptomic signatures of Aspergillus niger to uncover key genes important for high protein traffic through its secretory pathway. MinJin Kwon 1,2 , Thomas Jørgensen 1 , Benjamin M Nitsche 1,3 , Mark Arentshorst 1 , Joohae Park 1 , Arthur F.J. Ram 1,2 , Vera Meyer 1,3 . 1) Molecular Microbiology,Institute of Biology Leiden, Leiden, Netherlands; 2) Kluyver Centre for Genomics of Industrial Fermentation, P.O. Box 5057, 2600 GA Delft, TheNetherlands; 3) Institute of Biotechnology, Department Applied and Molecular Microbiology, Berlin University of Technology, Gustav-Meyer-Allee 25,13355 Berlin, Germany.The filamentous fungus Aspergillus niger is well known for its exceptional high capacity to secrete proteins. However, system-wide insights into itssecretory capacities are sparse and rational strain improvement approaches are thus limited. To gain a global view on the transcriptional basis of thesecretory pathway of A. niger, we have investigated its transcriptomic fingerprint when specifically forced to overexpress the hydrolytic enzymeglucoamylase (GLA). An A. niger wild-type strain and an GLA over-expressing strain where cultivated under maltose-limited chemostat conditions. ElevatedglaA mRNA and extracellular GLA levels in the over-expressing strain were accompanied by reinforced transcription of 772 genes and down-regulation of815 genes when compared to the wild-type situation. Using GO term enrichment analysis, four higher order categories were identified in the up-regulatedgene set: i) translocation, ii) protein glycosylation, iii) vesicle transport and iv) ion homeostasis. Among these, about 130 genes have predicted functionsfor the protein passage through the endoplasmaticum reticulum including well-known target genes of the HacA transcription factor, e.g. bipA, clxA, prpA,tigA and pdiA. To identify those genes, which are generally important for high-level secretion in A. niger, we compared the GLA transcriptome with sixother secretion stress transcriptomes of A. niger, including a constitutive active HacA transcriptome, several UPR stress transcriptomes and a carbonsourceinduced secretion transcriptome. Overall, 40 genes were commonly up-/down-regulated under these three conditions (36 genes up-regulated, 4down-regulated), thus defining the core set of genes important for ensuring high protein traffic through the secretory pathway.103. Identification of two Golgi-localized putative UDP-galactofuranose transporters with overlapping function in Aspergillus niger. Joohae Park 1 , BorisTefsen 2 , Ellen Lagendijk 1 , Irma van Die 2 , Arthur Ram 1,3 . 1) Molecular Microbiology, Institute of Biology Leiden, Leiden, Netherlands; 2) Department ofMolecular Cell Biology and Immunology, VU University Medical Center, van den Boechorststraat 7, 1081 BT Amsterdam, The Netherlands,; 3) KluyverCentre for Genomics of Industrial Fermentation, P.O box 5057, 2600 GA Delft, The Netherlands.Galactofuranose-containing glycoconjugates are present in numerous microbes, many of which are pathogenic for humans. Metabolic aspects of themonosaccharide have proven difficult to elucidate, because galactofuranose metabolites and glycoconjugates are relatively unstable during analyses.Recent advances with genetic approaches have facilitated a better understanding of galactofuranose biosynthesis. Galactofuranose (Galf) the five-ringisomer of galactopyranose (Galp), is an essential component of the cell wall and required for a structural integrity [1-2]. Recently, it has been postulatedthat UDP-Galp, is converted to Galf by a UDP-galactopyranose mutase (UgmA) and subsequently transported into the Golgi by a putative UDP-Galftransporterfor the further biosynthesis of cell wall polymers such as galactomannan, galactoaminogalactan and cell wall glycoproteins (galactomannoproteins)[3-4]. Based on homology searches, we have identified two putative UDP-Galf-transporters in A. niger. We have studied the function of thetransporters by making deletions mutants (either single or double mutants) and by studying their localization by making GFP fusions. We conclude that thetwo putative UDP-Galf-transporters (named (UgtA and UgtB) have an overlapping function in UDP-Galf-transport and that both proteins are localized inGolgi equivalents. References: [1] Damveld, R.A. et al., 2008. Genetics 178 (2), 873-81; [2] Schmalhorst, P.S. et al. 2008, Euk. Cell 7 (8), 1268-77; [3] Engel, J.et al., 2009. J. Biol. Chem. 284; [4] Bernard, M., Latge, J. P., 2001. Med. Myc. 39, 9-17;.146

FULL POSTER SESSION ABSTRACTS104. Maltose permease-encoding mRNA is cleaved under induction condition of amylolytic gene expression in Aspergillus oryzae. Mizuki Tanaka,Takahiro Shintani, Katsuya Gomi. Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi, Japan.Eukaryotic mRNA is degraded by two degradation pathways: the 5' to 3' degradation pathway by Xrn1 and the 3' to 5' degradation pathway by exosome-Ski complex. To investigate the mRNA degradation mechanism in filamentous fungi, we generated the disruptions of orthologous genes encoding mRNAdegradation machinery in Aspergillus oryzae. Interestingly, the disruptants of ski2 and ski3, which encode the components of Ski complex, showed theremarkable growth defect on minimal medium containing maltose or starch as a sole carbon source, whereas they normally grew on the medium withglucose or fructose as a sole carbon source. Northern blot analysis showed that the 3'-truncated fragment of mRNA encoding maltose permease (malP)was accumulated in Ski complex deficient mutants. Circularized RT-PCR analysis revealed that the malP mRNA was cleaved at a large stem-loop structuresituated within the coding region. These results suggested that the malP mRNA is cleaved by endonuclease and the resultant 3'-truncated malP mRNA isdegraded rapidly by 3' to 5' degradation pathway. In higher eukaryotes, it has been reported that the mRNAs encoding secreted and membrane proteinswere cleaved by endoplasmic-reticulum (ER) endonuclease Ire1 during ER stress. Since A. oryzae produces copious amounts of amylolytic enzymes in thepresence of maltose, we presumed that malP mRNA is cleaved by Ire1 with the induction of amylolytic gene expression. Therefore, we generated thedouble deficient mutant for Ski complex and AmyR, the regulator of amylolytic genes expression. The resultant double mutant showed normal growth onmaltose medium, and 3'-truncated fragment of malP mRNA was not detected by Northern blot analysis. This result clearly indicated that malP mRNA iscleaved under induction condition of amylolytic gene expression in A. oryzae.105. Functional characterisation of Rac GTPase in Botrytis cinerea reveals impact on polarity, cell cycle and pathogenicity. Anna Minz-Dub 1 , LeonieKokkelink 2 , Paul Tudzynski 3 , Amir Sharon 1 . 1) Department of Plant Sciences, Britannia 536, Tel-Aviv University, Tel-Aviv 69978; 2) Universität zu Köln,Biozentrum, Institut für Botanik, Zülpicher Str. 47 b, 50674 Koeln, Germany; 3) Institut für Biologie und Biotechnologie der Pflanzen ,WestfaelischeWilhelms-Universitaet Muenster, Schlossplatz 8, D-48143 Muenster, Germany.Small GTPases of the Ras superfamily are involved in regulation of different cellular mechanisms including cell cycle and differentiation. Furthermore,small GTPase proteins are interconnected with many different signalling pathways. In this study we describe functional characterization of a Rho-typeGTPase BcRac from the necrotrophic plant pathogen Botrytis cinerea. Role of this protein in cell cycle, development and pathogenicity is described.Expression of a constitutively active (CA) version of the BcRac protein, or deletion of the gene had a severe impact on fungal growth and differentiation.The mutant strains have polarity defects, they do not produce conidia, disease symptoms on plants are delayed, and they produce and accumulateincreased amounts of ROS in culture. In addition, nuclear content and actin localization were altered in the CA-BcRac strain as compared to wild type. Aneffect of Rac-specific inhibitor NSC23766 on spore germination of wild type strain indicated that BcRac might be necessary for spore germination duringG2/M phase. Based on our observations, BcRac is an important regulator of development in B. cinerea, and alteration of its activity disrupts themorphogenetic program and influences fungal infection.106. Light matters: The transcription factor LTF1 regulates virulence and light responses in the necrotrophic plant pathogen Botrytis cinerea. JuliaSchumacher 1 , Adeline Simon 2 , Kim Cohrs 1 , Muriel Viaud 2 , Paul Tudzynski 1 . 1) IBBP, WWU Muenster, Schlossplatz 8, 48143 Muenster, Germany; 2) INRA,BIOGER, Avenue Lucien Brétignières, 78850 Grignon, France.The lifecycle of Botrytis cinerea/ Botryotinia fuckeliana includes the formation of white mycelia generating pigmented conidiophores with macroconidiafor propagation, pigmented sclerotia for over-wintering and sexual reproduction, microconidia for spermatization of the sclerotia, and the formation ofapothecia as fruiting bodies on spermatized sclerotia. Full-spectrum light induces the differentiation of conidia and apothecia, while sclerotia areexclusively formed during incubation in constant darkness. The relevance of light for virulence of the fungus is not that clear, however, infections areobserved under natural illumination conditions as well as in constant darkness. By a T-DNA insertional mutagenesis approach, we identified a novelvirulence-related gene encoding a GATA-type transcription factor (TF) with homologues in A. nidulans (NsdD) and N. crassa (SUB-1). As transcription isinduced by light (2.5-fold), it is called BcLTF1 for ‘Light-regulated TF 1’. By deletion and over-expression of BcLTF1, we confirmed the predicted role of theTF in virulence, and discovered furthermore its extraordinary functions in regulating light-dependent differentiation processes (growth defect of Dbcltf1 inlight, loss of sclerotia formation in darkness), the equilibrium between production and scavenging of reactive oxygen species (ROS), and secondarymetabolism. Hence, microarray analyses (WT, Dbcltf1; dark vs. exposure to light for 1h) revealed that the expression levels of 206 out of 313 lightdependentgenes are modulated by BcLTF1, including the genes of the putative carotenoid gene cluster and six out of eleven genes encoding TFs. Inaddition, the mutation of bcltf1 affects the expression of 1,616 genes irrespective of the light conditions, including the over-expression of known and so faruncharacterized secondary metabolism gene clusters. The over-expression of the gene encoding the alternative oxidase (AOX) and the under-expression ofgenes involved in oxidative stress responses are in accordance with the observed phenotypes of the deletion mutant, i.e. the hypersensitivity toexogenously applied oxidative stress even in the absence of light and the restoration of growth rates in continuous light by offering antioxidants, indicatingthat BcLTF1 is required to cope with oxidative stress that is caused by the exposure to light.107. Functional analysis of genes in the mating type locus of Botrytis cinerea. Razak Bin Terhem, Joost Stassen, Jan van Kan. Laboratory ofPhytopathology, Wageningen University, Wageningen, The Netherlands.Botrytis cinerea is a heterothallic ascomycete with two mating types, MAT1-1 and MAT1-2, each containing two genes. Besides the archetypal genesencoding the MAT1-1-1 (alpha-domain) protein and the MAT1-2-1 (HMG-box) protein, each idiomorph contains one additional gene, designated MAT1-1-5and MAT1-2-4, respectively. Homologs of these genes are only found in closely related taxa, and their function is as yet unknown. Knockout mutants weregenerated in all four genes in the B. cinerea MAT locus, either in the MAT1-1 strain SAS56 or in the MAT1-2 strain SAS405. Mutants were crossed with astrain of the opposite mating type, either the wild type or a knockout mutant, in all possible combinations. Knockout mutants in the MAT1-1-1 gene andthe MAT1-2-1 gene fail to show any sign of primordial outgrowth and are entirely sterile. This confirms the essential role of the alpha-domain protein andthe HMG-box protein in the mating process. By contrast, mutants in the MAT1-1-5 gene and the MAT1-2-4 gene do produce stipes, but these fail todevelop further into an apothecial disk. The MAT1-1-5 and MAT1-2-4 mutants show identical phenotypes, suggesting that these two genes jointly controlthe transition from stipe to disk development. RNA-seq data were obtained from a cross between two wild type strains and from a cross involving a MAT1-1-5 knockout mutant, from tissue at the stage of transition from stipe to disk. Differential gene expression analysis was performed to identify genes thatare possibly involved in development of the apothecial disk.27th Fungal Genetics Conference | 147

FULL POSTER SESSION ABSTRACTS108. The role of hydrophobins in sexual development of Botrytis cinerea. Razak Bin Terhem 1 , Matthias Hahn 2 , Jan van Kan 1 . 1) Laboratory ofPhytopathology, Wageningen University, Wageningen, The Netherlands; 2) Department of Biology, University of Kaiserslautern, Kaiserslautern, Germany.Hydrophobins are small secreted proteins that play a role in a broad range of developmental processes in filamentous fungi, e.g. in the formation ofaerial structures. Hydrophobins allow fungi to escape their aqueous environment and confer hydrophobicity to fungal surfaces. In Botrytis cinerea(teleomorph Botryotinia fuckeliana), one class I and two class II hydrophobin genes have been identified, as well as a number of hydrophobin-like genes.Previous studies showed that hydrophobins are not required for conferring surface hydrophobicity to conidia and aerial hyphae. We investigated the roleof hydrophobins in sclerotium and apothecium development. RNA seq analysis of gene expression during different stages of apothecium developmentrevealed high expression of the Bhp1 (class I hydrophobin) gene and of the Bhl1 (hydrophobin-like) gene in certain stages, whereas Bhp2 and Bhp3 (class IIhydrophobin) genes were always expressed at very low levels. We characterized different hydrophobin mutants: four single gene knockouts, three doubleknockouts as well as a triple knockout. Sclerotia of DBhp1/DBhp3 (double knock out) and DBhp1/DBhp2/DBhp3 (triple knock out) mutants showed easilywettable phenotypes. These results indicate that hydrophobins Bhp1 and Bhp3 are important for normal development of sclerotia of B. cinerea. Foranalyzing apothecial development, a reciprocal crossing scheme was set up. Morphological aberrations were observed in crosses with some hydrophobinmutants. When the DBhp1/DBhp2 (double knock out) and DBhp1/DBhp2/DBhp3 (triple knockout) mutants bearing a MAT1-1 mating type were used asmaternal parents (sclerotia), and fertilized with microconidia of a wild type MAT1-2 strain, the resulting apothecia were swollen, dark brown in color andhad a blotted surface. Instead of growing upwards, the apothecia in some cases fell down. This aberrant apothecial development was not observed in thereciprocal cross, when the same mutants were used as paternal parent (microconidia). These results indicate that the presence of hydrophobins Bhp1 andBhp2 in maternal tissue is important for normal development of apothecia of B. cinerea.109. The pescadillo homolog, controlled by Tor, coordinates proliferation and growth and response in Candida albicans yeast. Tahmeena Chowdhury 1 ,Niketa Jani 1 , Folkert J. Van Werven 2 , Robert J. Bastidas 3 , Joseph Heitman 3 , Julia R. Köhler 1 . 1) Division of Infectious Diseases, Boston Children'sHospital/Harvard Medical School, Boston, MA; 2) Institute for Integrative Cancer Research, MIT, Cambridge, MA; 3) Dept. of Genetics and MolecularBiology, Duke University, NC.Candida albicans has evolved as a colonizer and opportunistic pathogen of mammals. Among fungi infecting humans, it is unique in the frequency withwhich it switches between growth as budding yeast and growth as pseudohyphal and hyphal filaments. In vitro and presumably in vivo, filamentsconstitutively produce yeast from their sub-apical compartments. This developmental step is required for dispersal of planktonic yeast from biofilms. TheC. albicans pescadillo homolog PES1 is required for this lateral yeast growth. In eukaryotes, pescadillo homologs are involved in cell cycle progression andribosome biogenesis, processes that respond to nutrient availability. This work investigated the potential role of C. albicans PES1 in the Tor signalingpathway, which is a major nutrient signaling cascade. Results show that Tor signaling controls Pes1 expression and localization. C. albicans yeast but nothyphae require Pes1 for proliferation, and for proliferation arrest upon Tor1 inhibition with rapamycin. Pes1 inactivation via a temperature-sensitive alleleleads to defective exit of starved cells from the cell cycle. Pes1 inactivation also leads to rapid loss of phosphorylation of ribosomal protein S6, a marker oftranslational activity, as does Tor1 inhibition and genetic perturbation of Tor1 activation. These data support a role for Pes1 downstream of Tor1 incoordinating cell cycle progression with protein synthesis. As all cells must coordinate proliferation and growth, investigating why the requirement forPes1 in this role is yeast-specific will inform our understanding of morphogenesis and Tor signaling in C. albicans.110. Uncovering the mechanisms of thermal adaptation in Candida albicans. Michelle Leach 1,2 , Susan Budge 2 , Louise Walker 2 , Carol Munro 2 , AlistairBrown 2 , Leah Cowen 1 . 1) Department of Molecular Genetics, University of Toronto, Medical Sciences Building, 1 Kings College Circle, Toronto, Ontario,Canada, M5S 1A8; 2) Aberdeen Fungal Group, School of Medical Sciences, University of Aberdeen, Institute of Medical Sciences, Foresterhill, Aberdeen,AB25 2ZD, UK.The heat shock response is governed by one of the most highly conserved networks in eukaryotic cells. Upon sensing a sudden temperature upshift, theheat shock transcription factor (Hsf1) is rapidly phosphorylated and activated, leading to the induction of numerous genes that mediate thermaladaptation, including heat shock genes that encode molecular chaperones. We have shown that the major fungal pathogen of humans, Candida albicans,has retained a bona fide heat shock response even though it is obligatorily associated with warm-blooded animals [Molec. Micro. 74, 844]. Furthermore,this thermal adaptation is essential for the virulence of C. albicans [Fungal Gen. Biol. 48, 297]. To identify signalling pathways that contribute to long-termthermal adaptation resistance in C. albicans we performed unbiased genetic screens for protein kinase mutants that display temperature sensitivity. Thisscreen reproducibly highlighted several key signalling pathways associated with cell wall remodelling: the Hog1, Mkc1 and Cek1 pathways. None of thesepathways are essential for Hsf1 phosphorylation and activation; each pathway contributing to heat shock adaptation independently of Hsf1. Wedemonstrate that these pathways are differentially activated during heat shock, and that there is crosstalk between these pathways, with hightemperatures contributing to increased resistance to cell wall stress in the long term, and oxidative stress in the short term. Critically, this crosstalkbetween thermotolerance and other types of stress adaptation is mediated by the molecular chaperone Hsp90, whose down-regulation reduces theresistance of C. albicans to proteotoxic stresses. Hsp90 depletion also affects cell wall biogenesis by impairing activation of these signalling pathways.Furthermore, we show that Hsp90 interacts with and down-regulates Hsf1 thereby modulating short-term thermal adaptation. Therefore, Hsp90 lies at theheart of heat shock adaptation, modulating the short-term Hsf1-mediated activation of the classic heat shock response, coordinating this response withlong term thermal adaptation via Mkc1- Hog1- and Cek1-mediated cell wall remodelling.111. Characterisation of contact-dependant tip re-orientation in Candida albicans hyphae. Darren Thomson, Silvia Wehmeier, Alex Brand. AberdeenFungal Group, Aberdeen University, Aberdeen, United Kingdom.Candida albicans is a pleiomorphic fungus that lives as a commensal yeast in the human body but can become pathogenic in susceptible patient groups.Virulence is strongly linked with the production of penetrative hyphae that can adhere to and invade a wide range of substrates, including blood vessels,organ tissue, keratinised finger-nails and even soft medical plastics. Using live-cell imaging and nanofabricated surfaces, we are characterising the spatiotemporaldynamics of contact-induced hyphal tip behaviour (thigmotropism). To test whether tip re-orientation responses positively correlate with levelsof hyphal adhesion, we generated substrates with increasing adhesive force. Hyphal tip re-orientation was absent in poorly-immobilised hyphae and athreshold adhesive force was required sub-apically to generate the hyphal tip pressure required for re-orientation. Interestingly, sub-threshold adhesionresulted in sub-apical hyphal bending. Localization of fluorescent protein markers for the Spitzenkörper and the Polarisome (Mlc1-YFP and Spa2-YFP,respectively) showed that C. albicans hyphal tips grow in an asymmetric, ‘nose-down’ manner on a surface. Additionally, hyphal tips can detect surfacestiffness and show a distinct preference for nose-down growth on the softer of two substrates. Localisation of fluorescent cell-cycle reporter proteins overtime revealed that hyphal tip contact slowed the cell-cycle, suggesting that tip-contact perturbs cell-cycle mechanics. Finally, we examined the role ofcytoskeleton regulators in thigmotropism and determined the force that can be generated by the hyphal tip. Our results suggest that C. albicans hyphae148

FULL POSTER SESSION ABSTRACTScan exert sufficient force to penetrate human epithelial tissue without the need for secreted enzyme activity. This is consistent with the observed hyphalpenetration of medical-grade silicone, which has a similar Young’s modulus to human cartilage.112. Cdc14 association with basal bodies in the oomycete Phytophthora infestans indicates potential new role for this protein phosphatase. AudreyM.V. Ah-Fong, Howard S. Judelson. Plant Pathology & Microbiology, University of California, Riverside, CA.The dual-specificity phosphatase Cdc14 is best known as a regulator of cell cycle events such as mitosis and cytokinesis in yeast and animal cells.However, the diversity of processes affected by Cdc14 in different eukaryotes raises the question of whether its cell cycle functions are truly conservedbetween species. Analyzing Cdc14 in Phytophthora infestans should provide further insight into the role of Cdc14 since this organism does not exhibit aclassical cell cycle. Prior study in this organism already revealed novel features of its Cdc14. For example, instead of being post-translationally regulatedlike its fungal and metazoan relatives, PiCdc14 appears to be mainly under transcriptional control. It is absent in vegetative hyphae where mitosis occursand expressed only during the spore stages of the life cycle which are mitotically quiescent, in contrast to other systems where it is expressedconstitutively. Since transformants overexpressing PiCdc14 exhibit normal nuclear behavior, the protein likely does not play a critical role in mitoticprogression although PiCdc14 is known to complement a yeast Cdc14 mutation that normally arrests mitosis. Further investigation into the role of PiCdc14uncovered a novel role. Subcellular localization studies based on fusions with fluorescent tags showed that PiCdc14 first appeared in nuclei during earlysporulation. During the development of biflagellated zoospores from sporangia, PiCdc14 transits to basal bodies, which are the sites from which flagelladevelop. A connection between Cdc14 and flagella is also supported by their phylogenetic distribution, suggesting an ancestral role of Cdc14 in basalbodies and/or flagellated cells. To help unravel the link between PiCdc14 and the flagella apparatus, searches for its interacting partners using both yeasttwo hybrid and affinity purification are underway. Together with colocalization studies involving known basal body/centrosome markers such as centrinand gamma-tubulin, the location and hence the likely roles of PiCdc14 will be revealed.113. Colletotrichum orbiculare Bub2-Bfa1 complex, a spindle position checkpoint (SPOC) component in Saccharomyces cerevisiae, is involved in properprogression of cell cycle. Fumi Fukada 1 , Ayumu Sakaguchi 2 , Yasuyuki Kubo 1 . 1) Laboratory of Plant Pathology, Graduate School of Life and EnvironmentalSciences, Kyoto Prefectural University, Kyoto, Japan; 2) National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan.Colletotrichum orbiculare is an ascomycete fungus that causes anthracnose of cucumber. In Saccharomyces cerevisiae, the orientation of the mitoticspindle with respect to the polarity axis is crucial for the accuracy of asymmetric cell division. A surveillance mechanism named spindle position checkpoint(SPOC) prevents exit from mitosis when the mitotic spindle fails to align along the mother-daughter polarity axis. BUB2 is a component of SPOC andconstitutes the main switch for the mitotic exit network (MEN) signaling. We identified and named this homolog as CoBUB2 in C. orbiculare and generatedgene knock-out mutants. First, we observed morphogenesis and pathogenesis of the cobub2 mutants. The cobub2 mutants formed abnormal appressoriaand penetration hyphae on model substrates, and the cobub2 mutants also showed attenuate pathogenesis to cucumber leaves. Second, we observedmitosis based on mitotic spindle behavior and nuclear DAPI staining during appressorium development. In the wild type, mitosis occurred in appressoriumdeveloping conidia after 4h incubation, whereas interestingly, in the cobub2 mutants, mitosis occurred in pre-germinated conidia after 2h incubation.After development of appressorium, in some germlings the daughter nucleus was delivered from conidia to appressoria, and the others perform secondround of mitosis in appressorium developing conidia after 4h incubation. Third, we evaluated the timing of S phase and M phase during appressoriumdevelopment in wild type and the cobub2 mutants by cell cycle specific inhibitors. In the cobub2 mutants, it was shown that the transition period from G1phase to S phase accelerated about 2h than that of the wild type. Last, in S. cerevisiae, Bub2 forms GTPase activating protein (GAP) complex with Bfa1, andBub2-Bfa1 GAP complex constitutes SPOC. Then we named homolog of BFA1 as CoBFA1 in C. orbiculare and generated cobfa1 mutants. From observationof nuclear division, the cobfa1 mutants showed similar behavior of nuclear division to the cobub2 mutants. Therefore, it is assumed that CoBub2 formsGAP complex with CoBfa1, however, CoBub2-CoBfa1 GAP complex has different function from that in S. cerevisiae maintaining G1 phase duration orsetting up the proper time of S phase.114. Metazoan-like mitotic events in the basidiomycetous budding yeast Cryptococcus neoformans - a human fungal pathogen. L. Kozubowski 1,2 , V.Yadav 3 , G. Chatterjee 3 , M. Yamaguchi 5 , I. Bose 4 , J. Heitman 2 , K. Sanyal 3 . 1) Department of Medicine, Division of Infectious Diseases, Duke UniversityMedical Center, Durham, NC, USA; 2) Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC, USA; 3)Molecular Mycology Laboratory, Molecular Biology and Genetics Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore, India; 4)Department of Biology, Western Carolina University, Cullowhee, NC, USA; 5) Medical Mycology Research Center, Chiba University, Chiba, Japan.Mitosis in ascomycetous budding yeasts is characterized by several features that are distinct from those of metazoans. In Saccharomyces cerevisiae,centromeres are always clustered in a single spot, the kinetochores are fully assembled for the majority of the cell cycle, and the nuclear envelope (NE)does not break down (closed mitosis). Currently it is not clear how these mechanisms evolved or whether these features are a universal characteristichallmark of the budding mode of cellular division. Here we report an analysis of key mitotic events in the basidiomycetous human fungal pathogenCryptococcus neoformans. The dynamics of microtubules, the kinetochore, NE and the nucleolus were analyzed by time-lapse microscopy usingfluorescently tagged proteins. In striking contrast to ascomycetous budding yeast, centromeres in C. neoformans were not clustered in non-dividing cells.Prior to mitosis, centromeres underwent gradual clustering, eventually forming a single spot, which then migrated into the daughter cell where thechromosomal division occurred. One set of chromosomes migrated back to the mother cell and subsequent de-clustering of centromeres occurred in bothcells. Analysis of individual components of the kinetochore indicated that kinetochores assemble in a step-wise manner in C. neoformans. While the innerkinetochore (Cse4, Mif2) was present throughout the entire cell cycle, the middle kinetochore (Mtw1) assembled prior to mitosis when centromeresunderwent clustering, and this was then followed by assembly of the outer kinetochore (Dad1, Dad2). Formation of the outer kinetochore during mitosis,as observed in metaozoans that undergo an open mitosis, prompted us to examine the fate of the NE at various cell cycle stages. Several lines of evidencesuggested that C. neoformans undergoes a semi-open mitosis. The nuclear pore marker GFP-Nup107, and a nucleolar marker GFP-Nop1 dispersed into thecytoplasm during metaphase, a nuclear membrane marker Ndc1 exhibited a localization pattern that also suggests a partial opening of the NE duringmitosis. A semi-open mitosis was further confirmed by transmission electron microscopy. In summary, our data demonstrate that key mitotic events in C.neoformans are similar to that of metazoan cells. This study sheds new light on the evolution of mitosis during fungal speciation.115. Distinctive Mitotic Localization of a Novel Suppressor of nimA1 Provides New Insight into NIMA Function. Jennifer R. Larson, Stephen A. Osmani.Department of Molecular Genetics, The Ohio State University, Columbus, OH.The NIMA kinase is an essential regulator of mitotic events in Aspergillus nidulans. Not only is NIMA essential for initiating mitosis its overexpression canprematurely induce mitotic events including DNA condensation and nuclear pore complex (NPC) disassembly in A. nidulans and human cells. One of thekey roles for NIMA at the onset of mitosis is its regulation of NPCs. A previous study aimed at identifying suppressors of the temperature-sensitive nimA127th Fungal Genetics Conference | 149

FULL POSTER SESSION ABSTRACTSallele isolated two NPC proteins, which were named SONA and SONB for Suppressors Of NimA1. Although NIMA is essential for mitotic entry there is alsoevidence that NIMA and conserved related kinases have functions later in mitosis and in the DNA damage response. To further characterize the roles ofNIMA we designed a genetic screen to isolate additional suppressors of nimA1 that also cause conditional temperature-dependent DNA damagesensitivity. Our expectation was the identification of additional genes involved in NIMA regulation and in the DNA damage response. Here we describe onesuch gene, which we have named sonC. SonC contains a unique Zn(II)Cys6 binuclear DNA binding domain, which is highly conserved among theAscomycota. Deletion of sonC results in swollen, ungerminated spores, suggesting it is essential for a core growth process. As expected for a DNA bindingprotein, SonC localizes to nuclei during interphase. Interestingly, dual fluorescence imaging of SonC with histone H1 during mitosis revealed that a portionof SonC localizes with histone H1 along a distinct projection of chromatin that juts away from the main, condensed chromatin mass, which we hypothesizemay be the NOR. Supporting this hypothesis, the region of DNA that likely forms the projection is cradled by the nucleolus prior to mitosis as seen bycolocalization studies of SonC with the nucleolar protein Bop1. As mitosis proceeds, the H1 histones are evicted from the middle region of this projectionbut not at its distal end. This indicates that the chromatin in this region of the genome is altered during mitotic progression and we are testing the ideathat SonC might be important for NOR condensation and/or nucleolar disassembly during its mitotic segregation. Because SonC was identified as asuppressor of NIMA we propose that NIMA may have a function in regulating nucleolar disassembly during mitosis.116. Investigating Cell Cycle-Regulated Control of Appressorium Morphogenesis in the Rice Blast Fungus Magnaporthe oryzae. Wasin Sakulkoo, NicholasJ. Talbot. School of Biosciences, University of Exeter, Exeter EX4 4QD, United Kingdom.The rice blast fungus Magnaporthe oryzae elaborates specialized infection structures called appressoria to gain entry into rice plant tissue. The initiationof appresssorium morphogenesis has previously been shown to require a single round of mitosis in the germ tube, shortly after spore germination. Ondaughter nucleus migrates to the incipient appressorium at the germ tube tip and the other daughter nucleus moves back to the conidial cell from whichthe germ tube originates. We reasoned that an S-phase checkpoint mediates the apical-isotropic switch leading to swelling of the germ tube tip.Perturbation of DNA synthesis by hydroxyurea (HU) blocks the initiation of appressorium formation, but only when applied within 3-4h of sporegermination, prior to S-phase. Here, we report investigations regarding the interplay between cell cycle control and operation of the Pmk1 Mitogenactivatedprotein kinase cascade, which is essential for appressorium morphogenesis in M. oryzae. Furthermore we report changes in the global pattern ofgene expression of HU-treated conidia which has been carried out in order to determine the identity of morphogenetic genes that are controlled by the S-phase checkpoint. Progress on understanding the genetic control of early appressorium development will be presented.117. THE ROLE AND TRAFFIC OF CHITIN SYNTHASES IN Neurospora crassa. R. Fajardo 1 , R. Roberson 2 , B. Jöhnk 3 , Ö. Bayram 3 , G.H. Braus 3 , M. Riquelme 1 . 1)Department of Microbiology, CICESE, Ensenada, Mexico; 2) School of Life Sciences, Arizona State University, Arizona, USA; 3) Molecular Microbiology andGenetics, Georg-August University, Göttingen, Germany.Chitin is one of the most important carbohydrates in the cell wall in filamentous fungi. Chitin synthases (CHS) are involved in the addition of N-acetylglucosamine monomers to form chitin microfibrils. The filamentous fungus Neurospora crassa has one representative for each of the seven CHSclasses described. Previous studies have shown that in N. crassa, CHS-1, CHS-3 and CHS-6, are concentrated at the core of the Spitzenkörper and informing septa and seem to be transported in different populations of chitosomes. In this study we have endogenously tagged chs-2, chs-4, chs-5 and chs-7with gfp to study their distribution in living hyphae of N. crassa. CHS-5 and CHS-7 both have a myosin motor-like domain at their amino termini, suggestingthat they interact with the actin cytoskeleton. CHS-2 and CHS-7, appeared solely involved in septum formation. As the septum ring developed, CHS-2-GFPmoved centripetally until it localized exclusively around the septal pore. CHS-4 and CHS-5 were localized both at nascent septa and in the core of the Spk.We observed a partial colocalization of CHS-1-mCherry and CHS-5-GFP in the Spk. Total internal reflection fluorescence microscopy (TIRFM) analysisrevealed putative chitosomes containing CHS-5-GFP moving along wavy tracks, presumably actin cables. Collectively our results suggest that there aredifferent populations of chitosomes, each containing a class of CHS. Mutants with single gene deletions of chs-1, chs-3, chs-5, chs-6, or chs-7 grew slightlyslower than the control strain (FGSC#9718 and FGSC#988); only chs-6D displayed a marked reduction in growth. Both chs-5D and chs-7D strains producedless aerial hyphae and conidia. The double mutant chs-5D; chs-7D showed less growth, aerial hyphae production and conidiation than the single mutantchs-5D, but not than the chs-7D single mutant. A synergic effect was observed in double mutant chs-1D; chs-3D, in which growth, aerial hyphae productionand conidiation were significantly decreased. During the sexual cycle, after homozygous crosses, chs-3D and chs-7D strains did not produce perithecia andchs-5D produced less perithecia. We are analyzing chitin and glucan synthase activities in these single and double mutants. Additionally, we are conductingpulldown assays, and mass spectrometry to identify putative proteins that are interacting with CHS.118. DFG5 and DCW1 cross-link Cell Wall Proteins into the Cell Wall Matrix. Abhiram Maddi, Jie Ao, Stephen J. Free. Dept Biological Sci, SUNY Univ,Buffalo, Buffalo, NY.The cell wall is an essential organelle for the growth and survival of a fungus. The cell wall structure consists of matrix of cross-linked chitin, glucans, andcell wall glycoproteins. In Neurospora crassa, we have shown that the DFG5 and DCW1 proteins function in cross-linking the cell wall proteins into the cellwall matrix. We have also shown that the Candida albicans DFG5 and DCW1 proteins are required for the cross-linking of cell wall proteins into the cellwall. The DFG5 and DCW1 proteins are predicted to have a-1,6-mannanase activity. Our results suggest that they function in transglycosylation reactionsbetween a-1,6-mannans, which are found in galactomannan and the outer chain mannan structures present as modifications on cell wall proteins, and cellwall glucans. These galactomannans and outer chain mannans are modifications to the N-linked oligosaccharides attached to cell wall glycoproteins. As aresult of these transglycosylation reactions, the cell wall proteins are effectively cross-linked into the cell wall. The DFG5 and DCW1 enzymes are excellenttargets for the development of anti-fungal agents that could disrupt cell wall biosynthesis.119. Cell wall biology to illuminate mechanisms of pathogenicity in Phytophthora infestans. Laura Grenville-Briggs, Stefan Klinter, Francisco Vilaplana,Annie Inman, Hugo Mélida, Osei Ampomah, Vincent Bulone. Division of Glycoscience, Royal Institute of Technology, (KTH), Stockholm, Sweden.The cell wall is a dynamic extracellular compartment protecting the cell, providing rigidity, and playing an essential role in the uptake of molecules andsignalling. In pathogenic organisms, the cell wall is at the forefront of disease, providing contact between the pathogen and host. Using a multidisciplinaryapproach, we seek to understand the role of the cell wall in oomycete disease, both as a communication centre with the host organism and as acompartment that is continually reshaped and strengthened throughout the lifecycle, to penetrate and colonise the host. Understanding thesemechanisms in more detail will pave the way for better control of oomycete diseases. We are combining novel chemical genomics approaches with stateof-the-artbiochemistry and biophysics to study the cell wall and to develop new anti-oomycete drugs. P. infestans produces a variety of spores andinfection structures that are essential for disease development throughout its lifecycle. In particular thick-walled sporangia release wall-less motilezoospores that rapidly synthesise a cell wall upon contact with host plant cells. These cysts further differentiates to produce appressoria which build up150

FULL POSTER SESSION ABSTRACTSturgor pressure and act as a focal point for cell wall degrading enzymes to penetrate the host cell. A highly strengthened cell wall is thus essential for theonset of infection. Here we present the results of our detailed biochemical analyses, using GC-MS and methylation analysis to determine the neutral sugarcomposition and glycosidic linkages of the cell wall structural carbohydrates present at these key points in the lifecycle. Having previously established anessential role for a cellulosic cell wall in appressorium production and infection of potato by P. infestans (Grenville-Briggs et al 2008), we are now workingto elucidate the precise functions of the individual cellulose synthase (CesA) genes. Silencing each CesA using RNAi reveals overlapping functions withsubtle differences in phenotype. These results will be presented. Since the genome of P. infestans also contains a putative chitin synthase, but hyphal cellwalls are devoid of measurable chitin we are also investigating the role of this gene in the P. infestans cell wall and in pathogenicity and here we presentthe latest findings of this work.120. Analysis of the cell wall integrity (CWI) pathway in Ashbya gossypii. Klaus B Lengeler, Lisa Wasserström, Andrea Walther, Jürgen Wendland.Carlsberg Laboratory, Yeast Biology, DK-1799 Copenhagen V, Denmark.Fungal cells are constantly exposed to rapidly changing environmental conditions, in particular considering their osmotic potential. The cell wall takes onan important function in protecting the fungal cell from external stresses and controlling intracellular osmolarity, but it is also required to maintain regularcell shape. At the same time, cells must still be able to remodel the rigid structure of the cell wall to guarantee cell expansion during cell differentiationprocesses. While several signaling pathways contribute to the maintenance of the cell wall, it is the cell wall integrity (CWI) pathway that is most importantin regulating the remodeling of the cell wall structure during vegetative growth, morphogenesis or in response to external stresses. To characterize theCWI pathway in the filamentous ascomycete Ashbya gossypii we generated deletion mutants of several genes encoding for the most importantcomponents of the CWI pathway including potential cell surface sensors (e.g. AgWSC1), the following downstream protein kinases including a MAPKsignaling module (AgPKC1, AgBCK1, AgMKK1 and AgMPK1), and transcription factors known to be involved in CWI signaling (e.g. AgRLM1). An initialcharacterization of the corresponding mutants is presented. While a mutant in Agpkc1 shows a strong general growth defect, mutants in several othercomponents of the CWI pathway, in particular in the MAPK module, show a noticeable colony lysis phenotype. Finally, we show that the colony lysisphenotype may be useful to easily isolate recombinant proteins from A. gossypii.121. Dynamics of exocytic markers and cell wall alterations in an endocytosis mutant of Neurospora crassa. Rosa R. Mouriño-Pérez, Ramón O. Echauri-Espinosa, Arianne Ramírez-del Villar, Salomón Bartnicki-García. Microbiology Department, CICESE, Ensenada, B.C., Mexico.Morphogenesis in filamentous fungi depends principally on the establishment and maintenance of polarized growth. This is accomplished by the orderlymigration and discharge of exocytic vesicles carrying cell wall components. We have been searching for evidence that endocytosis, an opposite process,could also play a role in morphogenesis. Previously, we found that coronin deletion (Neurospora crassa mutant, Dcrn-1) causes a decrease in endocytosis(measured by the rate of uptake of FM4-64) together with marked alterations in normal hyphal growth and morphogenesis accompanied by irregularitiesin cell wall thickness. The absence of coronin destabilizes the cytoskeleton and leads to interspersed periods of polarized and isotropic growth of thehyphae. We used CRIB fused to GFP as an exocytic reporter of activated Cdc-42 and Rac-1. By confocal microscopy, we found that CRIB-GFP was present Inwild-type hyphae as a thin hemispherical cap under the apical dome, i. e. when growing in a polarized fashion and with regular hyphoid morphology. In theDcrn-1 mutant, the location of CRIB-GFP shifted between the periods of polarized and isotropic growth, it migrated to the subapical region and appearedas localized patches. Significantly, cell growth occurred in the places where the CRIB-GFP reporter accumulated, thus the erratic location of the reporter inthe Dcrn-1 mutant correlated with the morphological irregularity of the hyphae. We found that the Dcrn-1 mutant had a higher proportion of chitin thanthe WT strain (14.1% and 9.1% respectively). We also compared the relative cell wall area (TEM images) and we found a different ratio wall/cytoplasmbetween the Dcrn-1 mutant and the WT strain. In conclusion, we have found that the mutant affected in endocytosis has an an altered pattern ofexocytosis as evidenced by its distorted morphology and displaced exocytic markers. A direct cause-effect relationship between endocytosis andexocytosis remains to be established.122. Comprehensive genome-based analysis of cell wall biosynthesis in the filamentous phytopathogen Ashbya gossypii. R. Capaul 1 , M. Finlayson 1 , S.Voegeli 1 , A. I. Martinez 2 , Q. Y. Yin 3 , C. de Koster 3 , F. M. Klis 3 , P. Philippsen 1 , P. W. J. de Groot 2 . 1) Biozentrum, Molecular Microbiology, University of Basel,Klingelbergstr. 50-70, CH 4056 Basel, Switzerland; 2) Regional Center for Biomedical Research, Albacete Science & Technology Park, University of Castilla-La Mancha, Spain; 3) Swammerdam Institute for Life Sciences, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, The Netherlands.The filamentous ascomycete Ashbya gossypii and the yeast Saccharomyces cerevisiae are phylogenetically closely related. It is not known how A. gossypiihas evolved an exclusively hyphal growth mode with very rapid apical extension requiring cell wall expansion rates that are up to 40-fold faster comparedto S. cerevisiae. The genome of A. gossypii encodes 44 putative cell wall-associated GPI proteins, 10 without a homolog in S. cerevisiae. This analysis alsorevealed amplification of several cell wall protein-encoding genes, notably CWP1. Transcriptome studies showed that one third of the CWP-encodinggenes are expressed at higher levels than ribosomal protein genes. Mass spectrometric analysis of protein extracts from purified walls of rapidly growinghyphae resulted in the identification of 14 covalently bound cell wall proteins (CWPs). Some CWPs that are common in hemiascomycetes are missing in A.gossypii. On the other hand, the chitin deacetylase Cda1/Cda2 was identified in addition to three novel proteins (Agp1, Awp1, and Sod6), all withouthomologs in baker's yeast (NOHBYs). Phenotypic analysis confirmed the importance of these NOHBYs for cell wall integrity. Interestingly, hyphal walls of A.gossypii contain very little chitin and orthologs of genes required for cell wall remodeling and degradation of septa during cell division in S. cerevisiae showlow expression or are absent. Conclusions: Loss of distinct cell wall genes, acquisition of novel genes, and amplification as well as increased expression ofevolutionary conserved fungal cell wall genes led to the evolution of fast polar surface expansion of A. gossypii hyphae.123. cAMP regulation in Neurospora crassa conidiation. Wilhelm Hansberg, Sammy Gutiérrez, Itzel Vargas, Miguel-Ángel Sarabia, Pablo Rangel. Institutode Fisiología Celular, Universidad Nacional Autónoma de México, México D.F., México.In N. crassa, conidiation is started when an aerated liquid culture is filtered and the resulting mycelial mat is exposed to air. Three morphogenetictransitions take place: hyphae adhesion, aerial hyphae growth and conidia development [1]. Each transition is started by an unstable hyperoxidant state(HO) and results in growth arrest, autophagy, antioxidant response and an insulation process from dioxygen [2,3]. These responses stabilize the systemand growth can restart in the differentiated state. We found that ras-1 bd has increased ROS formation during conidiation resulting in increased aerialmycelium growth and increased submerged conidiation. Different ras-1 point mutations were generated that affected growth and conidiation. Only threeproteins have a predicted RAS association domain: NRC-1, the STE50p orthologue (STE50) and adenylate cyclase (AC). The Dncr-1 was more resistantwhereas the Dste50 more sensitive to added H 2O 2. The AC mutant strain cr-1 affects vegetative growth and aerial hyphae formation. Oxidative stress andRAS-1 determined partially cAMP levels during the first two HOs of the conidiation process. Higher cAMP levels than Wt were observed in ras-1 bd . In bothstrains, [cAMP] decreased within minutes at the start of the first two HOs and thereafter, as rapidly, levels recover to initial values. N. crassa has a high27th Fungal Genetics Conference | 151

FULL POSTER SESSION ABSTRACTS(PDE H) and a low affinity (PDE L) phosphodiesterases. The Dpde H strain grows slow and does not conidiate; no evident phenotype was reported for Dpde L.We found that PDE L was mainly responsible for the cAMP decrease during the first HO and that hyphal adhesion was retarded in Dpde L. Both PDE H andPDE L were responsible for cAMP decrease during the second HO. H 2O 2 and low Ca ++ activated PDE L and inhibited PDE H. This opposite regulation can explainthe cAMP decrease during the HOs of the N. crassa conidiation process. [1] Toledo I et al. (1986) Aerial growth in Neurospora crassa: characterization of anexperimental model system. Exp Mycol. 10:114-125. [2] Hansberg W; Aguirre J (1990) Hyperoxidant states cause microbial cell differentiation by cellisolation from dioxygen. J Theoret Biol 142:201-221. [3] Hansberg W et al. (2008) Cell differentiation as a response to oxidative stress. In: Stress in Yeasts &Filamentous Fungi (Ed. Avery et al.) Elsevier IBSN 978-0-12-374184-4.124. Ste12 is a negative regulator of conidiation and cell wall lytic enzymes production in response to nitrogen deprivation and light in Trichodermaatroviride. Maria Fernanda Nieto-Jacobo 1 , Alfredo Herrera-Estrella 2 , Alison Stewart 1 , Artemio Mendoza-Mendoza 1 . 1) Bioprotection Research Centre,Lincoln University, Lincoln, Canterbury, New Zealand; 2) Laboratorio Nacional de Genómica para la Biodiversidad, Centro de Investigación y de EstudiosAvanzados del IPN Sede Irapuato, Irapuato 36821, Guanajuato, Mexico.Ste12 is a transcription factor found exclusively in the fungal kingdom. In Saccharomyces cerevisiae, Ste12 regulates mating and invasive/pseudohyphalgrowth, while in saprophytic and parasitic filamentous fungi Ste12-like proteins control mating, plant penetration and invasive growth. Ste12 and Ste12-like proteins are downstream components of the MAPK PMK1 pathway which are capable of regulating several genes encoding fungal virulence factorsinvolved in both plant and animal infection. Among the virulence factors are diverse range of lytic enzymes and cell surface components. Several membersTrichoderma genus are mycoparasites of plant fungal pathogens; so they are widely used as biocontrol agents. In addition, Trichoderma spp. penetrateplant roots and establish beneficial relationships with their host. One crucial element in biocontrol activity and root colonization of Trichoderma is thesynthesis of lytic enzymes. Several lytic enzymes in Trichoderma are regulated by nitrogen metabolite repression. Here we observed that the ste12-liketranscription factor gene is highly up-regulated when Trichoderma is grown on nitrogen depleted medium. To find the role of ste12 in Trichoderma, aste12-like orthologue gene was deleted in T. atroviride and the effects on fungal development and response to different biotic and abiotic stimulievaluated. Our results demonstrate that growth and conidiation of a T. atroviride Ste12-like mutant was only slightly altered in complete media. Weevaluated the ability of the Dste12-like mutant to use a variety of nitrogen sources using Biolog microtiter plates. We noticed that when essential aminoacids are used as the sole nitrogen source, the deletion mutant grew faster than the wild type, however this situation did not occur when the same aminoacids were used as the sole carbon source. In addition, induction of conidiation in response to light or mechanical injury was stronger in the Dste12-likemutants than in the wild type but only when a secondary nitrogen source was used in the medium. Finally we observed that some lytic enzymes aredifferently produced between the wild type and Dste12-like mutants under nitrogen deprivation conditions. We propose that the T. atroviride Ste12-likeorthologue regulates lytic enzymes and conidiation by a mechanism that involves nitrogen catabolite repression.125. Black holes in fungal virulence: loss of RNAi in C. gattii outbreak strains reveals a novel RNAi factor. Marianna Feretzaki, Xuying Wang, BlakeBilmyre, Joseph Heitman. Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC.Genome instability and mutations provoked by transposon movement are counteracted by novel defense mechanisms in organisms as diverse as fungi,plants, and mammals. In the human fungal pathogen Cryptococcus neoformans we have previously characterized an RNAi silencing pathway that defendsthe genome against mobile elements and artificially introduced repeats of homologous DNA. Repetitive transgenes and transposons are silenced by anRNAi-dependent pathway during sexual development (sex-induced silencing, SIS) and during vegetative mitotic growth (MIS). RNAi silencing pathways areconserved in the Cryptococcus pathogenic species complex and are mediated by core RNAi components, including an RNA-dependent RNA polymerase(Rdp1), Argonaute (Ago1) and Dicer (Dcr1 and Dcr2). Surprisingly, all of the canonical known RNAi components are missing from all C. gattii VGII strains,the molecular type responsible for the North American Pacific Northwest outbreak. To identify novel components of the RNAi pathway, we surveyed thegenome of the C. gattii R265 isolate for missing genes. One of the most interesting is ZNF3. In previous studies we found that Znf3, a protein with threezinc finger domains, is required for opposite- and same-sex mating in C. neoformans var. neoformans. Surprisingly, in C. neoformans var. grubii ZNF3 is notessential for sexual development. However, it is required for mitotic- and sex-induced silencing via RNAi. SIS is less efficient in znf3D unilateral matings andis abolished in znf3D x znf3D bilateral matings, similar to the phenotypes of rdp1D mutants. Znf3 is also required for transgene-induced mitotic silencing;znf3D mutations abrogate silencing of repetitive transgenes during vegetative growth. Znf3 tagged with mCherry is localized in the cytoplasm in bright,distinct foci. Co-localization of Znf3 with the P-body marker Dcp1-GFP further supports the hypothesis that Znf3 is a novel element of the RNAi pathwayand operates to defend the genome during sexual development and vegetative growth.126. The Crz1/Sp1 transcription factor of Cryptococcus neoformans is activated by calcineurin and regulates cell wall integrity. Sophie Lev 1 , DesmariniDesmarini 1 , Methee Chayakulkeeree 2 , Tania Sorrell 1 , Julianne Djordjevic 1 . 1) Centre for Infectious Diseases and Microbiology, Sydney Medical School andWestmead Millennium Institute, University of Sydney, Westmead 2145 NSW, Australia; 2) Faculty of Medicine, Siriraj Hospital, Mahidol University,Bangkok, Thailand.Cryptococcus neoformans survives host temperature and regulates cell wall integrity via a calcium-dependent phosphatase, calcineurin. However,downstream effectors of C. neoformans calcineurin are largely unknown. In S. cerevisiae and other fungal species, a calcineurin-dependent transcriptionfactor Crz1 translocates to nuclei upon activation and triggers expression of target genes. We now show that the C. neoformans Crz1 ortholog (Crz1/Sp1),previously identified as a protein kinase C target during starvation, is a bona fide target of calcineurin under non-starvation conditions, during cell wallstress and growth at high temperature. Both the calcineurin-defective mutant, Dcna1, and a CRZ1/SP1 mutant (Dcrz1) were susceptible to cell wallperturbing agents. Furthermore, expression of the chitin synthase encoding gene, CHS6, was reduced in both mutants. We tracked the subcellularlocalization of Crz1-GFP in WT C. neoformans and Dcna1 in response to different stimuli, in the presence and absence of the calcineurin inhibitor, FK506.Exposure to elevated temperature (30-37°C vs 25°C) and extracellular calcium caused calcineurin-dependent nuclear accumulation of Crz1-GFP.Unexpectedly, 1M salt and heat shock triggered calcineurin-independent Crz1-GFP sequestration within cytosolic and nuclear puncta. To our knowledge,punctate cytosolic distribution, as opposed to nuclear targeting, is a unique feature of C. neoformans Crz1. We conclude that Crz1 is selectively activatedby calcium/calcineurin-dependent and independent signals depending on the environmental conditions.127. A Fungal Adhesin Guides Community Behaviors by Autoinduction and Paracrinal Signaling. Linqi Wang, Xunyun Tian, Rachana Gyawali, Xiaorong Lin.Biology, Texas A&M University, College Station, TX.Microbes live mostly in a social community rather than in a planktonic state. Such communities have complex spatiotemporal patterns that requireintercellular communication to coordinate gene expression. Here, we demonstrate that Cryptococcus neoformans, a model eukaryotic pathogen, respondsto an extracellular signal in constructing its colony morphology. The signal that directs this community behavior is not a molecule of low molecular weight152

FULL POSTER SESSION ABSTRACTSlike pheromones or quorum sensing molecules, but a secreted protein. We successfully identified this protein as the conserved adhesin Cfl1 in theextracellular matrix. The released Cfl1 acts as an auto-induction signal to stimulate neighboring cells to phenocopy Cfl1-expressing cells. We propose thatsuch adhesin/matrix-initiated communication system exists in divergent microbes and our work represents the first adhesin/matrix-mediated signalingmechanism in simple eukaryotes.128. The PacC Signal Transduction Pathway regulates Sexual Development in Neurospora crassa. Chinnici Jennifer, Arnold Jason, Stephen J. Free. DeptBiological Sci, SUNY Univ, Buffalo, Buffalo, NY.As is common in the ascomycetes, the Neurospora crassa life cycle has both asexual and sexual developmental phases. Sexual development in N. crassais characterized by the formation of a protoperithecium, fertilization, and the maturation of the protoperithecium to form a perithecium. In a screeningexperiment, we identified over 600 isolates from the N. crassa single gene deletion library that are unable to complete sexual development. Many of theseare affected in the process of anastomosis, and we have previously reported on these mutants. We now report on the other female developmentdefective mutants identified in our screening experiments. Co-segregation and complementation experiments were carried out on these mutants and weidentified 80 genes that are required for female development (in addition to the 24 genes needed for anastomosis). We find that these genes fall into 5general classes: 1) signal transduction pathway genes (25 genes), 2) transcription factor genes (7 genes), 3) chromatin remodeling genes (17 genes), 4)genes required for autophagy (11 genes), and 5) miscellaneous genes (20 genes). The PacC pathway genes are among the identified signal transductionpathway genes needed for female development. The activation of the PacC signal transduction pathway is a key signaling event in sexual development.Our experiments also suggest that autophagy and anastomosis are important for the movement of nutrients from the hyphal tissues supporting thedeveloping perithecium.129. Aspergillus flavus MAP kinase AflMpkB positively regulates developmental process but not aflatoxin production. Sang-Cheol Jun 1,2 , Dong-Soon Oh 1 ,Jong-Hwa Kim 1 , Kwang-Yeop Jahng 2 , Kap-Hoon Han 1 . 1) Dept. of Pharmaceutical Engineering, Woosuk Univ, Wanju, Korea; 2) Div. of Biological Sciences,Chonbuk National University, Jeonju, Korea.Developmental process of eukaryotes is controlled by the multiple regulatory systems including signal transduction pathways and transcription factors.One of the central signaling mechanisms includes mitogen-activated protein kinase (MAPK) pathway that transfer extracellular signals into nucleus,generating cellular responses. Previously, we have showed that Aspergillus nidulans MpkB, the yeast Fus3 MAP kinase ortholog, regulates sexualdevelopment and secondary metabolism. Here, we identified and characterized the ortholog of the A. nidulans mpkB gene in Aspergillus flavus, AflmpkB,to understand whether the AflmpkB gene has conserved function with A. nidulans mpkB. Deletion of AflmpkB did not affect hyphal growth but showedreduced conidia production Furthermore, AflmpkB null strain didn’t produce any sclerotia while WT and recipient strain produced a lot of sclerotia innormal conditions. However, loss of AflmpkB resulted in normal aflatoxin biosynthesis, suggesting that the major function of AflmpkB is positive regulationof conidiation, sclerotia development but not mycotoxin production. These results indicate that A. nidulans and A. flavus MpkB have conserved anddivergent roles in development and secondary metabolism.130. Subcellular localization and kinase activity of GK4, a Phytophthora infestans GPCR-PIPK involved in actin cytoskeleton organisation. Chenlei Hua 1 ,Harold Meijer 1 , Kiki Kots 1,2 , Tijs Ketelaar 2 , Francine Govers 1 . 1) Laboratories of Phytopathology, Wageningen University, Droevendaalsesteeg 1, 6708 PBWageningen, The Netherlands; 2) Laboratories of Cell Biology, Wageningen University, Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands.For dispersal and host infection plant pathogens largely depend on asexual spores. Pathogenesis and sporulation are complex processes that aregoverned by various cellular signaling networks including G-protein and phospholipid signaling. Oomycetes possess a family of novel proteins called GPCR-PIPKs (GKs) that are composed of a seven trans-membrane spanning (7-TM) domain fused to a phosphatidylinositol phosphate kinase (PIPK) domain.Based on this domain structure GKs are anticipated to link G-protein and phospholipid signalling pathways. Our studies in the potato late blight pathogenPhytophthora infestans revealed involvement of one of twelve GKs (i.e. PiGK4) in spore development, hyphal elongation and infection. Moreover, ectopicexpression in P. infestans of subdomains of PiGK1 and PiGK4 fused to a fluorescent protein showed that the GPCR domain targets the GKs to membranessurrounding different cellular compartments. To further elucidate the function of the PIPK domain we tested kinase activity of PiGK4 both in vivo and invitro and analysed the relationship between PiGK4, phosphoinositide signaling and the organisation of the actin cytoskeleton using complementation inyeast combined with various live-cell markers.131. External calcium ions and deletion of per-1 gene suppressed the abnormal morphology of och-1 and frost mutants in Neurospora crassa. MasayukiKamei, Yuko Tsukagoshi, Shinpei Banno, Masakazu Takahashi, Akihiko Ichiishi, Makoto Fujimura. Faculty of Life Sciences, Toyo University, ORA-GUN,GUNMA, Japan.Calcium ions play important roles in the growth and development in filamentous fungi. The frost mutant show slow growth and hyperbranchingphenotypes that can be corrected by Ca 2+ addition in Neurospora crassa. The frost gene is an ortholog of S. cerevisiae cdc1 which encodes putative Mn 2+ -dependent lipid phosphatase. We found that the abnormal morphology of the och-1 mutant was quite similar to that of the frost mutant and itsabnormality was also corrected by external Ca 2+ . The och-1 gene encodes an alpha-1,6-mannosyltransferase that is probably involved in sugar processingfor GPI-anchor proteins. In yeast, the mutation of per1 gene, encoding a protein is required for GPI remodeling pathway, suppresses the abnormal growthphenotype of cdc1 mutant. To examine the effect of per-1 gene, an ortholog of per1 in S. cerevisiae, on the phenotypes of the frost and och-1 mutants, weisolated two double mutants, frost; Dper-1 and och-1; Dper-1. Although per-1 gene disruptant showed the normal growth phenotype, per-1 gene deletionsignificantly suppressed the slow growth and hyperbranching phenotypes of frost and also och-1 mutants. Addition of Ca 2+ did not affect the growth andmorphology of the two double mutants. These results suggest the connection between FROST and OCH-1 may participate in lipid remodeling or calciumsignaling in Neurospora crassa.132. Functional analysis of carbonic anhydrases from the filamentous ascomycete Sordaria macrospora. Ronny Lehneck 1 , Piotr Neumann 2 , AchimDickmanns 2 , Ralf Ficner 2 , Stefanie Pöggeler 1 . 1) Institute of Microbiology and Genetics, Department of Genetics of Eukaryotic Microorganisms, Georg-August-University Göttingen; 2) Institute of Microbiology and Genetics, Department of Molecular Structural Biology, Georg-August-University Göttingen.Carbonic anhydrases (CA) are widely distributed enzymes, which catalyzes the reversible hydration of carbon dioxide to bicarbonate and protons. Basedon their amino acid sequence and structure, they can be divided into five distinct groups (a, b, g, d, x) which share no sequence similarity and havesupposable evolved independently. All known fungal CAs belong either to the a-class or to the b-class. Our model organism Sordaria macrospora encodesat least four carbonic anhydrases: three of the b type, termed cas1, cas2 and cas3 (carbonic anhydrase of Sordaria) and one a-type, termed cas4.Previously, the functions of CAS1, CAS2 and CAS3 have been intensively studied (Elleuche and Pöggeler 2009) and displayed an involvement in fruiting-27th Fungal Genetics Conference | 153

FULL POSTER SESSION ABSTRACTSbody development and ascospore germination. Here, we present a functional characterization of the secreted a-CA CAS4. CAS4 seems to be involved inammonium metabolism but not in ascospore germination. The Dcas4 mutant displayed a slightly reduced vegetative growth rate and a delayed fruitingbodydevelopment. Based on real time PCR analysis cas4 is upregulated during the sexual development. Moreover, we present the phenotype of aquadruple mutant without any CAS genes. The complete CAS deletion strain (Dcas1/2/3/4) is able to grow under ambient air but the vegetative growthrate is drastically reduced and the mutant is only able to form thin hyphae. The mutant is even under elevated CO 2 levels (5 %) not able to form fruitingbodies. Heterologous expression in Saccharomyces cerevisiae demonstrated that CAS1 and CAS2 are active enzymes, but only CAS1 displays considerablein vitro activity. Furthermore, X-ray and gel filtration analyses revealed a tetrameric structure of CAS1 with a conserved histidine and two cysteine residuesin the active center.Elleuche and Pöggeler 2009: b-Carbonic anhydrases play a role in fruiting body development and ascospore germination in the filamentous fungusSordaria macrospora; PLoS ONE. 2009; 4(4): e5177.133. The Coprinopsis cinerea cag1 (cap-growthless1) gene, whose mutation affects cap growth in fruiting body morphogenesis, encodes the buddingyeast Tup1 homolog. H. Muraguchi, K. Kemuriyama, T. Nagoshi. Dept Biotechnology, Akita Prefectural Univ, Akita, Japan.We have mutagenized a homokaryotic fruiting strain, #326, of Coprinopsis cinerea and isolated a mutant that fails to enlarge the cap tissue on theprimordial shaft in fruiting. Genetic analysis of this mutant, cap-growthless, indicated that the mutant phenotype is brought about by a single gene,designated as cag1. The cag1 locus was mapped on chromosome IX by linkage analysis using RAPD markers mapped to each chromosome. The cag1 genewas identified by transformation experiments using BAC DNAs and their subclones derived from chromosome IX, and found to encode a homolog ofSaccharomyces cerevisiae Tup1. The Coprinopsis genome includes another Tup1 homologous gene, designated Cc.tupA. Expression levels of these twotup1 paralogs were examined using a real-time quantitative PCR method. Cc.tupA is predominantly expressed in vegetative mycelium. In contrast, in thecap tissue, transcript levels of cag1 are similar to that of Cc.tupA. Since it is known that S. cerevisiae Tup1 forms homotetramer, interactions of Cag1 withitself and Cc.TupA were examined using yeast two-hybrid system. Cag1 interacts with itself through the N-terminal region and with Cc.TupA. Like Tup1,which interacts with Cyc8, the N-terminal region of Cag1 also interacts with the N-terminal region of Cc.Cyc8, which contains tetratricopeptide repeats.Based on expression and yeast two-hybrid analyses of Cag1 and Cc.TupA, combined with information on S. cerevisiae Tup1, we speculate that, invegetative mycelium, Cc.TupA represses expression of genes required for cap growth, and Cag1, which might become expressed at the top of primordialshafts to produce the cap tissue and continue to be expressed in the cap tissue, might derepress and activate the expression through interaction withCc.TupA.134. Adaptation of the microtubule cytoskeleton to multinuclearity and chromosome number in hyphae of Ashbya gossypii as revealed by electrontomography. R. Gibeaux 1 , C. Lang 2 , A. Z. Politi 1 , S. L. Jaspersen 3 , P. Philippsen 2 , C. Antony 1 . 1) European Molecular Biology Laboratory, Heidelberg,Germany; 2) Biozentrum, Molecular Microbiology, University of Basel, CH 4056 Basel, Switzerland; 3) Stowers Institute for Medical Research, Kansas City,USA.The filamentous fungus Ashbya gossypii and the yeast Saccharomyces cerevisiae evolved from a common ancestor based on the high level of gene orderconservation. Interestingly, A. gossypii lost the ability of cell divisions and exclusively grows as elongating multinucleated hyphae. Using electrontomography we reconstructed the cytoplasmic microtubule (cMT) cytoskeleton in three tip regions with a total of 13 nuclei and also the nuclearmicrotubules (nMTs) of four mitotic bipolar spindles. Each spindle pole body (SPB) nucleates three cMTs on average, similarly to S. cerevisiae SPBs. 80% ofcMTs were growing as concluded from the structure of their plus-ends. Very long cMTs closely align for several microns along the cortex to generatedynein-dependent pulling forces on nuclei. The majority of nuclei carry duplicated side-by-side SPBs, which together emanate an average of six cMTs, inmost cases in opposite orientation with respect to the hyphal growth axis. Such cMT arrays explain why many nuclei undergo short-range back and forthmovements. Following mitosis, daughter nuclei carry a single SPB. The increased probability that all three cMTs orient in one direction explains the highrate of long-range nuclear bypassing observed in these nuclei. These results demonstrate how cMT arrays, despite a conserved number of microtubules,could successfully adapt to the demands of multinuclearity during evolution from mono-nucleated budding yeast-like cells to multinucleated hyphae. Themodelling of A. gossypii mitotic spindles revealed a very similar structure to mitotic spindles of S. cerevisiae in terms of nMT number, length distributionand three-dimensional organisation even though A. gossypii carries 7 and S. cerevisiae 16 chromosomes per haploid genome. Our results suggest that thenMT cytoskeleton remained largely unaltered during the evolution and that two nMTs attach to each kinetochore in A. gossypii in contrast to only one in S.cerevisiae.135. High resolution proteomics of spores, germlings and hyphae of the phytopathogenic fungus Ashbya gossypii. L. Molzahn 1,2 , A. Schmidt 2 , P.Philippsen 1 . 1) Biozentrum, Molecular Microbiology, University of Basel, CH4056 Basel, Switzerland; 2) Biozentrum, Proteomics Facility, University of Basel,CH4056 Basel, Switzerland.Growth of the filamentous ascomycete A. gossypii is regulated by a genome very similar to the Saccharomyces cerevisiae genome even though thegrowth modes of both organisms differ significantly. During the previous decade progress was made to better understand some of these differences. 1.Cytokinesis in A. gossypii is not coordinated with mitosis and cell separation does not occur due to loss of specific genes which most likely led to theevolution of multinucleated hyphae. 2. Short nuclear cycle times and dynein-dependent pulling forces excerted on nuclei by autonomous cMT arrays withfast growing microtubules maintain a high nuclear density also in fast growing hyphae. 3. Polar growth sites once established support permanent andconstantly accelerating polar surface expansion at hyphal tips at rates of up to 40mm2/min compared to 1mm2/min of yeast buds. Very efficientexocytosis and endocytosis could be documented in hyphal tips of A.gossypii. We want to understand on a system level the differences between bothorganisms and have started a proteomic approach. Total protein extracted from spores and developing A. gossypii hyphae was digested with trypsin,mixed with heavy isotope-labeled reference peptides and subjected to high resolution tandem MS analyses. We could identify 3900 proteins at eachdevelopmental stage. Significant quantitative changes of these proteins with respect to clusters of orthologous groups (COG) or gene ontology (GO) termswere identified during A.gossypii development and between log-phase growing S. cerevisiae cells and fast growing A. gossypii hyphae. Importantdifferences concern ribosome biogenesis and translation, mitochondria biogenesis and respiration, glycolysis and gluconeogenesis, chromatin remodeling,chaperones, cell wall biosynthesis and the first reaction in several biosynthetic pathways.136. Indoor Fungal Growth and Humidity Dynamics. Frank J.J. Segers 1 , Karel A. van Laarhoven 2 , Henk P. Huinink 2 , Olaf Adan 2 , Jan Dijksterhuis 1 . 1) Appliedand Industrial Mycology, CBS-KNAW Fungal Biodiversity Centre, Utrecht, Netherlands; 2) Department of Applied Physics, Eindhoven University ofTechnology, Eindhoven, Netherlands.154

FULL POSTER SESSION ABSTRACTSIndoor fungi are present in a considerable part of the European dwellings and cause cosmetic and structural damage. The presence of indoor fungi posesa potential threat to human health as a result of continuous exposure as they are able to form allergens and mycotoxins. Indoor fungal growth does notexist without the presence and availability of water. Not much is known on the response of fungi to humidity dynamics during different stages of theirdevelopment. Relative humidity (RH) and water activity (aw) are used in many studies for the amount of water available for the fungus. A RH of 80% orhigher is thought to be required for fungal growth to occur. On average the RH is below 50% in normal buildings, suggesting a crucial role of humiditydynamics for fungal growth. In order to study the fungal response to humidity dynamics, two indoor fungal species, Cladosporium halotolerans andPenicillium rubens, were dried in controlled humidity vessels to stop growth and are rehydrated under high humidity conditions after a week. Non-linearSpectral Imaging Microscopy (NSIM) is a non-intrusive method to follow the response of fungal cells under varying relative humidity conditions by lookingat the metabolic activity of separate cells. The different developmental stages of C. halotolerans and P. rubens before and after periods of a certain level ofhumidity are determined by using Cryo Scanning Electron Microscopy (CryoSEM). A different response to humidity dynamics was seen between severaldevelopmental stages and both fungi used. More in depth research will be done on the specific cellular response of the fungi to humidity dynamics.137. Essentiality of Ku70/80 in Ustilago maydis is related to its ability to suppress DNA damage signalling at telomeres. Carmen de Sena-Tomas 1 , EunYoung Yu 2 , Arturo Calzada 3 , William K. Holloman 2 , Neal F. Lue 2 , Jose Perez-Martin 1 . 1) IBFG (CSIC-USAL), Zacarias Gonzalez 3, 37007 Salamanca, Spain; 2)Cornell University Medical College, 1300 York Avenue, 10021 New York; 3) CNB (CSIC), Darwin 3, 28049 Madrid, Spain.Ku heterodimer is formed of two subunits Ku70 and Ku80 that bind with high affinity to DNA ends in a sequence independent manner. Ku has a role inseveral cellular processes including DNA repair, telomere maintenance, transcription and apoptosis. Ku heterodimer is essential in human cells as well as inUstilago maydis, a well-characterized fungal system used in DNA repair studies. We found that depletion of Ku proteins in U. maydis elicits a DNA damageresponse (DDR) at telomeres resulting in a permanent cell cycle arrest, which depends on the activation of the Atr1-Chk1 signalling cascade. Aconsequence of this inappropriate activation is the induction of aberrant homologous recombination at telomeres manifested by the formation ofextrachromosomal telomere circles, telomere lengthening and the accumulation of unpaired telomere C-strand. Abrogation of the DDR response bydeleting either chk1 or atr1 genes alleviates much of these aberrant recombination process suggesting that one of the roles of Ku proteins at telomeres inUstilago maydis is related to the suppression of unscheduled DNA damage signalling at telomeres, in addition to the protection of telomeres.138. Magnaporthe oryzae effectors with putative roles in cell-to-cell movement during biotrophic invasion of rice. Mihwa Yi 1 , Xu Wang 2 , Jung-Youn Lee 2 ,Barbara Valent 1 . 1) Department of Plant Pathology, Kansas State University, Manhattan, Kansas 66506, USA; 2) Department of Plant and Soil Sciences,University of Delaware, Newark, Delaware 19711, USA.Previous studies implicated rice plasmodesmata in two different aspects of rice blast disease caused by the hemibiotrophic ascomycetous fungus,Magnaporthe oryzae. First, effectors that are translocated into the cytoplasm of living rice cells move ahead into uninvaded host plant cells by amechanism that depends on effector protein size and rice cell type. This suggested that these effectors move through plasmodesmata to preparesurrounding host cells for fungal infection. Second, biotrophic invasive hyphae (IH) search for locations to move into neighboring rice cells and theyundergo extreme constriction when crossing the host cell wall. These findings and additional evidence suggested that IH manipulate host pit fieldscontaining plasmodesmata for cell-to-cell movement. Our goals are to test these hypotheses, and to understand the molecular mechanisms responsiblefor cell-to-cell movement in blast disease. We have identified six biotrophy-associated secreted (Bas) proteins that accumulate around IH at the pointwhere they have crossed the rice cell wall to invade neighboring rice cells. We designated these effectors as putative fungal movement proteins (fMPs).When imaged as fluorescently labeled fusion proteins, the fMPs show unique localization patterns at the cell wall crossing points. Functional analysis ofthe fMPs is underway. Precise microscopic characterization with correlative light and electron microscopy (CLEM) and time-course, live-cell imaging isbeing performed to decipher how the fungus manipulates the rice cell wall junction area for effector trafficking and its own cell-to-cell spread. The fMPswill be localized relative to each other and to plasmodesmata-specific fluorescent markers. We will compare the structure and function of riceplasmodesmata in invaded versus non-invaded rice cells. Our results will identify novel host targets exploited by the fungus and related infectionmechanisms at the wall crossing sites to facilitate colonization in planta.139. Functional characterization of autophagy genes Smatg8 and Smatg4 in the homothallic ascomycete Sordaria macrospora. Stefanie Poeggeler,Oliver Voigt. Genetics of Eukaryotic Microorganisms, Georg-August University, Göttingen, Germany.Autophagy is a degradation process involved in various developmental aspects of eukaryotes. However, its involvement in developmental processes ofmulticellular filamentous ascomycetes is largely unknown. Here, we analyzed the impact of the autophagic proteins SmATG8 and SmATG4 on the sexualand vegetative development of the filamentous ascomycete Sordaria macrospora. A yeast complementation assay demonstrated that the S. macrosporaSmatg8 and Smatg4 genes can functionally replace the yeast homologs. By generating homokaryotic deletion mutants, we showed that the S. macrosporaSmATG8 and SmATG4 orthologs were associated with autophagy-dependent processes. Smatg8 and Smatg4 deletions abolished fruiting-body formationand impaired vegetative growth and ascospore germination, but not hyphal fusion. We demonstrated that SmATG4 was capable of processing theSmATG8 precursor. SmATG8 was localized to autophagosomes and SmATG4 was distributed throughout the cytoplasm of S. macrospora. Furthermore, wecould show that Smatg8 and Smatg4 are not only required for nonselective macroautophagy, but for selective macropexophagy as well. Our resultssuggest that in S. macrospora autophagy seems to be an essential and constitutively active process to sustain high energy levels for filamentous growthand multicellular development even under nonstarvation conditions. (Voigt O, Pöggeler S Autophagy genes Smatg8 and Smatg4 are required for fruitingbodydevelopment, vegetative growth and ascospore germination in the filamentous ascomycete Sordaria macrospora. Autophagy. 2012 Oct 12;9(1).[Epub ahead of print]).140. Laser microdissection and transcriptomics of infection cushions formed by Fusarium graminearum. Marike Boenisch 1 , Stefan Scholten 2 , SebastianPiehler 3 , Martin Münsterkötter 3 , Ulrich Güldener 3 , Wilhelm Schäfer 1 . 1) Molecular Phytopathology and Genetics, Biocenter Klein Flottbek, University ofHamburg, Germany; 2) Developmental Biology and Biotechnology, Biocenter Klein Flottbek, University of Hamburg, Germany; 3) Institute of Bioinformaticsand Systems Biology, Helmholtz Zentrum Münich (GmbH), Neuherberg, Germany.The fungal plant pathogen Fusarium graminearum Schwabe (teleomorph Gibberella zeae (Schwein) Petch) is the causal agent of Fusarium head blight(FHB) of small grain cereals and cob rot of maize worldwide. Trichothecene toxins produced by the fungus e.g. nivalenol (NIV) and deoxynivalenol (DON)contaminate cereal products and are harmful to humans, animals, and plants. We demonstrated recently, that F. graminearum forms toxin producinginfection structures during infection of wheat husks, so called infection cushions (Boenisch and Schäfer, 2011). The aims of the presented study were tofurther clarify the penetration mechanism of infection cushions by histological studies and to identify molecular characteristics of infection cushions byexpression analysis. Structural characteristics of infection cushions were visualized by 3D images following laser scanning microscopy. We observed27th Fungal Genetics Conference | 155

FULL POSTER SESSION ABSTRACTSmultiple penetration events underneath infection cushions by scanning electron microscopy. Colonization of the underlying plant tissue was studied bybright field microscopy and transmission electron microscopy of LR-White serial sections. To understand the molecular basis of initial colonization of theleaf surface followed by infection cushion development, a laser capture microdissection (LCM) approach was established to isolate specifically runnerhyphae and infection cushions. Several hundred runner hyphae and infection cushions grown on wheat glumes were collected using the PALM system(Zeiss) avoiding contamination with plant tissue. Total mRNA of runner hyphae and infection cushions were isolated and amplified. The cDNA library ofeach developmental stage was used for next generation sequencing with Illumina HiSeq 2000. Quantitative expression analysis show marked differences ingene expression patterns between runner hyphae and infection cushions. Different functional pathways specific for each infection stage were identified.Thereby new insights in the initial infection process of FHB disease are gained. To our knowledge, we provide the first transcriptome data of runnerhyphae and infection cushions from a fungal plant pathogen obtained under in planta conditions. In summary, the power of combined microscopic andmolecular approaches to analyze cell type-specific gene expression during fungal-plant-interactions is demonstrated.141. Biochemical and biophysical analysis of the CarO rhodopsin of Fusarium fujikuroi. Jorge García-Martínez 1 , Marta Castrillo 1 , Javier Avalos 1 , UlrichTerpitz 2 . 1) Departamento de Genética, Universidad de Sevilla, Sevilla, Spain; 2) Lehrstuhl für Biotechnologie und Biophysik, Julius-Maximilians-UniversitätWürzburg, Biozentrum / Am Hubland, Würzburg, Germany.Light controls many substantial processes in filamentous fungi, such as reproduction and pathogenicity. Fungi naturally possess light sensors, which reactto a broad range of wavelengths with absorption maxima in the blue, green or red regions of the spectrum. Rhodopsins are green light-absorbingmembrane-integrated photoreceptors consisting of seven transmembrane helices forming an interior pocket for the chromophore, either all-trans or 11-cis retinal, covalently bound to the protein via a protonated Schiff-base. Type I rhodopsins, predicted to bind all-trans retinal, are widespread inascomycota and basidiomycota. Upon light-activation, type I rhodopsins act as proton pumps or sensory proteins; however, detailed knowledge of theirphysiological function and biological role in fungi is still missing. The gibberellin-producing fungus Fusarium fujikuroi contains two rhodopsin encodinggenes, carO and opsA, whose mutations produce no external phenotypic alterations. The carO gene is linked and co-regulated with genes coding forenzymes for retinal-synthesis, whose expression is strongly induced by light. To gain information on CarO biological role, we have combined biophysicalmethods to analyse the localisation and function of this rhodopsin in F. fujikuroi mycelia. We established a strain expressing CarO fused to a yellowfluorescent protein (YFP) under control of the carO promoter. This strain was investigated with confocal laser scanning microscopy (cLSM) and superresolutionfluorescence imaging (dSTORM) to reveal the subcellular localisation of CarO. Protein-localisation was compared with data recorded from a S.cerevisiae DSY5 strain overexpressing CarO-YFP. Additionally, the carO-YFP gene fusion was expressed in neuroblastoma cells, where it exhibited anefficient ion pump-activity, as demonstrated by Patch-clamp techniques. The results suggest a light dependent ion-pumping role in the fungus,nonessential under standard laboratory conditions.142. Roles of membrane and organellar calcium channels and transporters in controlling pulsatile [Ca 2+ ] c signatures. Hye-Seon Kim 1 , Jung-Eun Kim 2 , KirkCzymmek 1 , Robert Cirino 1 , Randall Duncan 1 , Hokyoung Son 3 , Yin-Won Lee 3 , Seogchan Kang 2 . 1) Department of Biological Sciences, University of Delaware,Newark, DE 19711; 2) Department of Plant Pathology & Environmental Microbiology, The Pennsylvania State University, University Park, PA 16802; 3)Department of Agricultural Biotechnology and Center for Fungal Pathogenesis, Seoul National University, Seoul 151-921.Calcium ions translate diverse environmental stimuli into many different physiological and developmental functions in fungi via an evolutionaryconserved cell-signaling pathway. Using the expression of Yellow Cameleon YC3.60, a fluorescent protein-based, ratiometric Ca 2+ sensor in Magnaportheoryzae, Fusarium oxsyporum, and F. graminearum, we reported that cytoplasmic tip high Ca 2+ signatures exhibited distinct species-specific and agedependentpulsatile patterns (FGB 49:589). We successfully expressed a new circularly permuted Ca 2+ sensor, GCaMP5, in F. graminearum and F.oxysporum and GCaMP3 in Neurospora crassa. The improved sensitivity, photostability, and fast kinetics of GCaMP5 enabled us to image smaller Ca 2+changes in hyphae tips with high-speed imaging that showed that the tip high Ca 2+ gradient has multiple origins. Disruption of F. graminearum genesencoding plasma membrane Ca 2+ channels (Mid1, Cch1, and Fig1), vacuole/ER Ca 2+ pumps (Pmc, Pmr), calcineurin transcription factor (Crz1), and vacuoleH + /Ca 2+ exchanger (Vcx1 and Vcx2) significantly altered the amplitude, interval, and origin of Ca 2+ pulses and also affected growth. Additional phenotypesassociated with these mutants are currently being characterized. The combination of molecular genetics, genomics, live cell imaging, and correlativemicroscopy will help us study the mechanism underpinning fungal Ca 2+ signaling at multiple scales ranging from the function and mode-of-action ofindividual genes to nano-scale dynamics of individual proteins and subcellular machineries.143. Characterization of positive regulator for asexual and sexual reproduction in the cereal head blight pathogen Gibberella zeae. Jungkwan Lee 1 ,Boknam Jung 1 , Hokyoung Son 2 , Yin-Won Lee 2 . 1) Department of Applied Biology, Dong-A University, Busan 604-714, Republic of Korea; 2) Department ofAgricultural Biotechnology and Center for Fungal Pathogenesis, Seoul National University, Seoul 151-921, Republic of Korea.Gibberella zeae is an important plant pathogen that causes cereal head blight and produces mycotoxins that are harmful to animals and humans.Ascospores and conidia contribute to the primary inoculums and propagation for disease epidemics. In this study, we identified one putative C2H2 zincfinger transcription factor (prd1) that is required for both conidiation and sexual reproduction, as screening transcription factor mutant collection wepreviously generated. prd1 deletion mutants impaired conidial production and lost both self-fertility and female fertility, but retain male fertility. Theoverexpression of the gene increased the amount of conidial production and resulted in earlier maturation of fruiting body formation than the wild-typestrain. The vegetative growth of deletion and overexpression mutants was increased and decreased on nutrient-rich mediua, respectively, but was notdifferent from the wild-type strain on nutrient-poor media. This study was the first report for transcription factor which positively regulates bothconidiation and sexual reproduction, and the characterization of genes regulated by this gene will be further studied.144. Functional analysis of Elongator complex protein 3 in Gibberella zeae. Y. J. Lee 1 , H. Son 1 , J.-C. Kim 2 , G. J. Choi 2 , Y.-W. Lee 1 . 1) Department ofAgricultural Biotechnology and Center for Fungal Pathogenesis, Seoul National University, Seoul 151-921, Republic of Korea; 2) Eco-friendly New MaterialsResearch Group, Research Center for Biobased Chemistry, Division of Convergence Chemistry, Korea Research Institute of Chemical Technology, Daejeon305-343, Republic of Korea.Gibberella zeae (anamorph: Fusarium graminearum) is a causal agent of Fusarium head blight (FHB) which causes huge economic losses in cereal cropssuch as wheat and barley. In addition to yield reduction, mycotoxin contamination of grain presents a threat to human safety. We examined one ofRestriction-Enzyme-Mediated Integration (REMI) mutants Z43R9282 showing defects in virulence and sexual development and identified a gene encodingElongator complex protein 3 (ELP3). ELP3 is a catalytic subunit of Elongator complex and contains histone acetyltransferase (HAT) domain. The biologicalfunction of ELP3 gene was studied by targeted deletion in G. zeae. Deletion of ELP3 resulted in retarded growth and delay of sexual developmentcompared to the wild-type strain. Most of the ascospores had two cells in the ELP3 deletion mutants, while wild-type ascospores usually had four cells. The156

FULL POSTER SESSION ABSTRACTSlength of the mutant conidia was approximately 25% longer than the wild type. Deletion mutants of ELP3 were sensitive to stress conditions, such as highsaltstress (NaCl and KCl), suggesting a role in adaptation to environmental condition. Virulence on wheat heads was greatly reduced in the ELP3 deletionmutants. These results demonstrate that ELP3 is required for normal sexual and asexual development and ELP3 could be involved in cell size regulation inG. zeae.145. Functional analyses of regulators of G protein signaling (FgRGS) and GzGPA proteins in Gibberella zeae. A.R. Park 1 , A.-R. Cho 1 , J.-A. Seo 2 , K. Min 1 , H.Son 1 , J. Lee 3 , G.J. Choi 4 , J.-C. Kim 4 , Y.-W. Lee 1 . 1) Department of Agricultural Biotechnology and Center for Fungal Pathogenesis, Seoul National University,Seoul 151-921, Republic of Korea; 2) Science and Technology Division, Ministry for Food, Agriculture, Forestry and Fisheries, Gyeonggi-Do 427-712,Republic of Korea; 3) Department of Applied Biology, Dong-A University, Busan 604-714, Republic of Korea; 4) Eco-friendly New Materials Research Group,Research Center for Biobased Chemistry, Division of Convergence Chemistry, Korea Research Institute of Chemical Technology, Daejeon 305-343, Republicof Korea.G protein signaling pathways play key roles in the regulation of fungal development, secondary metabolism, and virulence. Regulators of G proteinsignaling (RGS) proteins make up a highly diverse and multifunctional protein family that plays a critical role in controlling heterotrimeric G proteinsignaling. The genome of the plant pathogenic fungus Gibberella zeae contains seven RGS genes (FgFlbA, FgFlbB, FgRgsA, FgRgsB, FgRgsB2, FgRgsC, andFgGprK). Here we functionally characterized the function of these genes in various cellular processes. Mutant phenotypes were observed for deletionmutants of FgRgsA and FgRgsB in vegetative growth, FgFlbB and FgRgsB in conidia morphology, FgFlbA in conidia production, FgFlbA, FgRgsB, and FgRgsCin sexual development, FgFlbA and FgRgsA in spore germination and mycotoxin production, and FgFlbA, FgRgsA, and FgRgsB in virulence. Furthermore,FgFlbA, FgRgsA, and FgRgsB acted pleiotropically, while FgFlbB and FgRgsC deletion mutants exhibited a specific defect in conidia morphology and sexualdevelopment, respectively. Site-directed Ga subunits mutagenesis and overexpression of the FgFlbA gene revealed that deletion of FgFlbA and dominantactive GzGPA2 mutant, gzgpa2 Q207L , had similar phenotypes in cell wall integrity, perithecia formation, mycotoxin production, and virulence, suggestingthat FgFlbA may regulate asexual/sexual development, mycotoxin biosynthesis, and virulence through GzGPA2-dependent signaling in G. zeae. Especially,GzGPA2 might activate trichothecene production in a FgFlbA-dependent manner.146. A novel gene, GEA1, is required for ascus cell wall development in the ascomycete fungus, Gibberella zeae. H. SON 1 , J. Lee 2 , Y.-W. Lee 1 . 1)Department of Agricultural Biotechnology and Center for Fungal Pathogenesis, Seoul National University, Seoul 151-921, Republic of Korea; 2) Departmentof Applied Biology, Dong-A University, Busan 604-714, Republic of Korea.The ascomycete fungus Gibberella zeae is a devastating plant pathogen for major cereal crops. Ascospores are produced via sexual reproduction andforcibly discharged from mature perithecia, which function as the primary inocula. Perithecium development involves complex cellular processes and isunder polygenic control. In this study, a novel gene, GEA1, was found to be required for ascus wall development in G. zeae. GEA1 deletion mutantsproduced normal-shaped perithecia and ascospores, yet ascospores were observed to precociously germinate inside of perithecium. Moreover, GEA1deletions resulted in abnormal ascus walls that collapsed prior to ascospore discharge. Based on localization of GEA1 to the endoplasmic reticulum (ER),GEA1 may be involved in protein export from the ER to the ascus wall biogenesis. This is the first report to identify a unique gene required for ascus walldevelopment in G. zeae.147. A systems-biology approach to build gene-regulatory network models connecting osmotic stress responses and asexual development in Fusariumgraminearum. A. Thompkins, M. Sexton, S. Atkinson, B. Bass, E. Delancy, J. Rhodes, J. Flaherty. Science and Mathematics, Coker College, Hartsville, SC.Fusarium graminearum is a notorious fungal plant pathogen and causes head blight disease in small grain cereals and ear rot disease in maize. Infectionwith F. graminearum leads to yield losses and mycotoxin contamination. Mycotoxin formation and asexual development are thought to share commonnodes of genetic regulation. However, the regulatory networks connecting salt/osmotic stress to either is limited or undefined. Salt tolerance is a complextrait that remains poorly understood. Very few genes have been identified that are required for salt tolerance in plants, animals, or fungi. To address this,we screened >5,000 insertional mutants of F. graminearum (PH-1) for gain-of-function or loss-of-function phenotypic classes specific to both asexualdevelopment (conidiation) and osmotic stress responses. These screens yielded strains representing all classes and one outlier from each were chosen foradditional analyses. Mutant 9E1 exhibits an “osmotic hyper-tolerant” phenotype when cultured on growth media containing either NaCl or glycerol. Incontrast, mutant 11B1 displays an “osmotic-overly sensitive” phenotype, where growth is severely limited on concentrations of solute that have anegligible effect on growth by control strains. Both 9E1 and 11B1 grow normally on non-osmotically adjusted media and were subsequently chosen fortranscription-profiling experiments. Additionally, mining gene expression data of developmental mutants 8B5 (aconidial) and 8E8 (hyperconidial) haverevealed coordinately expressed, putative candidate regulatory genes. Based on a transcriptomics framework, we applied a bioinformatics approach toidentify shared gene regulatory networks involved in osmotic stress responses and conidiation.This project was supported in part by grants from the National Science Foundation (MCB 0845324), the National Center for Research Resources (5 P20RR016461), and the National Institute of General Medical Sciences (8 P20 GM103499) from the National Institutes of Health.148. Starvation enhances heterokaryon formation between incompatible strains of Fusarium oxysporum. Shermineh Shahi, Martijn Rep. Molecular PlantPathology, Swammerdam Institute for Life Sciences, Amsterdam, Nordholland, Netherlands.Fusarium oxysporum (Fo) is a pathogenic species complex with a broad host range. Comparative genomics revealed lineage-specific (LS) genomic regionsin Fusarium oxysporum f. sp. lycopersici (Fol) that account for more than 25% of the genome. At least two LS chromosomes can be transferred horizontallyto non-pathogenic Fo strains, resulting in acquired pathogenicity in the recipient [1]. Here we want to elucidate the chromosome transfer pathway and themechanisms by which the incompatibility reaction between strains is avoided. It has been suggested that heterokaryon formation is necessary forhorizontal chromosome transfer in Colletotrichum gloeosporioides [2] and that heterokaryon incompatibility is suppressed after conidial anastomosis tube(CAT) fusion [3]. To study nuclear dynamics during formation of heterokaryotic cells in Fo, we observed green or red fluorescent protein labeled nuclei ofFol and a non-pathogenic Fo strains in a vegetatively incompatible interaction.While in rich medium co-cultivation of both strains revealed no heterokaryotic cells, co-cultivation under starvation conditions led to up to ~30%heterokaryotic colonies (red and green nuclei). We were able to distinguish between different types of heterokaryotic conidia. In some cases aftergermination only one of the nuclei was able to propagate, which always originated from the pathogenic strain. In other cases both nuclei were able topropagate and these colonies in turn produced uninucleate conidia (yellow nuclei). Another intriguing finding was that the pathogenic strain used faredbetter under starvation conditions (higher germination and growth rate). We conclude that under starvation condition Fol is the dominant/fitter strain andthat heterokaryon formation in Fo is greatly enhanced, possibly by further suppressing non-self recognition machinery in CATs and/or increased hyphal27th Fungal Genetics Conference | 157

FULL POSTER SESSION ABSTRACTSfusion. We hypothesize that starvation might be a driving force for horizontal chromosome transfer in order to increase the chance of survival.Reference:[1] Ma, L.-J. et al; Nature 464, 367-373 (2010) [2] Manners, JM & He,C; Mycol Progress 10:383-388 (2011) [3] Ishikawa, FH et al; PLoS ONE7(2): e31175. doi:10.1371/journal.pone.0031175 (2012).149. Requirements for horizontal chromosome transfer in the plant pathogenic fungus Fusarium oxysporum. Ido Vlaardingerbroek, Martijn Rep. FNWI,University of Amsterdam, Amsterdam, Netherlands.Strains within the Fusarium oxysporum species complex are clonal and diverse. A number of them are pathogenic to plants but rarely can they infectmore than one host. Host specificity is determined by the presence of a set of secreted effector genes. These genes typically reside on Lineage Specific (LS)chromosomes that can be transferred between strains, even if they are vegetatively incompatible. These extra chromosomes typically carry nohousekeeping genes and have many more transposable elements then the non-LS or core chromosomes. If a strain receives one of these chromosomes itcan acquire the ability to infect a new host, compatible with the effector genes the chromosome harbours. Our main interests at this moment are (1)determining which chromosomes are amenable for transfer, and (2) which cellular processes are involved in transfer. To determine which chromosomescan be transferred, we created a bank of random insertional mutants carrying an antibiotic resistance marker. These have been tested for chromosometransfer. A few of these showed consistent transfer of the chromosome tagged with the marker. By screening a large number of transformants we shouldcover the entire genome. In addition to the screen we tagged an LS chromosome that we know can be transferred, as well as the smallest of the corechromosomes, with GFP. In this way we can directly compare transfer capability of these chromosomes. We hypothesize that the LS chromosomes’ uniquemake-up is required for transfer. We will also test these strains for stability of the tagged chromosome by screening spores for the loss of GFP expressionusing FACS. In this way we can test whether transferrable (LS) chromosomes differ in stability from core chromosomes under varying conditions. Toidentify cellular processes involved in chromosome transfer, we are making deletion mutants for genes required for cellular processes we suspect might beinvolved in chromosome transfer. These will be tested for transfer efficiency compared to the wild-type strains. We are currently investigating hyphalfusion, heterochromatin formation and programmed cell death. By combining the results from these two research lines we should be able to discoverwhich chromosomes can be transferred as well as the chromosomal features and processes involved.150. Characterization of the endocytotic proteins Yel1-Arf3-Gts1 in Ashbya gossypii and the role of Gts1 in endocytosis, actin localization andfilamentous growth. Therése Oskarsson, Klaus Lengeler, Jürgen Wendland. Carlsberg Laboratory, Copenhagen, Denmark.Endocytic vesicle formation and regulation thereof is performed by a complex protein machinery, coordinating every detail of the endocytic process frominitiation and pit formation to vesicle scission and uncoating.We have used the filamentous fungi Ashbya gossypii to study three proteins that are involved in uncoating of vesicles in clathrin-mediated endocytosis.We deleted the corresponding genes encoding the GTP-binding protein Arf3 and its regulators; the Guanine nucleotide Exchange Factor Yel1 and theArfGAP protein Gts1, using PCR-based gene targeting methods. We then characterized these mutant strains under various conditions.While no deletion-specific phenotypes could be observed in the Darf3 and Dyel1, the Dgts1 strain shows several severe mutant phenotypes. Deletion ofGTS1 results in a strong growth defect and renders mycelia with severe endocytotic deficiencies indicated by distinctly reduced endocytic rates, and largeimmobile vacuoles. Other phenotypic observations in A. gossypii Dgts1 strains indicate that Gts1 may have additional functions other than regulating theactivity of Arf3. We have observed effects of Gts1 on temperature stress resistance, actin localization and polar- as well as filamentous growth.The importance of GTS1 for polarized hyphal growth leads us to studying the GTS1 homolog of the human fungal pathogen Candia albicans in an effortto elucidate its role for the yeast-to-hyphal transition in this dimorphic fungi.151. A Late Embryogenesis Abundant (LEA) protein in Neosartorya fischeri confers protection against desiccation. Martin Richard van Leeuwen, Timon TWyatt, Tineke M van Doorn, Jan Dijksterhuis. Applied and Industrial Mycology, CBS-KNAW Fungal Biodiversity Centre, Utrecht, Netherlands.Late Embryogenesis Abundant (LEA) proteins were first characterized in cotton and wheat and are synthesized in abundance during the late maturationstage of seed development. As the seed matures, water content decreases greatly inducing severe desiccation stress. Expression of LEA proteins is linkedto the acquisition of desiccation tolerance. Using BLAST to search for LEA like proteins in various filamentous fungal genomes (Aspergillus niger, Aspergillusflavus, Emericella nidulans, Penicillium chrysogenum, Talaromyces stipitatus and Neosartorya fischeri) resulted in orthologs in each mentioned species,indicating the wide spread appearance of LEA proteins in fungi. Ascospores produced by N. fischeri are able to survive long periods under various stressors.However, deletion of the LEA gene resulted in diminished tolerance against desiccation and high temperatures. In addition, heterologous expression ofLEA in Escherichia coli conferred increased tolerance against osmotic- and salt stress. Interestingly, LEA was able to function as protectant for enzymes thatnormally lose activity under influence of stress. Lactate dehydrogenase (LDH) was inactivated by heat stress and freeze-thaw cycles. In the presence ofLEA, LDH activity was maintained. Our results show that LEA are wide spread in filamentous fungi and function in tolerance against stressors like heat,freeze-thaw and desiccation. LEA could play an important role in stress tolerance of survival propagules like ascospores and conidia.152. Coordination of polarized secretion by the exocyst complex is critical for filamentous growth and cytokinesis in Ustilago maydis. Michaela Wehr 1 ,Kay Oliver Schink 2 , Michael Bölker 1 . 1) Philipps University, FB Biologie, AG Boelker Marburg, Hessen, Germany; 2) Department of Biochemistry, Institute forCancer Research The Norwegian Radium Hospital, Montebello, N-0310 Oslo, Norway.To establish and sustain their polarity, cells have to transport proteins and membrane lipids to defined locations at the growing tip. This is achieved bydirectional transport of vesicles that fuse with the plasma membrane. Vesicle fusion and active exocytosis requires the presence of an octameric proteincomplex, the exocyst. In S. cerevisiae, two proteins of the exocyst complex, Sec3 and Exo70, were shown to serve as landmark proteins for exocytosis. Theother components of the exocyst tether secretory vesicles carrying the Rab GTPase Sec4 to the membrane. Fusion of secretory vesicles occurs viainteraction of the exocyst with SNARE proteins. To elucidate the function and regulation of the exocyst complex and its associated proteins in Ustilagomaydis, we have characterized the Rab GTPase Sec4 and the exocyst proteins Sec3, Exo70 and Sec15 by genetic, cell biological and biochemicalapproaches. We found that of the two landmark proteins, only one is important for polar growth in U. maydis. Interestingly, this gene is not essential,suggesting that in U. maydis exocytosis sites can be also marked by alternative mechanisms. Another essential player for polar growth in U. maydis is theexocyst subunit Sec15, which mediates the interaction of the exocyst with incoming secretory vesicles. Conditional mutants of sec15 are defective inhyphal tip growth and are affected in long-distance transport of secretory vesicles. In contrast to S. cerevisiae where Sec4 vesicles are transported alongthe actin cytoskeleton, long distance transport of vesicles depends in U. maydis on the microtubule cytoskeleton. Furthermore, we studied mutants ofdifferent motor proteins to get insights into the molecular mechanisms of secretory vesicle trafficking.153. Localization of Neurospora crassa Cell Fusion Proteins. Ci Fu, Stephen J. Free. Biological Sciences, University at Buffalo, Buffalo, NY.158

FULL POSTER SESSION ABSTRACTSA screen of mutants in Neurospora crassa single gene deletion library identified 24 cell fusion genes. Bioinformatics studies indicate that 14 of thesegenes are likely to function in signal transduction pathways, 4 genes are transcription factors, 3 genes are likely to be involved in the process of vesiculartrafficking, and 3 genes are highly conserved in fungal species with unknown functions. GFP and RFP fusion proteins were constructed for 2 vesiculartrafficking proteins AMPH-1 and HAM-10, and 1 conserved hypothetical protein HAM-8 to study their functions during cell fusion process. Fluorescentprotein markers for cellular organelles (including nucleus, mitochondria, golgi apparatus, endoplasmic reticulum, vacuole and vesicle), and for cytoskeleton(including actin filament and microtubule) were obtained from Fungal Genetic Stock Center. Strains expressing individual fluorescent protein marker wereused to study the cellular localizations of AMPH-1, HAM-10 and HAM-8 by using fluorescent confocal microscopy. The fluorescent protein marker strainswere also used to study the dynamics of organelle movements during cell fusion by using time-lapse fluorescence microscopy. Fluorescent signals fromAMPH-1, HAM-10 and HAM-8 were compared with two signaling molecules MAK-2 and SO to study their potential involvement in signal transduction.Results shown AMPH-1, HAM-8 and HAM-10 all colocalize with vesicle marker. One of the conserved hypothetical proteins, HAM-6, was modified with aFLAG tag to study its functions during cell fusion.154. Identification of novel Neurospora crassa genes involved in hyphal fusion by tanscriptomic analysis. Wilfried Jonkers, Abigail C. Leeder, N. LouiseGlass. Department of Plant and Microbial Biology, University of California, Berkeley, CA.Hyphal fusion of Neurospora crassa germlings is a highly regulated process involving -among others- the conserved MAP kinase MAK-2 and the SOprotein of unknown biochemical function. During chemotrophic interactions between two genetically identical germlings, MAK-2 and SO alternatelylocalize at the conidial anastomosis tubes (CATs) every 4 minutes, perfectly out of phase of each other. How this process is initiated, maintained and whatother proteins are involved is still unknown. One conserved fungal target of MAK-2 is the yeast Ste12-like transcription factor, named PP-1. Similar to mak-2, pp-1 is also required for hyphal fusion and normal mycelial growth. To identify downstream targets of MAK-2 and PP-1 that may play a role in germlingfusion, micro-array and RNAseq analyses were performed on wild type (WT) and Dpp-1 strains. Combining the micro-array and RNAseq data, 32 geneswere identified that showed at least 2-fold differential expression in WT as compared to Dpp-1. These include six genes, which are homologs of yeastSte12 targets. To test the involvement of these genes in hyphal fusion, a deletion strain was obtained or constructed and assayed for germling fusionphenotype. Three deletion strains were completely devoid of fusion: Dham-7, Dasm-1 and Dham-11, and one deletion strain, Dham-12 showed reducedfusion frequencies when compared to WT. ham-7 was previously identified as fusion gene while asm-1 was shown to be involved in meiosis. When Dham-7 + Dham-7 or Dham-7 + WT germlings are confronted which each other, chemotropic interactions are not initiated, CATs are not observed and MAK-2 andSO are localized predominantly to the cytoplasm. ham-11 is a newly identified gene involved in germling fusion; Dham-11 + Dham-11 germlings do notshow chemotropic interactions or cell fusion. However, in contrast to Dham-7, Dham-11 germling fuse normally with WT germlings. MAK-2 and SO alsoshow normal oscillation in WT and Dham-11 germlings undergoing chemotropic growth. The observations suggest that HAM-11 might be involved in theproduction or proper release of a signal capable of inducing cell recognition and the germling fusion process.155. N-acetylglucosamine (GlcNAc) Triggers a Morphogenetic Program in Systemic Dimorphic Fungi. Sarah A. Gilmore 1 , Shamoon Naseem 2 , James B.Konopka 2 , Anita Sil 1 . 1) Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA; 2) Department ofMolecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY.Cellular differentiation is an essential process for the development and growth of multicellular eukaryotic organisms. Similarly, many unicellularorganisms undergo a program of cellular differentiation to produce a new cell type specialized for survival in a distinct environmental niche. Systemicdimorphic fungal pathogens, such as Histoplasma capsulatum (Hc) and Blastomyces dermatitidis (Bd), can switch between a unicellular parasitic yeast formadapted for growth within mammals and an infectious soil-growing filamentous form as part of their natural life cycles. Temperature is thought to be thepredominant environmental cue that promotes cellular differentiation of systemic dimorphic fungi; however, work with other fungi indicates thatadditional environmental cues including CO2, light, and nutrient availability can influence how an organism responds to its environment. Recent worksuggests that the ubiquitous monosaccharide N-acetylglucosamine (GlcNAc) can play a role in cell signaling in fungi. We identified GlcNAc as a potentinducer of the yeast-to-filament transition in Hc and Bd. Micromolar concentrations of exogenous GlcNAc were sufficient to induce a robust morphologicaltransition of Hc yeast cells to filamentous cells at room temperature, indicating that dimorphic fungal cells may be sensing GlcNAc, or one of its catabolicbyproducts, to promote filamentation. Using GlcNAc as a tool to induce a robust and more synchronous phase transition of Hc yeast cells to filaments, weexamined the temporal regulation of the Hc transcriptome during morphogenesis to reveal candidate genes involved in establishing the filamentousgrowth program. Two genes we identified during transcriptome analysis included NGT1 and NGT2, which encode GlcNAc major facilitator superfamilytransporters. RNAi depletion of NGT1 or NGT2 rendered Hc cells unable to respond to exogenous GlcNAc. Furthermore, wild type levels of NGT1 and NGT2transcripts were important for efficient Hc yeast-to-filament conversion even in the absence of exogenously added GlcNAc. These data suggest that Ngt1and Ngt2 may monitor endogenous GlcNAc as part of an autoregulatory system that allows Hc to regulate its filamentous growth.156. How water influences fungal growth on "real" materials. H.P. Huinink 1 , K.A. Laarhoven, van 1 , M. Bekker 1 , J. Dijksterhuis 2 , O.C.G. Adan 1 . 1) AppliedPhysics, Eindhoven University of Technology, Eindhoven, Netherlands; 2) CBS - KNAW, Utrecht, Netherlands.Understanding fungal growth on construction materials is important to control problems with mould growth in buildings. The indoor environment isgenerally a harsh environment for a fungus. The climate is relatively dry and only during certain events at specific locations in the building (cooking,showering, etc.) there are peaks in the humidity. The porous nature of construction materials seems to play an important role in the survival of organisms,because it buffers the climate at the surface of materials by storing water. A model has been developed that describes the thermodynamic state and flowof water inside porous materials in connection to the growth of the organism. The model shows that the activity of water in a material is the keyparameter controlling growth. However, the model also proves that growth cannot be predicted on the basis of experiments performed on idealizedmicrobiological media (agar) with a well defined water activity. In those media water is always abundantly present irrespective of the activity. In porousmaterials however the amount of water dramatically reduces with the water activity. It is shown that porous materials with small pores in general containmore water than materials with big pores. A drop in the amount of water due to a decreasing activity has direct consequences for the food supply.Whereas in idealized media the amount of water is very high and therefore the mobility of nutrients, in porous materials the mobility of nutrients willdecrease with decreasing water activity. To understand the behavior of a fungus on materials, its growth has to be really studied on these materials.157. Identification and characterization of two genes required in the control of a cell degeneration in the filamentous fungi Podospora anserina. HerveLalucque 1,2 , Fabienne Malagnac 1,2 , Pierre Grognet 1,2 , Philippe Silar 1,2 . 1) Univ Paris Diderot, Sorbonne Paris Cité, Laboratoire Interdisciplinaire des Energiesde demain (LIED), 75205 Paris France; 2) Institut de Génétique et Microbiologie (IGM), UMR 8621 CNRS Univ Paris Sud, 91405 Orsay France.For several years, we use the coprophilous fungus Podospora anserina to study a cell degeneration called Crippled Growth (CG) triggered by an27th Fungal Genetics Conference | 159

FULL POSTER SESSION ABSTRACTSepigenetic and cytoplasmic element. In the wild-type strain, this element is produced during stationary phase and eliminated at growth renewal. However,in some particular growth conditions, the element is not eliminated in growing hyphae triggering CG. Previous results showed that CG is controlled by twoMAPK modules, the PaNox1 NADPH oxidase and IDC1, a protein with unknown activity. Here, we describe the identification and characterization of twonew partners involved in the control of CG, IDC2 and IDC3. Data show that IDC2 and IDC3 likely act downstream of PaNox1 to regulate the paMpk1 MAPK.We will present a thorough analysis of the phenotypic of the IDC2 and IDC3 mutants and the phylogenetic studies of the IDC2 and IDC3 proteins.158. Dynein drives oscillatory nuclear movements in the phytopathogenic fungus Ashbya gossypii and prevents nuclear clustering. S. Grava, M. Keller, S.Voegeli, S. Seger, C. Lang, P. Philippsen. Biozentrum, Molecular Microbiology, University of Basel, CH 4056 Basel, Switzerland.In the yeast Saccharomyces cerevisiae the dynein pathway has a specific cellular function. It acts together with the Kar9 pathway to position the nucleusat the bud neck and to direct the pulling of one daughter nucleus into the bud. Nuclei in the closely related multinucleated filamentous fungus Ashbyagossypii are in continuous motion and nuclear positioning or spindle orientation is not an issue. A. gossypii expresses homologues of all components of theKar9/Dyn1 pathway, which apparently have adapted novel functions. Previous studies with A. gossypii revealed autonomous nuclear divisions and,emanating from each MTOC, an autonomous cytoplasmic microtubule (cMT) cytoskeleton responsible for pulling of nuclei in both directions of the hyphalgrowth axis. We now show that dynein is the sole motor for bidirectional movements. Surprisingly, deletion of Kar9 shows no phenotype. Dyn1, thedynactin component Jnm1, the accessory proteins Dyn2 and Ndl1, and the potential dynein cortical anchor Num1 are involved in the dynamic distributionof nuclei. In their absence, nuclei aggregate to different degrees, whereby the mutants with dense nuclear clusters grow extremely long cMTs. Like inbudding yeast, we found that dynein is delivered to cMT +ends, and its activity or processivity is probably controlled by dynactin and Num1. Together withits role in powering nuclear movements, we propose that dynein also plays (directly or indirectly) a role in the control of cMT length. Those combineddynein actions prevent nuclear clustering in A. gossypii and thus reveal a novel cellular role for dynein.159. Quantification of the thigmotropic response of Neurospora crassa to microfabricated slides with ridges of defined height and topography. KarenStephenson 1 , Fordyce Davidson 2 , Neil Gow 3 , Geoffrey Gadd 1 . 1) Division of Molecular Microbiology, College of Life Sciences, University of Dundee, Dundee,United Kingdom; 2) Division of Mathematics, University of Dundee, Dundee, United Kingdom; 3) Institute of Medical Sciences, University of Aberdeen,Aberdee, United Kingdom.Thigmotropism is the ability of an organism to exhibit an orientation response to a mechanical stimulus. We have quantified the thigmotropic responseof Neurospora crassa to microfabricated slides with ridges of defined height and topography. We show that mutants that lack the formin BNI-1 and theRho-GTPase CDC-42, an activator of BNI-1, had an attenuated thigmotropic response. In contrast, null mutants that lacked cell end-marker protein TEA-1and KIP-A, the kinesin responsible for its localisation, exhibited significantly increased thigmotropism. These results indicate that vesicle delivery to thehyphal tip via the actin cytoskeleton is critical for thigmotropism. Disruption of actin in the region of the hyphal tip which contacts obstacles such as ridgeson microfabricated slides may lead to a bias in vesicle delivery to one area of the tip and therefore a change in hyphal growth orientation. This mechanismmay differ to that reported in Candida albicans in so far as it does not seem to be dependent on the mechanosensitive calcium channel protein Mid1. TheN. crassa Dmid-1 mutant was not affected in its thigmotropic response. Although it was found that depletion of exogenous calcium did not affect thethigmotropic response, deletion of the spray gene, which encodes an intracellular calcium channel with a role in maintenance of the tip-high calciumgradient, resulted in a decrease in the thigmotropic response of N. crassa. This predicts a role for calcium in the thigmotropic response. Our findingssuggest that thigmotropism in C. albicans and N. crassa are similar in being dependent on the regulation of the vectorial supply of secretory vesicles, butdifferent in the extent to which this process is dependent on local calcium-ion gradients.160. Specificity determinants of GTPase recognition by RhoGEFs in Ustilago maydis. Britta A.M. Tillmann 1 , Kay Oliver Schink 2 , Michael Bölker 1 . 1) Philipps-Universität Marburg FB Biologie, AG Bölker Karl-von-Frisch-Str. 8 35032 Marburg, Germany; 2) Department of Biochemistry, Institute for Cancer ResearchThe Norwegian Radium Hospital, Montebello, N-0310 Oslo, Norway.Small GTPases of the Rho family act as molecular switches and are involved in the regulation of many important cellular processes. They are activated byspecific guanine nucleotide exchange factors (Rho-GEFs). Rho-GTPases interact in their active, GTP-bound state with downstream effectors and triggervarious cellular events. The number of Rho-GEFs and downstream effectors exceeds the number of GTPases. This raises the question how signallingspecificity is achieved. In recent years it became evident that correct signalling depends on both the specificity of the activating Rho-GEF and on scaffoldingproteins that connect the activators with specific downstream effectors. Here, we analysed the Cdc42-specific U.maydis Rho-GEFs Don1, Its1 and Hot1andthe Rac1-specific Rho-GEF Cdc24 for their role in Cdc42 and Rac1 signalling both in vivo and in vitro. We observed that the recognition mechanisms forCdc42 differ between Hot1 and the other Cdc42-specific Rho-GEFs. While a single amino acid at position 56 of Cdc42 and Rac1 is critical for specificrecognition by Don1, Its1 and Cdc24, Hot1 is insensitive to changes at this position. Instead, Hot1 relies on a different set of amino acids to bind its specifictarget Cdc42. We could demonstrate that this unusual mechanism to discriminate between different Rho-type GTPases is also used by the mammalianorthologue of Hot1, TUBA1. These data allowed us to generate a chimeric Cdc42/Rac1 GTPase which can be activated by both Cdc42- and Rac1-specificRho-GEFs with comparable efficiency. Importantly, such a chimeric GTPase was able to complement the morphological phenotypes of Cdc42 and Rac1deletion mutants in vivo.161. Moisture dependencies of P. Rubens on a porous substrate. K.A. van Laarhoven 1 , F.J.J. Segers 2 , J. Dijksterhuis 2 , H.P. Huinink 1 , O.C.G. Adan 1 . 1)Eindhoven University of Technology, Eindhoven, Netherlands; 2) CBS - KNAW, Utrecht, Netherlands.Fungal growth indoors can lead to both disfigurement of the dwelling and medical problems such as asthma. It is generally accepted that the primarycause for mould growth is the presence of moisture. Strategies to prevent fungal growth are therefore often based on controlling indoor humidity. Still,mould is often encountered in ventilated buildings that are considered to be relatively dry. Preliminary experiments showed that fungi can survive onporous materials due to short intervals of favorable circumstances; even when - on average - conditions for growth are not met. This suggests that theinteractions between porous materials and the fluctuating indoor humidity play an important role in a colony’s survival. We study this interplay betweenindoor climate, substrate water household and fungal growth. A property of water that is crucial for fungal growth is water activity (a w). This propertydetermines a fungus’s ability to take up water. The effect of a w on fungal growth has been determined in the past by extensive growth experiments onagar, and many previous studies of growth on building materials take this parameter into account. Up till now, however, little attention has been paid tothe water content (q) of a substrate, which represents the amount of water that is physically present in a system. In most porous materials, even when awis relatively high, only little water is present. We suspect therefore that growth on porous substrates is limited by water content (whereas on agar, q isalways close to 100% and will therefore be of little concern). We performed growth experiments with P. rubens inoculated on gypsum while separatelycontrolling q and a w. Video microscopy was used to monitor the germination and subsequent growth of hyphae. The early development of the fungus was160

FULL POSTER SESSION ABSTRACTSthen quantified by determining parameters such as germination time and growth speed from the movies. The experiments show that the germinationrate, growth speed and growth density of P. rubens on gypsum increase with q while aw is constant, and increase with a w while q is constant. We concludefrom this that q and aw have separate effects on growth on porous substrates. An explanation for the effect of q could be that it limits a fungus’s access toboth water and nutrients. Follow up research will focus on modeling and explaining these effects.162. Localization of Ga proteins during germination in the filamentous fungus, Neurospora crassa. Ilva Esther Cabrera 1 , Carla Eaton 2 , Jacqueline Servin 1 ,Katherine Borkovich 1 . 1) Plant Pathology and Microbiology, University of California, Riverside, Riverside, CA; 2) Institute of Molecular BioSciences, MasseyUniversity, Palmerston North, New Zealand.Heterotrimeric G protein signaling is essential for normal hyphal growth in the filamentous fungus Neurospora crassa. We have previously demonstratedthat the non-receptor guanine nucleotide exchange factor RIC8 acts upstream of the Ga proteins GNA-1 and GNA-3 to regulate hyphal extension.Germination assays revealed essential roles for RIC8 and GNA-3 during this crucial developmental process. Localization of the three Ga proteins duringconidial germination was probed through analysis of cells expressing fluorescently tagged proteins. Functional TagRFP fusions of each of the three Gasubunits were constructed through insertion of TagRFP in a conserved loop region of the Ga subunits. The results demonstrated that GNA-1 localizes tothe plasma membrane and vacuoles, and also to septa throughout conidial germination. GNA-2 localizes to both the plasma membrane and vacuolesduring early germination, but is then found in vacuoles later during hyphal outgrowth. Interestingly, in addition to y plasma membrane and vacuolarlocalization, GNA-3 was found in distinct patches on the plasma membrane of the original conidium during early germination. This distinct localization ofGNA-3 supports the hypothesis that GNA-3 is needed for proper conidial germination, and this specific localization may be required for development.Further investigation is under way to determine the consequence of this localization. Colocalization of RIC8-GFP with GNA-1-TagRFP or GNA-3-TagRFP wasnot detected in cells expressing two fluorescent proteins. This finding suggests that their interaction may be transient not able to be captured via thismethod. A more sensitive microscopic approach is being implemented to better test for colocalization.163. Deciphering the roles of the secretory pathway key regulators YPT-1 and SEC-4 in the filamentous fungus Neurospora crassa. E. Sanchez, M.Riquelme. Center for Scientific Research and Higher Education of Ensenada (CICESE). Carretera Ensenada-Tijuana No. 3918, Zona Playitas, C.P. 28860,Ensenada-B.C.-Mexico.The transport of proteins through different compartments of the secretory pathway is mediated by vesicles. It is well known that vesicular trafficking isregulated by Rab GTPases, which in their active state interact with the membrane of the vesicles. Subsequently, through protein-protein interactions, theycoordinately associate with factors involved in transport and/or tethering to the receptor organelle. In contrast to other eukaryotic model systems, mostfilamentous fungi contain a Spitzenkörper (Spk), which is a multi-vesicular complex found at the hyphal apex to which cargo-carrying vesicles arrive beforebeing redirected to specific cell sites. The exact regulatory mechanisms utilized by the hyphae to ensure the directionality of the secretory vesicles thatreach the Spk are still unknown. Hence, we have analyzed the N. crassa Rab-GTPases YPT-1 and SEC-4, key regulators of the secretory pathway rather wellcharacterized in S. cerevisiae. YPT-1 regulates ER-Golgi and late endosome-Golgi traffic steps, while SEC-4 regulates post-Golgi vesicle traffic en route tothe plasma membrane. Laser scanning confocal microscopy of strains expressing fluorescently tagged versions of the proteins revealed that YPT-1 localizesat the Spk microvesicular core and at cytoplasmic pleomorphic punctate structures, suggesting its participation in different traffic steps. YPT-1accumulation at the Spk might suggest its function in mediating the traffic of vesicles from early endosomes as a recycling process. The pleomorphicstructures could correspond to late Golgi equivalents. The localization of SEC-4 at the Spk, suggests the participation of this Rab in late traffic steps ofGolgi-derived vesicles previous to exocytic events. The relative distribution of both Rabs compared to the molecular motor MYO-2 (presumably involved insecretory vesicle transport), the long coiled-coil protein USO-1 (tethering factor), the secreted protein INV-1, and proteins involved in cell wall biosynthesisis being analyzed and will provide better clues on the nature of the identified compartments.164. Functional characterization of CBM18 proteins, an expanded family of chitin binding genes in the Batrachochytrium dendrobatidis genome. PengLiu, Jason Stajich. Plant Pathology & Microbiology, Univ California, Riverside, Riverside, CA.Batrachochytrium dendrobatidis (Bd) is the causative agent of chytridiomycosis, one of the major causes of worldwide decline in amphibian populations.Little is known about the molecular mechanisms of its pathogenicity. Our previous work 1 from the initial analysis of the Bd genome revealed a uniqueexpansion 18 copies of the carbohydrate-binding module family 18 (CBM18), specific to Bd, and evolving under positive directional selection. CBM18 ispredicted to be a sub-class of chitin recognition domains. Our hypothesis is that some of these copies of CBM18 can bind chitin, a major component offungal cell walls, in vitro. In order to investigate CBM18’s intracellular localization, four CBM18 genes, representing tyrosinase-like, deacetylase-like andlectin-like groups, were cloned into a yeast GFP expression vector. Only two genes from lectin-like group fused with GFP, showing cell boundarylocalization. Furthermore, intracellular signals were observed on both GFP fusion proteins. According to the TargetP database, both proteins are predictedto have the secretion signal peptide. When co-stained with FM4-64, a dye to label vacuole membranes, the FM4-64 and GFP signals were mutuallyexclusive, indicating that the GFP fusion proteins were not destined for degradation. Expression of the proteins from the pHIL-S1 vector in the Pichiasystem will enable purification and characterization of binding properties of these molecules and affinity for chitin and other substrates. 1. Abramyan andStajich, mBio 2012; 3(3): e00150-12.165. The exocyst complex is necessary for secretion of effector proteins during plant infection by Magnaporthe oryzae. Yogesh K. Gupta 1 , MarthaGiraldo 2 , Yasin Dagdas 1 , Barbara Valent 2 , Nicholas J. Talbot 1 . 1) School of Biosciences, University of Exeter, EX4 4QD, UK; 2) Department of Plant Pathology,Kansas State University, Manhattan, Kansas, USA.Magnaporthe oryzae is a devastating plant pathogenic fungus, which causes blast disease in a broad range of cereals and grasses. A specialized infectionstructure called the appressorium breaches the leaf cuticle and subsequently the fungus colonizes host epidermal cells. Colonization of host tissue isfacilitated by small secreted proteins called effectors, that suppress plant immunity responses and may also mediate invasive growth. Some of theseeffectors have been shown to localize at the appressorium pore prior to plant infection, at the tips of primary invasive hyphae and in a specialized plantderived,membrane-rich structure called the Biotrophic Interfacial Complex (BIC). However the underlying mechanism controlling polarized secretion is notwell defined in M. oryzae. The exocyst is an octameric protein complex (composed of Sec3, Sec5, Sec6, Sec8, Sec10, Sec15, Exo70 and Exo84) that appearsto be evolutionary conserved in fungi and to play a crucial role in vesicle tethering to the plasma-membrane. The exocyst plays an important role inpolarized exocytosis and interacts with various signaling pathways at the apex of fungal cells. We are currently characterizing components of exocystcomplex during infection related development of M. oryzae. We have shown that the exocyst localizes to hyphal tips as in other fungi during hyphalgrowth in culture. Interestingly, exocyst components also localize around the appressorium pore, which suggests the pore is an active site for secretion atthe point of plant infection. We have recently shown that organization of the appressorium pore requires a hetero polymeric septin network and we show27th Fungal Genetics Conference | 161

FULL POSTER SESSION ABSTRACTShere that localization of the exocyst at the appressorium pore is septin dependent. The exocyst is furthermore involved in secretion of symplastic (hostcell-delivered) effectors but not apoplastic effectors. Targeted gene deletion of exocyst components Exo70 and Sec5 causes significant virulence defectsbecause of impaired secretion. We will present new information on the role of the exocyst during invasive growth of M. oryzae.166. Functional analysis of protein ubiquitination in the rice blast fungus Magnaporthe oryzae. Yeonyee Oh, Hayde Eng, William Franck, DavidMuddiman, Ralph Dean. Dept Plant Pathology, NCSU, Raleigh, NC.Rice blast is the most important disease of rice worldwide, and is caused by the filamentous ascomycete fungus, Magnaporthe oryzae. Proteinubiquitination, which is highly selective, regulates many important biological processes including cellular differentiation and pathogenesis in fungi. Geneexpression analysis revealed that a number of genes associated with protein ubiquitination were developmentally regulated during spore germination andappressorium formation. We identified an E3 ubiquitin ligase, MGG_13065 is induced during appressorium formation. MGG_13065 is homologous tofungal F-box proteins including Saccharomyces cerevisiae Grr1, a component of the Skp1-Cullin-F-box protein (SCFGrr1) E3 ligase complex. Targeted genedeletion of MGG_13065 resulted in pleiotropic effects on M. oryzae including abnormal conidia morphology, reduced growth and sporulation, reducedgermination and appressorium formation and the inability to cause disease. Our study suggests that MGG_13065 mediated ubiquitination of targetproteins plays an important role in nutrient assimilation, morphogenesis and pathogenicity of M. oryzae.167. The role of autophagy in Cryphonectria hypovirus 1 (CHV1) infection in Cryphonectria parasitica. M. Rossi, M. Vallino, S. Abba', M. Turina. Instituteof Plant Virology, National Research Council (CNR), Torino, Italy.The interaction between Cryphonectria parasitica, the causal agent of chestnut blight, and Cryphonectria hypovirus 1 (CHV1) results in fungalhypovirulence associated with alterations of fungal development, reduced sporulation and pigmentation, accumulation of cytosolic vesicles. The role ofthese vesicles is to support CHV1 maintenance and replication, but the origin of these compartments is still under debate. Due to the phylogeneticproximity between CHV1 and poliovirus, which induces autophagosome proliferation in infected cells, we decided to explore the involvement ofautophagy in vesicle accumulation and virus replication in CHV1-infected mycelium. We are studying the autophagy dynamic in CHV1-infectedCryphonectria expressing GFP-CpAtg8. Atg8 is the fungal orthologue of the mammalian LC3, an essential protein for autophagosome formation which isconsidered a reliable autophagosome marker. In CHV1-free hyphae, GFP-CpAtg8 distribution was mostly cytosolic, but in presence of CHV1 we observed apunctate distribution of fluorescence which is compatible with the binding of GFP-CpAtg8 with autophagosome membranes. The induction of autophagy isalso supported by the observed increase of accumulation of GFP-CpAtg8 in presence of CHV1 compared with virus-free mycelium which could be due to anactivation of gene transcription and/or to protein stabilization. Overall our results seem to confirm the activation of autophagy by CHV1. We are nowtesting through various approaches if CHV1 is able to induce autophagosomes proliferation to support its own replication or if this is an effect of fungaldefense against hypovirus infection.168. Neurospora crassa protein arginine methyl transferases are involved in growth and development and interact with the NDR kinase COT1. D.Feldman, C. Ziv, M. Efrat, O. Yarden. Dept of Plant Pathology and Microbiology, Faculty of Agricutlure, The Hebrew University of Jerusalem, Rehovot, Israel.The protein arginine methyltransferaseas (PRMTs) family is conserved from yeast to human, and regulates stability, localization and activity of proteins.We have characterized deletion strains corresponding to genes encoding for PRMT1/3/5 (designated prm-1, prm-3 and skb-1, respectively) in N. crassa.Deletion of PRMT-encoding genes conferred reduced growth rates and altered Arg-methylated protein profiles (as determined immunologically). Dprm-1exhibited reduced hyphal elongation rates (70% of wild type) and increased susceptibility to the ergosterol biosynthesis inhibitor voriconazole. In Dprm-3,distances between branches were significantly longer than the wild type, suggesting this gene is required for proper regulation of hyphal branching.Deletion of skb-1 resulted in hyper conidiation (2-fold of the wt) and increased tolerance to the chitin synthase inhibitor polyoxin D. Inactivation of twoPRMTs responsible for asymmetric dimethylation (Dprm-1;Dprm-3) conferred changes in both asymmetric as well as symmetric protein methylationprofiles, suggesting either common substrates or cross-regulation of different PRMTs. Taken together, all N. crassa PRMTs are involved in fungal growth,hyphal cell integrity and affect asexual (but not sexual) reproduction. The PRMTs in N. crassa apparently share cellular pathways which were previouslyreported to be regulated by the NDR (Nuclear DBF2-related) kinase COT1, whose dysfunction leads to a pleiotropic change in hyphal morphology. Usingco-immunpercipitation experiments, we have shown that SKB1 and COT1 can physically interact. To date, two isoforms of COT1 (67 and 73KDa) have beenidentified and studied. We have now identified a third, 70kDa, isoform of COT1, whose abundance was increased in a Dskb-1 background. This isoform, aswell as the two others, are Arg-methylated, as determined on the basis of immunological detection and results indicate that the methylation observedinvolves the activity of more than one PRMT enzyme. The fact that environmental suppression of the cot-1 phenotype is more pronounced in prm-3 andskb-1 backgrounds links these PRMTs to the environmental response associated with COT1 function. Based on the highly conserved structure of the PRMTsand the NDR kinases in eukaryotes, it is likely that these proteins undergo similar interactions in other organisms.169. Role of tea1 and tea4 homologs in cell morphogenesis in Ustilago maydis. Flora Banuett, Woraratanadharm Tad, Lu Ching-yu, Valinluck Michael.Biological Sciences, California State University, Long Beach, CA.We are interested in understanding the molecular mechanisms that govern cell morphogenesis in Ustilago maydis. This fungus is a member of theBasidiomycota and exhibits a yeast-like and a filamentous form. The latter induces tumor formation in maize (Zea mays) and teosinte (Zea mays subsp.parviglumis and subsp. mexicana). We used a genetic screen to isolate mutants with altered cell morphology and defects in nuclear position. One of themutants led to identification of tea4. Tea4 was first identified in Schizosaccharomyces pombe, where it interacts with Tea1 and other proteins thatdetermine the axis of polarized growth. Tea4 recruits a formin (For3), which nucleates actin cables towards the site of growth, and thus, polarizessecretion (Martin et al., 2005). Tea1 and Tea4 have been characterized in Aspergillus nidulans and Magnaporthe oryzae (Higashitsuji et al., 2009; Patkar etal., 2010; Takeshita et al., 2008; Yasin et al., 2012). Here we report the characterization for the first time of the Tea4 and Tea1 homologs in theBasidiomycota. The U. maydis tea4 ORF has coding information for a protein of 1684 amino acid residues that contains a Src homology (SH3) domain, aRAS-associating domain, a phosphatase binding domain, a putative NLS, and a conserved domain of unknown function. All Tea4 homologs in theBasidiomycota contain a RA domain. This domain is absent in Tea4 homologs in the Ascomycota, suggesting that Tea4 performs additional functions in theBasidiomycota. We also identified the Umtea1 homolog, which codes for a putative protein of 1698 amino acid residues. It contains three Kelch repeats.The Tea1 homologs in the Ascomycota and Basidiomycota contain variable numbers of Kelch repeats. The Kelch repeat is a protein domain involved inprotein-protein interactions. The tea1 gene was first identified in S. pombe and is a key determinant of directionality of polarized growth (Mata and Nurse,1997). To understand the function of tea1 and tea4 in several cellular processes in U. maydis, we generated null mutations. We demonstrate that tea4 andtea1 are necessary for the axis of polarized growth, cell polarity, normal septum positioning, and organization of the microbutubule cytoskeleton. We alsodetermined the subcellular localization of Tea1::GFP and Tea4::GFP in the yeast-like and filamentous forms.162

FULL POSTER SESSION ABSTRACTS170. Sex determination directs uniparental mitochondrial inheritance in Phycomyces blakesleeanus. Viplendra P.S. Shakya, Alexander Idnurm. School ofBiological Sciences, University of Missouri-Kansas City, MO.Uniparental inheritance (UPI) of mitochondria is common among eukaryotes. Various mechanisms have been suggested for UPI, but the underlyingmolecular basis is yet to be fully explained. We used a series of genetic crosses to establish that the sexM and sexP genes in the mating type locus controlthe UPI of mitochondria in the Mucoromycotina fungus Phycomyces blakesleeanus. Inheritance is from the (+) sex type, and is associated with degradationof the mitochondrial DNA from the (-) parent in the developing zygospore. Hence, the UPI of mitochondria in Phycomyces shows that this process can bedirectly controlled by genes that determine sex identity, independent of cell size or the complexity of the genetic composition of a sex chromosome.171. Exploring the role of a highly expressed, secreted tyrosinase in Histoplasma capsulatum mycelia. Christopher F. Villalta 1 , Dana Gebhart 2 , Anita Sil 1 .1) Microbiology and Immunology, UCSF, San Francisco, CA; 2) AvidBiotics Corporation, South San Francisco, California, United States of America.The human pathogen Histoplasma capsulatum is a dimorphic ascomycete that resides in the soil at ambient temperature as a mycelium. Infection ofimmunocompetent individuals with H. capsulatum occurs when mycelial fragments and associated conidia are inhaled. These fungal cells undergo aconversion to a budding-yeast form in response to mammalian body temperature. We are interested in genes that specify the biological attributes ofeither the infectious form (mycelia or conidia) or the parasitic form (yeast). Previous work from our lab compared the gene expression profiles of mycelia,conidia, and yeast cells to determine genes that were preferentially expressed in each developmental form. We determined that the TYR1 gene, whichencodes a putative polyphenol oxidase, or “tyrosinase”, is highly differentially expressed in the mycelial form of H. capsulatum. Notably, the H. capsulatumgenome contains seven tyrosinases, all of which are more highly expressed in mycelia and conidia compared to yeast. These enzymes contain a conservedtyrosinase domain, but their function in pathogenic fungi has not been investigated. Our expression data suggest that tyrosinases play a specific role in thebiology of H. capsulatum filaments and spores. Strains that either lack TYR1 or express deregulated TYR1 display altered growth properties during themycelial phase. Interestingly, our preliminary results indicate that Tyr1 is secreted into the media during mycelial-phase growth. We are currentlyinvestigating whether Tyr1 affects mycelial growth by modifying a cell-surface or secreted molecule. Additionally, we are determining if Tyr1 is importantin the production of infectious spores.172. Hypobranching induced by both anti-oxidants and ROS control gene knockouts in Neurospora crassa. Michael K. Watters, Jacob Yablonowski, TaylerGrashel, Hamzah Abduljabar. Dept Biol, Valparaiso Univ, Valparaiso, IN.Wild-type Neurospora grows with the same branch density (statistical distribution of physical distances between branch points along a growing hypha) ata wide range of incubation temperatures. Previous work highlighted the impact of reactive oxygen species (ROS) control on branch density. Here we reportthe branching effects of selected ROS control gene knockout mutants; the impact of exogenously added anti-oxidants. In all ROS control mutants tested,growth was shown to branch tighter when grown at higher temperatures and looser when grown at lower temperatures. The branch density displayed bythe ROS mutants at low temperature is measurably hypobranched. In tests on wild type Neurospora, added Ascorbic Acid and Glutathione producedunusual branching patterns. Hypha exposed to Ascorbic Acid or Glutathione display a distribution of branching with two distinct maxima. They show anincrease in both very closely spaced branching as well as an increase in more distantly spaced branching. At lower doses however, hypobranching, again, isobserved with average branch density being linearly related to the dose of added anti-oxidants. We also report on the interaction between ROS mutantsand added anti-oxidants.173. Septum formation starts with the establishment of a septal actin tangle (SAT) at future septation sites. Diego Delgado-Álvarez 1 , S. Seiler 2 , S.Bartnicki-García 1 , R. Mouriño-Pérez 1 . 1) CICESE, Ensenada, Mexico; 2) Georg August University, Göttingen, Germany.The machinery responsible for cytokinesis and septum formation is well conserved among eukaryotes. Its main components are actin and myosins, whichform a contractile actomyosin ring (CAR). The constriction of the CAR is coupled to the centripetal growth of plasma membrane and deposition of cell wall.In filamentous fungi, such as Neurospora crassa, cytokinesis in vegetative hyphae is incomplete and results in the formation of a centrally perforatedseptum. We have followed the molecular events that precede formation of septa and constructed a timeline that shows that a tangle of actin filaments isthe first element to conspicuously localize at future septation sites. We named this structure the SAT for septal actin tangle. SAT formation seems to bethe first event in CAR formation and precedes the recruitment of the anillin Bud-4, and the formin Bni-1, known to be essential for septum formation.During the transition from SAT to CAR, tropomyosin is recruited to the actin cables. . Constriction of the CAR occurs simultaneously with membraneinternalization and synthesis of the septal cell wall.174. Characterization of the Neurospora crassa STRIPAK complex. Anne Dettmann 1 , Yvonne Heilig 1 , Sarah Ludwig 1 , Julia Illgen 2 , Andre Fleissner 2 , StephanSeiler 1 . 1) Institute for Biology II, Molecular Plant Physiology, Freiburg, Germany; 2) Biozentrum, Technische Universität Braunschweig,Germany.The majority of fungi grow by polar tip extension, branching and intercellular fusion to generate a supra-cellular, syncitial mycelium. This hyphal networkformation increases the fitness of the organisms and is central to the organization and function of the fungal colony. Multiple mutants deficient in hyphalfusion and/or intercellular signaling were characterized in Neurospora crassa, the currently best understood model for interhyphal signaling. Among themare components of the two MAK1 and MAK2 MAP kinase cascades and a cell fusion-specific phosphatase 2A termed the STRIPAK complex. While theMAK2 cascade is central for signaling through oscillatory recruitment of the MAK2 module to opposing tips of communicating cells, the MAK1 cell wallintegrity pathway is assumed to play a critical role in the cell wall rearrangement after the physical contact of the two partner cells. The mechanisticfunction of the STRIPAK complex and the functional relationship of the three modules is not resolved. By a combination of genetic, biochemical and life cellimaging techniques, we present the characterization of the STRIPAK complex of N. crassa that consists of HAM2/STRIP, HAM3/striatin, HAM4/SLMAP,MOB3/phocein, PPG1/PP2AC and PP2AA. We further describe that the fungal STRIPAK complex localizes to the nuclear envelope and regulates the nuclearaccumulation of the MAP kinase MAK1 in a MAK2-dependent manner.175. Does the CENP-T-W-S-X tetramer link centromeres to kinetochores? Jonathan Galazka, Mu Feng, Michael Freitag. Biochemistry and Biophysics,Oregon State University, Corvallis, OR.In vertebrates, the centromeric proteins, CENP-T, -W, -S and -X, form a tetramer (CENP-T-W-S-X) in vitro that binds DNA [1]. Furthermore, theunstructured N-terminus of CENP-T interacts with the Ndc80 complex at kinetochores [2]. This suggests that CENP-T-W-S-X has a central role in linkingcentromeric DNA to kinetochores. Despite the appeal of this model, there is no evidence that this complex forms in vivo, no information of the DNAsequences it may bind at centromeres and little understanding of how it interacts with canonical nucleosomes. CENP-T, -W, -S, and -X are conserved infungi, including Neurospora [1-3]. Neurospora is an attractive model in which to understand the function of the CENP-T-W-S-X complex as its centromeric27th Fungal Genetics Conference | 163

FULL POSTER SESSION ABSTRACTSDNA is nearly completely assembled, allowing ChIP-seq reads to be mapped unambiguously [4]. Here, we report on our investigations of the NeurosporaCENP-T-W-S-X complex, including its interactions with centromeric DNA and canonical centromeric nucleosomes.[1] Nishino, T. et al. 2012. CENP-T-W-S-X Forms a Unique Centromeric Chromatin Structure with a Histone-like Fold. Cell 148, 487-501.[2] Schleiffer, A. et al. 2012. CENP-T proteins are conserved centromere receptors of the Ndc80 complex. Nat. Cell Biol. 14, 604-613.[3] Smith, K. M. et al. 2012. Centromeres of filamentous fungi. Chrom. Res. 20, 635-656.[4] Smith, K. M. et al. 2011. Heterochromatin is required for normal distribution of Neurospora crassa CenH3. Mol. Cell. Biol. 31, 2528-2542.176. Proper actin ring formation and septum constriction requires coordination of SIN and MOR pathways through the germinal centre kinase MST1.Yvonne Heilig, Anne Dettmann, Stephan Seiler. Institute for Biology II, Molecular Plant Physiology, Freiburg, Germany.The highly conserved nuclear Dbf2p-related (NDR) kinases control polar morphogenesis and cell proliferation. In fungi, NDR kinases function as effectorsof the morphogenesis (MOR) and septation initiation (SIN) networks and are activated by germinal centre (GC) kinases. The Neurospora crassa SIN kinasesSID1 and DBF2 are essential for septum formation. In contrast, the MOR kinases POD6 and COT1 promote apical tip growth and function as negativeregulators of septation. We identified a third GC kinase MST1 that functions as promiscuous enzyme, activating DBF2 and COT1. As typical for SINcomponents, MST1 localized to spindle pole bodies and constricting septa. Moreover, Dmst-1 displayed synthetic interactions with sin, but not mormutants, placing MST1 in parallel to the central SIN kinase cascade CDC7-SID1-DBF2. Consistent with these genetic data, we determined that the two GCkinases MST1 and SID1 are regulated by CDC7 in an opposite manner. Lifeact- and formin-GFP reporter constructs revealed the formation of aberrantcortical actin rings in Dmst-1, which resulted in mispositioned septa and irregular spirals in the mutant. In summary, our data identify an antagonisticrelationship between the SIN and MOR during septum formation that is, at least in parts, coordinated through the GC kinase MST1.177. Regulatation of the BUD3-BUD4 landmark complex by the NDR kinases DBF2 and COT1 during septum formation in Neurospora crassa. YvonneHeilig, Stephan Seiler. Institute for Biology II, Molecular Plant Physiology, Freiburg, Germany.Cytokinesis is essential for cell proliferation, yet the mechanisms for determining the site of cell division are poorly understood. Our data indicate thatthe anillin BUD4 marks septum placement by organizing the RHO4-BUD3-BUD4 GTPase module and that this complex is controlled through two NDRkinase signaling cascades, the septation initiation network (SIN) and the morphogenesis network (MOR). Epistasis analysis of sin and mor mutants placesthe SIN upstream of the MOR. DBF2 functions as competitive inhibitor of COT1 by forming hetero-dimers, thereby replacing the COT1 co-activatorsMOB2A/B. In turn, COT1 functions as negative regulator of septum formation. We demonstrate that COT1, but not DBF2, binds to and phosphorylatesBUD3 and BUD4. Mutational analysis of BUD3 identifies Ser798, located within an amphiphatic helix of BUD3 that is phosphorylated by COT1. Localizationof this amphiphatic helix at septa is only possible in its nonphosphorylated form. In summary, our data suggest a model, in which the MOR kinase COT1phosphorylates BUD3 and BUD4 and that this modification inhibits cortical localization and function of the BUD complex. Interference of the SIN with MORactivity at the septum relieves this inhibition and allows initiation of septation.178. Development of a Protein-Protein Interaction Platform in Neurospora Crassa. Shouqiang Ouyang, Katherine Borkovich. Plantn Pathology andMicrobiology, University of California, Riverside, Riverside, CA.The objective of this study is to generate a protein-protein interaction platform for Neurospora crassa. We have constructed Dmus-51::nat and Dmus-52::nat strains that also carry the Drid::nat mutation to eliminate RIP. These strains are used as recipients for transformation. Ten genes were solicited ascandidates from the N. crassa community, including SAD-1/SAD-2, WC-1/WC-2, FRQ/FRH, OS-4/ RRG-1and GNB-1/GNG-1. We construct vectors for eachprotein by amplifying the ORFs from wild type N. crassa genomic DNA using gene-specific primers. Protein constructs are expressed with a V5-GFP or S-tag-RFP tag from the pan-2 or inl locus, respectively in N. crassa. Protein complexes can be isolated by immunoprecipitation using antibody to the GFP/V5or RFP/S-tag epitope. Both immunoprecipitation and the overlap localization of fluorescent proteins (GFP and RFP) data will streamline our ability tomonitor protein-protein interactions and co-localization in vivo in N. crassa.179. Specific Structural Features of Sterols Affect Cell-Cell Signaling and Fusion in Neurospora crassa. Martin Weichert 1 , Ewald Priegnitz 1 , RaphaelBrandt 1 , Thorben Nawrath 2 , Stefan Schulz 2 , André Fleissner 1 . 1) Institut für Genetik, Technische Universität Braunschweig, Spielmannstrasse 7, 38106Braunschweig, Germany; 2) Institut für Organische Chemie, Technische Universität Braunschweig, Hagenring 30, 38106 Braunschweig, Germany.Sterols are major constituents in the plasma membrane of eukaryotic cells. They modulate the physical properties of the lipid bilayer, e.g. fluidity. Byinteracting with certain lipids and proteins in the plasma membrane, sterols cluster into microdomains which might act as platforms for many biologicalfunctions, such as signal transduction. In the early stages of colony formation in Neurospora crassa, germinating spores direct their growth towards eachother, establish physical contact, and fuse. Cell-to-cell signaling requires the coordinated dynamic recruitment of the MAP kinase MAK-2 and thecytoplasmic protein SO to the tips of interacting cells. Subsequent plasma membrane fusion is facilitated by the transmembrane protein PRM1. Here, wereport that mutants affected in the biosynthesis of ergosterol, the major sterol in most fungal species, show distinct defects during germling fusion.Deletion of erg-2, which encodes an enzyme mediating the last step in the pathway, strongly impairs both directed growth and cell fusion. Interestingly,both MAK-2 and SO mislocalize at the tips of interacting Derg-2 germlings. In contrast, the absence of ERG-10a and ERG-10b, two enzymes with redundantfunction that act upstream of ERG-2, does not affect cell-to-cell communication. However, Derg-10a Derg-10b germling pairs show DPrm1-like deficienciesin plasma membrane merger. By relating the sterol composition and fusion competence of several erg mutants, we find that not the absence of ergosterolbut the accumulation of sterol intermediates specifically impairs distinct steps of germling fusion. While the presence of two double bonds in the sterolside chain provokes Derg-2-like deficiencies, an altered double bond arrangement in the sterol ring system causes DPrm1-like defects. During sexualdevelopment, cell fusion precedes the fertilization of fruiting bodies. Unlike the defects during germling fusion, female and male mating partners of Derg-2and Derg-10a Derg-10b efficiently fuse, suggesting that alterations in the sterol composition specifically impair signaling mechanisms mediating vegetativecell fusion. These data suggest that specific structural features of sterols differentially affect membrane properties and functions, such as the membranerecruitment of proteins, the assembly of signaling complexes, and plasma membrane fusion.180. The role of NADPH oxidases in Neurospora crassa cell fusion. Nallely Cano-Dominguez 1 , Ernestina Casto-Longoria 1 , Jesus Aguirre 2 . 1) Departamentode Microbiologia, CICESE, Ensenada, Baja California, Mexico; 2) Departamento de Biologia Celular y Desarrollo. Instituto de Fisiologia Celular UNAM,Mexico City, D.F. Mexico.Hansberg and Aguirre proposed that reactive oxygen species (ROS) play essential roles in cell differentiation in microorganisms. ROS are generatedmainly during mitochondrial electron transport and by the action of certain enzymes. The NADPH oxidases (NOX) are enzymes that catalyze the production164

FULL POSTER SESSION ABSTRACTSof superoxide by transferring electrons from NADPH to oxygen. Neurospora crassa contains the NADPH oxidases NOX-1 and NOX-2 and a commonregulatory subunit NOR-1. NOX-2 is essential for ascospore germination, while NOX-1 is required for sexual and asexual development, polar growth andcell fusion. NOR-1 is essential for all these NOX functions. We have found that a functional NOR-1::GFP fusion is localized throughout the cytoplasm,enriched at the hyphal tip and sometimes in aggregates. This suggests that the functional NOX complexes are probably not localized at the plasmamembrane. Up to now NOX function in fungi has been evaluated in mutants that completely lack NOX proteins. We generated nox-1 alleles that result inNOX-1 proteins carrying substitutions of proline 382 by histidine or cysteine 542 by arginine, which affect NADPH-binding. Equivalent mutations inphagocytic Nox2/gp91phox do not affect protein stability but completely lack oxidase activity. P382H and C542R mutants did not produce sexual fruitingbodies and showed a decreased growth and differentiation of aerial mycelia, without affecting production of conida. These results indicate that sexualdevelopment depends on ROS production by NOX-1, whereas during asexual differentiation NOX-1 plays an important role independently of its catalyticactivity. Dnox-1, Dnor-1, P382H NOX-1 and C542R NOX-1 mutants were all able to produce some conidial anastomosis tubes (CATs) but they were unableto complete cell-cell fusion. All these mutants are also impaired in vegetative hyphae-hyphae fusion, which might explain the growth defects in Dnox-1 andDnor-1 strains. CATs production is delayed in the presence of antioxidant N- acetyl cystein (NAC) and Dsod-1 strains show an increase in CATs fusions. Theresults suggest that some ROS may be implicated in signaling CATs homing and vegetative fusion.181. DYNAMICS OF THE PROTEINS BUD-2 AND BUD-5 DURING CELL POLARIZATION IN NEUROSPORA CRASSA. E. Castro-Longoria, C. Araujo-Palomares, N.Cano-Domínguez. Microbiology Department, CICESE. Carretera Ensenada-Tijuana No 3918 Zona Playitas, C.P. 22860. Ensenada, B.C. México.Polarized growth in filamentous fungus requires an excellent and precise machinery to select specific sites where multiple protein complexes assembleto ensure the generation of highly polarized hyphae. One of these protein complexes is the Rsr1p/Bud1p-Bud2p-Bud5p module, which, in Saccharomycescerevisiae, has the function of selecting the proper site of budding. However, in filamentous fungi the function of this module is unknown. In this study, wecharacterized the intracellular localization and dynamics of protein homologues for BUD-2 and BUD-5 in the filamentous fungus Neurospora crassa.Preliminary results of in vivo confocal microscopy analysis shows that both BUD-2 and BUD-5 display distinct localization patterns in both mature hyphaeand germlings. In mature hyphae, BUD-2 localization is confined to the apical cytosol, occupying the core of the Spitzenkörper (Spk), while BUD-5 wasobserved in the apical region of the cells as a bright spot with higher intensity at the center base adopting a hand fan shape, partially colocalizing with theSpk. In contrast, BUD-2 in germlings was associated with the cell membrane and organized as a cap shape covering the apex of the cells, while BUD-5localization was observed in three different ways: as a bright spot at the apex of germinating spores, then as a cytosolic crescent-shape in longer germtubes and finally adopting a similar localization pattern as in mature hyphae. BUD-2 and BUD-5 also display distinct localization patterns during branchingand septum formation. BUD-2 participates in septum formation while BUD-5 was only involved during the initiation of lateral branches. The distinctcellular localization patterns of BUD-2 and BUD-5 suggest that although both proteins may be important for cell polarity establishment, they alsoparticipate in other morphogenetic processes in N. crassa.182. The role of calcium and calmodulin during cell fusion and colony initiation in Neurospora crassa. Chia-Chen Chang, Nick Read. Fungal Cell BiologyGroup, Institute of Cell Biology, University of Edinburgh, Edinburgh EH9 3JH.Calcium is an ubiquitous signalling molecule which regulates many important processes in filamentous fungi including spore germination, hyphal growth,mechanosensing, stress responses, circadian rhythms, and virulence. Transient increases in cytosolic free calcium ([Ca 2+ ] c) act as intracellular signals. As theprimary intracellular Ca 2+ receptor, calmodulin (CaM) converts these Ca 2+ signals into responses by regulating the activity of numerous target proteins. Wehave found that both Ca 2+ -free medium and two CaM antagonists (calmidazolium and trifluoperazine) selectively inhibit a form of cell fusion called conidialanastomosis tube (CAT) fusion that occurs during colony initiation in the fungal model Neurospora crassa. GFP labelled CaM localized as dynamic particlesassociated with the plasma membrane and moved around within the cytoplasm in both germ tubes and CATs. In particular, CaM showed a dynamicaccumulation at two growing tips of CATs that exhibit chemoattraction towards each other. CaM also localized at developing septa in germ tubes. The b-tubulin inhibitor, benomyl, reduced the movement of CaM in the cytoplasm. Moreover, the absence of extracellular Ca 2+ inhibited the recruitment of CaMto CAT tips as well as inhibiting CAT chemoattraction. The deletion of the myosin-5 (myo-5) gene caused the mis-localization of CaM in tips of growinggerm tube and CATs. This suggests that the movement of cytoplasmic CaM involves transport along microtubules, and the recruitment of CaM to tipsinvolves myosin-5 along F-actin and is dependent on extracellular Ca 2+ .183. Deletion of cAMP phosphodiesterase pde-2/acon-2 gene causes the enhanced osmotic sensitivity in os-1 and os-2 mutants of N. crassa. C. Kurata,M. Kamei, S. Banno, M. Fujimura. Dept Life Sci, Toyo Univ, Gunma, Japan.N. crassa has two putative cyclic nucleotide phosphodiesterases, PDE-1 (NCU00237) and PDE-2/ACON-2 (NCU00478). The pde-2 disruptants showed thenormal mycelial growth but lacked the ability to produce conidia, these phenotypes resembled those of the hah mutant which has a point mutation in thePKA (protein kinase A) regulatory subunit gene. The phenotypes of double mutants, pde-2;pkac-1 and hah;pkac-1 mutants, resembled those of the pkac-1mutant which shows slow growth and hyperconidiation. In contrast, hyperconidiation of the adenylyl cyclase cr-1 mutant was suppressed by the hahmutation but not by the pde-2 mutation. These results indicate that PDE-2 act as a major cAMP phosphodiesterase in cAMP-PKA pathway, its deletionleads to the hyper-activation of this pathway. Any mutants in cAMP-PKA pathway including pde-2 and hah mutants, did not show osmotic sensitivity.However, both pde-2 and hah mutations caused the enhanced osmotic sensitivity in os-1 (histidine kinase) and os-2 (MAP kinase) mutants, suggesting ofcross-talk between cAMP-PKA pathway and OS-2 MAP kinase pathway.184. Genetic analysis of GNB-1 and CPC-2 with the G alpha subunits in Heterotrimeric G protein signaling in Neurospora crassa. AMruta Garud. PlantPathology, UC, Riverside, Riverside, CA.Heterotrimeric G protein signaling is mediated by Gabg subunits. Neurospora crassa has three Ga subunits (GNA-1, GNA-2 and GNA-3), one Gb (GNB-1)and one Gg (GNG-1). The GNB-1 protein contains seven tryptophan-aspartate (WD) repeats, suggesting it assumes a beta propeller form. Genetic epistasishas been demonstrated between gnb-1 and the three Ga subunit genes. gna-3 is epistatic to gnb-1 for submerged culture conidiation, while gna-1 andgna-2 are epistatic to gnb-1 during aerial conidiation. In contrast, gnb-1 is epistatic to gna-2 and gna-3 during aerial hyphae development. Additionalproteins that have a 7-WD repeat structure have been implicated as Gb subunits in other fungi. The Cross Pathway Control (CPC-2) protein has a seven WDrepeat structure, and shares 70% similarity to the mammalian protein RACK-1. In Neurospora, CPC-2 was previously shown to play a role in general aminoacid control. Genetic epistasis with CPC-2 and the Ga proteins is being studied, using strains lacking cpc-2 and one Ga gene, as well as cpc-2 deletionmutants carrying constitutively activated, GTPase-deficient Ga alleles. It is seen that gna-3 is epistatic to cpc-2 during apical extension, aerial hyphae heightand asexual sporulation in submerged cultures. gna-1 and gna-2 demonstrate some functional independence. Yeast two hybrid assays show that CPC-2interacts with GNA-1 and GNA-3. Additional interactions are being examined using additional in vivo and in vitro methods to validate whether CPC-2 acts27th Fungal Genetics Conference | 165

FULL POSTER SESSION ABSTRACTSas another Gb subunit in Neurospora.185. Communication Interference during Cell Fusion in Neurospora crassa is controlled by a Region under Balancing Selection in the HeterokaryonIncompatibility Locus het-c. Jens Heller, Javier Palma-Guerrero, N. Louise Glass. Department of Plant and Microbial Biology, University of California atBerkeley, Berkeley, CA.Vegetative hyphal fusion events are associated with establishment of a fungal colony. However, non-self recognition during fusion events is important toprevent hybrids between genetically dissimilar individuals that might spread mycoviruses, debilitated organelles, and others throughout a fungalpopulation. In filamentous fungi, the ability of two individuals to form a productive heterokaryon via hyphal fusion is controlled by specific loci termed hetloci. Stable heterokaryons will only form if the individuals involved have identical alleles at all het loci. Accordingly, heterokaryotic cells formed betweenstrains that differ in allelic specificity at one or more het loci are rapidly destroyed (programmed cell death) or strongly inhibited in their growth. InNeurospora crassa, three allelic specificity groups were identified for het-c, which is one of the eleven genetically identified het loci in this species. Weobserved that strains with different haplotypes at het-c not only show heterokaryon incompatibility (HI) after cell fusion, but also show reducedchemotrophic interactions and cell fusion between conidial germlings (communication interference). These data indicate that N. crassa germlings candistinguish both self and nonself at a distance, and which presumably involves diffusible ligands. Two regions of the glycine-rich single-pass plasmamembrane protein HET-C were shown to be under balancing selection and both have different functions. By analyzing different chimeras of het-c, wedemonstrate that the HET-C specificity domain (amino acids 194-236; region I), is required for inducing programmed cell death during HI, but does notaffect communication interference during germling fusion. In contrast, the second region of het-c that is also under balancing selection (amino acids 521-599; region II) is responsible for communication interference during germling fusion. To understand the mechanism underlying communicationinterference, we are identifying which amino acids in HET-C region II are responsible for this trait. In addition, we are determining the cellular localizationof HET-C during germling fusion and whether the HET-C region II is a processed form, resulting in a diffusible peptide that is responsible for communicationinterference during chemotropic interactions and cell fusion of conidial germlings.186. The N. crassa Bem46 protein: alternative splicing and eisosomal association. Krisztina Kolláth-Leib, Frank Kempken. Department of Botany,Christian-Albrechts University, Kiel, Germany.The bud emergence (BEM) 46 proteins are evolutionarily conserved members of the a/b-hydrolase super family. The exact function(s) of the proteinremain unknown. Vegetative hyphae, perithecia and ascospores of Neurospora crassa RNAi and over-expressing transformants develop normally, buthyphal germination from ascospores is impaired. These results indicate a role of BEM46 in maintaining cell type-specific polarity in N. crassa. In an attemptto further analyse BEM46 function, alternative splicing was observed in the bem46 RNAi line. We present evidence that alternative splice products impairascospore germination. The BEM46 protein is localized in the perinuclear endoplasmatic reticulum and also forms spots near to the plasma membrane(Mercker et al. 2009). The use of Lifeact-TagRFP (Lichius & Read pers. comm.) and Bem46-eGFP in heterokaryons of N. crassa indicated that the Bem46protein is not interacting with actin. Likewise, the use of the lipid raft-stainer TexasRedTM showed no co-localization with Bem46-eGFP. We analyzed thepotential co-localization of Bem46 with the eisosomal protein LSP1. To that end we cloned the corresponding N. crassa ortholog of lsp1 and fused it toRFP. Indeed we were able to demonstrate a co-localization of LSP1 and BEM46. A yeast two-hybrid approach was undertaken using a previouslyestablished N. crassa two-hybrid library (Seiler pers. comm.). We identified one interacting protein, the anthranylate synthase component II (Walker &DeMoss 1986). Further investigation showed that the BEM46 protein is likely to interact with the F domain of that protein, which is a N-(5’-phosphoribosyl) anthranylate isomerase. The interaction was confirmed in vivo by employing bimolecular fluorescence complementation assays.References:Mercker M, Kollath-Leib K, Allgaier S, Weiland N, Kempken F (2009) Curr Genet 55:151-161Margaret S. Walker & John A. DeMoss (1986) J Biol Chem 261:16073-16077.187. The alternative oxidase induction pathway is involved in senescence associated with over-replication of a mitochondrial plasmid in Neurosporacrassa. Nicolette Dutken, Jonathon Gutzeit, Maze Ndonwi, John Kennell. Biology, Saint Louis University, St Louis, MO.Senescence in Neurospora crassa is caused by dysfunctional mitochondria, which is most often due to the effects of mitochondrial plasmids. Variantforms of the Mauriceville plasmid cause senescence by integrating into the mitochondrial genome or by over-replicating, disrupting essentialmitochondrial genes or their synthesis. Genetic analysis of plasmid-containing strains that escape senescence indicate that two nuclear mutations arerequired for longevity. One of the mutations associated with a long lived (LL) strain involves the regulation of Alternative Oxidase (AOX). AOX is induced bymitochondrial dysfunction and is among several nuclear encoded genes involved in mitochondrial function and/or biogenesis that are upregulated duringsenescence. A model of senescence is proposed in which dysfunctional mitochondria stimulate mitochondrial biogenesis resulting in an accumulation ofdefective mitochondria. Here we show that the LL strain fails to induce AOX due to a mutation in aod-2 that encodes a zinc cluster transcriptional regulatorof the structural alternative oxidase gene, aod-1. Surprisingly, a functional AOX is not required for senescence. This implies that other genes controlled bythe AOX induction pathway play a critical role in mitochondrial function in N. crassa. Homologs of AOD2 in other fungal species have been shown toregulate gluconeogenesis as well as genes involved in mitochondrial function including subunits of the electron transport chain. Mutations in the AOXinduction pathway are not sufficient to overcome plasmid induced senescence and a second nuclear mutation is required. This mutation interferes withthe integrative form of senescence and is hypothesized to be associated with either mitochondrial recombination or the selection of mitochondrialrearrangements. The studies of senescence in N. crassa provide insights into how fungi respond to mitochondrial damage.188. Relationship among mutagen sensitivity, senescence and mitochondrial morphology in the ultraviolet sensitive-5 mutant of Neurospora crassa.Kiminori Kurashima, Michael Chae, Hirokazu Inoue, Shin Hatakeyama, Shuuitsu Tanaka. Laboratory of Genetics, Saitama Univercity, Saitama, Japan.The uvs-5 mutant of Neurospora crassa had been isolated that showed high sensitivity to mutagens (Schroeder, 1970 Mol. Gen. Genet. 107:291-304).This mutant also has phenotypes such as senescent, i.e. shortened life span, and progressive accumulation of mitochondrial DNA deletions (Hausner et al.,1995 Fungal Genet. Newsl. 42A: 59). These phenotypes were quite similar to the mus-10 mutant that we reported previously (Kato et al., 2010 Genetics185:1257-1269). Further, mus-10 and aged uvs-5 strains showed fragmented mitochondrial feature although tubular shape was observed in wild typestrain. Since we found that the uvs-5 mutation had been mapped very closely to fzo1, which encoded homologue of dynamin-like GTPase mitofusin, thesequence of the fzo1 gene in the uvs-5 mutant was determined. A single mutation was found as a deducing amino acid substitution of Gln to Arg in the386 th position locating in the conserved GTPase domain. Forced expression of wild-type FZO1 in the uvs-5 strain suppressed the defect in mitochondrialmorphology and the mutagen sensitivity, but did not in the case of expressing mutated FZO1. Moreover, introduction of this mutation into theendogenous fzo1 gene of the wild-type strain resulted in showing phenotypes of the uvs-5 mutant. Thus, we concluded that the responsible gene of uvs-5166

FULL POSTER SESSION ABSTRACTSis fzo1. Attempting to disruption the fzo1 gene was failed, so the fzo1 gene were suggested to be essential for viability as showing in almost all eukaryotesexcept for yeast. Therefore the mutation of uvs-5 may be a useful model for studying the relation between mitochondrial fusion and early senescence inhigher eukaryotes.189. Localization of EGL-1 and EGL-2, two GPI anchored cell wall b (1-3) endoglucanases, at hyphal apices and septa, and in interconidial septa inNeurospora crassa. Leonora Martinez, Meritxell Riquelme. Microbiology Department, Center for Scientific Research and Higher Education at Ensenada,Baja California.The unitary model of cell wall growth suggests that the polarized extension of hyphae in filamentous fungi results from the coordinated synthesis anddischarge of new cell wall polymers, the action of hydrolytic enzymes that provide plasticity to the wall and turgor pressure to drive cell expansion.Currently, there is limited information on enzymes capable of hydrolyzing cell wall polymers and that could be contributing to cell wall remodeling. EGL-1and EGL-2 are putative b (1-3) endoglucanases in Neurospora crassa, with potential binding sites for a glucosyl phosphatidylinositol group (GPI), whichwould allow them to get anchored into the plasma membrane. To investigate whether these proteins participate in key morphogenetic events during thedevelopment of N. crassa, EGL-1 and EGL-2 were labeled with the green fluorescent protein (GFP). For egl-1, the gfp gene was inserted within the egl-1encoding sequence, just after the signal peptide sequence. For egl-2, the insertion took place right before the GPI-binding site. Both endoglucanases werelocalized in the hyphal apical plasma membrane and in septa, however, EGL-2-GFP was strongly and more definite localized at the apical dome and EGL-1-GFP showed less intensity with increasing fluorescence from the subapex to the tip. EGL-1-GFP was mostly found at hyphal septa and interconidial septaand EGL-2-GFP was faintly present in a few old septa. Our results suggest that lytic activity of enzymes, such as the endoglucanases EGL-1 and EGL-2 in N.crassa, is present in critical areas during vegetative morphogenesis, where these enzymes probably play a role in cell wall remodeling, as postulated by theunitary model of cell wall growth.190. Stability of a G protein alpha subunit in genetic backgrounds lacking the G beta subunit or a cytosolic guanine nucleotide exchange factor.Alexander V. Michkov, Katherine A. Borkovich. Plant Pathology and Microbiology, University of California, Riverside, Riverside, CA.Heterotrimeric G proteins consist of alpha, beta and gamma subunits. Regulation is accomplished through the alternation between binding of GDP(inactive form) and GTP (active form) by the alpha subunit and dissociation of the alpha subunit and beta-gamma dimer. GDP/GTP exchange is facilitatedby both cell surface G protein coupled receptors and cytosolic guanine nucleotide exchange factors (GEFs), such as RIC8. Neurospora crassa has three Galpha subunits (GNA-1, GNA-2 and GNA-3), one G beta (GNB-1), and one G gamma (GNG-1). Interestingly, mutants lacking gnb-1 or the cytosolic GEF ric8exhibit some defects in common with the gna-1 deletion mutant, which may be explained by the reduced GNA-1 protein levels observed in these mutants.Previous studies in our laboratory showed that levels of gna-1 mRNA are similar in wild type and mutants lacking gnb-1 or ric8, consistent with a posttranscriptionalmechanism. Using genetic and biochemical approaches, this study investigated the mechanism underlying regulation of GNA-1 stability inregards to GTP/GDP bound state and amount of protein (normal or overexpressed). The results demonstrate that levels of GNA-1 protein are not visiblyreduced over 36 hours in a wild-type background after halting translation using cycloheximide, suggesting GNA-1 is very stable in wild type. To checkstability of GDP or GTP bound GNA-1 in different backgrounds, we transformed mutants lacking the gna-1 gene and gnb-1 or ric8 with a wild type (gna-1 WT ) or constitutively active, GTPase-deficient gna-1 allele (gna-1 Q204L ). Overexpressing gna-1 WT (GDP bound) in a wild-type background increased the levelof GNA-1 protein ~ 3 fold, while overexpression in a gnb-1 mutant gave a nominal increase (~ 1.6x). Overexpressing gna-1 Q204L (GTP bound) in the Dgnb-1or Dric8 backgrounds led to ~ 2 fold higher levels of GNA-1 compared to wild type. In summary, GNA-1 is very stable in wild type, but stability decreasesdramatically in gnb-1 and ric8 deletion mutants. The GTP-bound G alpha protein is more stable in a gnb-1 mutant background than GDP-bound GNA-1protein.191. Functional analysis the Saccharomyces cerevisiae Ste20, Cla4 homologue in Neurospora crassa. Yuhei Nogami, Makoto Fujimura, Akihiko Ichiishi.Faculty of Life Sciences, Toyo University, ORA-GUN, GUNMA, Japan.Signal transduction pathways are important for a variety of features of fungal development. Small GTPases of Rho family act as molecular switchesregulating cell signalling, cytoskeletal organization and vesicle trafficking in eukaryotic cells. The Rho family GTPase Cdc42 was first identified in the yeastSaccharomyces cerevisiae, where it is essential for initiation of bud formation and the subsequent switch from apical to isotropic growth. The activation ofCdc42 is catalyzed by Cdc24 guanine nucleotide exchange factors (GEFs), which convert Cdc42 from an inactive GDP-bound form to the active GTP boundform. Bem1 functions as a scaffold connecting Cdc42 with its activator Cdc24. The GTP-bound Cdc42 can activate p21-activated kinase (PAK), Ste20 andCla4. Neurospora crassa has two PAK family kinases Cla4 and Ste20 homologs. We have few knowledge of the function of their PAK kinases in N. crassa. Inthis study, we performed functional analysis of stk-4 (Ste20 homolog) and vel (Cla4 homolog) in N. crassa. The stk-4 deletion mutant showed slow growththan wild type strain, and vel deletion mutant showed more severe growth defects. To determine the subcellular localization and dynamics of STK-4protein, we constructed GFP-STK-4 fusion constructs. The gfp encoding sequence was fused to the 3’ end of the stk-4 open reading frame. We alsoconstructed the GFP-BEM-1 fusion protein. These constructs were introduced into his-3 locus, and observed using confocal fluorescence microscopy (LSM-510). Both fusion proteins were accumulated at growing hyphal tips and septa. From there results, we consider that STK-4 and BEM-1 are function at thesame site.192. Dissecting the Pathway of Cellulase Secretion in Neurospora crassa. Trevor Starr, Timo Schuerg, Louise Glass. Plant and Microbial Biology, UCBerkeley, Berkeley, CA.Due to their capacity to secrete large amounts of proteins, particularly hydrolytic enzymes, filamentous fungi are of great interest for high-level proteinproduction in various industries, such as the textile, pharmaceutical, and biofuels industries. Although the basic components of the eukaryotic secretionpathway characterized in yeast and higher organisms are also conserved in filamentous fungi, the highly polarized and compartmentalized growth mode offilamentous fungal hyphae mandates pathways of secretion that are specific to these fungi. While certain aspects of filamentous fungal secretion areunder active study, a basic characterization of the entire pathway from start to finish remains to be performed. Such a characterization may provideinsights into how filamentous fungi are able to secrete large amounts of enzymes and how these fungi can be engineered to produce even more enzymesin the future. This is particularly of interest to the process of biofuels production, in which the inexpensive production of large amounts of cellulases is amajor bottleneck to the efficient and cost-effective production of cellulosic biofuels. In nature the model fungus Neurospora crassa secretes a host ofcellulases to allow it to grow on burnt vegetation. The tractability of N. crassa makes it an excellent model to characterize protein secretion in filamentousfungi, particularly the secretion of industrially relevant cellulases. To achieve this goal we are characterizing the cellulase secretion pathway in N. crassa byfollowing the trafficking of fluorescently tagged Endoglucanase 2 (EG-2), a major secreted endocellulase. To determine the compartments through whichcellulases traffic we are co-localizing EG-2-GFP with fluorescently-tagged markers of the ER, Golgi, endosomes, and the Spitzenkorper and are assaying the27th Fungal Genetics Conference | 167

FULL POSTER SESSION ABSTRACTSconsequences to EG-2-GFP trafficking of blocks to secretion imposed by pharmacological or mutational insults. Our initial results indicate that EG-2-GFPshows localization to the ER and is mostly absent from the Spitzenkorper, suggesting trafficking through a classical ER to Golgi secretory pathway andterminal secretion along lateral hypahl walls. Additonally, targeted blocks to the secretory pathway indicate a potential role of endosomes in EG-2-GFPtrafficking.193. Towards understanding the endoplasmic reticulum associated degradation process of misfolded glycoproteins in Neurospora crassa. GeorgiosTzelepis 1 , Hiroto Hirayama 2 , Tadashi Suzuki 2 , Akira Hosomi 2 , Mukesh Dubey 1 , Magnus Karlsson 1 . 1) Uppsala BioCenter, Department of Forest Mycology andPlant Pathology, Swedish University of Agricultural Sciences, Box 7026, 75007, Uppsala, Sweden; 2) Glycometabolome Team, Systems GlycobiologyResearch Group, RIKEN Advanced Science Institute, 2-1 Hirosawa, Wako Saitama 351-0198, Japan.N-glycosylation is an important post-translational modification of proteins, which occurs in the Endoplasmic Reticulum (ER). These N-linked glycans arereported to play an important role in correct protein folding. Glycoproteins that are unable to fold properly are subjected to destruction by an ERassociateddegradation process (ERAD). Degradation of these glycoproteins generates free oligosaccharides (fOs). In animal and plant cells mainly threetypes of hydrolytic enzymes are involved in the ERAD pathway. First, PNGases which cleave the sugar chain from the protein releasing fOs with N,N'-diacetylchitobiose moieties (fOs-GN2). Secondly, ENGases which catalyse the glycosidic bonds in N,N'-diacetylchitobiose moieties, generating fOs with asingle N-acetylglucoseamine at their reducing ends (fOs-GN1), and thirdly, a-mannosidases responsible for trimming the mannose chains before finaldegradation in lysosomes. The existence of this pathway in filamentous ascomycetes is unknown. In this study we investigate the function of ENGases in N.crassa by analysing the phenotype of deletion strain Dgh18-10 and quantifying the content and type of fOs (fOs-GN1 or fOs-GN2), using dual gradient highperformance liquid chromatography. Since cytosolic PNGase is enzymatically inactive in N. crassa, ENGases possibly have a crucial role in the ERADpathway. We found that deletion of an intracellular ENGase results in severe phenotypic effects. This deletion strain shows significantly slower growth ratein carbon-rich media but grows faster in abiotic stress conditions, indicating a more resistant cell wall. Moreover, the conidiation rate is higher in Dgh18-10compared to WT. Sexual reproduction is also affected, since no ascospores were observed in Dgh18-10. Additionally, the total amount of extracellularproteins was significantly lower in this deletion strain compared to WT. Finally, this mutation causes repression of three chitin synthase genes in N. crassa.Similar results were also observed in the mycoparasitic ascomycete Trichoderma atroviride. These data may suggest that deletion of cytosolic ENGaseleads to accumulation of misfolded glycoproteins in the fungal cytosol, which somehow affects its protein secretion/structure of cell wall. This is the firststudy of the ERAD pathway in filamentous ascomycetes.194. Saccharomyces cerevisiae spore development and protection against reactive oxygen species. Steve Gorsich, Tricia Stokes, Michelle Steidemann,Kyle Kern. Dept Biol, Central Michigan Univ, Mt Pleasant, MI.The generation of spores in S. cerevisiae is essential for sexual reproduction and survival of the organism. When diploid S. cerevisiae cells undergomeiotic division to produce four spores it is important for each spore to not only get a haploid copy of nuclear chromosomes, but also a completecomplement of organelles and potentially RNP granules. For instance mitochondria undergo temporally regulated fusion and fission events to assure thatmitochondria are represented equally in each of the resulting spores. When this network is not maintained, due to mutations in mitochondrial fissiongenes (e.g. dnm1/dnm1), it has been shown that fewer spores survive and the ones that do survive have reduced respiratory fitness. In addition tomutations affecting spore production we hypothesized that environmental factors could also influence spore development. In the present study, wedemonstrated that hydrogen peroxide can phenocopy the mitochondrial fission mutant’s phenotypes. Wild-type S. cerevisiae exposed to hydrogenperoxide have mitochondrial morphology and distribution defects during spore development, reduced spore viability, and decreased respiratorycompetency just as seen in dnm1/dnm1 fission mutants. We next hypothesized that the phenotypes associated with dnm1/dnm1 mitochondrial fissionmutants were caused by increased sensitivity to reactive oxygen species (ROS). To support this we demonstrated that dnm1/dnm1 mutants have anincrease in ROS during spore development. In addition, sporulation defects associated with dnm1/dnm1 or wild-type cells exposed to hydrogen peroxidewere rescued when we overexpressed oxidative stress protection genes. These findings suggest that the ability of S. cerevisiae to produce optimalnumbers of fit spores is heavily influenced by their ability to protect themselves from exogenous or endogenous ROS.195. Genetic analysis of the role of peroxisomes in the virulence and survival in Fusarium graminearum. K. Min 1 , H. Son 1 , J. Lee 2 , G. J. Choi 3 , J.-C. Kim 3 , Y.-W. Lee 1 . 1) Department of Agricultural Biotechnology and Center for Fungal Pathogenesis, Seoul National University, Seoul 151-921, Republic of Korea; 2)Department of Applied Biology, Dong-A University, Busan 604-714, Republic of Korea; 3) Eco-friendly New Materials Research Group, Research Center forBiobased Chemistry, Division of Convergence Chemistry, Korea Research Institute of Chemical Technology, Daejeon 305-343, Republic of Korea.Peroxisomes are single-membrane-bound organelles that are required for diverse biochemical processes, including b-oxidation of fatty acids anddetoxification of reactive oxygen species (ROS). In this study, the role of peroxisomes was examined in Fusarium graminearum by functional analysis ofthree genes (PEX5, PEX6, and PEX7) encoding peroxin proteins required for peroxisomal protein import. PEX5 and PEX7 deletion mutants failed to localizethe fluorescently-tagged peroxisomal targeting signal type 1 (PTS1)- and PTS2-containing proteins to peroxisomes, respectively, whereas the PEX6 mutantwere unable to localize both fluorescent proteins. Deletion of PEX5 and PEX6 triggered reduced growth on long chain fatty acids and butyrate, while thePEX7 deletion mutants utilized fatty acids other than butyrate. Virulence on wheat heads was greatly reduced in the PEX5 and PEX6 deletion mutants,because they were impaired in spreading from inoculated florets to the adjacent spikelets through rachis. Disruption of PEX5 and PEX6 droppedsurvivability of aged cells in planta and in vitro due to the accumulation of ROS followed by necrotic cell death. We suggest that PTS1-type peroxisomalcatalases are responsible for ROS scavenging. These results demonstrate the functions of peroxisomes in survival and ROS detoxification of filamentousfungi.196. roGFP and anti-oxidant defences in the rice blast fungus Magnaporthe oryzae. Marketa Samalova, Sarah Gurr, Mark Fricker. Plant Sciences,University of Oxford, Oxford, United Kingdom.The ascomycete fungus Magnaporthe oryzae causes rice blast disease. Germination and development of its infection structure, the appressorium on thehost surface is orchestrated by a complex set of signals from within the fungus, and later between the fungus and resistant or susceptible plant that,respectively, either triggers host defence or leads to infection. Host defences involve localised production of reactive oxygen species (ROS), which eitherkill the pathogen directly or block fungal invasion by oxidative cross-linking of cell wall glycoproteins. By contrast, infection suggests that the invadingfungus can tolerate or, indeed, bypass such defences. Here, we report rice blast fungus’ capacity to withstand transient oxidative stress during earlydevelopment. We determine the intrinsic cytoplasmic cell glutathione (GSH) concentration by confocal imaging of monochlorobimane, which becomesfluorescent when conjugated to GSH. The redox poise of the glutathione pool was measured by 4-D confocal excitation ratio imaging of GRX1-roGFP2. Wereveal that this fungus has an extraordinary ability to tolerate severe insults of H 2O 2, with rapid recovery of its reduced GSH pool and thence continued168

FULL POSTER SESSION ABSTRACTSgrowth. Exploring in vivo responses during infection of susceptible (S) and resistant (R gene) host plants reveals that pathogen penetration andproliferation is hugely restricted in the R plants, but surprisingly, there is no change in the roGFP ratio in planta. Thus the sparse infection hyphae within Rplants maintain a highly reduced cytoplasm at all times. This questions whether production of ROS by the host is the primary mechanism responsible forrestricting pathogen growth in resistant plants.197. Dimorphism and virulence in pathogenic zygomycetes. Soo Chan Lee, Alicia Li, Joseph Heitman. Molec Gen & Microbiol, Duke Med Ctr, Durham, NC.Fungal dimorphism evolved in multiple fungal lineages. Many pathogenic fungi are dimorphic, for example, switching between yeast and filamentousstates. This switch alters host-pathogen interactions and is critical for pathogenicity. However, in pathogenic zygomycetes, whether dimorphismcontributes to pathogenesis is a central unanswered question. The pathogenic zygomycete Mucor circinelloides exhibits multi-budded yeast growth underanaerobic/high CO 2 growth conditions, which Louis Pasteur discovered (Etudes sur la Biere. 1876). Interestingly, we found that in the presence of thecalcineurin inhibitor FK506, Mucor exhibits multi-budded yeast growth. We discovered that Mucor encodes three calcineurin catalytic A subunits (CnaA,CnaB, and CnaC) and one calcineurin regulatory B subunit (CnbR). Disruption of the cnbR gene results in mutants locked in yeast phase growth. Theseresults reveal that the calcineurin pathway governs the dimorphic transition from yeast to hyphae. In virulence tests, we found that the cnbR yeast-lockedmutants are less virulent than wild-type in a heterologous host system, providing evidence that hyphae are a more virulent form of this fungus. Proteinkinase A activity was elevated during yeast growth under anaerobic conditions, in the presence of FK506, or in the yeast-locked cnbR mutants, indicating anovel connection between PKA and calcineurin. The cnaA mutants are hypersensitive to calcineurin inhibitors and display a hyphal polarity defect. Themutants produce spores that are larger than wild-type. Notably, we found spore size is linked to virulence in previous studies (Li et al. PLoS Pathogens.2011). Interestingly, the cnaA mutants were found to be more virulent than wild-type. We also observed that the cnaA mutants germinate earlier insidemacrophages, providing a possible explanation for the greater virulence of the mutants. Another pathogenic zygomycete, Rhizopus delemar has three cnagenes. Phylogenetic analysis revealed that the triplicated cna genes might result from a whole genome and/or segmental gene duplications. Our resultsdemonstrate that the calcineurin pathway orchestrates the dimorphic transition, spore size dimorphism, virulence, and hyphal polarity in Mucor, and thecalcineurin pathway elements have been adapted in zygomycetes via variation in their evolutionary trajectory.198. Genetic analysis of the components of the ime-2 mediated signaling events during nonself recognition and programmed cell death (PCD) inNeurospora crassa. Joanna A. Bueche 1 , Elizabeth A. Hutchison1 1,2 , N. Louise Glass 1 . 1) Plant and Microbial Biology, UC Berkeley, Berekeley, CA, 94720; 2)Cornell University Microbiology Department, Ithaca, NY 14853.Recently, we revealed genetic and functional differences in meiotic initiation machinery between Neurospora crassa and Saccharomyces cerevisiae.While N. crassa is missing some meiotic genes identified in yeast, it has three homologs of the middle meiotic transcriptional regulator, Ndt80. None of theNDT80 homologs are required for meiosis in N. crassa. One of the NDT80 homologs, vib-1 is essential for heterokaryon incompatibility (HI) in N. crassa, anonself recognition mechanism in filamentous fungi. Mutations in vib-1 suppress cell death caused by HI as well as secretion of the extracellular proteasesduring the nitrogen starvation. Furthermore, deletion of a IME2 (a kinase involved in initiation of meiosis in S. cerevisiae) homolog in N. crassa, ime-2, doesnot affect sexual development, results in a significant elevation of secreted proteases in response to nitrogen starvation. Morever, a Dvib-1 Dime-2 mutantrestored wild-type levels of cell death during the HI and normal production of extracellular proteases; a deletion of ime-2 suppressed these vib-1phenotypes. Based on the evidence, we hypothesize that IME-2 negatively regulates a cell death pathway that functions in parallel to the VIB-1 HI pathwayand a protease secretion pathway positively regulated by VIB-1. We used a slightly modified yeast consensus sequence for Ime2 phosphorylation to scan(Scansite) the entire N. crassa genome for possible targets and obtained a list of 30 candidates including VIB-1. All targets were assessed for secretion ofthe extracellular proteases in absence of nitrogen. Strains containing deletions of 13 of the 30 genes identified in the screen were significantly affected inprotease secretion. Mutations in these candidate genes will be tested for the ability to alleviate cell death and Heterokaryon Inocpatibilty (HI) in thepresence and absence of ime-2 and vib-1 hence assessing their role in the parallel HI/PCD pathway redundant with VIB-1.199. PRO45 is a component of the conserved STRIPAK complex in Sordaria macrospora. Steffen Nordzieke 1 , Benjamin Fränzel 2 , Sandra Bloemendal 1 , DirkWolters 2 , Ines Teichert 1 , Ulrich Kück 1 . 1) General and Molecular Botany, Ruhr-University Bochum; 2) Analytical Chemistry, Ruhr-University Bochum,Universitätsstr. 150, 44801 Bochum, Germany.The complex formation of three-dimensional fruiting bodies in Sordaria macrospora is mediated by an interaction between developmental proteins andconserved signaling cascades and thus an excellent experimental system for developmental biology.We recently have characterized a STRIPAK complex in Sordaria macrospora that is involved in the regulation of fruiting body development. This complexcontains striatin (PRO11), a striatin-interacting protein (PRO22), the scaffolding subunit of protein phosphatase 2A (SmPP2AA) and a phocein homologue(SmMOB3) [1, 2].Here we describe PRO45, a novel subunit of the STRIPAK complex in filamentous fungi which is a homolog of the human sarcolemmal membraneassociated protein (SLMAP). We also present the functional characterization of PRO45: Strains lacking the gene for PRO45 show sterility together with asevere defect in hyphal fusion and vegetative growth rate. The primary structure of PRO45 contains a forkhead-associated (FHA) as well as atransmembrane domain. Complementation studies showed that a lack of the FHA domain is responsible for the described defects, whereas a missingtransmembrane domain does not affect development.Tandem affinity purification (TAP) followed by mass spectrometry and coimmunoprecipitation (Co-IP) showed subunits of the STRIPAK-complex asinteraction partners, confirming the homology of human and fungal STRIPAK. Further microscopic studies provide evidence for a localization of PRO45 inthe ER as well as in the nuclear envelope. Integrating these observations, we propose that PRO45 has a function in the physical and signaling connection ofSTRIPAK-complex and nucleus.[1] Pöggeler S, Kück U 2004. Eukaryot. Cell 3: 232-240[2] Bloemendal S, Lord KM, Rech C, Hoff B, Engh I, Read ND, Kück U. 2010. Eukaryot. Cell 9: 1856-1866[3] Bloemendal S, Bernhards Y, Bartho K, Dettmann A, Voigt O, Teichert I, Seiler S, Wolters DA, Pöggeler S, Kück U. 2012. Mol. Microbiol. 84: 310-323.200. Molecular Determinants of Sporulation in Ashbya gossypii. Jurgen W. Wendland, Lisa Wasserstroem, Klaus Lengeler, Andrea Walther. YeastGenetics, Carlsberg Laboratory, Copenhagen V, Kopenhagen V, Denmark.Previously we have analysed the pheromone response MAPK signal transduction cascade in A. gossypii. The major findings were (i) deletion of bothpheromone receptor genes STE2 and STE3 did not inhibit sporulation whereas (ii) deletion of the transcription factor STE12 resulted in hypersporulation(Wendland et al. 2011). Here we present our analysis of key A. gossypii homologs of Saccharomyces cerevisiae sporulation specific genes. We show that27th Fungal Genetics Conference | 169

FULL POSTER SESSION ABSTRACTSmutants in IME1, IME2, KAR4, and NDT80 are blocked in sporulation. Mutants in IME4, KAR4, and UME6 also confer a vegetative growth defect. IME4expression was found during vegetative growth while IME2 was not detected under these conditions. We performed transcriptional profiling of nonsporulatingstrains and determined a core set of about 50 down-regulated sporulation specific genes in these mutants. Interestingly, this set of downregulatedgenes is upregulated in the A. gossypii ste12 mutant providing regulatory evidence of the hypersporulation phenotype of this mutant. Othergenes identified in the RNAseq data indicated that during development of sporangia metabolic genes for nutrient uptake are active. Therefore weperformed Return-To-Growth assays with mutants inhibited in the sporulation pathway. These strains were kept under conditions in which the wild typeinitiates sporulation. This lead to induction of sporangium formation, a stage at which these strains remained. Supply of new nutrients resulted in hyphaloutgrowth in all mutants indicating that after initiation of the sporulation program A. gossypii can reverted to vegetative growth at different stages. Inaddition we identified differential regulation of two endoglucanases encoded by ENG1 and ENG2. While ENG1 was not differentially regulated, ENG2 wasdown-regulated in e.g. ime1 but strongly up-regulated in ste12. Deletion analysis of ENG2 showed that Eng2 is required for hyphal fragmentation intoindividual sporangia. We can thus provide a detailed overview of the genetic regulation of sporulation in A. gossypii. A comparison with S. cerevisiaehighlights the role of KAR4 in sporulation upstream of IME1. Finally, our study provides further evidence that the pheromone signaling response MAPKcascadein A. gossypii has a regulatory control function over sporulation alongside regulation of sporulation by nutritional cues.201. VELVET is regulated by ENV1 and impacts development of Trichoderma reesei. Hoda Bazafkan 1 , Doris Tisch 2 , Monika Schmoll 1 . 1) Health &Environment - Bioresources, AIT Austrian Institute of Technology GmbH, Tulln, Austria; 2) Vienna University of Technology, Institute of ChemicalEngineering, Vienna.In Trichoderma reesei (teleomorph Hypocrea jecorina), light is a crucial environmental factor for initiation of sexual development and considerablyinfluences expression of glycoside hydrolases. In both processes, the light regulatory protein ENV1 plays a key role. Transcriptome analysis revealed thatvel1 (encoding the VELVET orthologue) transcription is regulated by the carbon source in the medium. Moreover, ENV1 negatively regulates vel1 in light.Genes coregulated with vel1 are enriched in functions of amino acid metabolism as well as carbon metabolism and include three non ribosomal peptidesynthases (NRPS). This regulatory pattern supports a connection of vel1 with primary and secondary metabolism also in T. reesei. VELVET is known to be aregulator of sexual and asexual development in fungi. Also for T. reesei a function in development was likely, as several genes involved in sexualdevelopment are coregulated with vel1. Investigation of strains lacking the T. reesei orthologue vel1 under various nutritional, light- and temperatureconditions showed that VEL1 is essential for conidiation and growth of aerial hyphae. Moreover, in crossing assays, sexual development with strainslacking vel1 was delayed and in contrast to the wild-type never occurred in constant darkness. When vel1 was missing in both mating partners, no fruitingbodies were formed. Although male fertility was intact, female fertility was found to be dependent on the presence of vel1. Strains lacking the lightregulator gene env1 are able to undergo sexual development with wild-type strains, but in crosses of two strains lacking env1, no fruiting bodies areformed in light. This defect is assumed to be caused by a deregulation of the pheromone system in these strains. Interestingly, also strains lacking vel1 areunable to mate with env1 deletion mutants in light. Together with the regulatory connection between these genes, these findings support a function ofvel1 in the same pathway as env1. We conclude that VEL1 in T. reesei regulates sexual and asexual development and is connected to the light responsepathway via ENV1.202. Sexual reproduction and mating type function in the penicillin producing fungus Penicillium chrysogenum. Julia Böhm 1 , Birgit Hoff 1 , Simon Wolfers 1 ,Céline O'Gorman 2 , Paul Dyer 2 , Stefanie Pöggeler 3 , Ulrich Kück 1 . 1) Christian Doppler Laboratory for Fungal Biotechnology, Ruhr-Universität Bochum,Universitätsstr. 150, 44780 Bochum, Deutschland; 2) School of Biology, University of Nottingham, University Park, Nottingham, NG7 2RD, U.K; 3) AbteilungGenetik eukaryotischer Mikroorganismen, Institut für Mikrobiologie und Genetik, Georg-August Universität Göttingen, 37077 Göttingen, Deutschland.Penicillium chrysogenum is a filamentous fungus of major medical and historical importance, being the original and present day industrial source of theantibiotic penicillin with a world market value of about 600 million € per year. The species has been considered asexual for over 100 years and despiteconcerted efforts it has not been possible to induce sexual reproduction. However, we recently were able to detect mating type loci in different strains,indicating a sexual lifecycle. Isolates, carrying opposite mating types, were found in near-equal proportion in nature and we observed transcriptionalexpression of mating type loci as well as pheromone and pheromone receptor genes [1]. Utilising knowledge of mating-type (MAT) gene organization wenow describe conditions under which a sexual cycle can be induced leading to the production of cleistothecia and meiotic ascospores, which were similarto those described recently for Eupenicillium crustaceum [2]. Evidence of recombination was obtained using both molecular and phenotypic markers. Thenewly identified heterothallic sexual cycle was used for strain development purposes, generating offspring with novel combinations of traits relevant topenicillin production.Furthermore, the MAT1-1-1 mating-type gene, known primarily for a role in governing sexual identity, was also found to control transcription of a widerange of genes including those regulating penicillin production, hyphal morphology and conidial formation, all traits of biotechnological relevance. Forfunctional characterization MAT1-1-1 knockout and overexpression strains were generated and analyzed. These discoveries of a sexual cycle and MATgene function are likely to be of broad relevance for manipulation of other asexual fungi of economic importance.[1] Hoff B, Pöggeler S, Kück U (2008) Eighty years after its discovery, Fleming`s Penicillium strain discloses the secret of its sex. Eukaryot Cell 7: 465-470[2] Pöggeler S, O'Gorman CM, Hoff B, Kück U (2011) Molecular organization of the mating-type loci in the homothallic ascomycete Eupenicilliumcrustaceum. Fungal Biol. 115: 615-624.203. Exponentiate complexity: non-mating GPCRs in the basidiomycete Schizophyllum commune. Daniela Freihorst, Susann Erdmann, Erika Kothe.Institute for Microbiology, Dept. Microbial Communication, Friedrich Schiller University, Jena, Germany.The filamentous fungus S. commune is a model organism for sexual development of basidiomycetes. Numerous studies revealed the importance of twogene loci, A and B, responsible for tetrapolar mating and sexual development. Both occur in multiallelic subloci leading to a large number of differentspecificities in nature (9 to 32 depending on locus), which then control compatibility or abortion of mating. While A codes for homeodomain transcriptionfactors, B codes for a pheromone/receptor system. The B-receptors (Ste3-like, seven transmembrane domains, G-protein coupled) recognize pheromonesof non-self specificity and induce signal transduction pathways and specific gene regulation. After sequencing of strain H4-8 four new Ste3-like GPCRs,homologous to the known B-specific ones, were found. Three of the four B-receptor like genes (brls) are located close to the B-locus. Their function isunknown, because a B-locus defective strain without any interactions seen in B-dependent development still contains those four GPCRs, which obviouslydo not respond to any wild type pheromone. However, our results indicate their importance since sequence identity - analyzed by PCR and sequencing -between unrelated strains was found arguing for conservation of these genes. Gene expression was first observed by Reverse Transcriptase PCR as well asMicroarray analyses, which disproved the theory that brls are pseudo genes. Expression was then investigated by quantitative Real Time PCR duringmating interaction and in monokaryotic strains, which showed comparable results only between gene brl4 and the true mating receptor bar2. Also RNA170

FULL POSTER SESSION ABSTRACTSSeq data did not substantiate a mating dependent expression for those genes. To get more insights into the function an over expression of the gene brl2under control of tef1-promoter was performed. Phenotypes of independent mutants showed no hints for a faster mating, changes in clamp formation ornuclear distribution tested by mating experiments. Only an asymmetric distribution of fruiting bodies was visible, which seems to originate from thedifferent protein background respectively epigenetics of the two partners just before mating interaction and dikaryotization. Tagging of the receptors forvisualization is planned, which will lead to more knowledge about localization and putative interacting proteins.204. Characterization of new STRIPAK complex interaction partners in the filamentous ascomycete Sordaria macrospora. Britta Herzog, YasmineBernhards, Berit Habing, Eva Reschka, Sabine Riedel, Stefanie Pöggeler. Institute of Microbiology and Genetics, Department of Genetics of EurkaryoticMicroorganisms, Georg-August-University Göttingen, Germany.Using Sordaria macrospora as model organism we investigate the complex process of fruiting-body development and involved proteins in thisfilamentous ascomycete. This differentiation process is regulated by more than 100 developmental genes. Recently, we have shown that a homologue ofthe human STRIPAK (striatin-interacting phosphatase and kinase) complex engages a crucial role in sexual development in fungi. The S. macrospora striatinhomologue PRO11 and its interaction partner SmMOB3 are key components of this complex (Bloemendal et al., 2012). PRO11 contains a conserved WD40repeat domain and is supposed to function as scaffolding protein linking signaling and eukaryotic endocytosis (Pöggeler and Kück, 2004). SmMOB3(phocein) is a member of the MOB family (Bernhards and Pöggeler, 2011). Beside their important role in multicellular development and hyphal fusion bothproteins seem to be involved in vesicular trafficking and endocytosis.By means of yeast two-hybrid screens and GFP-Trap analysis we identified several new interaction partners of PRO11 and SmMOB3. Similar to PRO11and SmMOB3, a multitude of them are predicted to be involved in vesicular trafficking and are localized to the ER or to the Golgi. Here, we show theresults of a detailed analysis of the new STRIPAK complex interaction partners. Initially, we isolated the cDNA of the genes and confirmed the interactionby yeast two-hybrid. For further characterization and to get knowledge about their cellular functions we created knock-out strains and analyzed theirmorphological phenotypes. For localization and expression studies we constructed EGFP-tagged fusion proteins and expressed them in S. macrospora.Bernhards and Pöggeler, 2011; Curr Genet 57 (2): 133-49.Bloemendal et al., 2012; Mol Microbiol 84 (2): 310-23.Pöggeler and Kück, 2004; Eukaryot Cell 3 (1): 232-40.205. Hypocrea jecorina meiosis generates segmentally aneuploid progeny to enhance production of xylan-degrading hemicellulases. T.-F Wang, C.LChen, P. W.-C. Hsu, W.-C. Li, S.-Y. Tung, C.-L. Wang, H.-C. Kuo. Institute of Molecular Biology, Academia Sinica, Taipei, TaiwanI.Hypocrea jecorina is the sexual form of Trichoderma reesei, an industrially important cellulolytic filamentous fungus. We report that H. jecorina meiosisutilizes a novel Ku70-dependent duplication mechanism to generate segmentally aneuploid progenies, thus increasing the diversity of genotypes andensuring more efficient xylan degradation. H. jecorina sexual reproduction yields hexadecad asci with 16 linearly arranged ascospores. Our results indicatethat these ascospores are generated via two rounds of postmeiotic mitosis following the two meiotic divisions. Remarkably, the hexadecad asci frequently(>90%) contain four or eight inviable ascospores with an equal number of viable segmentally aneuploid ascospores. Array-based comparative genomichybridization revealed that all the viable segmental aneuploid progenies have a large chromosomal duplication (~0.5Mbp). Deletion of thenonhomologous end-joining gene ku70 restores canonical meiosis and 16 viable euploid ascospores. Segmental duplication contains genes involved inxylan degradation and enhances expression of several carbohydrate-active enzymes, particularly cell wall degrading hemicellulases.206. Deletion of MAT 1-2-1 gene results in mating type switching in Ceratocystis fimbriata. P. Markus Wilken 1 , Emma T. Steenkamp 2 , Mike J. Wingfied 1 , Z.Wilhelm de Beer 2 , Brenda D. Wingfield 1 . 1) Dept Genetics, University of Pretoria, Pretoria, Gauteng, South Africa; 2) Dept Microbiology and PlantPathology, University of Pretoria, Pretoria, Gauteng, South Africa.Sequencing of the Ceratocystis fimbriata genome has made it possible to consider the long standing question as to how uni-directional mating typeswitching functions in this fungal pathogen and its relatives. Uni-directional mating type switching was first observed in the homothallic ascomycete C.fimbriata in the 1960’s. Two forms of progeny arise after meiosis, some self-fertile and thus not requiring an opposite mating partner to complete thesexual cycle. Other isolates are self-sterile and unable to reproduce sexually. This loss of self-fertility has been shown to be associated with the loss of afragment of the mating specific gene, MAT1-2-1, in self-sterile strains. The aim of this study was interrogate the full genome sequence of C. fimbriata todetermine whether the full MAT1-2-1 gene is deleted and whether other MAT genes are affected during mating type switching. We were able todetermine that C. fimbriata has both the MAT1-2-1 gene and the MAT1-1 genes (MAT1-1-1 and MAT1-1-2). The self-sterile isolates had only lost theMAT1-2-1 gene and one copy of a 230 base pair perfect repeat which flanks this gene in the self fertile isolates. The loss of the entire MAT1-2-1 geneexplains the loss of fertility and the repeats are suggestive of the involvement of recombination during the deletion event. This study illustrates a uniquemating strategy in the fungi, not previously understood at the molecular level. The newly gained knowledge will also make it possible to consider themechanisms underpinning uni-directional switching in other species of Ceratocystis.207. Mannitol is essential for the development of stress resistant ascospores in Neosartorya fischeri. Timon T. Wyatt 1 , M.R. van Leeuwen 1 , H.A.B.Wösten 2 , J. Dijksterhuis 1 . 1) Applied and Industrial Mycology, CBS-KNAW Fungal Biodiversity Centre, Utrecht, the Netherlands; 2) Microbiology, UtrechtUniveristy, Utrecht, the Netherlands.The sugar alcohol mannitol is one of the main compatible solutes in Neosartorya fischeri and accumulates especially in conidia and ascospores. In fungi,mannitol has been implicated in a wide variety of functions including carbon storage, maintaining reduction potential, water absorption, heat stressprotection, protection against oxidative stress, and tolerance against osmotic stress. Biosynthesis of mannitol in ascomycota mainly depends on mannitol1-phosphate dehydrogenase (MPD). In our study a functional analysis was performed of the MPD encoding gene mpdA of N. fischeri. The fluorescenceproteins GFP and dTomato were put under control of the mpdA promoter. Expression of mpdA was observed in aerial hyphae and conidiophores, but wasespecially high in ascomata and ascospores. Disruption of mpdA reduced mannitol as much as 85% of the wild type and increased trehalose levels to morethan 400%. Decreased mannitol accumulation had no obvious effect on mycelium growth when exposed to temperature and oxidative stress, while anincreased stress sensitivity of conidia against heat and oxidative stress was observed. The most distinct phenotype of mpdA disruption was the completeabsence of ascospores. Formation of fruiting bodies (ascomata) and asci was not affected but the developmental defect was shown to occur after meiosis.Similar results were obtained by adding the MPD inhibitor nitrophenide to the wild-type strain. Our result suggest a role of mannitol as carbon storagemolecule during sexual development, but also its role as scavenger of hydroxyl radicals can be of importance for the formation of sexual spores. Mannitolmight regulate the Reactive Oxygen Species (ROS) levels induced by Nox (NADPH oxidases) family enzymes during sexual development. Taken together,27th Fungal Genetics Conference | 171

FULL POSTER SESSION ABSTRACTSthese results show a novel function for mannitol in fungal growth and sexual development.208. A small lipopeptide pheromone with limited proline substitutions can still be active. Thomas J. Fowler, Stephanie L. Link. Department of BiologicalSciences, Southern Illinois University Edwardsville, Edwardsville, IL.Mating in many fungi involves communication with lipopeptide pheromones. These signaling molecules activate G protein-coupled receptors located onthe surface of a compatible mating partner and initiate a mating response. Some of the mushroom fungi code for scores of different lipopeptidepheromones among the mating types. Despite their small predicted size of approximately eleven amino acids, these pheromones have very specificpheromone receptor targets for mate discrimination. In past heterologous expression and mating studies in Saccharomyces cerevisiae, we have maderandom amino acid substitutions in one pheromone, Bbp2(4), from the mushroom fungus Schizophyllum commune, and site-directed mutations in aclosely related pheromone, Bbp2(7). These studies indicated that the peptide portion of the pheromones can be more variable than expected. Within arandom mutagenesis study of Bbp2(4), it was noted that the imino acid proline could be substituted for several of the natural residues and an activemutant pheromone was still produced. In this study, the heterologous mating assay was employed to test the extent that proline residues might besubstituted into a pheromone before activity was no longer detected. Mature Bbp2(4) is predicted to be eleven amino acids with a farnesyl tail(DSPDGYFGGYC-farnesyl). Single substitutions of proline at several non-natural positions did not stop production of active pheromone, but substitutionswith proline at several previously identified critical amino acid positions led to negative results in the mating assays. Among the substitutions that do notdisrupt all activity are DSPPGYFGGYC-farnesyl and DSPDGYFGPYC-farnesyl. The three-dimensional conformations of proline-substituted peptides insolution were predicted with PEP-FOLD and viewed with JMOL. The conformational differences of small pheromones tolerated by one receptor aresurprising. Substitution of two or more prolines at adjacent non-natural positions in a single pheromone does inhibit production of an active pheromone inthe heterologous mating assay. At present, it cannot be determined if multiple proline substitutions inhibit pheromone processing, pheromone transport,or interaction with the receptor.209. Function of Ras proteins in fungal morphogenesis of Schizophyllum commune. E.-M. Jung, N. Knabe, E. Kothe. Department of Microbiology, FriedrichSchiller University, Jena, Germany.The white rot basidiomycete Schizophyllum commune has been used as a model organism to study mating and sexual development as well as analysis ofcell development. Subsequent to nutrient and pheromone recognition, intracellular signal transduction was regulated by different pathways and MAPKsignalling cascades. The S. commune genome encodes more than 30 putative signal transduction proteins of the Ras superfamily containing the Ras, Rho,Rab, Ran and Arf subfamilies. Phylogenetic investigation of Ras proteins from various basidiomycetes show that they cluster in two main groups. Highsequence similarities between these proteins in basidiomycetes suggesting an ancient duplication event. To investigate the function of the small G-proteins Ras1 and Ras2 mutants with constitutively active ras1 alleles as well as a DRasGap1 mutant were analyzed. They show phenotypes withdisorientated growth pattern, reduced growth rates and hyperbranching effects. The fungal cytoskeleton, composed of actin and microtubules has beeninvestigated by immunofluorescence microscopy to reveal whether Ras signaling influences the formation of cytoskeleton. The second Ras protein, Ras2,was detected by genome analysis. Its function is analysed in current studies.210. The developmental PRO40/SOFT protein participates in signaling via the MIK1/MEK1/MAK1 module in Sordaria macrospora. Ines Teichert 1 , EvaSteffens 1 , Nicole Schnab 1 , Benjamin Fränzel 2 , Christoph Krisp 2 , Dirk A. Wolters 2 , Ulrich Kück 1 . 1) General & Molecular Botany, Ruhr University Bochum,Bochum, Germany; 2) Analytical Chemistry, Ruhr University Bochum, Bochum, Germany.Filamentous fungi are able to differentiate multicellular structures like conidiophores and fruiting bodies. Using the homothallic ascomycete Sordariamacrospora as a model system, we have identified a number of developmental proteins essential for perithecium formation. One is PRO40 [1], thehomolog of Neurospora crassa SOFT, and this protein was employed for protein-protein interaction studies to gain insights into its molecular function.Data from yeast two hybrid experiments with PRO40 as bait show an interaction of PRO40 with the MAP kinase kinase (MAPKK) MEK1. MEK1 is a memberof the cell wall integrity (CWI) pathway, one of three MAP kinase modules present in S. macrospora. The S. macrospora CWI pathway consists of MAPkinase kinase kinase (MAPKKK) MIK1, MAPKK MEK1 and MAP kinase (MAPK) MAK1, with additional upstream components, protein kinase C (PKC1) andRHO GTPase RHO1. Data from tandem affinity purification - MS experiments with PRO40 and MEK1 as bait indicate that PRO40 forms a complex withcomponents of the CWI pathway. Analysis of single and double knockout mutants shows that PRO40, MIK1, MEK1 and MAK1 are involved in the transitionfrom protoperithecia to perithecia, hyphal fusion, vegetative growth, and cell wall stress response. Differential phosphorylation of MAPKs in a pro40knockout strain was detected by Western analysis. We propose that PRO40 modulates signaling through the CWI module in a development-dependentmanner. Further interaction studies and complementation analyses with PRO40 derivatives provide mechanistic insight into the function of PRO40domains during fungal development. [1] Engh et al. (2007) Eukaryot Cell 6:831-843.211. Map-based identification of the mad photosensing genes of Phycomyces blakesleeanus. Silvia Polaino Orts 1 , Suman Chaudhary 1 , Viplendra Shakya 1 ,Alejandro Miralles-Durán 2 , Luis Corrochano 2 , Alexander Idnurm 1 . 1) Cell Biology & Biophysics, University of Missouri-KC, Kansas City, MO; 2) Departamentode Genética, Universidad de Sevilla, Spain.Phycomyces blakesleeanus is a filamentous fungus, a member of the subphylum Mucoromycotina. The main reason for the presence of Phycomyces inlaboratories is its sensitivity to light. The fruiting bodies phototropism of Phycomyces has served as a model of response to blue light in fungi. In 1967, inthe laboratory of Nobel laureate Max Delbrück, the first sensory mutants were isolated. Analysis on these strains has enabled a proposed sensorytransduction pathway that describes the flow of information from the sensors to the effectors. There are ten mutants, called mad mutants, divided intotwo classes: those of type 1 are madA, madB, madC and madI, which are altered only in photoresponses but not in others tropisms of the sporangiophore.The mutants in the madA and madB genes are altered in all photoresponses (phototropism, photomorphogenesis, photocarotenogenesis andphotomecism). These two mad genes are the only ones that have been identified and their corresponding proteins interact to form the Mad complex, themain photoreceptor complex of Phycomyces. The mutants altered in the madC gene are only affected in the phototropism. The remaining mad mutantsare called type 2 and are altered in the phototropism and other responses of the sporangiophore, like gravitropism and avoidance. Phycomyces cannot bestability transformed with DNA. To identify the eight unknown mad mutants, a positional cloning approach was taken coupled to Illumina sequenceinformation. A genetic map was constructed between two wild type parents, and then mad mutants crossed to one of these parents. Through mapping,we have identified candidates for the madC, madD, madJ, madF and madI genes, with greatest follow up characterization in madC. The madC geneencodes a Ras GTPase-activating protein, implicating Ras in the light signal transduction pathway in fungi.172

FULL POSTER SESSION ABSTRACTS212. The C 2H 2 transcription factor HgrA promotes hyphal growth in the dimorphic pathogen Penicillium marneffei. Hayley E. Bugeja, Michael J. Hynes,Alex Andrianopoulos. Department of Genetics, University of Melbourne, Parkville, VIC, Australia.Penicillium marneffei (recently renamed Talaromyces marneffei) is well placed as a model experimental system for investigating fungal growth processesand their contribution to pathogenicity. An opportunistic pathogen of humans, P. marneffei is a dimorphic fungus that displays multicellular hyphal growthand asexual development (conidiation) in the environment at 25°C and unicellular fission yeast growth in macrophages at 37°C. We have characterised thetranscription factor hgrA (hyphal growth regulator), which contains a C 2H 2 DNA binding domain closely related to that of the stress-response regulatorsMsn2/4 of Saccharomyces cerevisiae. HgrA is not required for controlling yeast growth in response to the host environment, nor does it appear to have akey role in response to stress agents, but is both necessary and sufficient to drive the hyphal growth program. hgrA expression is specific to hyphal growthand its deletion affects multiple aspects of hyphal morphogenesis and the dimorphic transition from yeast cells to hyphae. Loss of HgrA also causes cellwall defects, reduced expression of cell wall biosynthetic enzymes and increased sensitivity to cell wall, oxidative, but not osmotic stress agents. As well ascausing apical hyperbranching during hyphal growth, overexpression of hgrA prevents conidiation and yeast growth, even in the presence of inductivecues. HgrA is a strong inducer of hyphal growth and its activity must be appropriately regulated to allow alternative developmental programs to occur inthis dimorphic pathogen.213. Involvement of a specific ubiquitin ligase in the assembly of the dynein motor. Ryan Elsenpeter, Robert Schnittker, Michael Plamann. Sch BiologicalSci, Univ Missouri, Kansas City, Kansas City, MO.Cytoplasmic dynein is a large, microtubule-associated motor complex that facilitates minus-end-directed transport of various cargoes. The dynein heavychain (DHC) is >4000 residues in length, with the last two-thirds of the heavy chain forming the motor head. Six domains within the dynein motor exhibitvarying degrees of homology to the AAA+ superfamily of ATPases. These domains form a ring-like structure from which a microtubule-binding domainprotrudes. Using a genetic assay, we have isolated over 30 DHC mutants of Neurospora that produce full-length proteins that are defective in function. Toexplore the mechanism by which mutations in the C-terminal region of the DHC affect function, we have identified both intragenic and extragenicsuppressors. Interestingly, analysis of the extragenic suppressors revealed that loss of function for a putative E3 ubiquitin ligase restored dynein function ina select set of C-terminal DHC mutants. Our results suggest that these C-terminal DHC mutations block assembly of the dynein motor and loss of activity ofa specific E3 ubiquitin ligase restores dynein assembly.214. Identification and characterization of new alleles required for microtubule-based transport of nuclei, endosomes, and peroxisomes. K. Tan, A. J.Roberts, M. Chonofsky, M. J. Egan, S. L. Reck-Peterson. Dept Cell Biology, Harvard Medical School, Boston, MA.Eukaryotic cells use the microtubule-based molecular motors dynein and kinesin to transport a wide variety of cargos. Cytoplasmic dynein is responsiblefor minus-end-directed microtubule transport (from the cell periphery towards the nucleus), while kinesins-1, -2 and -3 move cytoplasmic cargo in theopposite direction. While much is known about how these motors work in vitro, many questions regarding the mechanism and regulation of microtubulebasedcargo transport in cells remain. To identify novel alleles and genes required for microtubule-based transport, we have performed a genetic screen inthe filamentous fungus, Aspergillus nidulans. We fluorescently-labeled three different organelle populations that are known to be cargo of dynein andkinesin in Aspergillus: nuclei, endosomes, and peroxisomes. After mutagenesis we used a fluorescence microscopy-based screen to identify mutants withdefects in the distribution or motility of these organelles. Here, we report the identification and characterization of new alleles of kinesin, dynein and thedynein regulatory factors, Lis1 and Arp1 (a component of the dynactin complex). In vivo analysis of two new dynein alleles revealed that mutations in twoof dynein’s nucleotide binding sites (termed AAA1 and AAA3), led to the accumulation of endosomes and peroxisomes at the hyphal tip, with more subtledefects on nuclear distribution compared to dynein null alleles. In vitro studies of the AAA3 motor mutation showed dramatic reduction in velocity andprolonged binding to the microtubules in single molecule motility assays.215. Pheromone-induced G2 cell cycle arrest in Ustilago maydis requires inhibitory phosphorylation of Cdk1. Sónia M. Castanheira, José Perez-Martín.Centro Nacional de Biotecnología. CSIC. Darwin 3, Campus de Cantoblanco, 28049 Madrid, Spain.Ustilago maydis is a dimorphic basidiomycete that infects maize. In this fungus virulence and sexual development are intricately interconnected.Induction of pathogenicity program requires that two haploid compatible cells fuse and form an infective filament after pheromone signaling. Thepheromone signal is transmitted by a well-known MAPK cascade. Interestingly, Saccharomyces cerevisiae and Ustilago maydis use a similar MAPK cascadeto respond to sexual pheromone and in both cases a morphogenetic response is provided (shmoo and conjugative hypha, respectively). However, while S.cerevisiae arrests its cell cycle in G1 in response to pheromone, U. maydis does this by arresting at G2. The mechanisms and physiological reasons involvedin the distinct cell cycle response to pheromone in U. maydis are largely unknown. In this communication we will introduce our attempts to characterizethe molecular mechanisms behind pheromone-induced cell cycle arrest in U. maydis .Our results have indicated that inhibitory phosphorylation of Cdk1 ispart of the mechanism of the pheromone-induced G2 cell cycle arrest. This inhibitory phosphorylation depends on the essential kinase Wee1. We analyzedthe transcriptional pattern of cell cycle related genes in response to overactivation of pheromone pathway (using a constitutively activated allele of fuz7,the MAPKK of the cascade) and found that two main G2/M regulators -Hsl1, a kinase involved in downregulation of Wee1 and Clb2, the mitotic cyclinweredownregulated at transcriptional level. Using chimeric promoter fusions we found that transcriptional downregulation was not as important forpheromone-induced cell cycle arrest as expected and we are analyzing other possible regulatory options such as stability or subcellular localization ofthese regulators.216. Microtubule-dependent mRNA transport and mitochondrial protein import in Ustilago maydis. T. Langner 1 , T. Pohlmann 1 , C. Haar 1 , J. Koepke 2 , V.Goehre 1 , M. Feldbruegge 1 . 1) Institute for Microbiology, Heinrich-Heine University, Duesseldorf, Northrhine-Westfalia, Germany; 2) MARA, Philipps-University, Marburg, Hesse, Germany.Transport, subcellular localization, and local translation of mRNAs constitute a very important mechanism to ensure correct targeting of proteins todistinct subcellular domains. Although mRNA transport is well studied in various organisms, its function in regulating specific cellular processes likemitochondrial protein import is still ambiguous. We use the corn pathogen Ustilago maydis as a model system to study microtubule-dependent mRNAtransport during formation of infectious filaments. The key RNA-binding protein Rrm4 is an integral part of this long-distance transport machinery.Combining proteomics, in vivo UV cross-linking, and biochemical approaches, we uncovered that Rrm4 plays a crucial role in active transport of mRNAsencoding mitochondrial proteins. In Rrm4 loss-of-function mutants, mitochondrial proteins are altered in expression and localization, which correlateswith impaired production of reactive oxygen species (ROS). We propose that microtubule-dependent mRNA transport and local translation are crucial forcorrect import of mitochondrial proteins. This work is funded by iGRAD-plant graduate school (German research council, DFG/ GRK1525).27th Fungal Genetics Conference | 173

FULL POSTER SESSION ABSTRACTS217. “The vacuole” of Neurospora crassa may be composed of multiple compartments with different structures and functions. Barry J. Bowman 1 , EmmaJean Bowman 1 , Robert Schnittker 2 , Michael Plamann 2 . 1) MCD Biology, University of California, Santa Cruz, CA; 2) Department of Biology, University ofMissouri, Kansas City, KA.The structure of the “vacuole” in Neurospora crassa and other filamentous fungi is highly variable with cell type and position in the hypha. Largespherical vacuoles are typically observed in older hyphal compartments, but approximately 100 microns behind the hyphal tip, vacuolar markers are seenin a dynamic network of thin tubules. At the edge of this network nearest the tip, a few distinct round organelles of relatively uniform size (2-3 microns)have been observed (Bowman et al. Eukaryotic Cell 10:654 ). The function of these round organelles is unknown, although the vacuolar ATPase and avacuolar calcium transporter are strongly localized there. To help identify organelles we have tagged SNARE proteins and Rab GTPases with GFP and RFP.Several of these tagged proteins (sec-22, rab-7, rab-8) appear in the tubular vacuolar network and in the membrane of the round organelles. A uniqueaspect of the round organelles is their association with dynein and dynactin (Sivagurunathan et al. Cytoskeleton, 69:613). In strains with mutations in thetail domain of the dynein heavy chain the dynein is often seen in clumps. This aggregated dynein appears to be tightly associated with (and possibly inside)the round organelles, but not in the tubular vacuolar network. Further analysis of the location of SNARE and Rab proteins may help to identify the functionof the round organelles.218. Comparisons of two wild type A mating type loci and derived self-compatible mutants in the basidiomycete Coprinopsis cinerea. Yidong Yu, MonicaNavarro-Gonzaléz, Ursula Kües. Molecular Wood Biotechnology + Technical Mycology, University of Goettingen, Goettingen, Germany.The A mating type locus in Coprinopsis cinerea controls defined steps in the formation of a dikaryotic mycelium after mating of two compatiblemonokaryons as well as fruiting body formation on the established dikaryon. Usually, three paralogous pairs of divergently transcribed genes for twodistinct types of homeodomain transcription factors (termed HD1 and HD2) are found in the multiple alleles of the A locus. For regulation of sexualdevelopment, heterodimerization of HD1 and HD2 proteins coming from allelic gene pairs is required. In some A loci found in nature, alleles of gene pairsare not complete or one of the two genes have been made inactive. Functional redundancy allows the system still to work as long as an HD1 gene in oneand an HD2 gene in the other allele of one gene pair are operative (Casselton and Kües 2007). Here, we present the structures of two completelysequenced A loci, A42 (this study) and A43 (Stajich et al. 2010). Evidences for gene duplications, deletions and inactivations are found. The loci differ in thenumber of potential gene pairs (five versus three), in genes that have been duplicated in evolution, in genes that have been lost in evolution and in genesthat are still present but have been made inactive. Furthermore, self-compatible mutants of the A loci are found that due to fusions of an HD1 and an HD2gene can carry out sexual reproduction without mating with another compatible strain. The products of the fusion genes can take over the regulatoryfunctions normally executed by heterodimers of HD1 and HD2 proteins that come from different nuclei. In this study, we present a sequenced fusion genefrom a mutant A43 locus. The 5'-half of an HD2 gene was fused in frame to a complete HD1 gene through a linker made up from former promotersequence. An earlier described fusion protein (Kües et al. 1994) similarly contains the 5'-half of an HD2 gene that however was fused to the 3'-half of anHD1 gene. Comparison between the resulting fusion proteins indicates that presence of the HD2 homeodomain and the NLSs (nuclear localization signals)from the HD1 protein are likely essential for the function of the fusion proteins. Other domains required for function in the wild type proteins (such as forheterodimerization) are dispensable for fusion proteins that mediate a self-compatible phenotype.219. Transformation of an NACHT-NTPase gene NWD2 suppresses the pkn1 defect in fruiting body initiation of the Coprinopsis cinerea mutantProto159. Yidong Yu 1 , Pierre-Henri Clergeot 2 , Gwenäel Ruprich-Robert 3 , Markus Aebi 4 , Ursula Kües 1 . 1) Molecular Wood Biotechnology + TechnicalMycology, University of Goettingen, Goettingen, Germany; 2) Department of Botany, Stockholm University, Stockholm, Sweden; 3) Institute of Geneticsand Microbiology, University Paris-Sud, Orsay, France; 4) Institute of Microbiology, ETH Zurich, Zurich, Switzerland.Homokaryon AmutBmut is a self-compatible strain of the mushroom Coprinopsis cinerea which can carry out sexual reproduction without fusing withanother compatible strain. Due to its single nucleus, this strain allows easy induction of mutations in fruiting body formation. One such mutant is the strainProto159, which is defective in the first step of fruiting body initiation (primary hyphal knot formation; pkn1). This mutant has been isolated afterprotoplasting and regeneration of oidia (Granado et al. 1997). It has a reduced growth speed and a reduced rate of oidiation (asexual spore formation)compared to the wild type AmutBmut. In addition, with age, the mycelium of Proto159 produces a dark-brown pigment that diffuses into the medium.This pigmentation is not found in AmutBmut. Proto159 never makes any sclerotium nor initiates formation of any fruiting structure. Complementationtests have been made through transformations with a cosmid bank of the wild type AmutBmut (Bottoli et al. 1999) and the defect has beencomplemented after transformation with the wild type gene NWD2. This gene codes for a NACHT-NTPase (signal transduction protein with a NACHTdomain which is found in animal, fungal and bacterial proteins and named after four different types of P-loop NTPases NAIP, CIITA, HET-E and TP1).However, sequencing of this gene in the mutant Proto159 did not reveal any point mutations, deletions or insertions within this gene. One possibility toexplain the pkn1 defect in mutant Proto159 in connection with the transformation data is that insertion of further copies of gene NWD2 into the genomeof mutant Proto159 has a suppressor effect on the defect in the yet unknown gene pkn1. This situation is reminiscent to findings in Schizophyllumcommune where formation of fruiting bodies has been induced in monokaryons upon transformation with the gene Frt1 (Horton and Raper 1991). GeneFrt1 encodes another type of P-loop NTPase (Horton and Raper 1995) than NWD2. However, the proteins share a novel short motif of amino acid similarityat their C-terminal ends.220. Dynamics of the actin cytoskeleton in Phytophthora infestans. Harold Meijer 1 , Chenlei Hua 1 , Kiki Kots 1,2 , Tijs Ketelaar 2 , Francine Govers 1 . 1) LabPhytopathology, Wageningen University, Wageningen, Netherlands; 2) Lab Cell Biology, Wageningen University, Wageningen, Netherlands.The actin cytoskeleton is conserved among all eukaryotes and plays essential roles during many cellular processes. It forms an internal framework in cellsthat is both dynamic and well organised. The plethora of functions ranges from facilitating cytoplasmic streaming, muscle contraction, formation ofcontractile rings, nuclear segregation, endocytosis and facilitating apical cell expansions. Oomycetes are filamentous organisms that resemble Fungi butare not related to Fungi. The two groups show significant structural, biochemical and genetic differences. One prominent lineage within the class ofoomycetes is the genus Phytophthora. This genus comprises over 100 species that are all devastating plant pathogens threatening agriculture and naturalenvironments. The potato late blight pathogen Phytophthora infestans was responsible for the Irish potato famine and remains a major threat today.Previously the actin organization has been studied in several oomycetes. Next to the common F-actin filaments and cables, cortical F-actin containingpatches or plaques have been observed as in Fungi. However, only a static view was obtained. Here, we use an in vivo actin binding moiety labelled to afluorescent group to investigate the actin cytoskeleton dynamics in hyphae of P. infestans. Our results provide the first visualisation of the dynamicreorganization of the actin cytoskeleton in oomycetes. In the future, this line will provide insight in the role of the actin cytoskeleton during infection.174

FULL POSTER SESSION ABSTRACTSComparative and Functional Genomics221. A novel approach for functional analysis of genes in the rice blast fungus. Sook-Young Park 1 , Jaehyuk Choi 1 , Seongbeom Kim 1 , Jongbum Jeon 1 ,Jaeyoung Choi 1 , Seomun Kwon 1 , Dayoung Lee 1 , Aram Huh 1 , Miho Shin 1 , Junhyun Jeon 1 , Seogchan Kang 2 , Yong-Hwan Lee 1 . 1) Dept. of AgriculturalBiotechnology, Seoul National University, Seoul 151-921, South Korea; 2) Dept. of Plant Pathology & Environmental Microbiology, The Pennsylvania StateUniversity, University Park, PA 16802, USA.Null mutants generated by targeted gene replacement are frequently used to reveal function of the genes in fungi. However, targeted gene deletionsmay be difficult to obtain or it may not be applicable, such as in the case of redundant or lethal genes. Constitutive expression system could be analternative to avoid these difficulties and to provide new platform in fungal functional genomics research. Here we developed a novel platform forfunctional analysis genes in Magnaporthe oryzae by constitutive expression under a strong promoter. Employing a binary vector (pGOF), carrying EF1bpromoter, we generated a total of 4,432 transformants by Agrobacterium tumafaciens-mediated transformation. We have analyzed a subset of 54transformants that have the vector inserted in the promoter region of individual genes, at distances ranging from 44 to 1,479 bp. These transformantsshowed increased transcript levels of the genes that are found immediately adjacent to the vector, compared to those of wild type. Ten transformantsshowed higher levels of expression relative to the wild type not only in mycelial stage but also during infection-related development. Two transformantsthat T-DNA was inserted in the promotor regions of putative lethal genes, MoRPT4 and MoDBP5, showed decreased conidiation and pathogenicity,respectively. We also characterized two transformants that T-DNA was inserted in functionally redundant genes encoding alpha-glucosidase and alphamannosidase.These transformants also showed decreased mycelial growth and pathogenicity, implying successful application of this platform infunctional analysis of the genes. Our data also demonstrated that comparative phenotypic analysis under over-expression and suppression of geneexpression could prove a highly efficient system for functional analysis of the genes. Our over-expressed transformant library would be a valuable resourcefor functional characterization of the redundant or lethal genes in M. oryzae and this system may be applicable in other fungi.222. Distribution and evolution of transposable elements in the Magnaporthe oryzae/grisea clade. Joelle Amselem 1,2 , Ludovic Mallet 1,3 , HeleneChiapello 3,4 , Cyprien Guerin 3 , Marc-Henri Lebrun 2 , Didier Tharreau 5 , Elisabeth Fournier 6 . 1) INRA, URGI, Versailles, France; 2) INRA, UMR BIOGER, Thiverval-Grignon, France; 3) INRA, UR MIG, Jouy-en-Josas, France; 4) INRA, UR BIA, Castanet-Tolosan, France; 5) CIRAD, UMR BGPI, Montpellier, France; 6) INRA,UMR BGPI, Montpellier, France.Magnaporthe oryzae is a successful pathogen of crop plants and a major threat for food production. This species gathers pathogens of differentPoaceaes, and causes the main fungal disease of rice worldwide and severe epidemics on wheat in South America. The evolutionary genomics ofMagnaporthe oryzae project aims at characterizing genomic determinants and evolutionary events involved in the adaptation of fungus to different hostplants. Such evolution may rely on variations in Transposable Elements (TEs) and gene content as well as modification of coding and regulatory sequences.Indeed, TEs are essential for shaping genomes and are a source of mutations and genome re-organizations. We performed a comparative analysis of TEs in9 isolates from the M. oryzae/grisea clade differing in their host specificity using a reference TEs consensus library (Mg7015_Refs_TE) made from M. grisea70-15 reference genome. We used REPET pipelines (http://urgi.versailles.inra.fr/Tools/REPET) to detect ab initio and classify TEs in M. grisea 70-15according to functional features (LTR, ITR, RT, transposase, etc.). After manual curation on consensus provided by the TEdenovo pipeline, we used theresulting consensus of TE families (Mg7015_Refs_TE) to annotate the 9 genome copies including nested and degenerated ones using TEannot pipeline. Wewill present results obtained for Mg7015_Refs_TE classification, their annotation, distribution along the genome and preliminary results provided bycomparison in M. oryzae/grisea species studied regarding correlation with phylogeny and host specificity.223. Alternative structural annotation of Aspergillus oryzae and Aspergillus nidulans based on RNA-Seq evidence. Gustavo C Cerqueira 1 , Brian Haas 1 ,Marcus Chibucos 2 , Martha Arnaud 3 , Christopher Sibthorp 4 , Mark X Caddick 4 , Kazuhiro Iwashita 5 , Gavin Sherlock 3 , Jennifer Wortman 1 . 1) Broad Institute,Boston, MA; 2) Institute for Genome Sciences, University of Maryland School of Medicine, Baltimore, USA; 3) Department of Genetics, Stanford UniversityMedical School, Stanford, USA; 4) School of Biological Sciences, University of Liverpool, Liverpool, United Kingdom; 5) National Research Institute ofBrewing, Hiroshima, Japan.The correct structural annotation of genes is fundamental to downstream functional genomics approaches. Genes undetected by gene predictionalgorithms, incorrect gene boundaries, misplaced or missing exons and wrongly merged genes can jeopardize attempts to produce a comprehensivecatalog of an organism’s metabolic capabilities. We are currently working toward generating alternative and improved structural annotation of Aspergillusoryzae and Aspergillus nidulans. Our approach consists of assembling partial transcript sequences from RNA-Seq data, aligning transcript assemblies totheir respective genomic loci and finally adjusting the gene models according to the new trancript evidence. Novel putative genes were defined based ontranscriptionally active regions containing splice junctions and open reading frames. Gene loci having transcripts suggesting alternative splicing variantswere reported. The nucleotide composition in the vicinity of splicing sites was re-evaluated in the light of the newly defined exons-introns boundaries. Themodified structural annotation was compared to the original structural annotation of these genomes and alternative gene models derived fromapproaches similar to those presented here. The improved gene models are available through the Aspergillus genome database (http://http://www.aspergillusgenome.org).224. Improved Gene Ontology annotation for biofilm formation, filamentous growth and phenotypic switching in Candida albicans. Diane O. Inglis,Marek S. Skrzypek, Arnaud B. Martha, Binkley Jonathan, Prachi Shah, Farrell Wymore, Gavin Sherlock. Department of Genetics, Stanford University,Stanford, CA.The opportunistic fungal pathogen, Candida albicans, is a significant medical threat, especially for immunocompromised patients. Experimental researchhas focused on specific areas of C. albicans biology with the goal of understanding the multiple factors that contribute to its pathogenic potential. Some ofthese factors include cell adhesion, invasive or filamentous growth and the formation of drug resistant biofilms. The Candida Genome Database (CGD,http://www.candidagenome.org/) is an internet-based resource that provides centralized access to genomic sequence data and manually curatedfunctional information about genes and proteins of the fungal pathogen Candida albicans and other Candida species. The Gene Ontology (GO;www.geneontology.org) is a standardized vocabulary that the Candida Genome Database (CGD; www.candidagenome.org) and other groups use todescribe the function of gene products. To improve the breadth and accuracy of pathogenicity-related gene product descriptions and to facilitate thedescription of as-yet uncharacterized but potential pathogenicity-related genes in Candida species, CGD has undertaken a three-part project: first, theaddition of terms to the Biological Process branch of the GO to improve the description of fungal-related processes; second, manual recuration of geneproduct annotations in CGD to use the improved GO vocabulary; and third, computational ortholog-based transfer of GO annotations from experimentally27th Fungal Genetics Conference | 175

FULL POSTER SESSION ABSTRACTScharacterized gene products using these new terms to uncharacterized orthologs in other Candida species. Through genome annotation and analysis, weidentified candidate pathogenicity genes in seven non-albicans Candida species and in one additional C. albicans strain, WO-1. We also defined a set of theC. albicans genes at the intersection of biofilm formation, filamentous growth, pathogenesis and phenotypic switching and now, finger and tentacledevelopment, of this opportunistic fungal pathogen, which provide a compelling list of candidates for further experimentation.225. Genome sequencing of Verticillium albo-atrum pathotypes in order to understand wilt disease in hop production. J. Jakse 1 , G. Rot 2 , V. Jelen 1 , S.Radisek 3 , S. Mandelc 1 , A. Majer 1 , B. Zupan 2 , B. Javornik 1 . 1) Agronomy Department, Biotechnical faculty, University of Ljubljana, Ljubljana, Slovenia; 2)Bioinformatics Laboratory, Faculty of Computer and Information Science, University of Ljubljana, Ljubljana, Slovenia; 3) Slovenian Institute of Hop Researchand Brewing, Zalec, Slovenia.Verticillium wilt of hop is a vascular disease caused by V. albo-atrum, outbreaks of the lethal strains of which threaten current hop production in Europe.Fungal isolates differ in aggressiveness and have been classified by pathogenicity tests into mild and lethal pathotypes. In general, the mild strain infectionvaries in intensity from year to year and rarely causes the death of the whole plant, whereas lethal strain infection causes very severe symptoms, withrapid plant withering and dieback. Lethal strains with increased virulence in hop were first reported in the UK in 1933, followed by outbreaks in Slovenia in1997 and in Germany in 2005. Sequencing the genomes of mild and lethal V. albo-atrum hop isolates aimed at the dissection of the pathotype genomes, inorder to provide an insight into their genomic structure, which might explain the increased virulence of the lethal strain, enable the detection of virulenceassociatedfactors and elucidate the pathogenicity in Verticillium spp. Genomes of three mild and three lethal strains from three different geographicregions were sequenced by Illumina technology. The reference lethal strain, with a larger genome than the mild strains, as confirmed by flow cytometry,was sequenced using three different length libraries producing a total of 76.3 M reads. From 4.8 to 11.5 M reads were obtained for the other five strains.Additionally, 38.3 M RNA-seq reads of mild and lethal strain transcriptomes were produced for annotation of the transcribed regions. Bioinformaticsanalyses included de-novo assembly of the reference genome, followed by mapping of the other genomes for comparison of mild and lethal strains todetermine specific regions of the strains. The reference genome was assembled into 715 contigs, with a total length of 33.59 Mb. Comparison of lethalversus mild strains revealed that 0.5 Mb of DNA was only present in the lethal strains. Gene prediction tools supported by RNA-seq analysis revealed 9858gene models, 91 of which were present in the lethal unique region. Analysis of repetitive DNA based on prebuilt models masked 1.53% of the assembledgenome, while de-novo identification of repeats masked 5.86% of the genome. The presented sequencing study established a new genomic resource fornon-alfalfa V. albo-atrum strains and will enable their virulence to be studied.226. Aegerolysin proteins from Aspergillus species. Nada Krasevec 1 , Kristina Sepcic 2 , Sasa Rezonja 1,2 , Nina Sluga 1,2 , Peter Macek 2 , Gregor Anderluh 1 . 1) L11Laboratory for Molecular Biology and Nanobiotechnology, National Institute of Chemistry, Ljubljana, Slovenia; 2) Department of Biology, BiotechnicalFaculty, University of Ljubljana, Slovenia.Currently, Aegerolysin family (Pfam06355) comprises over 300 proteins, mostly assigned as putative hemolysins, however, their function and biologicalrole is unknown. Some of them, i.e. aegerolysin, ostreolysin, pleurotolysin A, erylysin A from the Basidiomycota mushrooms (Agrocybe aegerita, Pleurotusostreatus and P. eryngei), and their orthologues, Asp-hemolysin from the human pathogens Aspergillus fumigatus (Eurotiales, Ascomycota) and PA0122(rahU) from Pseudomonas aeruginosa (Proteobacteria), have been characterized as lipid- or membrane-binding proteins. Aegerlysins are specificallydistributed among certain fungal species belonging to both Ascomycota and Basidiomycota taxa, however, they could be also found in bacteria and plants.In 2004, it was reported that in addition to the aegerolysin component A (pleurotolysin A, PlyA), a 59 kDa component B (pleurotolysin B, PlyB) is obligatoryfor the observed hemolytic activity of these proteins. In contrast to aegerolysins (component A), that appear widely distributed among differentorganisms, initial bioinformatical search of component B homologues results in a much lower number of similar putative proteins, even more, bothcomponents combined could be found in a few of fungal species only. Joint Genome Institute (JGI) has recently sequenced eight Aspergillus species (A.tubingensis, A. brasiliensis, A. acidus, A. glaucus, A. versicolor, A. sydowii, A. wentii and A. zonatus) as a result of community sequencing proposal(CSP2011). The genome sequences are available at MycoCosm (JGI) (http://genome.jgi.doe.gov/programs/fungi/index.jsf) and at the Aspergillus GenomeDatabase (AspGD) (http://www.aspgd.org/). The task of EUFGEN (EURotiales Functional GENomics consortium, http://www.eufgen.org/) is to complementthese genome sequences to those already available for the Aspergillus species. The strains were provided by CBS-KNAW fungal biodiversity center(http://www.cbs.knaw.nl/collection/AboutCollections.aspx). Our aim is to clarify experimentally the relation between the genome context for the twocomponents and their presumed hemolytic activity.227. Assembly, Annotation, and Analysis of Multiple Mycorrhizal Fungal Genomes. Alan Kuo 1 , Igor Grigoriev 1 , Annegret Kohler 2 , Francis Martin 2 ,Mycorrhizal Genomics Initiative (MGI) Consortium. 1) Fungal Genomics Program, DOE Joint Genome Institute, 2800 Mitchell Dr., Walnut Creek, CA, 94598USA; 2) Lab of Excellence ARBRE, Department of Tree-Microbe Interactions, INRA-Nancy, 54280 Champenoux, France.Mycorrhizal fungi play critical roles in host plant health, soil community structure and chemistry, and carbon and nutrient cycling, all areas of intenseinterest to the US Dept. of Energy (DOE) Joint Genome Institute (JGI). To this end we are building on our earlier sequencing of the Laccaria bicolor genomeby partnering with INRA-Nancy and the mycorrhizal research community in the MGI to sequence and analyze dozens of mycorrhizal genomes of allBasidiomycota and Ascomycota orders and multiple ecological types (ericoid, orchid, and ectomycorrhizal). JGI has developed and deployed highthroughputsequencing techniques, and Assembly, RNASeq, and Annotation Pipelines. In 2012 alone we sequenced, assembled, and annotated 12 draft orimproved genomes of mycorrhizae, and predicted ~232831 genes and ~15011 multigene families, All of this data is publicly available on JGI MycoCosm(http://jgi.doe.gov/fungi/), which provides access to both the genome data and tools with which to analyze the data. Preliminary comparisons of thecurrent total of 14 public mycorrhizal genomes suggest that 1) short secreted proteins potentially involved in symbiosis are more enriched in some ordersthan in others amongst the mycorrhizal Agaricomycetes, 2) there are wide ranges of numbers of genes involved in certain functional categories, such assignal transduction and post-translational modification, and 3) novel gene families are specific to some ecological types.228. Comparative reannotation of 21 Aspergillus genomes. Asaf A. Salamov, Robert Riley, Igor Grigoriev. DOE Joint Genome Inst, Walnut Creek, CA.We used comparative gene modeling to reannotate 21 Aspergillus genomes from MycoCosm and AspGD.Initial automatic annotation of individualgenomes may contain some errors of different nature, for example, missing genes, incorrect exon-intron structures, 'chimeras', which fuse 2 or moregenes,or splitting genes into 2 or more models.The main premise behind the comparative modeling approach is that for closely related genomes mostorthologous families have the same conserved gene structure. The algorithm maps all gene models predicted in all individual Aspergillus genomes to eachgenomes and for each locus selects among the potentially many competing models the one, which most closely resembles the orthologous genes fromother genomes. This procedure is iterated until no change in gene models will be observed. For the 21 Aspergillus genomes we predicted a total of 4503new gene models ( ~2% per genome), supported by comparative analysis, additionally correcting ~18% of oldgene models. This resulted in total of 4065176

FULL POSTER SESSION ABSTRACTSmore genes with annotated PFAM domains(~3% increase per genome). Analysis of few genomes with transcriptomics data shows that new annotation setsalso have a higher number of EST-supported splice sites at exon-intron boundaries.229. Using the phenotypic information in the PHI-base database to explore pathogen genomes, transcriptomes and proteomes. Martin Urban 1 , JohnAntoniw 2 , Natalia Martins 3 , Artem Lysenko 2 , Jacek Grzebyta 2 , Elzbieta Janowska-Sedja 2 , Mansoor Saqi 2 , Kim Hammond-Kosack 1 . 1) Plant Biology and CropScience, Rothamsted Research, Harpenden, Hertfordshire, United Kingdom; 2) Computational and Systems Biology, Rothamsted Research, Harpenden,Hertfordshire, United Kingdom; 3) Embrapa - Genetic Resources and Biotechnology, Brasília, Brazil.The Pathogen-Host Interactions database (www.phi-base.org), called PHI-base, stores expertly curated molecular and biological information on genes forwhich the effect on pathogen-host interactions has been tested experimentally. Fungal, oomycete and bacterial pathogens which infect animal, plant, fish,insect and/or fungal hosts are included. Information is also given on the target sites of some anti-infective chemistries. This database, available since 2005,is used to analyse effectively the growing number of verified genes that mediate an organism's ability to cause disease and/or to trigger host responses.PHI-base is also used as a valuable resource for the functional annotation of novel genomes (http://phytopathdb.org), in comparative genomics studiesand for the discovery of candidate targets in medically and agronomically important microbial pathogens for intervention with synthetic chemistries andnatural products (fungicides). Each curated entry in PHI-base is checked by individual species experts and is supported by strong experimental evidence(e.g. gene deletion, complementation experiments) and literature references. This extensive manual curation aims to position PHI-base as a ‘goldstandard’ for researchers in the pathogen-host biology community. Genes are annotated using controlled vocabularies (Gene Ontology terms, ECNumbers, etc.), and links to other external data sources (for example, NCBI taxonomy, EMBL and UniProt) are provided. Here we describe a significantupdate of PHI-base (Version 3.4) in which the data content has more than doubled. PHI-base now provides information on more than 2,200 genesdescribed in 3000 pathogen-host interactions, which are associated with more than 106 pathogenic species. A Fusarium species case study is presented,where the database content has been used in an integrated network analysis (combining information from gene co-expression, predicted protein-proteininteractions and sequence similarity) to predict proteins in Fusarium graminearum that may be involved in pathogenicity. This approach has identified 215candidates including 29 proteins currently annotated as ‘hypothetical’. As the content of PHI-base grows, we expect this database to be an importantresource for exploring conserved and species-specific themes in pathogenicity.230. RNA-Seq analysis reveals new gene models and alternative splicing in Fusarium graminearum. Chunzhao Zhao 1,2 , Cees Waalwijk 1 , Pierre Wit 1 ,Dingzhong Tang 2 , Theo vanderLee 1 . 1) Wageningen-UR, Wageningen, Gelderland, Netherlands; 2) 3State Key Laboratory of Plant Cell and ChromosomeEngineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.The genome of Fusarium graminearum has been sequenced and annotated, but correct gene annotation remains a challenge. In addition,posttranscriptional regulations, such as alternative splicing and RNA editing, are poorly understood in F. graminearum. Here we took advantage of RNA-Seq to improve gene annotations and to identify alternative splicing and RNA editing in F. graminearum. In total 25,720,650 reads were generated fromRNA-Seq. Transcripts were detected for 84% of the genes predicted by machine annotation in the BROAD database, Of these reads, 74.8% matched toexonic regions, 10.6% to untranslated regions (UTRs), 12.9% to intergenic regions and only 1.7% to intronic regions. We identified and revised 655incorrectly predicted gene models (10% of the gene models that could be tested), including revisions of intron predictions, intron splice sites andprediction of novel introns. In addition, we identified 231 genes with two or more alternative splice variants, mostly due to intron retention. In-frameanalysis showed that the majority of these alternatively spliced transcripts lead to premature termination codons, PTCs. Apart from PTC isoforms, somealternatively spliced transcripts encoding proteins with diverse lengths were identified. The effects of the diversity in the transcript length on the biologicalfunction of proteins are still unknown, but several functions including binding properties, intracellular localization, enzymatic activity or stability may beaffected. Interestingly, the expression ratios between different transcript isoforms appeared to be developmentally regulated. Surprisingly, no RNA editingwas identified in F. graminearum. Moreover, 2459 novel transcriptionally active regions (nTARs) were identified and our analysis indicates that many ofthese could be genes that were missed in the automated annotation. A number of representative novel gene models and alternatively spliced genes werevalidated by reverse transcription polymerase chain reaction and sequencing of the generated amplicons. Our results demonstrate that posttranscriptionalregulation can be studied efficiently using our developed RNA-Seq analysis pipeline and may be important in adaptation of F. graminearum to changingenvironmental conditions that occur during different growth stages.231. Comparison of transcriptome technologies in the MpkA deletion mutant of Aspergillus fumigatus. Clara Baldin 1,3 , Sebastian Mueller 2 , Marco Groth 4 ,Konrad Gruetzmann 5 , Reinhard Guthke 2 , Olaf Kniemeyer 1,3 , Axel Brakhage 1,3 , Vito Valiante 1 . 1) Department of Molecular and Applied Microbiology, LeibnizInstitute for Natural Product Research and Infection Biology, Hans-Knöll-Institute, Beutenbergstr. 11a, 07745 Jena, Germany; 2) Department of SystemsBiology / Bioinformatics, Leibniz Institute for Natural Product Research and Infection Biology, Hans-Knöll-Institute, Beutenbergstr. 11a, 07745 Jena,Germany; 3) Department of Microbiology and Molecular Biology, Friedrich Schiller University Jena, Beutenbergstrasse 11a, 07745 Jena, Germany; 4)Genome Analysis, Leibniz Institute for Age Research - Fritz Lipmann Institute, Beutenbergstr. 11, 07745 Jena, Germany; 5) Department of Bioinformatics,Friedrich Schiller University Jena, Ernst-Abbe-Platz 2, 07743 Jena, Germany.RNA deep sequencing techniques are rising as powerfull strategy to analyze the transcriptome profile of different organisms. Especially, this approachwill be very helpful whenever a microarray platform has not been established yet or when different platforms show low reproducibility of the generateddata. In the present study, the expression profile of Aspergillus fumigatus has been analysed via different transcriptome analysis approaches. A. fumigatusis a saprophytic fungus that is emerging as one of the most important airborne fungal pathogens. The adaptation of this fungus to different environmentsstimulated research on the regulation of the cell-wall integrity pathway, which is mediated by the Mitogen Activated Protein Kinase (MAPK) MpkA.Previuos microarray analyses showed that MpkA is involved not only in the regulation of genes responsible for cell wall maintenance, but also inprotection against reactive oxygen species, iron starvation response and secondary metabolites production (Jain et al., Mol. Microbiol. 2011). Using thesame strains and lab conditions, we performed a transcriptome study using RNA deep sequencing to directly compare different transcriptome analysistechniques. The RNA-seq technique was found to be more sensitive than microarray analyses giving us the possibility to gain new insight into the role ofMpkA. We were able to identify a substantial number of novel transcripts, to detect new exons, untranslated regions, thousands of new splice junctions,and found evidence for widespread alternative splicing events. We could also identify a large group of genes belonging to known and unknown geneclusters, which are normally involved in secondary metabolite production. They are differentially regulated in the DmpkA mutant strain. Moreover, thetranscriptome data were compared to proteome data. Comparison between these two biological levels contributes to a better understanding of transcriptstability and of post-transcriptional regulatory mechanisms, giving a more global overview about MpkA regulatory circuits (Müller, Baldin et al., BMC2012).27th Fungal Genetics Conference | 177

FULL POSTER SESSION ABSTRACTS232. Spliceosome twintrons ( “stwintrons”) revealed by fungal nuclear genomes. Michel Flipphi 1 , Erzsébet Fekete 1 , Claudio Scazzocchio 2 , LeventeKaraffa 1 . 1) Department of Biochemical Engineering, University of Debrecen, H-4010, Debrecen, Hungary; 2) Department of Microbiology, Imperial CollegeLondon, London SW7 2AZ, UK.The spliceosome is an RNA/protein complex, responsible for intron excision in eukaryotic genes. In mitochondria and plastids intron excision does notinvolve the spliceosome. For a class of chloroplast introns (II and III) "introns within introns” (twintrons) have been described. The removal of the internalintron is necessary for the excision of the external intron, and thus RNA maturation. Analogous structures have not been described for splicesomal introns.We have predicted four putative instances of “introns within introns” in nuclear genes of fungi. We call these “swtintrons” for “spliceosomal twin introns”.Putative stwitrons show a variable phylogenetic distribution. The presence of the internal intron predicts specific splicing intermediates. We haveexperimentally confirmed the existence of the predicted intermediate for the splicing of an RNA encoding a putative cyclic imidine-hydrolase of Fusariumverticillioides (Sordariomycetes, Hypocreales), where the internal intron interrupts the donor sequence between the first and second nucleotide andpredicted an analogous structure for a gene encoding a sugar transporter in two Magnaportacea. In the bioDA gene (encoding an enzyme catalysing twosteps of biotin biosynthesis of the Sordariales, an internal intron, predicted to interrupt a donor sequence of an intron between the second and thirdnucleotide has been confirmed by isolation of the splicing intermediate. In the fourth instance the putative internal intron disrupts the donor sequencebetween the fourth and fifth nucleotide of the 5’ sequence. In this instance, the presence of the internal intron was disproved, revealing an unsuspectedcase of alternative splicing.233. NGS data revealed that the NSDA sterile mutant contains a mutation in the SCF ubiquitin ligase subunit gene, culC, in Aspergillus nidulans. Dong-Soon Oh 1 , Dong-Min Han 2 , Masayuki Machida 3 , Kap-Hoon Han 1 . 1) Dept Pharmaceutical Engineering, Woosuk Univ, Wanju, Korea; 2) Division of LifeScience, Wonkwang University, Iksan, Korea; 3) Bioproduction Research Institute, Hokkaido Center, National Institute of Advanced Industrial Science andTechnology (AIST), Sapporo, Japan.Sexual development and fruiting body production of fungi play pivotal roles in production of ascospores by meiosis as well as adaptation of variousenvironmental changes. In a homothallic fungus Aspergillus nidulans, many environmental factors and genes affecting sexual development have beenelucidated. One of the first and important attempts for understanding the sexual development of A. nidulans was isolation of NSD, BSD and ASD mutants,which are defective in the process. Among them, NSD mutants are divided into four different complementation groups, NSDA-D, and two of the mutants,NSDC and NSDD, have already been characterized about the responsible genes, nsdC and nsdD and their functions. However, nsdA and nsdB mutations areremained to be unveiled. Since classical complementation experiments were not successful, we analyzed the whole genome sequence of NSDA mutantobtained from Next Generation Sequencing (NGS) to identify the nsdA4 mutation. As a result, we found three NSDA mutant-specific mutations andconfirmed the mutations by PCR followed by sequencing analysis. One of the mutations was found in AN3939 locus which encodes SCF ubiquitin ligasesubunit CulC. The mutation was G to T transversion, making D468Y amino acid residue change. Since the COP9 signalosome and ubiquitin ligase playimportant roles in fungal development, this mutation could be the correct nsdA4 mutation responsible for the sterile NSDA mutant phenotype. However,since two more mutant-specific mutations were also found in NSDA, detailed genetic characterization and mutation analyses will have to be performed.This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (No.2012R1A1A4A01012864).234. Whole genome sequencing of two Aspergillus oryzae strains isolated from Meju, a traditional brick of dried fermented soybean, in Korea. Dong-Soon Oh 1 , Seung-Bum Hong 2 , Jong-Hwa Kim 1 , Goro Terai 3 , Hiroko Hagiwara 3 , Masayuki Machida 3 , Kap-Hoon Han 1 . 1) Dept of Pharmaceutical Engineering,Woosuk Univ, Wanju, Korea; 2) Korean Agricultural Culture Collection, NIAB, Korea; 3) Bioproduction Research Institute, Hokkaido Center, NationalInstitute of Advanced Industrial Science and Technology (AIST), Sapporo, Japan.In Korea, various Aspergillus oryzae-like fungi are generally regarded as one of causal agents of Korean Meju, a soybean brick for soybean paste,fermentation. Since the fungal strain plays important roles in Japanese fermented food, A. oryzae type strain of Japan, RIB40, has been sequenced andanalyzed in detail. Despite the importance of the A. oryzae strains in Korean fermented food, not many fungal strains have been isolated from fermentedfoods as well as Meju and the characteristics of the fungi isolated from Meju have not been elucidated so far, especially in molecular genetics andgenomics level. In this study, we tried to reveal the differences between Japanese and Korean A. oryzae strains by characterizing the whole genomestructure and their features. The whole genome sequence of two A. oryzae-like fungi, which were isolated from Korean Meju by Korean AgriculturalCulture Collection (KACC), were obtained by Next Generation Sequencing. Comparison of the genome sequences between RIB40 and Korean isolates byusing ortholog and homolog analyses revealed that, in one of the Korean isolates, about 50 kb subtelomeric region of chromosome III, where the aflatoxingene cluster located, was deleted, suggesting that chromosome deletion have been occurred inside the genome of the same species. Not only theaflatoxin gene cluster but also the other regions were modified in the Korean isolates. Gene annotation analysis and characteristics including those inrelation to closely related species Aspergillus flavus will be discussed.235. Systematic analysis of the uncharacterized genes, which widely conserved among filamentous fungi, in Aspergillus oryzae. N. Imaru 1,2 , F. Senoo 1,2 ,Y. Ikeda 1 , S. Terado 1,2 , K. Iwashita 1,2 . 1) National Research Institute of Brewing, Higashihiroshima, Hiroshima, Japan; 2) AdSM, Hiroshima Univ.,Higashihiroshima, Hiroshima, Japan.The genome sequences of Aspergillus oryzae revealed huge number of uncharacterized genes, which were occupied about 50% of A. oryzae genes. Mostof these genes were widely conserved among other Aspergillus species and filamentous fungi, but not found in other organisms. Moreover, severalgenome array analysis revealed that some of these genes were highly expressed in various conditions, such as liquid or solid-state cultivations. In thiswork, we designated these gene as cff (Conserved among Filamentous fungi and Function unknown genes) genes. The analysis of the functions of thesecffgenes will be important to reveal the novel molecular mechanisms which conserved among filamentous fungi. In this context, we constructed cff genesdisruptants library and analyzed the phenotype of these cff disruptants to examine the function of the genes and to identified new drug or breeding targetgenes. First of all, we isolated function unknown genes according to KOG category of A. oryzae genome database and further selected the genes that areconserved at least 7 species among 14 filamentous fungi as the cff candidate genes. Then we further examined several database, such as Swiss plot, AspGDetc., to verify the function-unknown then decided cff genes. From these cff genes, we performed the disruption of the highly expressed cff 147 genes andobtained 130 cff genes disruptants including 9 heterokaryon type disruptants. We observed the morphological phenotype of these cff genes disruptantson the minimal medium and natural medium using three serial powder plates, as a model assay of industrial conditions. As the result, some disruptantsshowed characteristic phenotypes in the hyphae growth and the conidiation. Furthermore, we examine the drug sensitivity of these disruptants usinghydroxyurea, camptothecin, micafungin et. al.. As the results, significant growth inhibition was observed in some disruptants, while some disruptantshown slight drug resistant. Now we are going to examine stress responses and second metabolite productions. We will further analyses the detail178

FULL POSTER SESSION ABSTRACTSmolecular function of the genes which shown significant phenotype in these analysis.236. Penicillium purpurogenum degrades lignocellulose. What can we learn of this process by analyzing the genome, transcriptome and secretome ofthe fungus? Wladimir Mardones 1 , Eduardo Callegari 2 , Jaime Eyzaguirre 1 . 1) Department of Biology, Universidad Andres Bello, Santiago, Chile; 2) SanfordSchool of Medicine, Division of Basic Biomedical Sciences, University of South Dakota, Vermillion, SD.Penicillium purpurogenum grows on a variety of natural carbon sources and secretes to the medium numerous cellulolytic and hemicellulolytic enzymes.Although some information on the lignocellulose biodegradation process has been obtained by the study of individual enzymes, a more comprehensiveapproach has been attempted by analysis of the genome, transcriptome and secretome of the fungus. A genome sequence draft has been attained bymeans of Illumina Hi-Seq 2000 analysis followed by assembly (Allpaths-LG) and partial annotation (MAKER pipeline): 36 Mb total length, 579 scaffolds, N50238 Kbp, 8984 genes predicted. Using the same sequencing technology and the Trinity assembler, a transcriptome of the fungus grown on sugar beet pulp(50% pectin, 20% cellulose) has been obtained. It includes 7,172 ESTs with mean length of 307 bp; 5195 ESTs were significantly identified in the genome.The secretome of the sugar beet pulp culture was analyzed by shotgun mass spectrometry (2D Nano-LC MS/MS) and 53 proteins were identified byMASCOT. An analysis of the genome draft for genes related to lignocellulose biodegradation enzymes (using dbCAN) showed 347 genes of putativeCAZYmes (38 carbohydrate esterases, 245 glycosyl hydrolases, 56 glycosyl transferases, 6 polysaccharide lyases and 2 carbohydrate binding modules). Thetranscriptome data (using BLASTX) showed that 111 CAZy genes were transcribed. In addition, 46 putative CAZymes were identified in the secretome.Among the 46 recognized, 6 are cellulases and 19 are pectinases, directly related to the degradation of sugar beet pulp. This is the first Penicillium genomesequenced using next generation technology and annotated for its lignocellulose biodegradation enzyme genes. Most of the identified genes correspondto putative non-characterized enzymes. This information will be of value for a better understanding of the lignocellulose biodegradation by filamentousfungi. Support: FONDECYT 1100084; UNAB DI-61-12/R.237. Functional genomics of lignocellulose degradation in the Basidiomycete white rot Schizophyllum commune. Robin A. Ohm 1 , Martin Tegelaar 2 , HanA. B. Wösten 2 , Igor V. Grigoriev 1 , Luis G. Lugones 2 . 1) US DOE Joint Genome Institute, Walnut Creek, CA, USA; 2) Department of Microbiology and KluyverCentre for Genomics of Industrial Fermentations, Utrecht University, Utrecht, The Netherlands.White and brown rot fungi are among the most important wood decayers in nature. Although more than 50 genomes of Basidiomycete white and brownrots have been sequenced by the Joint Genome Institute, there is still a lot to learn about how these fungi degrade the tough polymers present in wood. Inparticular, very little is known about how these fungi regulate the expression of genes involved in lignocellulose degradation. In Ascomycetes, severalconserved transcription factors involved in regulation of complex carbon source degradation have been identified, but there are no homologs of these inBasidiomycetes. Few Basidiomycete white or brown rots are genetically amenable, hindering a functional genomics approach to the study of lignocellulosedegradation. A notable exception is Schizophyllum commune, for which numerous genetic tools are available. S. commune was grown on several carbonsources (glucose, cellulose, lignin or beech wood) and gene expression was analyzed. Numerous genes are strongly up-regulated on the complex carbonsources, compared to on glucose. As expected, many of these encode CAZymes (notably glycoside hydrolase family 61) and FOLymes, but also several wellconserved proteins with unknown function. Interestingly, three transcription factor genes are up-regulated during growth on complex carbon sources,suggesting they may be involved in regulating this process. These transcription factors are highly conserved in Basidiomycetes, but not in Ascomycetes.The two laccase genes of S. commune are very lowly expressed on complex carbon sources, suggesting that their function in lignocellulose degradation islimited. A promoter analysis of up-regulated genes reveals a conserved putative transcription factor binding site, which is also present in related fungi.Experiments to validate these findings, as well as a proteomics analysis during growth on complex carbon sources, are currently in progress.238. Functional characterization of genes expressed in early infection stages by the phytopathogenic fungus Botrytis cinerea. J. Espino, N. Temme, A.Viefhues, B. Oeser, P. Tudzynski. Institut of Plant Biology and Biotechnology, Westf. Wilhelms University, Schlossplatz 8, 48143 Muenster, Germany.Botrytis cinerea is a phytopathogenic fungus that causes important economic losses in the agricultural field, due to its aggressiveness and ability toproduce the “grey mould disease” in more than 200 plant species. Nowadays, the main strategy of control consists in the use of fungicides, although somestrains are becoming resistant to these chemicals. Therefore, the knowledge of the molecular mechanisms during host-plant interaction could be a usefultool to develop new effective treatments against this organism. In microarray studies, we have identified more than 150 genes which are expressed duringthe early stages of infection, but not in conidia, suggesting an important role during fungal germination and penetration. Most of these genes codify forproteins with unknown function. By means of bioinformatic analyses, transmembrane domains and signal peptides were identified in some of theseproteins, suggesting a possible role in signaling pathways or as effectors in the interaction with the plant. We are currently focusing on 18 of these genes,and we validated their expression by real time PCR. In all cases the expression pattern observed in the microarrays studies could be confirmed byquantitative PCR results. Some of them showed an expression at 12 hours post inoculation even 10,000-fold compared to the expression in conidia. Inorder to elucidate the possible role of these genes, we have generated knock-out mutants of 9 single genes and 9 genes located in clusters. Pathogenicitystudies as well as further characterization of the different deletion mutants are now in progress.239. Regulation of biofilm formation in Candida parapsilosis. Linda Holland, Leona Connolly, Denise Lynch, Geraldine Butler. School of Biomolecular andBiomedical Science, Conway Institute, University College Dublin, Dublin, Ireland.Candida parapsilosis is a major cause of infection in premature neonates, particularly because of its tendency to grow as biofilms on indwelling medicaldevices. The biofilm architecture of C. parapsilosis biofilms is substantially different to that of Candida albicans, in particular because C. parapsilosis doesnot make true hyphae, suggesting that the regulation of biofilm formation may also be very different. To address this question we have adapted a fusionPCR method originally developed for C. albicans to construct gene deletions in the type strain C. parapsilosis CLIB214 (1). To date, we have generated 100homozygous deletion strains. We selected predicted protein kinase genes, transcription factors and also genes that are known to be important for biofilmformation in either C. albicans or C. parapsilosis. The collection was assayed for changes in biofilm formation using 24-well Nunc polystyrene plates and bymeasurement of the dry weight of mature biofilm. Eight deletion strains, efg1, czf1, mkc1, gzf3, ume6, ace2, cph2 and bcr1 have a defect in biofilmdevelopment. Only efg1 and bcr1 deletions of C. albicans have similar defects. C. albicans and C. parapsilosis therefore share some key regulators ofbiofilm formation, but there are also substantial differences. References: (1) Noble SM, French S, Kohn LA, Chev V, Johnson AD. Nat Genet. 2010Jul;42(7):590-8. Systematic screens of a Candida albicans homozygous deletion library decouple morphogenetic switching and pathogenicity.240. Functional analysis of the Mps1 MAP kinase pathway in the rice blast fungus Magnaporthe oryzae. E. Grund 1 , M.-J. Gagey 1 , V. Toquin 2 , R. Beffa 3 , N.Poussereau 1 , M.-H. Lebrun 1,4 . 1) MAP CNRS-UCB-INSA-Bayer CropScience, Lyon, France; 2) Biochemistry Dept, Bayer CropScience, Lyon, France; 3) BayerCropScience AG, Frankfurt/Main, Germany; 4) BIOGER INRA, Thiverval-Grignon, France.27th Fungal Genetics Conference | 179

FULL POSTER SESSION ABSTRACTSSignaling pathways are important in coordinating fungal cellular processes required for stress resistance, development and pathogenicity. The Mps1 MAPkinase pathway of Magnaporthe oryzae is involved in cell wall integrity, sporulation and pathogenicity. Dmps1 mutants displayed an abnormal mycelialgrowth (reduced aerial hyphae and melanisation), did not sporulate and were non-pathogenic on plants as reported (a). Sensitivity of M. oryzae to cell walldegrading enzymes (CWDE) and cell wall inhibitors (CWI) was found to be dependent on pH. Indeed, M. oryzae cell walls display a resistance to enzymaticdegradation at pH 5, while they are sensitive at pH 6. Dmps1 loses this pH 5 induced cell wall resistance, while it is as sensitive to CWDE as wild type at pH6. M. oryzae is highly resistant to calcofluor (cell wall disorganizing agent) at pH 5 (10x) compared to pH 6. Dmps1 loses this pH 5 induced calcofluorresistance, while it is as sensitive as wild type at pH 6. M. oryzae is more sensitive (20x) to Nikkomycin Z (chitine synthase inhibitor) at pH 5 than pH 6,while sensitivity to Aculeacin (glucan synthase inhibitor) is independent of the pH. However, Dmps1 is as sensitive as wild type to these inhibitors at bothpH. We conclude that the pH 5 induced resistance of fungal cell walls to CWDE and calcofluor requires the Mps1 pathway. This also suggests that the Mps1pathway is strongly activated at pH 5 compared to pH 6. To test this hypothesis, we are assaying the phosphorylation status of Mps1 at different pH as wellas under several stress conditions and developmental stages to know when this pathway is activated. Additionally we constructed an activated allele ofMkk1, the MAPKK upstream of Mps1, placed under the control of either its own promoter (b) or the repressible pNIA1 promoter. These transformants willbe used to assess the effect of controlled activation of the Mps1 pathway on M. oryzae cellular functions. The different conditions of Mps1 pathwayactivation will be used for a comparative transcriptomic analysis of wild type and Dmps1 mutants.(a) Xu, 2000. Fungal Genet. Biol. 31:137-152.(b) Fujikawa et al., 2009. Mol. Microbiol. 73(4):553-70.241. A RNA-Seq directed functional genomics screen to identify novel cell wall genes in the hyphal tip of Neurospora crassa. Divya Sain, Lorena Rivera,Jason Stajich. Plant Pathology & Microbiology, University of California, Riverside, Riverside, CA.The cell wall is one of the most important organelles of the fungal cell and differentiates pathogenic fungi from the plants and animals they infect. Thismakes cell wall biosynthesis an excellent target for anti-fungal drugs. To identify new targets we employed a functional genomics approach informed bygene expression patterns based on RNA-Seq of the filamentous fungus Neurospora crassa. The growing tips of fungal hyphae are enriched for cell wallbiosynthesis activity proteins and transcripts (1-2). Based on this idea we sequenced RNA from the tip (1 hr growth) and colony interior (20 hr growth) ofvegetative growing culture of N. crassa. 70 genes were up-regulated in the tip (at least 5 fold) and we supplemented this list with 42 tip expressed genesfrom a study of N. crassa colony development using microarrays, where mRNA transcripts in the colony tips were enriched in functional categories relatedto cell wall growth and morphogenesis (2). We used the N. crassa knockout collection (3) to identify developmental phenotypes and under chemical stressconditions to expose sensitivity in cell wall and growth defects. Almost 60 percent of the genes were found to be sensitive to cell wall stress agents,Caspofungin (cell wall integrity inhibition) and SDS (cell wall disruption) suggesting that our gene-set was enriched for genes having a cell wall defect. Wetested these genes for defects in the hyper-osmolar stress (NaCl & Glycerol) and oxidative stress pathways as well as sexual development pathway. Wefound 20 knockout strains having defects in all or nearly all of these pathways suggesting these cell wall genes are involved in multiple pathways of growthand development of filamentous fungi. This set includes Zn-Cys transcription factors (NCU04866 & NCU04663), Glycoside Hydrolase 13 family proteins(NCU08131 & NCU08132) and genes with no annotated function (NCU04826 & NCU01254). All of these genes possess homologs in other Peziomycotinafungi. Hence our approach using gene expression selected a candidate gene-set enriched for growth processes that may be useful as targets for anti-fungaldrug development against filamentous pathogenic fungi. 1) Bartnicki-Garcia & Lippman. Science 1969; 165(3890):302-4. 2) Kasuga & Glass. Euk Cell 2008;7(9):1549-64. 3) Colot et al. PNAS 2006; 103(27):10352-7.242. Identification of centromeres in the plant pathogen Zymoseptoria tritici (synonym Mycosphaerella graminicola). Klaas Schotanus 1 , Lanelle R.Connolly 2 , Kristina M. Smith 2 , Michael Freitag 2 , Eva H. Stukenbrock 1 . 1) MPRG Fungal Biodiversity, Max-Planck-Institute for Terrestrial Microbiology,Marburg, Germany; 2) Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR, USA.Several plant pathogenic fungi contain small, apparently dispensable chromosomes, and in several cases pathogenicity genes have been identified onthese chromosomes. The ascomycete Zymoseptoria tritici has up to eight dispensable chromosomes in addition to thirteen “core” chromosomes. Duringmeiosis dispensable chromosomes are lost at elevated rates, resulting in progeny with distinct novel chromosome sets. So far little is known about the roleof these chromosomes and their evolutionary dynamics. We hypothesize that loss of dispensable chromosomes during meiosis may be correlated tounstable centromeres. Thus, one goal was to identify and characterize centromeric regions on core and dispensable chromosomes to allow us toinvestigate the underlying genetics of chromosome instability. Both core and dispensable chromosomes in the Z. tritici reference isolate IPO323 have beensequenced from telomere to telomere, yielding a unique opportunity to identify the centromeric regions in the genome. We tagged the Z. triticicentromere-specific histone 3 (CenH3) with GFP and confirmed correct insertion by Southern analyses. We demonstrate expression of GFP-tagged CenH3by western blot and epifluorescence microscopy. ZtCenH3-GFP was localized in discrete foci in interphase nuclei, but in contrast to other fungi (e.g.,Neurospora, Fusarium, Saccharomyces and Schizosaccharomyces) there are several foci per nucleus instead of a single chromocenter. We also performedchromatin immunoprecipitation followed by DNA sequencing (ChIP-seq) on the CenH3-GFP strains. To confirm the CenH3-GFP results, we tagged twoadditional centromere proteins (CEN-B and CEN-S) with GFP. To assess stability of centromeres during mitosis and meiosis, we obtained evolved asexualprogeny of IPO323 after 50 and 100 generations and progeny from a cross. Comparison of centromeric positions in the genome of the founder strain(IPO323) versus sexual and asexual progeny will allow us to infer dynamics of centromeres on core and dispensable chromosomes, and aid in ourunderstanding of the evolutionary dynamics of dispensable chromosomes in fungal plant pathogens.243. Loss of the RNAi pathway in VGII Cryptococcus gattii sheds light on the intact system in Cryptococcus neoformans. R Blake Billmyre, Xuying Wang,Marianna Feretzaki, Joseph Heitman. Duke University, Durham, NC.Loss of RNAi in VGII Cryptococcus gattii sheds light on RNAi roles in Cryptococcus neoformans R. Blake Billmyre, Xuying Wang, Marianna Feretzaki, andJoseph Heitman RNAi is a broadly conserved homology-dependent silencing mechanism which functions to defend the genome by silencing transposonsand viral elements. The opportunistic human pathogen C. neoformans, utilizes an RNAi-dependent process to robustly silence repetitive elements duringthe sexual cycle. Interestingly, RNAi components have been broadly lost from the VGII subtype of the closely related sister species C. gattii. We have takena comparative genomics approach to compare the RNAi deficient genome of VGII C. gattii with the RNAi proficient genomes of VGI C. gattii and serotypesA and D C. neoformans. This approach has identified a total of fourteen gene losses or truncations of otherwise conserved genes in VGII, including three ofthe known canonical RNAi components. Two of the remaining eleven genes have been shown to have a role in the sex-induced silencing pathway in C.neoformans var. grubii, despite a lack of homology with previously identified RNAi components in other organisms. One of these genes CPR2, waspreviously studied in our lab as a constitutively active G-protein coupled pheromone receptor. cpr2D also confers a moderate defect in sex-inducedsilencing, but no defect in silencing during vegetative growth. Similarly, the zinc finger factor Znf3 was previously identified in our lab and here was180

FULL POSTER SESSION ABSTRACTSunexpectedly found to be important for both sex-induced and vegetative silencing. The nine remaining missing genes are being tested for roles in bothsex-induced and mitotic silencing (SIS, MIS).244. A chemical-genetic map of a human fungal meningitis pathogen. Jessica C. S. Brown 1 , Benjamin VanderSluis 2 , Raamesh Deshpande 2 , Arielle Butts 3 ,Sarah Kagan 4 , Itzhack Polacheck 4 , Damian J. Krysan 3 , Chad L. Myers 2 , Hiten D. Madhani 1 . 1) Biochemistry and Biophysics, U. California, San Francisco, SanFrancisco, CA; 2) Computer Science and Engineering, U. Minnesota, Minneapolis, MN; 3) Pediatrics, U. Rochester Medical Center, Rochester, NY; 4) ClinicalMicrobiology and Immunology, Tel Aviv University, Tel Aviv, Israel.The systematic profiling of the impact of small molecules on the growth rate of gene deletion mutants is termed chemogenomic profiling. This approachbeen extensively used in model organisms, primarily baker’s yeast, to functionally annotate genes and to obtain insights into mode-of-action (MOA) forchemical compounds (1). Here we describe the application of systematic chemical-genetics a significant human pathogen. Cryptococcus neoformans is anopportunistic basidiomycetes pathogen responsible for lethal meningitis in immunocompromised patients. Current therapies are inadequate due to apaucity of drugs and a poor understanding of pathogenesis. Our laboratory previously constructed a partial gene deletion collection and used it to identifynumerous genes required for infection as well as for the production of virulence factors (2). This work identified numerous novel infectivity genes, butmany did not have an identifiable molecular function. We have now used chemogenomic profiling to both bridge this gap in gene annotation and to obtaininsights into drug MOA. To accomplish this goal, we identified and utilized over 200 diverse chemical compounds that impact pathogen growth to create aunique phenotypic signature for ~1500 C. neoformans gene deletion strains. We used colony arrays, robotics, automated image analysis, and extensivedata normalization algorithms to analyze several million phenotypic measurements. We used these data to identify clusters of genes and compounds withrelated patterns of chemical-genetic interactions. Our analysis identified virulence genes that act through related mechanisms. For example, one gene setinvolves a number predicted to be involved in histone modification. Members of a second set of genes are required in production of the C. neoformanspolysaccharide capsule, a well-established virulence factor. We have also obtained new insights into the MOA of several antifungal compounds. TheCryptococcus chemical-genetic map will be a valuable resource for functional annotation of the genome of this meningitis pathogen, characterization ofnew drug targets, and the identification lead compounds for antifungal drug development.1. Hillenmeyer et al., Science 320 (2008). 2. Liu et al., Cell 135 (2008).245. Whole genome sequencing of high-mortality and low-mortality strains of Cryptococcus neoformans var. grubii to discover genetic determinants ofvirulence. Tami R. McDonald, Kirsten Nielsen. Department of Microbiology, University of Minnesota, Minneapolis, MN.In sub-Saharan Africa, meningitis caused by the fungus Cryptococcus neoformans var. grubii is a major cause of AIDS-related mortality. To investigate therole of fungal genotype in clinical disease, we sequenced 8 genes for 503 clinical isolates of Cryptococcus neoformans var. grubii. A phylogenetic analysis ofthese strains demonstrated that 501 isolates were VNI strains. Haplotype network analysis revealed three major groups (BURST groups 1 - 3). Patientmortality was associated with fungal strain genotype, with strains in BURST group 3 demonstrating low mortality. Whole genome sequencing of 13representative genotypes revealed SNPs unique to the high mortality strains, and SNPs unique to the low-mortality strains, pointing to possible targets forfuture gene deletion and allele swap experiments to determine the role of the genes in pathogenesis.246. Identification of high temperature-regulated genes controlled by Sch9 through comparative transcriptome analysis in Cryptococcus neoformans.Dong-Hoon Yang 1 , Kwang-Woo Jung 1 , Jang-Won Lee 1 , Min-Hee Song 1 , Anna Floyd 2 , Joseph Heitman 2,3 , Yong-Sun Bahn 1 . 1) Biotechnology Dept, YonseiUniversity, Seoul, South Korea; 2) Departments of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC, USA; 3)Departments of Medicine, and Pharmacology and Cancer Biology, Duke University Medical Center, Durham, NC, USA.Adaptation to temperature changes is one of crucial virulence factors for Cryptococcus neoformans during host infection. In the human fungal pathogen,diverse signal transduction pathways, such as Ras/Cdc24, calmodulin/calcineurin, Mpk1 and Hog1 MAPK pathways, are involved in the temperatureadaptation process. In addition to the pathways, the Sch9 protein kinase has been implicated in thermotolerence of C. neoformans, but its regulatorymechanism remains elusive. In this study we aimed to identify Sch9-dependent or -independent temperature-regulated genes in a genome scale and toelucidate the regulatory mechanism of Sch9 in thermotolerance of C. neoformans. For this purpose, we performed comparative transcriptome analysiswith the wild type serotype A H99 strain and sch9D mutant during temperature upshift from 25°C to 37°C or 40°C. The temperature upshift caused a globalscale of remodeling in gene expression profiles (1872 genes, P

FULL POSTER SESSION ABSTRACTSbinding module, are also increased relative to brown-rot fungi. Indeed, secretomic analysis identified GH6, GH7, CDH and PMO peptides only in white-rotfungi. Overall, these results show that, relative to brown rot fungi, white rot polyporales maintain greater enzymatic diversity supporting lignocelluloseattack.248. Genomic context and distribution of effector genes in Fusarium oxysporum. Sarah Maria Schmidt 1 , Peter van Dam 1 , Petra M. Houterman 1 , InesSchreiver 2 , Lisong Ma 1 , Stephan Amyotte 3 , Biju Chellappan 1 , Sjef Boeren 4 , Frank L.W. Takken 1 , Martijn Rep 1 . 1) Molecular Plant Pathology, SwammerdamInstitute for Life Sciences, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, the Netherlands; 2) Fachgebiet Medizinische Biotechnologie,Institut für Biotechnologie, Technische Universität Berlin, Gustav-Meyer-Allee 25, Germany; 3) Department of Plant Pathology, University of Kentucky,201F Plant Science Building, 1405 Veterans Drive, Lexington, KY 40546-0312, USA; 4) Laboratory for Biochemistry, Wageningen University, Dreijenlaan 3,6703HA, Wageningen, the Netherlands.Strains of the Fusarium oxysporum species complex (FOSC) are able to infect a wide range of mono- and dicotyledonous plants. Based on the hostspecificity of individual strains, the FOSC is divided into various formae speciales. All strains share a common core genome and possess additional lineagespecific(LS) chromosomes. The fungus secretes effector proteins into the host vascular system that presumably manipulate the host to promote infection.In the tomato pathogen F. oxysporum f. sp. lycopersici (Fol) these effectors are encoded by SIX (Secreted In Xylem) genes. Interestingly, all SIX genes arepresent on a single LS chromosome that can be transferred horizontally to a previously non-pathogenic Fo strain, resulting in gain of pathogenicity towardstomato. Upon close inspection of this tomato pathogenicity chromosome we discovered that a non-autonomous miniature transposable element (mite) ispresent in the promoters of all SIX genes. Promoter deletion analysis at two different SIX gene loci did not reveal a direct role of the mite for SIX geneexpression. However, we were able to use this genomic signature to predict novel effector gene candidates in the Fol genome. Expression of several ofthese novel candidates during infection was confirmed by mass spectroscopic analysis of the xylem sap of Fol-infected tomato plants. We also discovered asmall reservoir of ‘silent’ effector genes that are not expressed during infection. Next, we used our method to predict effector gene candidates in thegenomes of several other formae speciales and developed a more global picture of the effector gene complement in the FOSC. Effector genes in Foconsistently reside in repeat-rich enviroments. Some strains contain 2 or 3 paralogs of an effector gene. Additionally, many genomes feature truncatedeffector gene homologs. Overall, the effector gene distribution among different formae speciales is patchy, and there is no unique set of effectors that iscommon to all plant pathogenic strains of the FOSC.249. Whole genome sequencing reveals new links between diverse plant pathogens; an expanded AvrLm6-like gene family in Venturia species. JasonShiller 1 , Angela van de Wouw 2 , Dan Jones 1 , Joanna Bowen 3 , Carl Mesarich 3 , Matthew Templeton 3 , Kim Plummer 1 . 1) La Trobe University, Melbourne,Australia; 2) University of Melbourne, Melbourne, Australia; 3) Plant and Food Research, Auckland, New Zealand.Venturia inaequalis and V. pirina are hemi-biotrophic fungi that cause apple scab and pear scab, respectively. These diseases cause significant losses togrowers worldwide. In some cases, scab is controlled with resistant cultivars, but fungicides are more commonly used. Resistance to scab follows the genefor-genemodel, whereby a gene coding for a resistance protein in the host will have a cognate gene coding for an avirulence protein (or effector) in thefungus. No Venturia effectors have been characterised to date, but work is underway to identify effectors from the whole genome sequences and secretedproteins of V. inaequalis and V. pirina. Whole genome sequencing of Venturia inaequalis and V. pirina has revealed predicted proteins with some sequencesimilarity to AvrLm6, a Leptosphaeria maculans effector that triggers resistance in Rlm6 canola. The mechanism of action of AvrLm6 is unknown. Untilrecently, AvrLm6 was thought to be unique to L. maculans, with orthologues absent, even in closely related species. AvrLm6-like genes from Venturia sppwhose genomes are sequenced form large families containing up to 30 members. We have also identified orthologues in F. oxysporum (Fo5176) and C.higginsianum from public database searches. Gene expansions have also been observed for other effector-like genes in the Venturia genomes. TheAvrLm6-like predicted protein from V. inaequalis, with the highest sequence identity to AvrLm6, was unable to trigger a resistance response in Rlm6canola. However, this does not preclude the AvrLm6-like proteins from being functionally active in the Malus-Venturia pathosystem. Transcriptomeanalyses (RNA-seq) of in planta and in vitro samples of V. inaequalis have revealed that a number of AvrLm6-like genes are up-regulated during infection(compared to growth in vitro). These results were confirmed with qRT-PCR. The most highly up-regulated predicted protein, ALVi_149, was tagged withYFP. YFP expression was observed only in the sub-cuticular stromata (specialised, biotrophic infection structures). RNA silencing is currently underway todetermine the role of ALVi_149 in pathogenicity of V. inaequalis. The major question that remains is; what purpose do these genes serve for these diversefungi and what is driving the gene expansions in Venturia spp?250. Oömycetes Protein Array Project. Samantha Taylor 1 , Regina Hanlon 2 , Mandy Wilson 2 , Jean Peccoud 2 , Brett Tyler 1 . 1) Oregon State University,Corvallis, OR; 2) Virginia Tech, Blacksburg, VA.Oömycetes are eukaryotes that outwardly resemble fungi, but are related to brown and golden-brown algae. The most destructive oömycete genus isPhytophthora, with over 80 species that collectively attack a wide range of plant species, causing damage to crops that is estimated in billions of dollarsannually in the US. The goal of the Oömycetes Protein Array Project is to generate a collection of cloned proteins from 1440 predicted oömycete effectorsequences, to use Gateway® technology to facilitate the easy transfer of clones into expression vectors, and to make the resulting clones available to thescientific community for further research. When the project is over, the final collection should include 390 clones from P. sojae, 550 clones from P.infestans, 370 clones from P. ramorum, and 130 clones from H. arabidopsidis.251. Extensive chromosomal reshuffling drives evolution of virulence in an asexual pathogen. Ronnie de Jonge 1,2 , Melvin Bolton 3 , Anja Kombrink 1 , KosteYadeta 1 , Grardy van den Berg 1 , Bart Thomma 1 . 1) Laboratory of Phytopathology, Wageningen University, Droevendaalsesteeg 1, 6708 PB Wageningen, TheNetherlands; 2) VIB Department of Plant Systems Biology, Ghent University, Bioinformatics and Evolutionary Genomics Division, Technologiepark 927, B-9052 Gent, Belgium; 3) United States Department of Agriculture, Agricultural Research Service, Northern Crop Science Laboratory, Fargo, ND 58102-2765,United States.Sexual recombination drives genetic diversity in eukaryotic genomes, and fosters adaptation to novel environmental challenges. Although strictly asexualmicroorganisms are often considered as evolutionary dead ends, they comprise many devastating plant pathogens. Presently, it remains unknown howsuch asexual pathogens generate the genetic variation that is required for quick adaptation and evolution in the arms race with their hosts. Here we showthat extensive chromosomal rearrangements in the strictly asexual plant pathogenic fungus Verticillium dahliae establish highly dynamic ‘plastic’ genomicregions that act as a source for genetic variation to mediate aggressiveness. We show that these plastic regions are greatly enriched for in plantaexpressedeffector genes, encoding secreted proteins that enable host colonization including the previously identified race 1-specific effector Ave1 thatactivates Ve1-mediated resistance in tomato. The plastic regions occur at the flanks of chromosomal breakpoints and are enriched for repetitive sequenceelements, especially retrotransposons. Our results demonstrate that asexual pathogens may evolve by prompting chromosomal rearrangements, enabling182

FULL POSTER SESSION ABSTRACTSrapid development of novel effector genes. Likely, chromosomal reshuffling is a general mechanism for adaptation in asexually propagating organisms.252. Genomic census of transmembrane proteins of the marine fungus, Corollospora maritima. Derek Johnson, Joseph Spatafora. Botany and PlantPathology, Oregon State University, Corvallis, OR.The class Sordariomycetes (Ascomycota) presents a model phylogenetic system to study the genomic evolution of marine fungi, as there have been atleast four major independent transitions from terrestrial to marine environments. These marine fungi have adapted to an environment which requiresincreased control of the movement of water between the cell and the environment due to osmosis. Given this, the greatest adaptive pressure may befound working at the interface of the fungal cell with the environment in the form of transmembrane proteins and in biochemical pathways dealing inosmoregulation. We have initiated a comparative, phylogenomic study of the independent lineages of marine Sordariomycetes with the goal ofdetermining what pathways have been modified in the transition to the marine environment. Here we present preliminary data of the draft genome of themarine fungus Corollospora maritima, which is a member of the largest family of marine fungi, Halospheriaceae (Microascales). The draft genome of C.maritima is similar to other Sordariomycetes (37 MB, 10269 predicted gene models) but is characterized by an increased number of transmembraneproteins; both in raw number (2017), and as a percentage of protein coding genes (19.64%), than non-marine members of the Sordariomycetes. Thus, it ishypothesized that C. maritima has expanded osmoregulatory protein families and adapted novel transmembrane proteins as a consequence of thetransition to the marine environment. Transmembrane proteins belonging to C. maritima along with select species of fungi and Eukaryotes were predictedvia hidden markov modeling using the program TMHMM and clustered into orthologous groups using Ortho-MCL. Putative protein identities (e.g.,aquaporins, aquaglyceroporins, osmosensors and the sodium efflux ENA ATPases) were assigned to each cluster using the BLAST tool comparingorthologous protein cluster identities to proteins of the model organisms Neurospora crassa and Saccharomyces cerevisiae. Expansions and contractionsof transmembrane proteins will be presented in a phylogenetic context, and more complex patterns of evolution, such as more ancient lineage sorting vs.lineage specific expansions, will be tested for each orthologous group of proteins using gene tree/species tree reconciliation analyses.253. Fungal Calcium Signaling Database (FCSD). Venkatesh Moktali, Bongsoo Park, Seogchan Kang. Penn State University, University Park, PA 16802, USA.Calcium probably is one of the most versatile elements in biological systems. It serves as a pivotal signal in controlling diverse cellular and developmentalprocesses to ensure the healthy functioning of every organism ranging from microbes to humans. The mechanism of translating external stimuli to specificcellular and developmental responses via changes in calcium ions plays an essential role in the plant-microbe and microbe-environmental interactions.Accordingly, many genes of the calcium-signaling pathway have been found to be virulence factors of fungal pathogens. How this simple and ubiquitousion has evolved to control so many processes is one of the central questions in biology with many practical implications. Rapid advances in genomesequencing of many fungal and oomycete species have uncovered conserved core calcium signaling genes, as well as lineage-specific features. To supportsystematic studies on this evolutionary variability in fungi and oomycetes and the functional roles of individual genes, we built the Fungal Calcium SignalingDatabase (FCSD; http://fcsd.ifungi.org/), an online platform that categorizes and annotates key calcium signaling proteins from more than 120 publishedfungal and oomycete genomes. The database also archives experimental results from studies on mutants of calcium signaling genes and resulting calciumsignatures in both video and picture formats. The calcium signaling genes in FCSD are divided into five major groups namely, calcium-permeable channels,calcium pumps, calcium exchanger/antiporter, calcium signaling regulators, and calcium-binding proteins. Comparison of calcium signaling machineriesbetween fungi and oomycetes has been conducted to identify evolutionary changes that have shaped up this signaling pathway in these kingdoms. TheFCSD will greatly support the fungal community in studying and understanding calcium signaling.254. Evolutionary genomic analysis of cytochrome P450 proteins in the subphyla Pezizomycotina. Venkatesh Moktali, Seogchan Kang. The PennsylvaniaState University, University Park, PA 16802.The subphylum Pezizomycotina presents a vast diversity of ecological niches and biochemical processes observed in fungal subphyla. Changes inmembers of the cytochrome P450 (CYP) superfamily appear to have played key roles in fungal niche adaption and evolution. Availability of genomic datafrom many species in this subphylum has enabled comprehensive phylogenomic studies to understand the taxon-specific genetic changes that potentiallyunderpin the observed functional and ecological diversity. CYPs from 53 Pezizomycotina species were analyzed to study the gene birth and death patternsat the genus level. This analysis revealed niche- and class-specific CYP family expansions and contractions. Putative metabolic functions were assigned toindividual CYPs in each species based on sequence similarity to functionally characterized CYP proteins. Also, pathogenic Pezizomycotina fungi weredivided into three classes (hemibiotrophs, obligate biotrophs and necrotrophs) to identify CYP family expansions and innovations potentially associatedwith these classes. Large losses in CYP families were observed among obligate biotrophs whereas hemibiotrophs and necrotrophs showed gene gains aswell as functional innovation in the form of species-specific CYP families. Examination of the classes/divisions within Pezizomycotina suggested a numberof independent losses and gains in CYP families. These findings shall be presented in the poster.255. Uncovering the evolutionary pressures shaping the Glomeromycota-Glomeribacter endosymbiosis. Stephen J. Mondo, Teresa E. Pawlowska. PlantPathology, Cornell University, Ithaca, NY.Many eukaryotes interact with heritable endobacteria to satisfy diverse metabolic needs. Of the characterized fungal-bacterial endosymbioses, theassociation between Gigasporaceae (Glomeromycota) and Ca. Glomeribacter is one of the best described. Glomeribacter is a member of the Burkholderialineage of b-proteobacteria, and was shown previously to represent one of the few cases of an ancient, long-term non-essential endosymbiont. In order tofurther explore what adaptations have taken place to shape this unique bacterial lifestyle, we have sequenced three Glomeribacter genomes anddeveloped a computational pipeline to compare across bacteria engaging in different lifestyles using genome wide patterns of mutation accumulation. Weused PAML to identify gene orthologs that exhibited both over-accumulation and under-accumulation of amino acid substitutions and then used thesedata to compare across taxa at the level of functional gene categories. We found that bacteria can be grouped by lifestyle using this approach.Glomeribacter, as expected, appears most similar to other potentially long-term non-essential endosymbionts. Therefore, we were able to exploit thedifferences in mutation accumulation patterns between these taxa to identify processes, which may be relevant within the particular interaction betweenGlomeribacter and its host. While several of these processes, including vitamin synthesis and amino acid transport, have been identified previously, weadditionally discovered features related to lipid biosynthesis and energy metabolism to be of potential importance for this symbiosis. Interestingly, genesexhibiting an under-accumulation of nonsynonymous substitutions (indicative of purifying selection) in Glomeribacter tend to be involved inrecombination, cell division, and ribosome maintenance. While these processes are typically fast evolving in endosymbiotic organisms, they may representfeatures that increase the stability of Glomeribacter in their fungal host population and increase their resilience to genetic drift. We speculate that theseprocesses are unique to the Glomeribacter-Glomeromycota symbiosis and could partially explain why Glomeribacter has been successful as a nonessentialendosymbiont for over 400 million years.27th Fungal Genetics Conference | 183

FULL POSTER SESSION ABSTRACTS256. The Unique family of Telomere-Linked Helicases in Fungi. Olga Novikova 1,2 , Mark Farman 2 . 1) Department of Biological Sciences, University atAlbany, Albany, NY; 2) Department of Plant Pathology, University of Kentucky, Lexington, KY.Subtelomeres and telomeres are highly dynamic regions of eukaryotic chromosomes and their maintenance is crucial for cellular function. Severalhelicases are known to be involved in maintenance of telomere integrity, e.g. RecQ-like helicases such as human BLM or WRN helicases. Curiously, RecQlikehelicase genes are found in very close proximity to telomeres in several fungal species. In the present study we performed comprehensive survey ofthese Telomere-Linked Helicases (TLHs) in 101 fully-sequenced fungal genomes. The TLHs were widely yet sporadically distributed among fungal speciesbeing present in 46 species belonging to all investigated groups except Zygomycetes. The TLHs were also unique to fungi. Many of the TLH genes werefound either next to telomeres or at the ends of contigs, providing indirect evidence that they are telomere associated. To date, the TLH gene families arethe only examples where the chromosomal positions of member genes are absolutely conserved across a kingdom. Despite the seemingly conservativepositions of the TLH genes on the chromosomes, the genes themselves are clearly not well conserved because they were found in only in half of the fungalgenomes surveyed. The TLHs were highly divergent from one another and demonstrated complex evolutionary histories that reflect recurrent cycles oftelomere crisis and recovery. Their telomere association leads us to hypothesize that the TLHs are involved either in telomere maintenance, or in therecovery processes associated with telomere crisis.257. Evolution of proteins containing intein- and Hedgehog-like Vint domains in Fungi. Olga Novikova, Marlene Belfort. Department of BiologicalSciences and RNA Institute, University at Albany, Albany, NY.Inteins are protein sequences that autocatalytically splice themselves out of the protein precursors - analogous to introns - and ligate the flanking regionsinto a functional protein. Intein-containing genes are present in all three kingdoms of life. Moreover, it was shown that the C-terminal domain ofeukaryotic Hedgehog (Hh) proteins has sequence similarity to inteins. The Hh pathway is one of the fundamental signal transduction pathways in animaldevelopment and is also involved in stem-cell maintenance and carcinogenesis. Two distinct domains can be found in Hh - the N-terminal ‘Hedge’ domain(HhN), and the intein-like C-terminal ‘Hog’ or ‘Hint’ domain (HhC). The hedgehog pathway is absent from Fungi. However, other families of proteins werefound containing Hog/Hint-like domains. Representatives from one of these families carry the von Willebrand factor type A (vWA) domain in addition tothe Hog/Hint-like domain. These domains are called Vint (von Willebrand Hint-like). Vint-containing proteins were initially reported for plants, fungi andsome metazoa. We explored the diversity of Vint-containing proteins, their distribution and evolutionary history in fungi. Vint-containing proteins arewidely distributed among fungal lineages; however, they are absent from some of the fungal species and entire fungal groups (e.g. Saccharomyces). Basedon the evolutionary pattern, we propose a modular model of the evolution for Vint-containing proteins. While the vWA domain seems to be the corefunctional unit, an additional domain, the U-box, is a recent acquisition. The vWA and U-box combination was found exclusively in fungi. The Vint domainis highly conserved and is likely under purifying selection. The functional role of Vint-containing proteins and the Vint domain in particular is the subject forfurther studies. This research has been supported by NIH grant GM44844.258. The genome and development-dependent transcriptome of Pyronema confluens: a window into fungal evolution. Stefanie Traeger 1 , Jason Stajich 2 ,Stefanie Pöggeler 3 , Minou Nowrousian 1 . 1) Department of General & Molecular Botany, Ruhr University Bochum, 44780 Bochum, Germany; 2) Departmentof Plant Pathology and Microbiology, University of California Riverside, CA 92521, USA; 3) Institute of Microbiology and Genetics, Department of Geneticsof Eukaryotic Microorganisms, Georg-August University, 37077 Göttingen, Germany.In the last decade, genomes of many filamentous ascomycetes have been sequenced and are invaluable for the analysis of the evolution of species andfor understanding their physiological and morphological properties. However, while there are at least ten genome sequences available for each of themore derived groups of filamentous ascomycetes (Sordariomycetes, Leotiomycetes, Eurotiomycetes, Dothideomycetes), only one genome from the basalgroup of Pezizomycetes has been sequenced, namely that the black truffle, a fungus with a specialized life-style and fruiting body. Therefore, wesequenced the genome and transcriptome of the Pezizomycete Pyronema confluens, a saprobe with typical apothecia as fruiting bodies. The genome wasassembled from a combination of Roche/454 and Illumina/Solexa reads. It has a size of 50 Mb, and a predicted 13369 protein-coding genes. P. confluens ishomothallic, and we found two MAT loci that are not fused or in close proximity, and encode an alpha domain and an HMG domain transcription factor.Only the MAT1-2 locus is flanked by the conserved apn2 gene, whereas both MAT loci are flanked by a pair of paralogous genes not found in this locationin other ascomycetes. Thus, the P. confluens MAT loci might reflect an evolutionary transition state on the way towards the relatively conserved genomicarrangement of MAT loci in higher ascomycetes. Sexual development in P. confluens is light-dependent, and qRTPCR analyses of predicted photoreceptorgenes showed that all are upregulated by light. Fruiting body formation is stimulated only by part of the visible spectrum, and we are currentlyinvestigating the effect of different wavelengths on development and gene expression. For RNA-seq analysis, we used three conditions: growth in the light,and two different conditions allowing only vegetative growth. We analyzed expression levels for genes with different degrees of evolutionary conservationto find out if genes with different lineage-specificities are preferentially expressed under any of the conditions investigated. Interestingly, the highestpercentage of genes upregulated during sexual development is found among the P. confluens orphan genes. This might indicate that, similar to thesituation in animals, genes associated with sexual reproduction evolve more rapidly than genes with other functions.259. Genome and transcriptome analysis of the mycoparasite Clonostachys rosea. Kristiina Nygren 1 , Mikael Brandström Durling 1 , Chatchai Kosawang 2 ,Dan Funck Jensen 1 , Magnus Karlsson 1 . 1) Forest Mycology and Plant Pathology, Swedish Univesity of Agricultural Sciences, Uppsala, Sweden; 2)Department of Plant Biology and Biotechnology, University of Copenhagen, Denmark.The ascomycete Clonostachys rosea is an efficient antagonist against a range of fungal plant pathogens, presumably as a consequence from itsmycoparasitic lifestyle. C. rosea is therefore used as a biological control agent against pathogens threatening agricultural crops. Still, very little is knownabout the mechanisms behind the mycoparasitism in C. rosea. By comparative genomics using C. rosea and publically available genome sequences fromclosely related species exhibiting different lifestyles we aim at exploring evolution of genes important for the transition into a mycoparasitic lifestyle. Andby analyzing the transcriptome during interactions with different fungal plant pathogens we intend to get a deeper understanding on the gene expressionof secreted enzymes during initial attack but also to study potential specialization towards specific fungal prey species. For these purposes we havesequenced the genome of C. rosea strain IK726. In addition we have performed an RNAseq study to investigate the gene expression during interactionswith the two plant pathogenes Botrytis cinerea and Fusarium graminearum. Our draft genome of C. rosea (strain IK726) reveals that the species has alarger genome than it’s closest sequenced relatives (58 Mb, which is 40 - 75 % larger than the 7 closest sequenced species). Preliminary comparativegenome analyses indicate that the mycoparasitic function of C. rosea differs from earlier findings in mycoparasites of the closely related genusTrichoderma. For example, in C. rosea we find a significant expansion of ABC-transporter genes.184

FULL POSTER SESSION ABSTRACTS260. The mitochondrial genomes of Fusarium circinatum, F. verticillioides and F. fujikuroi are unexpectedly similar. G Fourie 1 , N.A. van der Merwe 2 , B.D.Wingfield 2 , B. Tudzynski 3 , M.J. Wingfield 1 , E.T. Steenkamp 1 . 1) Department of Microbiology and Plant Pathology, Forestry and Agricultural BiotechnologyInstitute (FABI), University of Pretoria. Pretoria, South Africa; 2) Department of Genetics, Forestry and Agricultural Biotechnology Institute (FABI),University of Pretoria, Pretoria, South Africa; 3) University of Munster, Germany.The Gibberella fujikuroi species complex consists of species that are of considerable agricultural, medical and veterinary importance. Nevertheless, manyof the relationships among species in this complex remain largely unresolved, irrespective of the markers employed. In this study, we considered thefeasibility of using mitochondrial genes to resolve the higher level evolutionary relationships of the species in the complex. Because there is limitedinformation available regarding the structure and evolution of mitochondrial genomes in the G. fujikuroi species complex, we fully characterized themitochondrial genomes of the representative species; Fusarium circinatum, F. verticillioides and F. fujikuroi. Overall, the mitochondrial genomes of thethree species displayed a high degree of synteny, with all the genes in identical order and orientation. This similarity also extended to the intergenicregions, as well as introns that share similar positions within genes. The results show genome similarity beyond the expected characters common to themitochondrial genomes of the Sordariomycetes. The intergenic regions and introns generally contributed significantly to the size differences and diversityobserved among these genomes. Phylogenetic analysis of the concatenated protein-coding data set separated members of the G. fujikuroi complex fromother Fusarium species. The individual mitochondrial gene trees did not always support the phylogeny of the concatenated data set and at least six distinctphylogenetic trees were recovered. This incongruence could arise from biased selection on some genes or recombination among mitochondrial genomes,potentially linked to a hybridization event. The results suggest that using individual genes for phylogenetic inference could mask the true relationshipsbetween species in this complex.261. Comparative pathogenomics: next generation dissection of mechanisms of pathogenesis on plants. Donald M Gardiner 1 , Jana Sperschneider 3 , PaulaMoolhuijzen 4 , Matthew Bellgard 4 , Kemal Kazan 1 , Jen Taylor 2 , John Manners 2 . 1) Plant Industry, CSIRO, St Lucia, Queensland, Australia; 2) Plant Industry,CSIRO, Canberra, ACT, Australia; 3) Plant Industry, CSIRO, Perth, WA, Australia; 4) Centre for Comparative Genomics Murdoch University Perth, WesternAustralia, 6150.Comparative analyses between fungal plant pathogens that share a common host have revealed important mechanisms of virulence in a number ofdifferent systems. We have an interest in understanding how Fusarium pathogens of wheat and barley cause head blight and crown rot diseases on thesehosts. With modern genome sequencing technologies the power to undertake comparative analyses to assist in the understanding of key molecularmechanisms involved in pathogenesis has undoubtedly increased. We have recently sequenced an additional seven Fusarium isolates that are associatedwith wheat including isolates of F. equiseti, F acuminatum, F. culmorum and F. pseudograminearum and are using these in comparative analyses. Sequencebased homology searching between limited numbers of selected species have been particularly powerful in identifying signatures of horizontal transferbetween phylogenetically diverse species which have been an important force in the evolution of virulence and examples of these that we have shown tobe involved in virulence will be discussed. We are also developing methodologies to predict genes important in virulence that consider more remotehomologies between species that may represent structural and/or functional conservation which also consider phylogenetic distributions and enrichmentin species with particular lifestyles or shared plant hosts.262. Characterisation of stuA homologue in Fusarium culmorum. Matias Pasquali 1 , Francesca Spanu 2 , Virgilio Balmas 2 , Barbara Scherm 2 , Kim HammondKosack 3 , Lucien Hoffmann 1 , Marco Beyer 1 , Quirico Migheli 2,4 . 1) Environment and Agrobiotechnology Dept, CRP GABRIEL LIPPMANN, Belvaux, Luxembourg;2) Dipartimento di Agraria - Sezione di Patologia vegetale ed entomologia and Unità di ricerca Istituto Nazionale di Biostrutture e Biosistemi, Universitàdegli Studi di Sassari, Viale Italia 39, I-07100 Sassari, Italy; 3) Wheat Pathogenomics, Department of Plant Biology and Crop Sciences , RothamstedResearch, Harpenden, Herts AL5 2JQ, UK; 4) Centro interdisciplinare per lo sviluppo della ricerca biotecnologica e per lo studio della biodiversità dellaSardegna e dell'area mediterranea, Università degli Studi di Sassari, Viale Italia 39, I-07100 Sassari, Italy.Fusarium culmorum is one of the most harmful pathogens of durum wheat and the causal agent of foot and root rot (FRR) disease. F. culmorum producesdifferent trichothecene mycotoxins that are involved in the pathogenic process. The role of the gene FcStuA, a stuA ortholog protein with an APSESdomain sharing 98.5% homology to the FgStuA protein (FGSG10129), was determined by functional characterisation of deletion mutants obtained fromtwo F. culmorum wild-type strains, namely FcUK99 (a highly pathogenic trichothecene producer) and Fc233B (unable to produce toxin and with a mildpathogenic ability). The DFcStuA mutants originating from both strains showed common phenotypic characters including stunted vegetative growth, lossof mycelium hydrophobicity, altered pigmentation, decreased production of polygalacturonases, altered and reduced conidiation, delayed sporegermination patterns and complete loss of pathogenicity towards wheat stem base/root tissue. Toxin production in mutants originating from FcUK99strain was significantly decreased in vitro to 5% of the original production. Moreover, both sets of mutants were unable to colonise non-cereal planttissues, i.e. apple and tomato fruits and potato tubers. No differences between mutants, ectopic and wild-type strains were observed concerning the levelof resistance towards four fungicides belonging to three classes, the demethylase inhibitors epoxiconazole and tebuconzole, the succinate dehydrogenaseinhibitor isopyrazam and the cytochrome bc1 inhibitor trifloxystrobin. StuA is a global regulator in F. culmorum and is a potential target for novelfungistatic / fungicidal molecules.263. Comparative analysis of noncoding sequences in the Gibberella fujikuroi species complex. Christian Sieber 1 , Ulrich Güldener 1 , MartinMünsterkötter 1 , Karsten Suhre 1,2 . 1) Helmholtz-Zentrum München, Neuherberg, Bayern, GermanyHelmholtz Zentrum München, German Research Centerfor Environmental Health, Institute of Bioinformatics and Systems Biology, 85764 Neuherberg, Germany; 2) Department of Physiology and Biophysics,Weill Cornell Medical College, Education City, Qatar.Initially genome analysis methods focused on the coding part and its resulting proteins, therefore the role of noncoding DNA features was widelyunregarded for a long time. Due to the luxuriance of new sequencing technologies a plurality of genome sequences are now available and many differentrepeat families could been identified already, which exhibit a diversity of functions such as mRNA stabilization or the control of translation (Khemici 2004,Espeli 2001). Interspersed repeats account for a considerable amount of noncoding sequence in fungal genomes. The proportion of repetitive elementsand the compositions of repeat families differ from species to species whereas little is known about their origin and impact on the genomic functionality.The availability of new Fusarium genome sequences facilitate an extensive comparative approach across the Gibberella fujikuroi species complex. Besidesknown transposable elements and satellite repeats, a diversity of previously unknown interspersed repeat families are prominent in F.fujikuroi and closelyrelated species. While some of them are distributed through the whole complex, others can exclusively be found in only one genome. Interestingly thepredicted secondary structures of the elements exhibit a stable fold in terms of free energy and complementary base pairing. Moreover gene-chip andRNA-seq experiments reveal that some elements are part of the transcriptome and still seem to propagate further in the genome. Main questions are:What influence do interspersed repeats have on genome structure and the speciation process of fungi? How can repeats contribute to host-pathogen27th Fungal Genetics Conference | 185

FULL POSTER SESSION ABSTRACTSinteraction and specification?264. Virulence of Fusarium circinatum on Pinus species. S. L. Slinski 1,2 , B. D. Wingfield 1 , T. R. Gordon 2 . 1) Genetics, University of Pretoria, Pretoria,Gauteng, South Africa; 2) Plant Pathology, University of California, Davis, California.Fusarium circinatum causes pitch canker, an important disease of Pinus species worldwide. Little is know of the genetic determinants of virulence in thispathogen although virulence-related genes have been identified in other Fusarium species. The purpose of this work was to assess the heritability ofvirulence in F. circinatum and to identify genomic regions associated with virulence. Virulence was altered through a series of sibling crosses pushing thepopulations towards high or low virulence. Crossing high and low virulence parents from the F 3-High and F 3-Low generations, respectively, generatedprogeny (HL-F 1) with a nearly continuous distribution of virulence phenotypes. One hundred progeny were evaluated for polymorphic markers segregatingfor virulence by AFLP-PCR. Four markers were found to have a strong association with the long lesion length phenotypes. The AFLP-PCRs containing themarkers were sequenced using Illumina HiSeq technology and the genomic regions associated with virulence were located on the F. circinatum genome.These regions are being studied to determine the changes resulting in loss of virulence.265. Understanding the remodeling of the wheat grain genome expression during infection, a gate to get new insights on the molecular cross-talkcontrolling the development of the interaction between the wheat and Fusarium graminearum.. Chetouhi Cherif 1,2 , Bonhomme Ludovic 1,2 , CambonFlorence 1,2 , Lecomte philippe 1,2 , Biron G-David 3 , Langin Thierry 1,2 . 1) INRA, UMR 1095 GDEC, F-63039 Clermont-Ferrand cedex 2, France; 2) UBP, UMR 1095GDEC, F-63100 Clermont-Ferrand, France; 3) CNRS-UBP 6023 LMGE, F-63171 Aubière, France.Despite numerous progresses in understanding the molecular plant defences against pathogens, the molecular mechanisms used by a fungal pathogento counter the plant defences and to optimize the host cell environment for fungal growth remain largely unknown. One of the main goals of our group isto better understand resistance and susceptibility mechanisms in wheat to Fusarium graminearum. This fungal pathogen represents the main causal agentof Fusarium head blight (FHB), an important worldwide disease in wheat reducing grain yield and quality. Contamination of grain by trichothecenemycotoxins produced by F. graminearum during infection is the primary causes of reduced grain quality. Until now, very little is known on the molecularcross-talk between this fungus and its host during a compatible interaction. In this study, we characterize the impact of F. graminearum infection on thewheat cellular processes and identify wheat genes, so called susceptibility genes (S-gene), required for disease development. Until now, very little is knownon the transcriptional changes induced by F. graminearum in a susceptible wheat cultivar. A whole genome expression analysis was performed on theFrench susceptible wheat cultivar Récital challenged with a pathogenic and mycotoxigen F. graminearum strain. Using a microarray analysis, we haveidentified 1,453 differentially expressed genes while proteome comparative analysis showed 80 differentially regulated proteins between healthy andFusarium-damaged kernels at different development stages of the grain (flowering at 450 °Cd). These disease-associated genes and proteins belong tothree main functional groups including (i) plant defense, (ii) primary, secondary and energy metabolism and (iii) regulation and signaling. These resultsdemonstrate that the F. graminearum infection strategy associates (i) suppression of plant defense, and (ii) subtle changes in nutrient availability relatedprocesses.These preliminary results strongly suggest that F. graminearum manipulates the functioning of wheat kernel cells to optimize its nutrition, andtherefore that the disease susceptibility of wheat relies on a parasite manipulation by this pathogenic fungi. This is the first exhaustive study of themolecular mechanisms associated with FHB development in a susceptible wheat cultivar.266. Genetic and epigenetic changes in Fusarium graminearum following serial subculture. Rhaisa Crespo 2 , Heather E. Hallen-Adams 1 . 1) Food Scienceand Technology, Univ of Nebraska-Lincoln, Lincoln, NE; 2) University of Puerto Rico, Mayaguez, Mayaguez, PR.Fusarium isolates are notably unstable in culture and given to degradation unless certain precautions are taken. After only a few rounds of serialsubculture, isolates can irreversibly lose the ability to form sporodochia, followed by conidia. The mechanism of these changes is unknown. To understandthe nature of Fusarium morphological changes in culture, we began subjecting the sequenced strain of F. graminearum to serial subculture in July, 2011.Multiple lineages were begun from an initial soil stock, and each lineage is subcultured weekly, and a sample stored under glycerol at -80 C. For this study,we have performed shotgun pyrosequencing using 454 FLX-Plus on five of the lineages from one year after the study began; we have also sequenced onelineage from the beginning of the study (when all lineages should have been identical, and not significantly different from the published F. graminearumgenome), and compared all to the published genome. Finally, we have used Illumina for bisulfite pyrosequencing to obtain methylation profiles.267. Evolutionary and functional analysis of mitosis-related kinase genes in Fusarium graminearum. Huiquan Liu 1 , Jiwen Ma 1 , Shijie Zhang 1 , DaweiZheng 1 , Juanyu Zhang 1 , Chenfang Wang 1 , Jin-Rong Xu 1,2 . 1) College of Plant Protection, Northwest A&F University, Yangling, Shaanxi, China; 2) Departmentof Botany and Plant Pathology, Purdue University, West Lafayette, Indiana, United States of America.Eukaryotic cell cycle is a series of recurrences of a defined set of events; during which, nuclear DNA is replicated in the S phase and segregated into twodaughter nuclei during mitosis. To date, many protein kinases important for the onset and progression through mitosis have been identified. Most of thesemitosis-related kinases are thought to be conserved from yeast to humans. In the model fungi used for cell cycle studies, including Saccharomycescerevisiae, Schizosaccharomyces pombe, and Aspergillus nidulans, the single copy cdc2 (CDC28) CDK gene is a key regulator of cell cycle essential forgrowth. However, the filamentous ascomycete Fusarium graminearum, the causal agent of wheat and barley head blight disease, has two putative Cdc2orthologs. Either one of them is essential but deletion of both may be lethal. Whereas the cdc2B mutant has only minor defects in germination andgrowth, deletion of cdc2A had no obvious defects in growth but resulted in significant reductions in virulence. Ascosporogenesis but not peritheciumformation or ascogenous hyphal growth was blocked in the cdc2A mutant. Mutants deleted of the single CDK kinase gene FgCAK1 had similar phenotypeswith the cdc2A mutant. Both cdc2A and cdc2B interacted with itself and each other, and with FgCAK1. Therefore, cdc2A and cdc2B must have independentand overlapping functions in F. graminearum. It is likely that cell cycle regulation involves different cdc2 kinases and CDK activation mechanisms betweenvegetative and in plant growth. Infectious growth and ascosporogenesis may require only cdc2A, which is activated by FgCAK1. In addition, we found thatF. graminearum has two Aurora protein kinase genes that are orthologous to yeast IPL1. The two F. graminearum Aurora kinases differ at the amino acidresidue that is known to be related to different functions of Aurora A and Aurora B in humans, further indicating that yeast and F. graminearum differ insome key protein kinases involved mitosis. In addition, we systematically identified orthologs of other mitosis-related kinase genes in representative fungi.Although most of them are conserved across fungal tree of life, some mitosis-related kinase genes are lost or duplicated in certain lineages.268. Functional analysis of A MADS-box transcription Mcm1 in Fusarium graminearum . Cui Yang 1 , Guotian Li 2 , Qian Zheng 1 , Meigang Liu 1 , Jin-Rong Xu 1,2 ,Chen fang Wang 1 . 1) NWAFU-PU Joint Research Center, College of Plant Protection, Northwest A&F University, Yangling, Shanxi, China; 2) Department ofBotany and Plant Pathology, Purdue University, West Lafayette, Indiana, United States of America.186

FULL POSTER SESSION ABSTRACTSFusarium head blight, an important disease of wheat and barley is primarily caused by Fusarium graminearum in many parts of the world. In thishomothallic ascomycete, sexual reproduction and the mating type locus play a critical role in its infection cycle because ascospores are the primaryinoculum. In this study we identified and characterized the FgMCM1 gene in F. graminearum that is orthologous to yeast MCM1 MADS-box transcriptionfactor. Deletion of FgMCM1 resulted in the loss of perithecium production and pathogenicity. The Fgmcm1 mutant rarely produced conidia with abnormalmorphology and germination and was defective in response to various stresses. The FgMcm1-GFP fusion proteins localized to the nucleus and fullycomplemented the Fgmcm1 mutant. Interestingly, approximately half of the sub-cultures of the Fgmcm1 mutant often were significantly reduced ingrowth rate. These spontaneously occurred stunted subcultures had similar or more severe defects than the original Fgmcm1 mutant in most of thephenotypes. In yeast two-hybrid assays, FgMcm1 interacted with Mat1-1-1, Fst12, and Tup1 but not with Mat1-2-1. The Fgmcm1 mat1-1-1 double mutantwas stable, suggesting that defects of the Fgmcm1 mutant may be related to the interaction of FgMCM1 with the other MAT TF genes. The Fvmcm1mutants of F. verticillioides had similar defects but were not unstable. To further understand the instability of the Fgmcm1 mutant and slow growthsubcultures, RNA samples isolated from the wild type, original Fgmcm1 mutant, and a stunted subculture were sequenced. RNA-seq data and analyses willbe presented. Overall, our data indicate that FgMcm1 may interact with MAT locus and other transcription factor genes to regulate cell identity and fungaldevelopment and pathogenesis in F. graminearum.269. Genome sequencing of the Fusarium graminearum species complex in Korea. Haeyoung Jeong 1 , Ulrich Güldener 2 , Hee-Kyoung Kim 3 , SeunghoonLee 3 , Theresa Lee 4 , Sung-Hwan Yun 3 . 1) Systems & Synthetic Biology Research Center, KRIBB, Daejeon 305-806, South Korea; 2) Institute of Bioinformaticsand Systems Biology, Helmholtz Zentrum München - German Research Center for Environmental Health, Ingolstädter Landstrabe 1, 85764 Neuherberg,Germany; 3) Dept Med Biotech, Soonchunhyang Univ, Asan, Chungnam 336-745, South Korea; 4) Microbial Safety Team, National Academy of AgriculturalScience, RDA, Suwon 441-707, South Korea.The Fusarium graminearum (Fg) species complex, the causal agent of Fusarium head blight of small grain cereals, comprises at least 15 lineages, orphylogenetically distinct species. Among these, lineages 6 (F. asiaticum) and 7 (F. graminearum sensu stricto) are major populations of the Fg complexrecovered from rice and corn, respectively in Korea; lineages 3 (F. boothii) and 2 (F. meridonale) were also recovered from corn. The F. asiaticumpopulation is clearly different from the F. graminearum population in terms of self-fertility, trichothecenes production, and host preference. We havesequenced the genomes of 19 Fg complex isolates belonging to the 4 clades or species found in Korea using 454 pyrosequencing or Illumina Hiseqtechnologies. As a representative genome sequence for F. asiaticum, we reconstructed five linear replicons from the F. asiaticum SCKO4 strain, whichconsist of four chromosomes, each corresponding to those of the previously sequenced F. graminearum strain PH-1 along with a separated small segment(451kb). By integrating multiple gene models that included the results obtained by ab initio gene prediction tools which incorporated RNA-seq data wemanually identified a set of 12,448 protein-coding genes in SCKO4. Genome-wise comparison between SCK01 and PH-1 revealed a remarkable level ofgenomic synteny throughout the four chromosomes, but several rearrangements including inversions being located on chromosomes II and III in SCK04.Interestingly, the 451-kb fragment in SCK01 showed little sequence relatedness with the PH-1 genome. Similarly, we were able to assemble into 7 largecontigs from the genome of a representative strain (GWS2-6-3) of F. boothii. Using these three representative genomes, we have intensively analyzed andcompared the genomes of the Fg complex field isolates to provide insights into understating of evolutionary relationship among the Fg complex in Korea.270. Identification and functional analysis of virulence genes in different host-pathogenic forms of Fusarium oxysporum. P. van Dam, S.M. Schmidt, M.Rep. Molecular Phyopathology, University of Amsterdam - SILS, Amsterdam, the Netherlands.The species complex Fusarium oxysporum (Fo) represents one of the most abundant and widespread microbes of the soil microflora, including plantpathogenicstrains that, together, are able to infect a broad host range. In the tomato-pathogen Fusarium oxysporum f. sp. lycopersici (Fol), 11 smallSecreted In Xylem (SIX) effector proteins were identified. These were later shown to be encoded on Fol's mobile ‘pathogenicity-chromosome’ that can betransferred horizontally to non-pathogenic Fo strains, resulting in acquired pathogenicity. The goal of this project is to identify host-specific virulencegenes in other formae speciales of Fo.A representative set of isolates from different formae speciales, vegetative-compatibility groups and races will be selected for genome sequencing. Thedevelopment of a comparative genomics bioinformatics pipeline, relying on sequence homology and specific patterns in the promoter regions (e.g.transposable elements) for identification of putative effector genes in newly sequenced F. oxysporum genomes takes a central position in our strategy.Using this approach, several putative effector genes were already identified in a strain of Fo f. sp. melonis, which infects muskmelon.In combination with functional analysis of candidate effector genes, we want to compare the sets of virulence genes of formae speciales that infectmembers of the Cucurbitaceae and Solanaceae families, such as cucumber, muskmelon, watermelon, tobacco, sweet potato and eggplant. Newlydiscovered virulence genes can be used as molecular markers for diagnostic purposes. Additionally, phylogenetic analysis of virulence genes across formaespeciales will help to reconstruct the evolution of host-specific pathogenicity in Fo, and the dynamics of the mobile accessory genome.271. Protocol for generating gene knock-out transformants of the fungal pathogen Verticillium albo-atrum. M. Flajsman, S. Mandelc, B. Javornik. Univ ofLjubljana, Ljubljana, Slovenia.A protocol for generating knock-outs of Verticillium albo-atrum, which is a destructive soilborne fungal pathogen that causes vascular wilt diseases, wassuccessfully established. V. albo-atrum, along with V. Dahliae, is a significant source of crop plant disease, since between them they infect a broadspectrum of host species, from ornamental trees to major crops such as potato, tomato, cotton, tobacco and hop. The genom of V. albo-atrum has alreadybeen sequenced. Translation of genome sequence information into biological functions is therefore possible. One of the most powerful approaches fordissecting the gene function in phytopathogens is the study of the phenotypes of mutants in which a genomic locus has been altered by insertion (genedisruption) or replacement (gene replacement) with heterologous DNA. This is a high throughput reverse genetics approach, which greatly contributes tounderstanding the gene function of fungal pathogens. Our protocol for generating knock-outs of the fungal pathogen V. albo-atrum comprises twomethods: the creation of knock-out plasmids by the USER Friendly cloning technique and transformation of the fungal pathogen by Agrobacteriumtumefaciens-mediated transformation (ATMT). Knock-out strains of V. albo-atrum were made by site directed modifications of the pathogen genome bymeans of homologous recombination and achieved by introducing a DNA fragment containing two homologous recombination sequences flanking aselection marker. pRF-HU2 plasmid, containing a hygromycin resistance gene, was used for USER Friendly cloning of knock-out plasmids. Two PCRamplicons, containing homologous recombination sequences flanking a deletion gene, were inserted into the vector, which was used to transform E. colicells and isolated plasmids were electroporated into A. tumefaciens. V. albo-atrum knock-outs were generated by ATMT. Knock-out strains for two genesfound to be highly expressed at the protein level in the xylem of infected hop plants, were generated. V. albo-atrum knock-out transformants were verifiedby PCR testing and Southern blot analysis, which confirmed that deletion of the target gene had been successful. This is the first report, to our knowledge,of the creation of V. albo-atrum gene knock-outs. It demonstrates that knock-out transformants of this fungal pathogen can be efficiently made.27th Fungal Genetics Conference | 187

FULL POSTER SESSION ABSTRACTS272. Functional analysis of catalase-peroxidase encoding genes in the fungal wheat pathogen Zymoseptoria tritici. A. Mirzadi Gohari 1,3 , R. Mehrabi 1,2 , P.J.G.M. de Wit 2 , G. H.J. Kema 1 . 1) Wageningen University and Research Centre, Plant Research International, P.O. Box, 16, 6700 AA, Wageningen, TheNetherlands; 2) Wageningen University, Laboratory of Phytopathology, Droevendaalsesteeg 1, 6708 PB, Wageningen, The Netherlands; 3) Department ofPlant Protection, College of Agriculture, University of Tehran, Plant Pathology Building, Karaj, Iran.Zymoseptoria tritici is the new name of the foliar wheat pathogen cereal Mycosphaerella graminicola (teleomorph)/Septoria tritici (anamorph) thatcauses septoria tritici blotch particularly in regions with high rainfall and moderate temperatures during the production season. Similar to many plantpathogens, Z. tritici possesses three catalase-peroxidase genes that are known to detoxify H2O2 accumulated in the foliage during colonization. In thecurrent study, we functionally analysed these three catalase-peroxidase genes and found that MgCatD-1, encoding a secreted catalase-peroxidase, playsan important role in the pathogenicity of Z. tritici. MgDCat-1 mutants hardly induced any disease symptoms and expression analysis of MgDCat-1 in plantarevealed that it is up-regulated during pathogenesis, particularly at 8 dpi (days post inoculation). This coincides with an important switch from a biotrophicto a necrotrophic lifestyle during pathogenesis suggesting that this gene is likely required to overcome H2O2-dependant defence responses duringcolonization. Furthermore, the MgDCat-1 strain is hypersensitive to H2O2 as the spore germination dropped to 50% at 4mM H2O2 and to completeinhibition at 6 mM H2O2 compared to the WT IPO323 strain. These results show that secreted catalase-peroxidase is an important pathogenicity factor forsuccessful pathogenesis of Z. tritici.273. Efficient recycling of selective marker genes with the Cre-loxP recombination system via anastomosis in Cryphonectria parasitica. Dong-Xiu Zhang,Donald Nuss. Institute for Bioscience and Biotechnology Research, University of Maryland, Rockville, MD.Reverse genetic analysis has played a significant role in advancing fungal biology, but is also limited by the number of available selectable marker genes(SMGs). The Cre-loxP recombination system has been adapted for use in filamentous fungi to overcome this limitation. Expression of the Cre recombinaseresults in excision of an integrated SMG that is flanked by loxP sites allowing subsequent rounds of transformation with the same SMG. However, currentprotocols for regulated expression or presentation of Cre require multiple time consuming steps. During efforts to disrupt four independent RNAdependentRNA polymerase genes in a single strain of the chestnut blight fungus Cryphonectria parasitica, we tested whether Cre could successfully exciseloxP-flanked SMGs when provided in trans via anastomosis. Stable Cre-producing donor strains were constructed by transformation of wild-type C.parasitica strain EP155 with the Cre coding domain under the control of a constitutive promoter. Excision of multiple loxP-flanked SMGs was efficientlyachieved by simply pairing the Cre donor strain and the loxP-flanked SMGs transformed recipient strain and recovering mycelia from the recipient colonyjust above the anastomosis zone. This method was shown to be as efficient as and much less time consuming than excision by transformation-mediatedexpression of Cre and should be applicable for optimizing reverse genetics analysis in a broad range of filamentous fungi.274. RNA-seq reveals the pleiotropic regulating functions of the transcription factor XYR1 in Trichoderma reesei. Liang Ma 1 , Lei Zhang 1 , Ling Chen 1 , GenZou 1 , Chengshu Wang 2 , Zhihua Zhou 1 . 1) Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for BiologicalSciences, Chinese Academy of Sciences, Shanghai, China; 2) Key Laboratory of Insect Developmental and Evolutionary Biology, Institute of Plant Physiologyand Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.Trichoderma reesei is a well-known cellulase producer and widely applied in enzyme industry. XYR1 has been demonstrated to be the main transcriptionactivator of cellulase and hemicellulase gene expression in T. reesei. To comprehensively investigate the genes regulated by XYR1, RNA-seq was used todetermine the transcriptional profile of T. reesei Rut-C30 and the Dxyr1 strain cultured on either cellulose or glucose. A total of 467 distinct genes wereclassified as differentially expressed comparing parent strain with the xyr1-deleted strain when induced by cellulose, among which were mainly thoseinvolved in carbohydrate metabolism, lipid metabolism, protein fate and solute transport. Almost all the functional cellulase genes were found to bedownregulated in the Dxyr1 strain while all the differentially expressed protease genes were upregulated. Furthermore, the expression of 84 genesidentified as cellulose-induced were significantly impaired when xyr1 was deleted. In contrast, 281 genes showed expression difference in the Dxyr1 strainwhen cultured on glucose as carbon source, and most of these were involved in solute transport, lipid metabolism, secondary metabolism and amino acidmetabolism. In addition, we also found that the functional restoration of XYR1 in the Dxyr1 strain required the homologous insertion of xyr1 at its originallocus. Our study provides the global insight into the genes regulated by XYR1 beyond the hydrolase-encoding genes in T. reesei and might aid futurestudies to improve its cellulase production.275. Tyrosinase an important enzyme for melanin production in the oomycete Saprolegnia parasitica. Marcia Saraiva 1* , Irene de Bruijn 2 , Laura Grenville-Briggs 3 , Debra McLaggan 1 , Vincent Bulone 3 , Pieter van West 1 . 1) School of Medical Sciences, University of Aberdeen, Aberdeen, United Kingdom; 2)Laboratory of Phytopathology, Wageningen University, 6709 PD Wageningen, Netherlands; 3) KTH - Royal Institute of Technology, School of BiotechnologyAlbaNova University Center SE-106 91 Stockholm Sweden.Filamentous fungi posses a pigment that often is considered an indirect virulence factor giving the fungi leverage. The pigment, melanin, acts as aprotective agent against several threats and hazards that these organisms may encounter such as oxidants, killing by macrophages, UV and alsoantimicrobial compounds. In the biosynthetic pathway of melanin Tyrosinase is the first enzyme involved. Tyrosinases are widely distributed in nature,where they have been found in prokaryotes, eukaryotic microorganisms, invertebrates, plants and mammals. Here we describe the first functionalcharacterisation of a tyrosinase (SpTyr) from the fish pathogenic oomycete Saprolegnia parasitica. This aquatic water mould infect a wide range of fish,amphibians and crustaceans that are relevant to the aquaculture industry and aquatic ecosystems, causing a disease called Saprolegniosis. This disease isresponsible for millions of losses in aquacultures. Previously we found that SpTyr is highly expressed in sporulating mycelium. After developing andimplementing a transient gene silencing method (RNAi), based on the delivery of in vitro synthesized dsRNA into protoplasts of this water mould, weobtained SpTyr-silenced lines with a significantly decreased tyrosinase activity of 40-60% compared to control lines. The tyrosinase activity correlateddirectly with the level of SpTyr-silencing in the transient lines, which ranged from 68.7-37.5%. Furthermore, the melanin content was measuredspectrophotometrically in the SpTyr-silenced lines and found to be significantly reduced to 2-70%. Microscopic observations of SpTyr-silenced linesresulted in aberrant zoosporangium formation, with less pigment and abnormal morphology. Moreover, our results demonstrate that transient genesilencing can be successfully used to functionally characterise genes in S. parasitica and can provide a high-throughput tool for S. parasitica functionalgenomics.276. Comparative transcriptomics of Cordyceps bassiana to understand expression levels of its NRPS related genes. B. Shrestha 1 , J. Oh 2 , M.-W. Hyun 3 , J.-G. Han 1 , H.-W. Kwon 3 , S.-H. Hyun 2 , S. H. Kim 3 , H.-K. Choi 2 , G.-H. Sung 1 . 1) Mushroom Research Division, Rural Development Administration, Suwon 441-707,Republic of Korea; 2) College of Pharmacy, Chung-Ang University, Seoul 156-756, Republic of Korea; 3) Department of Microbiology and Institute of Basic188

FULL POSTER SESSION ABSTRACTSSciences, Dankook University, Cheonan 330-714, Republic of Korea.Cordyceps bassiana (= Beauveria bassiana) is a widely distributed entomopathogenic fungus, particularly important in the industry of biological controlagents against agricultural pests and considered as a model organism for understanding fungal entomopathogenicity. We conducted a genome datamining of C. bassiana C101 isolate for nonribosomal peptide synthetase (NRPS) and PKS-NRPS hybrid gene clusters. To identify NRPS related gene clusters,whole genome sequence of C. bassiana C101 was subjected to antiSMASH program with default settings. Their domain sequences were predicted basedon fungal-specific HMMER models using databases of Pfam and InterPro. A domain specificity signatures were derived by NRPSpredictor2 while substratespecificity prediction of the acyltransferase (AT) domains was based on web server SBSPKS. We identified 16 NRPS, 5 NRPS-like and 3 PKS-NRPS hybridgene clusters, of which three are known for their metabolites (i. g., beauvericin, bassianolide and tenellin). For transcriptome analysis of NRPS relatedgenes using Illumina RNA-Seq, C. bassiana C101 isolate was inoculated on SDAY, iron-, nitrogen- and lipid-rich media at 25°C and harvested to obtain itsmycelia after two weeks of incubation. The stromata and perithecia of C. bassiana were also cultivated on brown rice medium and harvested from 4 weeksto 8 weeks old cultures at weekly intervals. As a result, beauvericin synthetase, a NRPS synthesizing insecticidal compound beauvericin, was highlyexpressed at mycelium grown in dark on SDAY but not in stromata except very low expression at 8 weeks old stromata. Conversely, another NRPSbassianolide synthetase, synthesizing insecticidal compound bassianolide, was not detected under any condition except very low detection in 8 weeks oldstromata. Tenellin synthetase, a PKS-NRPS hybrid, was expressed only in stromata. Some NRPSs showed expression in both mycelia and stromata whileothers were expressed either in mycelia or stromata. In particular, the expression levels of beauvericin synthetase are correlated with the metabolicprofiling of beauvericin in mycelia and stromatal conditions, indicating that the detailed information of the expression levels of NRPSs can be used todiscover the metabolic diversity of C. bassiana with the further study of gene function and comparative metabolic profiling.277. Global analysis of the Colletotrichum gloeosporioides genome and transcriptome reveals a conserved role for pacC pH regulation in fungi. NoamAlkan 1 , Xiangchun Meng 3 , Eli Reuveni 1 , Gilgi Friedlander 1 , Serenella Sukno 4 , Michael Thon 4 , Robert Fluhr 1 , Dov Prusky 2 . 1) Plant Sciences, WeizmannInstitute of Science, Rehovot, Israel; 2) Department of Postharvest Science of Fresh Produce, Agricultural Research Organization, the Volcani Center, BetDagan, Israel; 3) Fruit Tree Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou, China; 4) Department of Microbiology andGenetics, University of Salamanca, Villamayor, Spain.Colletotrichum gloeosporioides, a widely distributed economically important agent in postharvest fruit disease, thrives by massive secretion of ammonia.The alkalized environment hijacks host response and activates fungal pathogenicity genes. In contrast, other types of fungi acidify the environment fortheir optimal growth. In both cases the pacC transcription factor, with homologs in yeasts and fungi, exerts critical pH control of gene expression. Toexplore global aspects of pH-dependent gene expression, we sequenced the C. gloeosporioides genome and compared transcriptomes of WT and a DpacCmutant to examine pacC regulation. The 54 Mb C. gloeosporioides genome comprises 16,603 transcripts. Transcriptome analysis of the mutant showedthat DpacC regulates more than 5% of the fungal genome including; transporters to maintain cellular homeostasis, cell wall degrading enzymes to optimizepathogenicity and GATA-like transcription factors. The predictions were verified by monitoring gene expression in different media and during fruitinfection a well as by pathogenicity assessment of selected deletion mutants. Analyses showed over-representation of pacC binding sites in the promotersof the pacC up-regulated genes. However, in the promoters of the down-regulated genes GATA-like binding sites were dominant. The results suggestduality in global pacC control; direct regulation of alkaline-induced transcripts but indirect regulation, by activation of other transcription factors to downregulateacid expressed transcripts. To establish the generality of this scenario, conservation of pacC distribution was examined in genes selected byhomology from 5 different fungi that have contrasting alkaline or acidifying pathogenicity strategies. The results showed that irrespective of pathogencolonization strategy, the homologs of up-regulated genes had over-representation of pacC binding sites. Significantly, the homologs of down regulatedgenes revealed cross-genome over-representation of GATA transcription factor binding sites. Thus, regulation by pacC is a phylogenetically conservedfungal mechanism exerting dual pH control for maintaining homeostasis and pathogenicity in changing environments.278 WITHDRAWN279. Anisogramma anomala: a unique fungus with a huge genome. Guohong Cai, Thomas Molnar, Bradley Hillman. Plant Biology and Pathology, RutgersThe State University of New Jersey, New Brunswick, NJ.Anisogramma anomala (Ascomycota: Diaporthales) is the causal agent of hazelnut eastern filbert blight (EFB). This obligate and biotrophic fungus onlyreproduces sexually through ascospore. EFB is the most important disease of commercially grown hazelnut (European hazelnut), Corylus avellana, in theUnited States. It only causes minor disease on its natural host, the American hazel C. Americana, which is an understory shrub commonly found indeciduous forests in Northeast region of North America. Despite quarantine efforts, EFB was found in Washington in the late 1960s and reached Oregon’sWillamette Valley, the main hazelnut production area, in 1986. Commercial cultivars carrying a single locus from cultivar Gasaway confer completeresistance to EFB in the west coast. However, certain strains from the east can break the resistance. We sequenced an Oregon strain using Illumina GA IIXplatform. Approximately 26M 146 bp paired-end reads were used to produce a draft assembly with contig N 50 of 10,384 bp and scaffold N 50 of 32,987 bp.Excluding gaps, total assembly was 333.6 Mb. This is huge for a fungus with single nucleus. The genome size produced by draft assembly was corroboratedby flowcytometry, which measured the genome size at approximately 370 Mb. The genome has low GC ratio of 32%. Approximately 85% of assembledgenome is repetitive, and AT-rich sequences are enriched in the repetitive regions. We are exploring the genome sequences looking for answers to theuniqueness of this fungus.280. Leptosphaeria maculans 'brassicae': "Transposable Elements changed my life, I feel different now". Jonathan Grandaubert 1 , Conrad Schoch 2 ,Hossein Borhan 3 , Barbara Howlett 4 , Thierry Rouxel 1 . 1) INRA-BIOGER, Thiverval-Grignon, France; 2) NCBI, National Institutes of Health, Bethesda, MD, USA;3) AAF Saskatoon, Canada; 4) School of Botany, University of Melbourne, Australia.The Dothideomycetes phytopathogens Leptosphaeria maculans and Leptosphaeria biglobosa form a complex of 8 species and putative subspeciessuggested to have diverged “recently”. In 2007, the sequencing of an isolate of Leptosphaeria maculans 'brassicae' (Lmb) provided the first referencegenome for this fungus. The 45-Mb genome has an unusual bipartite structure, alternating large GC-equilibrated and AT-rich regions. These AT-rich regionscomprise one third of the genome and are mainly composed of mosaics of truncated Transposable Elements (TEs) postulated to have “invaded” thegenome 5-10 MYA; they also comprise 5% of the predicted genes of which 20% encode putative effectors. In these regions, both genes and TEs areaffected by Repeat Induced Point mutation (RIP). To investigate when and how genome expansion took place in the evolutionary series, and theconsequences it had on fungal adaptability and pathogenicity, the genomes of five members of the species complex showing contrasted host range andinfection abilities were sequenced. In silico comparison of the reference genome with that of 30-32-Mb genome of L. maculans 'lepidii' (Lml), L. biglobosa'brassicae', L. biglobosa 'thlaspii' and L. biglobosa 'canadensis', showed these species have a much more compact genome with a very low amount of TEs27th Fungal Genetics Conference | 189

FULL POSTER SESSION ABSTRACTS(

FULL POSTER SESSION ABSTRACTS286. Genome sequencing, assembly and annotation of three marine fungal isolates using different next generation DNA sequencing methods forpharmaceutically important secondary metabolites. Abhishek Kumar, Frank Kempken. Dept. of Genetics & Mol. Bio, Institute of Botany, CAU Kiel, KIEL,SH, Germany.Next-generation sequencing (NGS) techniques have changed the facets of genomics and its application. Until now marine fungal isolated are neglected infungal genetics, largely due to fact that fungi have enormous diversity on land itself. We aimed to explore marine fungal isolates and their encodingnatural products as drugs against cancer under the EU-funded project marine fungi (www.marinefungi.eu). We have taken three marine isolates namely,Scopulariopsis brevicaulis, Pestalotiopsis sp. and Calcarisporium sp. We have established the genomic sequences from these marine isolates of using threedifferent next-generation sequencing methods (Roche 454, Illumina and ion-torrent) and predicted genes are presently in process of validation usingillumina based RNA-seq. We are also comparing wild type phenotypes with higher -yielding mutants of these fungi with special interest on specific naturalcompound. The assembled genome of Scopulariopsis brevicaulis is ~32 Mb in size with N50 equals to 88 kb and 935 contigs containing 16298 genes withaverage intron length equals to 129.4. During annotation process, we were able to annotate 9340 genes (57.31 %) while 6958 genes (43.69 %) remainednon-annotated in Scopulariopsis brevicaulis genome. This genome has 17 genes encoding for non-ribosomal peptide synthetases (NRPSs), 18 polyketidesynthases (PKSs) and one gene is hybrid NRPS-PKS. Similarly, the genome size for Pestalotiopsis sp. is~46 Mb with N50 equals to 71.9 kb and 4186 contigscontaining 23492 genes, which is surprisingly very high for a fungus. The average intron length and the average intron per gene are 126.8 and 2.2,respectively. During annotation process, we annotated 60% genes of Pestalotiopsis genome with 44 NRPSs, 62 PKSs and 7 hybrid NRPS-PKS genes. Theassembled genome size of as Calcariosporium sp. is about 35 Mb genome with N50 equals to 91.9 kb and 2464 contigs containing 15459 genes. Thepercentage GC% for this genome is 50.7%. The average intron length and the average intron per gene are 121 and 2.1, respectively. During annotationprocess, we annotated 72% genes, while 28% genes remained non-annotated for Calcariosporium genome with 52 NRPSs, 66 PKSs and 7 hybrid NRPS-PKSgenes.287. From Shiitake genomics to comparative mushroom genomics: Mushroomics. Hoi Shan Kwan, Chun Hang Au, Man Chun Wong, Jing Qin, Yung YungLee, Kin Sing Wong, Lei Li, Qianli Huang, Wenyan Nong, Man Kit Cheung, Jinhui Chang, Xuanjin Cheng. School of Life Sciences, The Chinese University ofHong Kong, Hong Kong, China.The era of mushroom "omics" has come amongst the mushrooming of "omics" in all life science fields. I propose to name “omics” of mushrooms"Mushroomics". My laboratory has been working on the genomics and transcriptomics of the wood-degrading fungus Lentinula edodes, Shiitakemushroom, one of the most important cultivated mushrooms. We sequenced the genome of the monokaryon L54A using Roche 454 and ABI SOLiDsequencing platforms. Over 13,000 protein-coding genes were predicted. We constructed a high-density genetic linkage map that was useful to linkscaffolds into super-scaffolds. The L. edodes genome assembly revealed a genome size of about 40 Mb. We performed RNA-Seq of multiple stages of L.edodes. For comparison, we also analyzed the transcription profiles of different stages of the model mushroom Coprinopsis cinerea using NimbleGenmicroarrays. Genes differentially expressed during fruiting body initiation and development in these mushrooms were identified. We conductedcomparative analyses on publicly available genome sequences of basidiomycetes and ascomycetes, and revealed genes expanded in genomes ofmushroom-forming fungi. The expanded genes included specific types of regulators, ubiquitin ligases, protein-binding proteins, protein kinases, andtranscription factors. In particular, F-box and paracaspase domain proteins were significantly expanded. The cataloging of the unique composition of plantbiomass-degrading enzymes in L. edodes genome revealed lignin-degrading laccases and manganese peroxidases, and multiple polysaccharide-degradingenzyme families, such as glycoside hydrolase families that target beta-glucans and pectin. We compiled the genome sequences of L. edodes and otherfungi into an Ensembl-based platform, equipped with a battery of genomic analysis tools, for comparative mushroom genomic analysis. Our works havegenerated rich resources for the analysis of genomics and transcriptomics of mushroom-forming fungi. Our analysis also provided insights into themolecular mechanisms of fruiting body development in fungi and the evolution of fungal complex multicellularity. Indeed, our works showed that the eraof "Mushroomics" has arrived.288. Comparative genomics of Ceratocystis polonica and Ophiostoma bicolor, two bark beetle-associated pathogenic fungi. Ljerka Lah 1 , Tom Hsiang 2 ,Colette Breuil 3 , Joerg Bohlmann 4 , Radovan Komel 1,5 . 1) Lab. for Mol. Biol, National Inst. of Chemistry, Ljubjana, Slovenia; 2) School of EnvironmentalSciences, University of Guelph, Guelph, Canada; 3) Dept. of Wood Science, Faculty of Forestry, University of British Columbia, Vancouver, Canada; 4)Michael Smith Laboratories, University of British Columbia, Vancouver, Canada; 5) Institute of Biochemistry, Faculty of Medicine, University of Ljubljana,Slovenia.Outbreaks of bark beetles and their pathogenic fungal associates are responsible for killing conifer trees and destroying forests worldwide. For example,in British Columbia, the mountain pine beetle and its major associated fungal pathogen Grosmannia clavigera have destroyed over 17 million hectares ofpine forests. The sequenced genome of G. clavigera has provided insight into fungal mechanisms involved in resistance to conifer defense compounds, andinto population genomics of this species. In Europe, where outbreaks of the European spruce bark beetle (Ips typographus) threaten Norway spruce (Piceaabies), genomic resources are lacking for pathogenic fungal associates, Ceratocystis polonica and Ophiostoma bicolor. The aim of our work is to createthese resources and to compare the genomes of these species with the annotated G. clavigera genome. The sizes of C. polonica and O. bicolor genomes,sequenced using Illumina and assembled using Abyss ver. 1.3.4, are 34.7 and 27 Mb, respectively. We identified (with Augustus ver. 2.6.1) and functionallyannotated 6405 genes in C. polonica and 7746 in O. bicolor. We report results from preliminary analyses on gene family evolution, and on genes that codefor enzymes important in eliminating and modifying conifer defense compounds, and those involved in specialized metabolism. The number of genescoding for ABC-transporters is approximately the same in all three species, while G. clavigera has more cytochrome P450 genes, polyketide synthases andribosomal peptide synthases than either European bark beetle associate. Our results lead us to hypothesize that despite the fact that these species occupyapparently similar ecological niches, there are differences in their metabolism of host defense compounds and specialized metabolism.289. The largest fungal mitochondrial genome of a basidiomycete contains signs of genetic flexibility and recombination events. Taina K. Lundell 1 , HeikkiSalavirta 1 , Ilona Oksanen 1 , Jaana Kuuskeri 1 , Miia Mäkelä 1 , Pia Laine 2 , Lars Paulin 2 . 1) Microbiology and Biotechnology, Department of Food andEnvironmental Sciences, Viikki Campus, University of Helsinki, FI-00014 Helsinki, Finland; 2) Institute of Biotechnology, DNA Sequencing and GenomicsLaboratory, Viikki Campus, University of Helsinki, FI-00014 Helsinki, Finland.Background and results. As the first part of de novo genome sequencing of the biotechnologically important, wood-decaying enzyme producing whiterotBasidiomycota species Phlebia radiata, we first assembled and gene annotated its mitochondrial genome (mtDNA) using 454 sequencing. The P.radiata mtDNA of 157 kb in size is the largest mitochondrial genome among fungi sequenced and characterized so far. The genome assembled as a singlecircular dsDNA molecule containing over 100 open reading frames. However, almost 80% of the mt genome is comprised of non-coding, intergenic andintronic sequence regions. Genes for mt SSU and LSU rRNA, 28 tRNAs, and fifteen genes encoding the conserved protein subunits in the mitochondrial27th Fungal Genetics Conference | 191

FULL POSTER SESSION ABSTRACTSinner membrane complexes I, III, IV and V are identified. Additional protein-coding ORFs (sum 39) are predicted including a gene for ribosomal protein(rps3), a viral RNA-directed DNA polymerase (reverse transcriptase), and a gene for bacterial-originated DNA-directed DNA polymerase II of family B(dpoB). Total of 57 intron-homing endonucleases with core LAGLIDADGD and GYI-YIG domains were recognized in over 30 group I and II type introns, up to3.4 kb in length, in ten of the fifteen conserved genes (cox1,2,3; cob; nad1,2,4,4L,5; rnl). Multigene phylogeny of the conserved proteins confirms currentfungal taxonomy and a common, single origin of the mtDNA within Basidiomycota. Conclusions. The exceptionally large mt genome is explained by longintergenic stretches of DNA carrying repetitive and partially overlapping sequence elements, presence of additional open reading frames with unknownfunction, existence of the 6.1 kb duplication-inversion, and due to frequent intron splicing of the coding sequences. A few of the qualities indicate plasmidor viral origin, such as the dpoB, and the cob gene-interrupting long group II intron with reverse-transcriptase ORF. These features together with theduplicated inversion and dense repeat stretches, and the long introns with intron-associated homing endonucleases are indications of genetic flexibility,not previously recognized to such extent in fungal mitochondrial genomes. Thus, it may be concluded that DNA recombination as well as regulation ofgene transcription are allowed and on-going events in the P. radiata mt genome.290. De Novo Assembly of Fungal Genomes and Detection of Structural Variation using Extremely Long Single-Molecule Imaging. Nicholas R. Rhind 1 ,Alex Hastie 2 , Ernest Lam 2 , Andy Nguyen 2 , Han Cao 2 . 1) Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, WorcesterMA; 2) BioNano Genomics, San Diego CA.De novo genome assemblies using only short read data are generally incomplete and highly fragmented due to the intractable complexity found in mostgenomes. This complexity, consisting mainly of large duplications and repetitive regions, hinders sequence assembly and subsequent comparativeanalyses. We have overcome these problems in four fungal genomes by assembling whole genome shotgun (WGS) contigs with a high resolution longrange physical-mapping strategy. We used a single molecule genome analysis system (Irys) based on NanoChannel Array technology that linearizesextremely long DNA molecules for observation. This high-throughput platform automates the imaging of single molecules of genomic DNA hundreds ofkilobases in size to measure sufficient sequence uniqueness for unambiguous assembly of complex genomes. High-resolution genome maps assembled denovo from the extremely long single molecules retain the original context and architecture of the genome, making them extremely useful for structuralvariation and assembly applications.We have built full genome assemblies of the four fission yeast genomes: Schizosaccharomyces pombe, S. japonicus, S. octosporus and S. cryophilus. The S.pombe assembly is similar to the published finished S. pombe genome, but reveals several complicated genomic features that were mis-assembled by aclone-based approached. In addition, we have resolved centromeric and telomeric repeats that had not been assembled by traditional approaches. The S.japonicus and S. octosporus assemblies identify a number of mis-assemblies in the published WGS genomes. The S. cryophilus genome is the first genomescaleassembly of what was a draft assembly of WGS contigs.Our strategy combining genome map-based scaffolding with deep sequencing offers an integrated pipeline for whole genome de novo assembly solvingmany of the ambiguities inherent when using sequencing alone. Additionally, genome maps serve as a much-needed orthogonal validation method toWGS assemblies. As a result, genome maps improve contiguity and accuracy of whole genome assemblies, permitting a more comprehensive analysis offunctional genome biology and structural variation.291. Discovering host specificity candidate genes of Sporisorium reilianum by genotyping mixed-variety offspring. T. Wollenberg 1,2 , J. Donner 2 , J.Schirawski 1,2 . 1) Microbial Genetics, RWTH Aachen University, Aachen, Germany; 2) Molecular Biology of Plant-Microbe Interaction, Georg-August-University Göttingen, Göttingen, Germany.The biotrophic plant pathogenic basidiomycete Sporisorium reilianum exists in two host-specific varieties, S. reilianum f. sp. zeae (SRZ) and S. reilianum f.sp. reilianum (SRS). SRS causes head smut of sorghum and leads to weak symptoms on maize, such as phyllody in the inflorescences. SRZ causes head smutof maize and leads to the formation of red phytoalexin-containing spots on inoculated sorghum leaves. Plant infection results after pairwise mating ofcompatible haploid cells that fuse to form an infective dikaryotic filament. The fungus persists in the host plant until flowering time, and in theinflorescences develops into diploid spores that germinate by meiotic division to haploid cells. Haploid cells of the two varieties are mating competent andare able to infect both maize and sorghum with a very low infection rate. To identify the genetic basis for the difference in host specificity, we analyzedvirulent and non-virulent segregants of a mixed-variety (SRZ x SRS) infection both phenotypically and genotypically by a PCR-based approach. Weidentified twelve chromosomal regions that were associated to the phenotypic behavior of the strains. This shows that host adaptation is a multi-genictrait. To identify associated genes we genotypically analyze a larger set of strains by genome-wide SNP comparison after Illumina re-sequencing. Genomecomparison of mixed-variety offspring is a powerful tool to discover candidate genes involved in host specificity.292. Comparing comparative “omics” in Coccidioides spp. Emily A. Whiston, John W. Taylor. Plant & Microbial Biology, U.C. Berkeley, Berkeley, CA.The mammalian pathogens Coccidioides immitis and C. posadasii are the only dimorphic fungal pathogens that form spherules in the host. Furthermore,all of Coccidioides’ closest known relatives are non-pathogenic. In this project, we are interested in genome changes between the Coccidioides lineage andits relatives, and how these changes compare to recently published comparative and population genomics, and transcriptomics studies in Coccidioides.Coccidioides and its closest sequenced relative, Uncinocarpus reesii, are estimated to have diverged 75-80 million years ago. Here, we have sequenced thegenomes of four species more closely related to Coccidioides than U. reesii: Byssoonygena ceratinophila, Chrysosporium queenslandicum, Amauroascusniger and A. mutatus. For each of these four species, we prepared genomic DNA Illumina sequencing libraries; the resulting genome assemblies rangedfrom 23-34Mb, with N50 of 90kb-205kb. Predicted genes were confirmed by RNAseq; the total number of genes ranged from 8,179-9,184. We assessedindividual gene gain/loss, and gene family expansion/contraction in Coccioides using these new genomes and other recently published genomes from theOnygenales order, including the yeast-forming dimorphic pathogens Histoplasma and Paracoccidioides, and the dermatophytes Microsporum andTrichopyton. We have compared these results to genes identified in recently published Coccidioides “omics” studies that show evidence of positiveselection, introgression and/or differential expression.293. The transcriptional response during cell-fusion incompatibility in Podospora anserina. Frédérique Bidart, Sven J. Saupe, Corinne Clavé. IBGC, CNRS,Bordeaux, France.Heterokaryon incompatibility is a form of non-self recognition common in filamentous fungi that occurs when filaments of different isolates of the samespecies fuse. Compatibility is controlled by so-called het loci and fusion of strains of unlike het genotype triggers a complex incompatibility reaction thatleads to the death of the fusion cell. Herein, we analyze the transcriptional changes during the incompatibility reaction in Podospora anserina. Theincompatibility response was found to be associated with a massive transcriptional re-programming: 2248 genes were up-regulated by a factor 2 of moreduring incompatibility. In turn, 2463 genes were down-regulated. HET and NACHT domains previously found to be involved in the control of heterokaryon192

FULL POSTER SESSION ABSTRACTSincompatibility were highly enriched in the up-regulated gene set. In addition, incompatibility was characterized by an up-regulation of secondarymetabolism clusters, toxins and effector-like proteins, proteolytic and other hydrolytic activities. Genes for ribosome synthesis and energy productionwere in contrast down regulated. There was a significant overlap between regulated genes during incompatibility in P. anserina and N. crassa indicatingcommunality in the incompatibility responses in these two species. In P. anserina, the up regulated set was found to be enriched for proteins lackingorthologs in other species and chromosomal distribution of the up-regulated genes was uneven with up regulated genes enriched in genomic islands andcertain chromosomes. Globally, this study shows that transcriptome changes occurring during cell fusion incompatibility in P. anserina are massive andpleiotropic and in several aspects related to host-pathogen interactions described in other fungal species.294. Comparative Analysis of Putative Rhodopsins in Early Diverging Fungal Lineages. Steven Ahrendt 1 , Edgar Medina 1,2,3 , Jason Stajich 1 . 1) PlantPathology & Microbiology, University of California, Riverside, Riverside, CA; 2) Departamento de Ciencias Biológicas, Universidad de Los Andes, Bogotá,Colombia; 3) University Program in Genetics & Genomics and Department of Biology, Duke University, Durham, NC.Species belonging to the early diverging zoosporic fungal lineages (Blastocladiomycota, Chytridiomycota, Cryptomycota, and Neocallimastigomycota)reproduce via motile uniflagellated spores. Previous work has shown that some of these zoosporic fungi are phototaxic [1]; however, light sensing inzoosporic fungi has not been fully explored. The opsins are a broad class of photosensitive, seven-transmembrane G-protein coupled receptor proteins.One sub-class of opsin, the type 2 Rhodopsins, has previously only been identified in metazoan lineages [2]. Here we describe the identification andstructural/functional analyses of a putative type 2 rhodopsin in several species of recently sequenced zoosporic fungi: Batrachochytrium dendrobatidis,Spizellomyces punctatus, Allomyces macrogynus, Rozella allomycis, Gonapodya prolifera, and Homolaphlyctis polyrhiza. Computational modeling of the B.dendrobatidis and S. punctatus proteins indicates that they both adopt the seven-transmembrane helix conformation typical of GPCRs. Additionalobserved motifs are the so-called “ion lock” and conservation of the retinal binding pocket. The B. dendrobatidis protein sequence is notably lacking theconserved lysine residue, however this residue is present in the S. punctatus sequence. The number of identified Ga proteins is roughly consistent amongthe basal lineages, the Zygomycetes, and the Dikarya. Comparative genomics analyses of rhodopsin and flagellar genes in the basal lineages, Zygomycetes,and Dikarya show a correlation of flagellum and rhodopsin presence across the fungi, suggesting an evolutionary linkage between light-sensing andmotility during the transition from aquatic to terrestrial lifestyles. [1] Saranak & Foster. Nature. 1997. [2] Spudich et al. Ann. Rev. Cell Dev. Biol. 2000.295. Molecular Tools to Silence and Confirm Genes in Phytophthora Sojae. Felipe R. Arredondo 1 , Brett M. Tyler 1 , Shiv D. Kale 2 . 1) Botany and PlantPathology, Oregon State University, Corvallis, OR; 2) Virginia Bioinformatics Institute, Virginia Tech. Blacksburg, VA.Bioinformatics analysis is a powerful tool that decreases the number of possible leads to confirm the function of an interesting gene. These leads are onlyvirtual and theoretical but still have to be proven correct and accurate. PEG/ Protoplast transformation and particle bombardment transient expressionhave been part of Phytophthora molecular research for many years and valuable tools in gene identification and confirmation of function. Possibly thereare many hundreds of genes in Phytophthora species involved during the infection mechanism; the function of the majority of these virulence genes islargely unknown. Introducing genes into P. sojae via PEG/Protoplast transformation is a reliable tool that can confirm gene function by over-expression orsilencing. Another reliable tool is particle bombardment with the PDS1000 to transiently express intact or modified genes into soybean leaves. In thisposter I will introduce the mechanics of these powerful tools.296. Functional Characterization of Transcription Factor Genes, MoNIT4 and MoLEU3, in Magnaporthe oryzae. Jaehyuk Choi 1 , Soyeon Yoo 2 , Yong-HwanLee 1,2,3 . 1) Center for Fungal Pathogenesis; 2) Department of Agricultural Biotechnology; 3) Center for Fungal Genetic Resources, Plant Genomics andBreeding Institute, and Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Korea.Rice blast disease caused by Magnaporthe oryzae is one of the most destructive threats to global rice production. Nitrogen metabolism has shown toplay an important role in pathogenicity of this fungus. Here, we characterized two genes encoding putative transcription factors belonging to theZn(II) 2Cys 6 family, MoNIT4 and MoLEU3, homologs of nit-4 in Neurospora crassa and leu3p in Saccharomyces cerevisiae, respectively. The DMonit4 mutantshowed reduction in conidiation and mycelial growth under nitrogen starvation compared to the wild-type. Addition of ammonium restored the growthdefects in the mutant. The expression of nitrate and nitrite reductase genes was significantly reduced in the mutants, suggesting that these genes areunder control of MoNIT4. The DMoleu3 mutant exhibits severe defects in conidiation and pathogenicity as well as mycelial growth under nitrogenstarvation. Addition of leucine complemented the defects in conidiation and mycelial growth in the DMoleu3 mutant. The decreased expression of 3-isopropylmalate dehydratase, 3-isopropylmalate dehydrogenase, and NADP-specific glutamate dehydrogenase genes supports that MoLEU3 functions as aregulator for leucine biosynthesis and ammonia assimilation pathways. Taken together, these findings will help to understand the nitrogen metabolismnetwork in M. oryzae and its role in the development of the rice blast disease.297. Comparative genomic analysis of world-wide Magnaporthe oryzae isolate collection. Jaeyoung Choi 1 , Gir-Won Lee 2 , Sook-Young Park 3 , JunhyunJeon 1 , Jaehyuk Choi 3 , Ki-Tae Kim 1 , Yong-Hwan Lee 1,3,4 . 1) Department of Agricultural Biotechnology, Seoul National University, Seoul 151-742, Korea; 2)Department of Bioinformatics and Life Science, Soongsil University, Seoul 156-743, Korea; 3) Center for Fungal Pathogenesis; 4) Center for Fungal GeneticResources, Plant Genomics and Breeding Institute, and Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul 151-921,Korea.Since the genome sequence of Magnaporthe oryzae 70-15 was released, functional and comparative genomic researches have been extensively carriedout. However, characteristics of essential genes in M. oryzae are largely unknown. In this study, we sequenced 39 M. oryzae isolates collected from worldwideand analyzed to find the set of core genes. The average assembly size and the total number of predicted genes in the 39 M. oryzae genomes were37Mb and 14,517 genes, respectively. We found that a total of 5,148 genes turned out to be conserved at the amino acid level (identity ³ 80%) among the40 M. oryzae genomes including strain 70-15. To investigate a set of the core genes, we applied three different databases: Fungal Transcription FactorDatabase (FTFD; http://ftfd.snu.ac.kr/), Fungal Cytochrome P450 Database (FCPD; http://p450.riceblast.snu.ac.kr/) and Fungal Secretome Database (FSD;http://fsd.snu.ac.kr/). In addition, three in-house databases were used for the prediction of cell wall-degrading enzymes (CWDEs), peroxidases andlaccases. As a result, we found a core set of genes in M. oryzae: 932 secreted proteins, 269 transcription factors, 33 cytochrome P450 genes, 18 CWDEs, 17peroxidases and 5 laccases. Furthermore, we discovered an overlap of 38 genes between the above groups which have secretory potential. To archive andmanage those newly sequenced genomes, we developed the Magnaporthe Atlas (http://www.magnaporthe.org/) as a web-based solution. Our core geneset of M. oryzae will facilitate discovery of lineage-specific innovations with implications in coevolution with specific host cultivars.298. Functional characterization of two genes encoding putative Zn(II) 2Cys 6 transcription factors, MoCOD1 and MoCOD2 in Magnaporthe oryzae.Hyunjung Chung 1 , Sook-Young Park 2 , Jeahyuk Choi 2 , Junhyun Jeon 1 , Yong-Hwan Lee 1,2,3 . 1) Department of Agricultural Biotechnology; 2) Center for Fungal27th Fungal Genetics Conference | 193

FULL POSTER SESSION ABSTRACTSPathogenesis; 3) Center for Fungal Genetic Resources, Plant Genomics and Breeding Institute, and Research Institute for Agriculture and Life Sciences,Seoul National University, Seoul 151-921, Korea.Transcription factors (TFs) play pivotal roles in regulation of gene expression during cellular processes. The rice blast fungus, Magnaporthe oryzaeundergoes a series of morphological changes during the infection. To elucidate the roles of TFs in development of rice blast disease, two Zn(II) 2Cys 6 TFgenes, MoCOD1 and MoCOD2, were characterized. Both DMocod1 and DMocod2 mutants showed defects in conidiation and pathogenicity. Reducedpathogenicity of the DMocod1 mutant was due to defects in invasive growth while the DMocod2 mutant exhibited no pathogenicity. Especially, restrictedinvasive growth and accumulation of dark brown granules around infection hyphae were frequently observed in the DMocod2 mutant. The granuleaccumulation is considered as a plant defense response. Genetic complementation with the wild type alleles restored the defects in conidiation andpathogenicity. Taken together, both MoCOD1 and MoCOD2 are responsible for conidia development and pathogenicity in the rice blast fungus. This is thefirst report of the involvement of Zn(II) 2Cys 6 TFs in pathogenesis of fungal plant pathogens.299. Comparative proteomics of monoclonal-antibody enriched haustoria from three races of Puccinia triticina. Christof Rampitsch 1 , Eva Beimcik 1 ,Aslihan Gunel 2 , Guus Bakkeren 3 , Tao Fan 1 . 1) Cereal Research Ctr, Agriculture & Agrifood Canada, Winnipeg, Canada; 2) Ahi Evran University, Departmentof Chemistry, Kirşehir, Turkey; 3) Pacific Agrifood Research Centre, Summerland BC, Canada.Puccinia triticina (Ptr) causes leaf rust on wheat and is a problem in most areas where wheat is grown. The host-pathogen interaction follows the genefor-genemodel, where an interaction between host resistance (R) genes and pathogen avirulence (avr) genes determines whether the plant will remainhealthy, or whether the pathogen can complete its life cycle and cause disease. Ptr is an obligate parasite which penetrates wheat leaves through stomata,colonizes the apoplastic space and forms haustoria inside host cells. Haustoria mediate nutrient up-take between the host and pathogen thus play a majorrole in pathology. Purification of milligram quantities of haustoria from Ptr to >95% homogeneity, determined visually by calcofluor white staining, hasbeen made possible through the development of specific monoclonal antibodies. The haustoria proteome is of great interest, since it likely containsproteins with potential roles in pathogenesis. We have purified haustoria from races 1, 9 and 161 for a comparative proteome analysis, because completegenomic sequences exists for these. This is an essential requirement for homology-based matching of mass spectral data. Haustoria are surrounded by theplant plasma membrane, an extrahaustorial matrix and extrahaustorial membrane. As a result, they are recalcitrant tissues that resist analysis byconventional proteomics approaches. We have designed a strategy for obtaining the maximum yield of tryptic peptides from purified haustoria, suitablefor LC-MS analysis. Preliminary results of the comparative proteome will be presented and discussed.300. RNA-seq analyses of gene expression in the microsclerotia of Verticillium dahliae. Dechassa Duressa 1 , Amy Anchieta 1 , Donquan Chen 2 , AnnaKlimes 3,4,5 , Katherine F. Dobinson 3,4 , Maria Garcia-Pedrajas 6 , Steven J. Klosterman 1 . 1) USDA-ARS, Salinas, CA; 2) Comprehensive Cancer Center & Divisionof Preventive Medicine, University of Alabama, Birmingham, AL; 3) Department of Biology, University of Western Ontario, London, ON, Canada; 4)Agriculture and Agri-Food Canada, London, ON, Canada; 5) Department of Physiological and Biological Science, Western New England University,Springfield, MA; 6) Estación Experimental La Mayora CSIC, Málaga, Spain.Verticillium dahliae is a soilborne fungus that causes wilt disease in plants. Verticillium wilt is difficult to control because the pathogen is capable ofpersisting in the soil for 10 to15 years as melanized microsclerotia, rendering crop rotation strategies for the control of this disease ineffective. Themicrosclerotia of V. dahliae produce infectious hyphae that give rise to primary infections. As such, the processes of microsclerotia formation,maintenance, and germination are critically important in the disease cycle of V. dahliae. To shed additional light on the molecular processes involved inmicrosclerotia biogenesis and melanin synthesis in V. dahliae, three replicate RNA-seq libraries were prepared from 10 day-old microsclerotia (MS)-producing cultures of V. dahliae (ave=52.23 million reads), and those not producing microsclerotia (NoMS, ave=50.58 million reads), and analyzed fordifferential gene expression. The comparisons revealed up-regulation of MS library genes involved in melanogenesis, including tetrahydroxynaphthalenereductase (344-fold increase) and scytalone dehydratase (231-fold increase), and of additional genes located in a 48.8 kilobase melanin biosyntheticcluster. Numerous hypothetical protein-encoding genes were also identified as differentially expressed in the MS library. For confirmation of differentialexpression, selected genes identified by RNA-seq as up- or down-regulated were analyzed by RT-qPCR of RNA from several MS and NoMs culture types,including MS cultures that were stored for 6 months at 4°C, and seven day old cultures having an intermediate number of melanized MS. These dataprovide further insight into gene expression during melanin biosynthesis and MS formation in V. dahliae, and the products encoded by these genes mayrepresent alternative disease control targets.301. Exploring the Genome Diversity of Mycorrhizal Fungi to Understand the Evolution and Functioning of Symbiosis. Francis M. Martin, MycorrhizalGenome Initiative Consortium. Tree-Microbe Interactions, Lab of Excellence ARBRE, INRA-Nancy, Champenoux, France.Genomics has introduced an important new dimension into mycorrhizal research by establishing data to serve as a new and fundamental resource forgenetics and molecular biology of the symbiosis formation. With the current genomic view of ectomycorrhizal (EM) fungi that we have, a possible scenariosuggests that (1) irreversible losses of lignocellulose decomposition pathways play a key role in the evolutionary stability of the ectomycorrhizalmutualisms and (2) that each major EM fungal clade has subsequently and independently designed symbiotic molecular toolboxes each time themycorrhizal lifestyle has arisen in the tree of life. This hypothesis would predict that symbiotic toolboxes are tailor made for each major fungal clade (e.g.,Agaricales, Boletales, Sebacinales) and may be tuned according to specific plant hosts. To further our understanding of the evolution of these symbioses,the Mycorrhizal Genomics Initiative targets a set of 30 fungal mycorrhizal species. Taxa have been selected for (1) their phylogenetic novelty, (2) theirability to establish different types of mycorrhizal symbioses and (3) their taxonomic relationships with already sequenced EM genomes. Several targetspecies are capable of forming different types of intracellular colonizing structures - and this plasticity depends on plant host. Several species are dominantfungus in their ecological settings and others are currently used in the commercial forestry industry to inoculate conifer or hardwood seedlings for lumber,bioenergy and landscape trees. I will discuss how the comparative analysis of mycorrhizal genomes has, and will continue, to shed light on the evolution ofmycorrhizal symbioses.302. Known unknown genes: evolution of eukaryotic BEM46. Abhishek Kumar, Krisztina Kollath-Leib, Frank Kempken. Dept. of Genetics & Mol. Bio,Institute of Botany, CAU Kiel, KIEL, SH, Germany.The bud emergence 46-like (BEM46) protein from Neurospora crassa belongs to the alpha/beta-hydrolase superfamily. Recently, we have reported thatthe BEM46 protein is localized in the perinuclear ER and also forms spots close by the plasma membrane. The protein appears to be required for cell typespecificpolarity in Neurospora crassa. Furthermore, initial studies suggested that the BEM46 amino acid sequence is conserved in eukaryotes and isconsidered to be one of the widespread conserved “known unknown” eukaryotic genes. To unravel origin and molecular evolution of these genes indifferent eukaryotes, we carried out a comprehensive sequence, structural functional and phylogenetic analyses of BEM46 orthologs. During this study, we194

FULL POSTER SESSION ABSTRACTSfound that all eukaryotes have at least a single copy of a bem46 ortholog. Upon scanning of these proteins from various spiecies, expansions leading intoseveral paralogs in vertebrates and plants were identified. We illustrate insertion/deletions (indels) in the conserved domain of BEM46 protein, whichallow differentiating fungal classes such as ascomycetes from basidiomycetes. Furthermore, we analyze several duplicates of this gene in different animaland plant genomes to understand possible mechanisms of evolution after separation from the fungal lineage. In addition, we unravel that BEM46 proteinfrom N. crassa possess a novel endoplasmic-retention signal (PEKK) using GFP-fusion tagging experiments, hinting there is need to re-define the motifs inconserved in various protein sequences as over a million of genome sequences will be available in next decade.303. Sugar ‘cubed’ - A Comparative Systems Analysis of Plant Cell Wall Polysaccharide Recognition and Degradation Using the Model FilamentousFungus Neurospora crassa. J.P. Benz, S. Bauer, N.L. Glass, C.R. Somerville. Energy Biosciences Institute, UC Berkeley, Berkeley, CA.Filamentous fungi are currently the primary source of plant cell wall degrading enzymes for the production of biofuels from lignocellulosic feedstocks.However, despite tremendous improvements of these enzyme cocktails over the last years, they are still rather inflexible and will not work optimally insituations such as complicated with a changing variety of feedstocks. Fungi have evolved with their host plants in a long and intricate relationship, and adetailed understanding of their responses to the various building blocks present in the plant cell wall material will also help to improve the industrialapplicability and versatility of these enzyme cocktails. In recent years, the ascomycete Neurospora crassa has been developed as a model system to studycellulose and xylan degradation by filamentous fungi. As a complement to these studies, here we performed a systems analysis of pectin degradation, thethird major plant cell wall polysaccharide. A combination of proteomics and transcriptomics was used to define the “toolbox” N.crassa uses to degrade thishighly complex heteropolysaccharide, and to identify new components that seem to work both synergistically and antagonistically in this process.Moreover, in combination with the data from two earlier studies, describing the responses to cellulose and xylan, the acquired knowledge allowed for thefirst time to put the individual responses to each of these three main plant cell wall polysaccharides into perspective. Central to this analysis was theconstruction of a co-expression matrix covering the most relevant carbon source-related inducing conditions. The applicability of this matrix could bedemonstrated by successfully guiding in the functional characterization of an unknown sugar transporter, which was identified to mediate L-arabinoseuptake. Only if we understand the building blocks of the carbon-related response pathways we can attempt to put them together into the “bigger picture”.The comparative approach presented here therefore is an important step towards a more profound understanding of the fungal degradation process ofcomplex biomass.304. Building upon whole genome resequencing in Neurospora. Kevin McCluskey, Aric Wiest, Robert Schnittker. Sch Biological Sci, Univ Missouri, KansasCity, Kansas City, MO.The availability of whole genome sequence allows immediate comparison between polymorphisms that have physiological impact and those that areneutral. We are exploiting this as we characterize genes responsible for Acriflavine resistance. Preliminary analysis showed that despite a wealth ofpolymorphisms among whole genome sequenced strains, the ORF NCU09975 encoding an abc3 transporter is not altered in the acriflavine resistant strainFGSC 1215. Additional analyses have pointed to the transcription factor gene NCU09974 and the polymorphism in this gene in the acriflavine resistantstrain is unique both in comparison to the reference genome strain and among the growing number of strains subject to whole sequencing. Continuingwork aims to test whether transfer of the NCU09974 allele from the acriflavine resistant strain to an otherwise sensitive strain will confer resistance.Additional studies will investigate whether the broad resistance seen in classical acriflavine resistant mutants can allow identification of a compound thatcan be used as a selectable agent in combination with the newly identified allele.305. Genome based phylogeny of early diverging fungal lineages. A. P. Gryganskyi 1 , G. Bonito 1 , M. Rodriguez-Carres 1 , T. M. Porter 2 , Y. Chen 3 , S. Robb 4 , H.-L. Liao 1 , I. M. Anishchenko 5 , O. V. Savytskyi 6 , R. Ortega 1 , J. E. Stajich 4 , J. Heitman 3 , A. P. Litvintseva 7 , T. Y. James 8 , S. Sekimoto 9 , J. Spatafora 10 , R. Vilgalys 1 . 1)Biology, Duke University, Durham, NC; 2) Ecology and Evolutionary Biology, McMaster University, Hamilton, ON, Canada; 3) Duke University MedicalSchool, Durham, NC; 4) Plant Pathology and Microbiology, University of California, Riverside, CA; 5) Institute of Botany, NASU, Kyiv, Ukraine; 6) Institute ofMolecular Biology and Genetics, NASU, Kyiv, Ukraine; 7) Centers for Disease Control and Prevention, Atlanta, GA; 8) Ecology and Evolutionary Biology,University of Michigan, Ann Arbor, MI; 9) Biological Sciences, The University of Alabama, Tuscaloosa, AL; 10) Botany and Plant Pathology, Oregon StateUniversity, Corvallis, OR.The phylogeny of the early diverging fungal lineages remains controversial in spite of a growing database of morphological, ultrastructural, biochemicaland molecular evidence. Here we present a comprehensive molecular phylogeny for the basal fungi using metagenomic data from 30 fungal taxa for whichwhole genomes or ESTs are available. Taxa include a dozen flagellated lineages, ten zygomycetous taxa, and key representatives of the Glomeromycota,Ascomycota, and Basidiomycota. Phylogenetic trees built from 434 orthologs (some missing data) and 29 orthologs (no missing data) are congruent andstatistically well supported. Our results show a clear separation of most flagellated fungi from terrestrial taxa. An analysis of the presence of the genesassociated with the flagellar apparatus supports the hypothesis that the flagellum was lost once concomitant with fungi transitioned to terrestrial habitats.Zygomycetous lineages occupy an intermediate position between flagellated fungi and the Dikarya with Entomophthoromycotina and Kickxellomycotinarepresentatives as a basal clade.306. Comparative analysis of 35 basidiomycete genomes reveals diversity and uniqueness of the phylum. Robert Riley 1 , Asaf Salamov 1 , Robert Otillar 1 ,Kirsten Fagnan 1 , Bastien Boussau 3 , Daren Brown 4 , Bernard Henrissat 5 , Anthony Levasseur 5 , Benjamin Held 6 , Laszlo Nagy 2 , Dimitris Floudas 2 , EmmanuelleMorin 7 , Gerard Manning 8 , Scott Baker 9 , Robert Blanchette 6 , Francis Martin 7 , David Hibbett 2 , Igor Grigoriev 1 . 1) Joint Genome Istitute, Lawrence BerkeleyNational Lab, Walnut Creek, CA; 2) Clark University, Worcester, MA; 3) UC Berkeley, Berkeley, CA; 4) USDA, Peoria, IL; 5) AFMB, Marseille, France; 6) UMN,St. Paul, MN; 7) INRA, France; 8) Salk Institute, La Jolla, CA; 9) Pacific Northwest National Lab, Richland, WA.Fungi of the phylum Basidiomycota (basidiomycetes), make up some 37% of the described fungi, and are important in forestry, agriculture, medicine,and bioenergy. This diverse phylum includes symbionts, pathogens, and saprobes including wood decaying fungi. To better understand the diversity of thisphylum we compared the genomes of 35 basidiomycete fungi including 6 newly sequenced genomes. The genomes of basidiomycetes span extremes ofgenome size, gene number, and repeat content. A phylogenetic tree of Basidiomycota was generated using the Phyldog software, which uses all availableprotein sequence data to simultaneously infer gene and species trees. Analysis of core genes reveals that some 48% of basidiomycete proteins are uniqueto the phylum with nearly half of those (22%) comprising proteins found in only one organism. Phylogenetic patterns of plant biomass-degrading genessuggest a continuum rather than a sharp dichotomy between the white rot and brown rot modes of wood decay among the members of Agaricomycotinasubphylum. There is a correlation of the profile of certain gene families to nutritional mode in Agaricomycotina. Based on phylogenetically-informed PCAanalysis of such profiles, we predict that that Botryobasidium botryosum and Jaapia argillacea have properties similar to white rot species, althoughneither has liginolytic class II fungal peroxidases. Furthermore, we find that both fungi exhibit wood decay with white rot-like characteristics in growth27th Fungal Genetics Conference | 195

FULL POSTER SESSION ABSTRACTSassays. Analysis of the rate of discovery of proteins with no or few homologs suggests the high value of continued sequencing of basidiomycete fungi.307. Functional analysis of roles of expanded genes in fruiting body development in Coprinopsis cinerea. Jinhui Chang, Hoi Shan Kwan. School of lifesciences, The Chinese University of Hong Kong, Hong Kong.We wish to study the relationship of expanded genes and evolutionary adaptation in mushroom-forming fungi. We planned to (1) detect the enrichmentof expanded genes specific for mushroom-forming fungi, (2) develop a pipeline to construct protein-protein interaction (PPI) networks and match themwith the established pathways in REACTOME database for fungal proteins, and (3) characterize the functions and action stage of the expanded kinases infruiting body development. The expanded genes in late evolved organisms contribute to adaptive functions and morphological characters. Mushroomformingfungi can be differentiated from the simple fungi by the extra morphological status of fruiting bodies. We hypothesize that the expanded genesunique to mushroom-forming fungi are critical for fruiting body development. By comparing 70 species from basidiomycota and ascomycota, we foundsignificant enrichment in some protein functional clusters in mushroom-forming fungi comparing to simple fungi. Among these clusters, we chose tofurther analyze the group of Posttranslational modification related genes. We predicted and compared the PTM sites density. We found that the greaterthe genome size the lower the ubiquitylation site density . We developed a novel pipeline to search the literature for interacting proteins and construct PPInetworks. With this pipeline, we proposed a light signal transduction phosphorylation cascade which involves some FunK1 kinases and components in PKAand MAPK pathways. To investigate the functional roles of these kinases in the putative cascade, we introduced the siRNA of corresponding genes intoCoprinopsis cinerea at five stages in the life cycle. We showed by transient knock down of expanded kinases that they play imported roles in light signaltransduction pathway and possess different functions in different developmental stages.308. Comparative analysis of fungal kinomes. Yousef Shbat, Abhishek Kumar, Frank Kempken. Dept. of Genetics & Mol. Bio, Institute of Botany, CAU Kiel,KIEL, SH, Germany.Many cellular processes are regulated by phosphorylation via protein kinases. To unravel the understanding of protein phosphorylation, normally theprotein kinase complements (known as ‘kinomes’) are examined genome-wide in eukaryotic species. About 2% of eukaryotic genes are protein kinases.Bioinformatics and comparative genomics were used to determine kinomes from eukaryotes and to explore in evolutionary and functional context.Kinases are major regulators of cellular processes in fungi, in similar fashion as they regulate other eukaryotes. For example, 77 viable mutants for ser/thrkinase genes (of 86 in total) were identified in N. crassa and 57% illustrated at least one growth or developmental phenotype. Given that there is over 100fungal genomes are known. Hence, there is a need of more comprehensive analysis of fungal kinomes. We have established kinomes of about 90 fungiwith 5604 kinases, which also included pseudokinases (~1%). Fungi have expansion of ser/thr kinases in comparisons to other classes of kinases. We thusreport how kinases in combination with ~80 other protein domains, evolved to perform their roles in different signalling cascades in these fungi.Furthermore, we annotated and analysed these kinases for evolutionary mechanisms operating in fungal kinomes.309. VeA, VelB and FluG affect conidiation and aflatoxin production of Aspergillus flavus. P-K. Chang, L. Scharfenstein, P. Li, K. Ehrlich, B. Mack.Agricultural Research Service, Southern Regional Research Center, New Olreans, LA.Asexual differentiation in Aspergillus nidulans involves complex control by a number of factors and is light-dependent. The VelB/VeA/LaeA complex in A.nidulans coordinates light signal with development and secondary metabolism. We investigated the roles of velvet family genes, veA, velB and velC, andfluG (fluffy phenotype in A. nidulans) in an aflatoxigenic Aspergillus flavus strain. Knockout strains of veA or velB conidiated poorly in the dark but not inthe light. Knockout strains of fluG also showed decreased conidiation but had increased sclerotial production. Deletion of fluG in the veA or velB knockoutresulted in a marked decrease in conidiation even in the light. Growth under stress (0.6 M potassium chloride) partially restored aforementioned defects inconidiation. The veA or velB knockout mutant but not the velC or fluG mutant was unable to produce aflatoxin. Overexpression of veA in the velB mutantonly restored conidiation while overexpression of velB in the veA mutant failed to restore either conidiation or aflatoxin production. Yeast two-hybridassays confirmed that VeA, VelB and LaeA form a complex but suggested that FluG is also likely to be an interacting partner. Concerted interactions of A.flavus VeA and VelB with LaeA are critical for conidiation in the dark and aflatoxin biosynthesis.310. The Environmental Molecular Sciences Laboratory molecular analysis capabilities for fungal biology. S. E. Baker. Environmental Molecular SciencesLaboratory, Pacific Northwest Natl Lab, Richland, WA.Tools for analysis of classical and reverse genetic mutants play an important role in fungal biology research. The Environmental Molecular SciencesLaboratory (EMSL) at the Pacific Northwest National Laboratory is a US Department of Energy national user facility. EMSL develops and utilizes cuttingedge mass spectrometry, NMR, imaging and computational capabilities to accelerate research in a number of areas. We have used EMSL’s massspectrometry capabilities to characterize glycosylation of secreted proteins of Aspergillus niger. In addition, we have explored the use of laser ablation andnano-DESI mass spectrometry for spatial localization of molecules associated with Trichoderma reesei mycelium. Finally, spores from wildtype and albinostrains of Aspergillus carbonarius were characterized using helium ion microscopy. As a national user facility, the EMSL is open to the fungal biologycommunity through a competitive, peer-reviewed proposal process.311. Comparative Genomics and Transcriptomics of Insect Pathogenesis. Kathryn E. Bushley, Joseph W. Spatafora. Dept Botany & Plant Pathology,Oregon State Univ, Corvallis, OR.We have sequenced the genome of Tolypocladium inflatum, the first sequenced representative of one of three major lineages of insect pathogens withinthe order Hypocreales. Comparisons of the gene space and transcriptome of T. inflatum with closely related plant pathogenic and endophytic fungi isproviding insights into secondary metabolite arsenals specific to insect pathogens as well as shedding light on shifts in primary metabolism associated witha transition to an insect host. We address the role of secondary metabolites in insect pathogenesis using a combination of comparative genomics to trackthe evolution of secondary metabolite clusters across the Hypocreales and transcriptomics to characterize patterns of gene expression within metaboliteclusters in media supplemented with insect cuticle (simulating insect infection) and hemolymph (simulating insect colonization). We also identify othergene families that are upregulated under these media conditions. GO enrichment analyses of upregulated genes showed that those involved in oxidationreductionreactions, iron-binding, and transport of iron and inorganic ions are important during both the infection and colonization phases. Genes withserine peptidase and serine hydrolase activity were uniquely upregulated in cuticle media while a large proportion of genes upregulated in hemolymphwere involved in transmembrane transport not only of iron, but also of sugars and other carbohydrates. We examine expansions and contractions of someof these gene families (e.g. proteases and P450s) that map to nodes in the phylogeny associated with shifts to insect hosts. We identify patterns that areshared across the three insect pathogenic lineages of Hypocreales versus those which have evolved independently in distinct lineages.196

FULL POSTER SESSION ABSTRACTS312. Integrated transcriptional profiling and analysis for identification of Cryptococcus neoformans genes regulated during human cryptococcalmeningitis. Y. Chen 1 , J. Tenor 1 , D. Toffaletti 1 , A. Litvintseva 2 , T. Mitchell 1 , J. Perfect 1 . 1) Duke University School of Medicine, Durham, NC; 2) Centers forDisease Control and Prevention, Atlanta, GA.Background: Cryptococcus neoformans is an opportunistic fungal pathogen that is the major cause of fungal meningitis in immunocompromisedindividuals worldwide. Accurate and comprehensive de novo transcriptome profiling of C. neoformans in the human host may allow a betterunderstanding of how it survives and produces disease. Methods and Results: To identify genes, whose expression is differentially regulated under in vivoand in vitro conditions, we selected two strains of C. neoformans var. grubii (serotype A), which were isolated from the cerebrospinal fluid (CSF) of twoAIDS patients from Uganda and the United States. Multilocus sequence typing (MLST) showed that one strain was from VNI clade and one strain from VNII.Next-generation sequencing (RNA-Seq) was used to determine transcriptional profiles of these strains under three conditions: fungal cells were directlytaken from CSF of the patients; fungal cells were grown in YPD at 37°C until the stationary phase; fungal cells that reached the stationary phase in YPDwere exposed to sterile human CSF for 9 hours. The sequencing results showed that there was no major difference in sequencing quality andcontaminations between in vivo and in vitro samples. Hierarchical clustering analysis revealed that the samples treated with same environment have moresimilarity in transcriptional profile. Comparative analysis of the expression pattern shows that 144 genes up-regulated in CSF when compared to YPD and87 genes were up-regulated in vivo compared to YPD and 39 genes overlapped between the CSF and in vivo condition. Some of the overlapping genes inCSF and in vivo have been reported to be related to the virulence composite of C. neoformans, such as Rim101 and ENA P-type ATPase 1. Furthermore, wesearched for the 100 most divergent expressed genes between the two strains. Gene Ontology (GO) term enrichment analysis showed an enrichment ofGO terms in transporter activity between the strains. Conclusion: We provide the first transcriptome profiling of C. neoformans taken directly from the CSFof two human patients. The comparisons between in vivo and in vitro samples helped us to identify a group of genes that may be important for surviving,adapting and proliferating of C. neoformans in the CSF of the human host.313. Structural and functional characterization of microRNA-like RNAs in the penicillin producing fungus Penicillium chrysogenum. Tim Dahlmann,Minou Nowrousian, Ulrich Kück. Christian Doppler Laboratory for Fungal Biotechnology, Ruhr-Universität Bochum, Universitätsstr. 150, 44780 Bochum,Deutschland, tim.dahlmann@rub.de.MicroRNAs are endogenous RNAs with a size of about 22 nt and post-transcriptionally regulate gene expression in metazoan and plants. MicroRNAs arederived from RNA hairpin precursors, which are usually transcribed by RNA polymerase II. Recent studies on small RNA binding components of the RNAinducedsilencing complex (RISC) show the existence of microRNA-like RNAs (milRNAs) in Neurospora crassa, which give a first hint of a post-transcriptionalregulatory mechanism based on microRNAs in fungi [1]. So far only little is known about microRNA-like molecules in other fungi, especially about their rolein fungal development and gene regulation.To investigate the occurrence of milRNAs and their involvement in gene regulation in the penicillin producing fungus Penicillium chrysogenum, weperformed predictions of putative microRNAs. Therefore small RNAs (19 - 50 nt) representing different growing conditions and developmental stages,were used for RNA next generation sequencing. The calculation of putative microRNA precursors was performed with the program miRDeep [2], and isbased on the distribution of RNA sequence reads in afore predicted RNA hairpin molecules. By this approach, we were able to identify structures, whichshow the typical characteristics of microRNA precursors. To confirm the in silico predictions, transcript analyses were performed. These analyses supportthe existence of small RNAs and their precursors and show various expression pattern of the putative milRNAs under different growing conditions. Toinvestigate the regulatory role of the identified milRNAs, strains lacking or overexpressing milRNAs were generated. In addition, we have constructedartificial microRNAs to investigate their use as molecular genetic tools to mediate gene specific RNA interference (RNAi). The results of this study provideevidence for milRNAs in P. chrysogenum and indicate a milRNA based silencing mechanism in this fungus.[1] Lee HC et al. (2010) Diverse pathways generate microRNA-like RNAs and Dicer-independent small interfering RNAs in fungi. Molecular Cell 38:803-814[2] Friedländer MR et al. (2008) Discovering microRNAs from deep sequencing data using miRDeep. Nat Biotechnol 26:407-415.314. Metatranscriptomic analysis of ectomycorrhizal root clusters in Pinus taeda: new methodologies for assessing functional gene expression in situ.H.-L. Liao 1 , Y. Chen 2 , T. D. Bruns 3 , K. G. Peay 4 , J. W. Taylor 3 , S. Branco 3 , J. M. Talbot 4 , R. Vilgalys 1 . 1) Department of Biology Duke University, Durham, NC; 2)School of Medicine, Duke University, Durham, NC; 3) Department of Plant and Microbial Biology, UC-Berkeley, Berkeley, CA; 4) Department of Biology,Stanford University, Stanford, CA.A highly diverse community of ectomycorrhizal (ECM) fungi are known to associate with members of the genus Pinus. Less is known about how diversefungal communities affect functional diversity within ECM roots. Here we present an optimized method for metatranscriptomic analysis of the ECM-pineroot interaction in a natural system. RNA was purified using a CTAB method from individual ECM root clusters collected at varying spatial scales across thedistribution range of P. taeda, and sequenced using Illumina HiSeq technology. About 35 million qualified reads were obtained. Sequences were initiallyassembled using reference based mapping (Bowtie) to sort the reads that represent rRNA from fungal and bacterial species. Reads from divergent regions(D1-D2) of fungal LSU rRNA were used to identify dominant ECM and other fungal community members. Subsequently, P. taeda genes and functionalgenes of dominant fungal species were sorted using public cDNA databases. The Trinity package was used for de novo assembly of un-mapped reads(mostly fungal genes). Blastx and Go packages were used for gene annotation. A typical ECM root cluster was found 45% P. taeda genes, 3% fungal rRNA,0.05% bacterial 16S rRNA, 30% fungal functional genes, 10% unknown sequences, and 12% unassembled reads. Analysis of D1-D2 LSU sequencesconfirmed that a single ECM fungal species usually dominates individual root clusters. De novo assemblies of fungal genes yielded 120 thousand contigsfrom 10 million reads representing 90 thousand unique genes with highly similarity to known ECM fungi. Functional analysis revealed that most of thetranscripts recovered were involved with translation, protein degradation, heat shock, superoxide metabolism, electron transfer, signaling, and C/Nmetabolism. Highly expressed transcripts recovered from Piloderma, which was abundant in our samples, included genes encoding a wide array ofmetabolic enzymes: chitosanase, phosphatase, glutamine synthetase, terpene synthases, b-glucanase; transporters for P+ and oligopeptides; cell signaling:calmodulin, cAMP-regulated phosphoprotein (Igo1); C/N related genes: lectin, cross-pathway control (cpc1); as well as several genes with unknownfunction. Future studies will seek to address how ECM metatranscriptomes change in response to different Pinus hosts and across different spatial scales.315. Transcriptomic response of Neurospora crassa germinating conidia to chitosan in sub-lethal dose. Federico Lopez-Moya 1 , David Kowbel 2 , N. LouiseGlass 2 , Luis Vicente Lopez-Llorca 1 . 1) Laboratory of Plant Pathology, Multidisciplinary Institute for Environment Studies (MIES) Ramón Margalef. Universityof Alicante, Alicante, SPAIN; 2) Department of Plant and Microbial Biology, University of California, Berkeley CA, 94720-3120 USA.Chitosan is a natural polymer able to permeabilize Neurospora crassa membranes, in an energy dependent manner. Plasma membrane permeabilisationby chitosan depends on membrane fluidity, with FFA unsaturated membrane fungi (N. crassa) being chitosan sensitive, the plasma membrane fluidity is an27th Fungal Genetics Conference | 197

FULL POSTER SESSION ABSTRACTSimportant factor in chitosan effect. Plasma membrane permeabilisation is also enhanced under starvation conditions, also causes intracellular ROSincreases and is involved in cell division. Conidial germination is the most sensitive step to chitosan in filamentous fungi. We have used Neurospora crassaconidia germinating in Vogels medium with chitosan at a sub-lethal concentration for evaluating the transcriptomic response of the fungus. Wedetermined chitosan IC 50 for N.crassa (4, 8 and 16 hours after inoculation, hai) and analyzed the effect on gene expression over time. Using RNA-seq wehave detected the genes involved in N. crassa response to chitosan, we analyzed with bioinformatic resources the expression involved on chitosanresponse in N. crassa conidia . We have also re-annoted the complete Neurospora crassa OR74A genome (Broad Institute) for allowing GO analyses(biological process, cell component and molecular function) of our RNA-seq data. Chitosan early induced (4-8 hai) some genes, involved in ROS protection(e.g. catalases, monooxigenase and SOD) and also in nitrogen compounds transporter and degradation. We also detected late expression of genes involvedin biosynthetic processes, nitrogen compounds (including proteins) metabolism and transport. Our strategy has allowed us to gain an important insight onchitosan mode of action. This will open new possibilities for application this versatile natural compound.316. Transcriptomic analysis of the interaction between Trichoderma harzianum and the phytopathogen Sclerotinia sclerotiorum using the RNA-seqapproach. Andrei S. Steindorff 1 , Marcelo H.S. Ramada 1 , Robert Miller 1 , Georgios J. Pappas 1 , Cirano J. Ulhoa 2 , Eliane F. Noronha 1 . 1) Cell Biology Dept,Brasilia University, Brasilia, DF, Brazil; 2) Biochemistry Dept, Federal University of Goias, Goiania, Brazil.The plant pathogen Sclerotinia sclerotiorum is the causal agent of the common bean’s (Phaseolus vulgaris) root rot disease, white mold, and itsoccurrence is responsible for the great yield losses in irrigated areas of the Southeast and Midwest regions of Brazil. Species of the fungi genusTrichoderma have been used in the biological control of this pathogen as an alternative to chemical control. To gain new insights into the biocontrolmechanism used by Trichoderma harzianum against the phytopathogenic fungus, Sclerotinia sclerotiorum, our research group performed a transcriptomicanalysis of this interaction using RNA-seq and quantitative real-time PCR (RT-qPCR) approaches. Six RNA-seq libraries from T. harzianum mycelium (isolate303/02) grown on cell walls of S. sclerotiorum (CWSS) and glucose during 12, 24 and 36 h were constructed, sequenced by Illumina HiSeq 2000 2x100pband analyzed by TopHat/Cufflinks pipeline. The T. harzianum CBS 226.95 v1.0 genome (http://genome.jgi.doe.gov/Triha1/Triha1.home.html) was used as areference for the bioinformatics analysis. Among the 13616 genes mapped, 1581 genes were found differentially expressed among the growth in thepresence of glucose or plant pathogen cell walls. Moreover, the expression pattern was also time course dependent. Transporters, fungal cell wallhydrolases, peptidases, transcriptional factors and proteins presenting role in the environmental interaction were found high expressed by theTrichoderma isolate in the three different times of growth and in the presence of the pathogen. Some genes with no known functions were also found.The role of cell wall hydrolases, peptidases and other hydrolytic enzymes in the mycoparasitism by Trichoderma species was strongly recorded, thereforethe description of the functions of those unknown functions genes in biocontrol will orientate future works. Indeed, the present work will contribute to aninitial mapping of the transcripts quite related to the interaction among these two fungi and for its further analysis under in vivo interaction.317. Genome evolution of the original Saccharomyces carlsbergensis lager yeast strain, Unterhefe No1, as revealed by whole genome sequencing.Andrea Walther, Ana Hesselbart, Jürgen Wendland. Carlsberg Laboratory, Copenhagen V, Denmark.The first lager beer yeast strain was purified by Hansen in 1883 who termed this original strain Unterhefe No1, also known as Saccharomycescarlsbergensis. It became evident that Unterhefe No1 is a hybrid between two closely related Saccharomyces species, particularly S. cerevisiae and a S.bayanus-like strain. We have compared key fermentation parameters including sugar utilization, ethanol and flavor production of Unterhefe No1 withcurrently used industrial lager yeast strains. We then sequenced the S. carlsbergensis genome using 454 next generation sequencing methods anddetermined its hybrid genome content. We found that the genome has evolved from a presumed ancestral tetraploid cell as a result of adaptation tobrewing conditions resulting in its current allotetraploid state. In contrast to a hypothetical tetraploid genome with 24Mb unique DNA contributed by itsparents and distributed on 32 chromosomes, the Unterhefe No1 genome consists of only 20.7Mb. It comprises chromosomes derived from S. cerevisiaeand a non-cerevisiae parental strain as well as a S. bayanus like mitochondrial genome. In total, we found 29 different chromosomes including evolvedchromosomes displaying several events of loss of heterozygosity and massive chromosomal rearrangements. Comparison of the genome of S.carlsbergensis with other yeast genomes provides insight into the evolution of this brewing strain as a consequence of adaptation to lager beerfermentation conditions.318. Retention of genes in a secondary metabolite gene cluster that has degenerated in multiple lineages of the Ascomycota. Daren W. Brown 1 , Hege H.Divon 2 , Erik Lysøe 3 , Robert H. Proctor 1 . 1) Bacterial Foodborne Pathogens and Mycology Research, USDA/ARS, Peoria, IL; 2) Section of Mycology,Norwegian Veterinary Institute, PO Box 750, Sentrum, 0106 Oslo, Norway; 3) Department of Plant Health and Plant Protection, Bioforsk - NorwegianInstitute of Agricultural and Environmental Research, 1432 Ås, Norway.Fungal secondary metabolite (SM) gene clusters encode proteins involved in SM biosynthesis, protection against SMs, and regulation of cluster genetranscription. RNA-Seq analysis of Fusarium langsethiae (class Sordariomycetes) revealed a cluster of six genes that were highly expressed during growth inoat-grain medium, but not in complete medium. All six genes share significant homology and synteny with genes in the Alternaria brassicicola (classDothideomycete) cluster responsible for production of the SM depudecin. HPLC analysis confirmed the presence of depudecin in oat-grain medium andabsence from complete medium cultures. A survey of publically available genome sequences identified eight complete and 14 partial depudecinbiosynthetic gene (DEP) cluster homologs in fungi across distantly related classes of Ascomycota. Most of the partial clusters included pseudogenes due tosingle nucleotide changes and/or multiple nucleotide deletions, indicating that the partial clusters are derived by degeneration of complete clusters. Mostof the partial clusters also included apparently functional homologs of the major facilitator superfamily (MFS) transporter (DEP3) and transcription factor(DEP6) genes. Retention of these two genes may provide a defense mechanism against depudecin produced by other fungi. Alternatively, DEP3 and DEP6in the partial clusters may have been repurposed to provide a selective advantage different from the advantage conferred by depudecin. The sharedsynteny of putative functional DEP3 and DEP6, as well as phylogenetic analysis of these genes, suggest that the DEP cluster has been transferredhorizontally between fungi multiple times.319. Functional characterization of unique non-ribosomal peptide synthetase genes in the cereal fungal pathogen Cochliobolus sativus. Yueqiang Leng,Shaobin Zhong. Department of Plant Pathology, North Dakota State University, Fargo, ND 58108.In filamentous fungi, nonribosomal peptide synthetases (NRPSs) are the major enzymes involved in biosynthesis of nonribosomal peptides (NRPs), someof which have been demonstrated to be involved in pathogenicity or virulence of fungal plant pathogens. However, the functions of many genes (NPS)encoding NRPSs are still not well understood. We identified 25 NPS genes from the genome sequence of the cereal fungal pathogen Cochliobolus sativus.Genome comparison among species in the genus of Cochliobolus identified 14 unique NPS genes in C. sativus with five (encoding protein ID# 130053,140513, 104448, 115356 and 350779, respectively) being unique to the pathotype 2 isolate ND90Pr of the fungus. Quantitative real time PCR revealed that198

FULL POSTER SESSION ABSTRACTSall these unique NPS genes of ND90Pr except the one for ID# 350779 were highly up-regulated in planta 12 hours post inoculation on barley cv. Bowman.Knockout mutants of the NPS gene for ID# 115356 and RNAi mutants of the NPS gene for ID# 140513 were significantly reduced in virulence on Bowman,but they had the same morphology and growth rate under the conditions of normal growth and oxidative/hyperosmotic stresses compared to the wildtype. These results indicate that these NPS genes are required for the high virulence of the pathotype 2 isolate on barley cv. Bowman. Functionalcharacterization of other unique NPS genes of ND90Pr will also be presented.320. Phylogenomics unveils secondary metabolites specific to mycoparasitic lineages in Hypocreales. C. Alisha Owensby, Kathryn E. Bushley, Joseph W.Spatafora. Botany & Plant Pathology, Oregon State University, Corvallis, OR.Hypocreales is an order characterized by a dynamic evolutionary history of interkingdom host jumping, with members that parasitize animals, plants, andother fungi. The monophyly of taxa attacking members of the same kingdom is not supported by molecular phylogenetics, however. For example,Trichoderma spp. and Elaphocordyceps spp. are both mycoparasitic, but are members of different families within Hypocreales, Hypocreaceae andOphiocordycipitaceae, respectively. In fact, both genera are more closely related to insect pathogens, than they are to each other. Multiple species ofTrichoderma have sequenced genomes, and recently genomes of several insect pathogens in Hypocreales have been completed (e.g. Metarhizium spp. andTolypocladium inflatum). The genus Elaphocordyceps represents a unique clade within Hypocreales, because whereas most species in the familyOphiocordycipitaceae are insect pathogens, most Elaphocordyceps parasitize truffles of the ectomycorrhizal genus Elaphomyces [Eurotiales, Ascomycota].To compare genes of a truffle pathogen with hypocrealean insect pathogens and mycoparasites, we sequenced the genome of Elaphocordycepsophioglossoides. Our draft assembly of the E. ophioglossoides genome is ~32 MB and has 10,779 gene models, 36 of which are predicted to producesecondary metabolites. We have identified three very large genes in E. ophioglossoides related to peptaibol producing nonribosomal peptide synthetase(NRPS) genes. Peptaibols, which disrupt osmoregulation by forming ion channels through lipid bilayers, have antibiotic and antifungal activity and are bestdescribed in Trichoderma spp. E. ophioglossoides and its beetle-pathogenic congener, T. inflatum, both possess three putative peptaibol synthetases whichwe identified through analysis of NRPS adenylation domains. Of the three peptaibol-specific domain clades, one is predicted to encode for thenonproteinogenic a-aminoisobutryic acid residues. We also show that, despite being very closely related, E. ophioglossoides and T. inflatum each possessthree different peptaibol-like genes, only two of which appear to be located in syntenic regions. The current distribution of fungi possessing peptaibolgenes is restricted to mycoparasitic lineages of Hypocreales and is generating hypotheses about the role of secondary metabolites in mycoparasitism.321. Genome and transcriptome sequence of the apomictic fungus Arnium arizonense (Podospora arizonensis). E. Coppin 1,2 , C. Drevet 3 , L. Peraza-Reyes 1,2 , D. Zickler 1,2 , E. Espagne 1,2 , J. Aït-Benkhali 1,2 , P. Silar 1,2,4 , A. E. Bell 5 , D. P. Mahoney 5 , R. Debuchy 1,2 . 1) Univ Paris-Sud, Institut de Génétique etMicrobiologie, Orsay, France; 2) CNRS, Institut de Génétique et Microbiologie, Orsay, France; 3) Univ Paris-Sud, eBio bioinformatics plateform, OrsayFrance; 4) UFR des Sciences du Vivant, Université Paris-7 Diderot, Paris, France; 5) Private Mycological Research, 45 Gurney Road, Lower Hutt, NewZealand.The homothallic fungus Arnium arizonense is closely related to the heterothallic Podospora anserina but displays several unique features. It is apomictic,i.e. dikaryotic croziers are formed inside the perithecia but neither karyogamy nor meiosis take place in the asci, although morphological changes in bothchromosomes and spindle pole bodies are reminiscent of those associated with meiosis in heteromictic Pezizomycotina. Instead of meiosis, the two nucleiundergo two mitoses and the resulting eight nuclei are enclosed in uninucleate ascospores, among which four mature normally, and four abort.Arrangement of the two ascospore types in individual asci is random (Mainwaring and Wilson, 1968, Trans Br mycol Soc, 51, 663). A. arizonense has twochromosomes, while most fungi in this group have seven chromosomes. Analysis of the genome sequence revealed that A. arizonense contained linkedcounterparts of the P. anserina mating-type genes, a structure that is typical of homothallic life style. Deletion of the mating-type locus resulted in the lossof perithecium formation, thus confirming the role of the mating-type genes in the fruit-body development. Genome annotation identified 11,165 genes,of which 476 undergo alternative splicing. Comparison of A. arizonense proteins with their orthologs in P. anserina revealed that A. arizonense genomecontains numerous pseudogenes. Direction for future work is to determine how apomixis takes place, as this process of asexual clonal reproductionthrough seeds has potential revolutionary applications in agriculture by allowing perpetuation of any important selected heterozygous genotype (reviewedby Ozias-Akins and van Dijk, 2007, Ann Rev Genet, 41, 509-537).322. Role of MAP kinase pathways in the pathogenicity of the wheat pathogen Mycosphaerella graminicola . Elisabetta Marchegiani 1 , Julie Vallet 1 , SiãnDeller 2 , Marc-Henri Lebrun 1 . 1) Bioger, INRA, Thiverval-Grignon, France; 2) Syngenta Limited, European Regional Centre, Priestley Road, Surrey ResearchPark, Guildford, Surrey, GU2 7YH, United Kingdom.Mitogen-activated protein kinases (MAPKs) are essential components of fungal signaling pathways involved in different developmental processes and arerequired for host plant infection. Mycosphaerella graminicola, the causal agent of Septoria tritici leaf blotch (STB) of wheat, has three MAPK pathways thatare all required for infection (MgFUS3 , MgHOG1, MgSLT2; Cousin et al., 2006; Mehrabi et al., 2006a, Mehrabi et al., 2006b). We showed that Mgfus3 nullmutants are non-pathogenic on intact wheat leaves (paint brush inoculation), but highly-reduced in pathogenicity when infiltrated into leaf tissues bysyringe injection (reduced necrosis, low number of pycnidia). This suggests that MgFUS3 is involved in fungal penetration, host colonization and pycnidiaformation. Mghog1 null mutants have pathogenicity defects similar to Mgfus3 null mutants. This result highlights that the role of HOG1 in pathogenicityon plants differs among fungi (Segmüller et al., 2007). Mgslt2 null mutants are fully non-pathogenic on inoculated wheat leaves either by paint brushinoculation or injection. This phenotype is unusual among slt2 null mutants from other fungi. Therefore, Mycosphaerella graminicola MAPK pathways mayhave evolved to control regulatory networks differing from other fungal plant pathogens. To identify which genes are under the control of the MgSLT2signaling pathway, we are developing different transcriptomics analyses. Expression profiling relies on the comparison of transcriptomes of Mgslt2 nullmutants and wild type strains grown under conditions corresponding to either an active or an inactive SLT2 pathway. Additional transcriptomics analyseswill be performed using an allele encoding a conditionally active MAPKK expressed under the control of an inducible/repressible promoter. Genes whoseexpression requires an active SLT2 MAPK will be further studied for their role in development and infection using reverse genetics. Cousin et al. (2006),Molecular Plant Pathology 7(4): 269-278. Mehrabi et al; (2006a), Molecular Plant-Microbe Interactions 19(4): 389-398; Mehrabi et al. (2006b), MolecularPlant-Microbe Interactions 19(11): 1262-1269; Segmüller et al. (2007), Eucaryotic Cell 6(2) 211-221.323. WITHDRAWN324. Ancient and abundant MITEs in epichloae genomes. Damien Fleetwood 1 , Chris Schardl 2 , Carolyn Young 3 . 1) Forage Improvement Section,AgResearch, Auckland, New Zealand; 2) Dept of Plant Pathology, University of Kentucky, Lexington, KY; 3) Forage Improvement Division, Samuel RobertsNoble Foundation, Ardmore, OK.27th Fungal Genetics Conference | 199

FULL POSTER SESSION ABSTRACTSThe Epichloë festucae genome contains thirteen known degenerate miniature inverted repeat transposable element (MITE) families that make up almost1% of the genome. Recent sequencing of a range of epichloae and related Clavicipitaceae family genomes revealed that every MITE family was active earlyin the evolution of the epichloid lineage although none are found in other closely related genera. Analysis of MITE integration sites showed that theseelements have a target integration site preference for 5’ genic regions of the E. festucae genome and are particularly enriched within alkaloid gene clustersand within 10-kb of other NRPS and PKS genes. Very few individual insertion sites are apparently shared among different species although one ancestralinsertion - three adjacent EFT-3m/Toru elements in the ergot alkaloid synthesis cluster - has mediated recombination events that in one strain may haveabolished synthesis of this bioprotective alkaloid. Overall these results suggest a potential role for MITEs in the evolution of the epichloae and theirsymbiotic associations with plants.325. Exploring the biomass modifying enzymes of new filamentous fungal isolates from Vietnam, using secretome and transcriptome analyses. GeorgeE Anasontzis 1,3 , Thanh Dang Tat 2 , Thuy Nguyen Thanh 2 , Hang Dinh Thi My 2 , Thanh Vu Nguyen 2 , Lisbeth Olsson 1,3 . 1) Industrial Biotechnology, ChalmersUniversity of Technology, Gothenburg, Västra Götaland, Sweden; 2) Department of Microbiology, FIRI - Food Industries Research Institute, Hanoi, Vietnam;3) Wallenberg Wood Science Center, Chalmers, Gothenburg, Sweden.In the bio-based economy concept, the current hydrocarbon fuels and non-biodegradable plastics will be replaced by new products which will derivefrom natural and renewable resources. The synthesis of such biofuels and biochemicals is still challenged by the difficulties to cost efficiently degradelignocellulosic materials to fermentable sugars or to isolate the intact polymers. Biomass degrading and modifying enzymes play an integral role both inthe separation of the polymers from the wood network, as well as in subsequent modifications, prior to further product development. The type ofapplication usually defines the conditions where the reactions should take place. Thus, novel enzymes with variable combined properties, such as differentthermotolerance, pH range of activity, substrate specificity and solvent tolerance, still need to be discovered and developed to achieve the highestpossible efficiency in each occasion. We took advantage of the rapidly evolving and high biodiversity of the tropics and have been screening variousisolates for their cellulases and hemicellulases activities. Promising strains were then cultivated in bioreactors with different carbon sources, such as wheatbran, spruce and avicel and their biomass degrading capacity was analysed through cross species protein identification of their secretome with iTRAQ.Information on the genes involved in the different stages of the fermentation and the carbon source are being acquired with next generation sequencingof the total transcriptome. Interesting transcripts will then be used to heterologously clone and express the respective genes and identify their role in thedegradation process.326. Fusarium Comparative Transcriptomics and Transcriptional Regulatory Network Reconstruction. L. Guo 1 , G. Zhao 2 , X. Zhao 3 , W. Jonkers 4 , L. Gao 2 , J.Xu 3 , C.H. Kistler 4 , L. Ma 1 . 1) Comparative Fungal Genomics Laboratory, University of Massachusetts Amherst, Amherst, MA; 2) Department of Electrical &Computer Engineering, University of Massachusetts Amherst, Amherst, MA; 3) Department of Botany and Plant Pathology, Purdue University, WestLafayette, IN; 4) USDA-ARS, Cereal Disease Laboratory, St Paul, MN.Genus Fusarium contains pathogens that infect hundreds of crop plants as well as humans and thus threatens global food safety and human health. As inother cellular organisms, diseases caused by this group of organisms are dynamically controlled through their transcriptional regulatory networks (TRNs).Reconstructing their TRNs will not only help us to comprehend the complexity of their cellular functions, but will also have broad implications for diseasemanagement and prevention. A robust searching algorithm using Bayesian networks model was developed based on nearly 200 gene expression datasetsof F. graminearum. The algorithm infers the relationship between candidate regulators (transcription factors and signaling proteins) and the target genesregulated by them. Preliminary validation of the inferred network using prior biological knowledge proofs the effectiveness of the program. Usingcomparative functional genomics approach, we have analyzed the microarray-based transcriptome data of F. graminearum (PH1), F. verticillioides (7600)and F. oxysporum f.sp. lycopersici (4287) in response to carbon (C) and nitrogen (N) starvation. In agreement with previous studies, under C and Nstarvation, fungal cells adjust to extreme environments via modulating expression of core orthologous genes to enhance cellular transport of lipid, peptideand carbohydrates but shut down unnecessary energy consumption such as protein synthesis. This analysis helps us to reach the understanding offunctional conservation of the orthologs, judging by their expression under the same biological condition in different species. Even though there is notequal amount of expression data for other Fusarium spp., the conservation of the regulatory modules will enable us to transfer the network knowledgefrom one system to improve the prediction of the other. The comparative functional analysis will also highlight critical pathways that constitute to speciesspecificphenotypes, such as pathogenicity in each species.327. The mycorrhizal genome initiative (MGI): Identification of symbiosis-regulated genes by using RNA-Seq. A. Kohler 1 , E. Tisserant 1 , E. Morin 1 , C.Veneault-Fourrey 1 , S. Abba 2 , F. Buscot 3 , J. Doré 4 , G. Gay 4 , M. Girlanda 2 , S. Herrmann 3 , T. Johansson 5 , U. Lahrmann 6 , E. Martino 2 , S. Perotto 2 , M. Tarrka 3 , A.Tunlid 5 , A. Zuccaro 6 , I. Grigoriev 7 , F. Martin 1 . 1) Lab of Excellence ARBRE, Tree-Microbes Department, INRA-Nancy, Champenoux, France; 2) Dipartimento diScienze della Vita e Biologia dei Sistemi, Università di Torino,Torino, Italy; 3) Department Soil Ecology, UFZ Centre for Environmental Research Leipzig-HalleLtd., Halle, Germany; 4) Ecologie Microbienne UMR CNRS 5557, USC INRA 1193, Universite Claude-Bernard LYON 1, Villeurbanne, France; 5) MicrobialEcology, Lunds University, Lund, Sweden; 6) Max-Planck Insitute for Terrestrial Microbiology, Marburg, Germany; 7) DOE Joint Genome Institute, WalnutCreek, California, USA.Genome and transcriptome analyses of Laccaria bicolor and Tuber melanosporum (Martin et al., 2008, 2010) revealed that the ectomycorrhizal symbiosisprobably developed several times during evolution by generating different ‘symbiosis molecular toolkits’. In L. bicolor a large set of small-secreted proteinsacts as putative effectors but not in T. melanosporum, while the up-regulation of transporter-coding genes seems to be a common feature of bothinteractions. To better understand the evolutionary origin of mycorrhizal symbiosis and to elucidate the molecular mechanisms involved, a largesequencing project of species from different taxa, phylogenetic clades and symbiotic lifestyles (ectomycorrhizae, ericoid and orchid mycorrhizae) wasstarted in 2011 by the Joint Genome Institute and the mycorrhizal genome initiative. To identify and to compare symbiosis-regulated genes large scaleIllumina transcriptome sequencing of mycelium and mycorrhizal roots from Paxillus involutus, Piloderma croceum, Hebeloma cylindrosporum, Sebacinavermifera, Tulasnella calospora and Oidiodendron maius was performed. Small-secreted proteins, transporters, CAZymes but also many lineage specificproteins were among the highly up-regulated transcripts.Martin, F., Aerts, A., Ahrén, D., Brun, A., Duchaussoy, F., Kohler, A., et al. 2008. The genome sequence of the basidiomycete fungus Laccaria bicolorprovides insights inot the mycorrhizal symbiosis. Nature 452 :88-92Martin, F., Kohler, A., Murat, C., Balestrini, R., Coutinho, P.M., Jaillon, O., Montanini, B., et al. 2010. Périgord black truffle genome uncovers evolutionaryorigins and mechanisms of symbiosis. Nature 464 :1033-1038.328. Transcriptome, secreted enzymes and systematics of the white rot basidiomycete Phlebia radiata. Jaana Kuuskeri 1 , Miia Mäkelä 1 , Kristiina Hildén 1 ,200

FULL POSTER SESSION ABSTRACTSPia Laine 2 , Lars Paulin 2 , Taina Lundell 1 . 1) Department of Food and Environmental Sciences, Division of Microbiology, Fungal Biotechnology Laboratory; 2)Institute of Biotechnology, DNA Sequencing and Genomics Laboratory, Viikki Campus, University of Helsinki, FINLAND.The efficient wood-degrading white-rot basidiomycete Phlebia radiata Fr. is able to degrade all the main components of lignocellulose and secretes arepertoire of lignin-converting peroxidases and oxidases as well as carbohydrate-acting enzymes. P. radiata is the second species of the genus Phlebia tobe genome sequenced, thus giving more insight in to the phlebioid clade of the order Polyporales, class Agaricomycetes, and comparative data forfunctional analyses on white-rot fungal mechanisms of wood decay and decomposition of lignin. We carried out transcriptome sequencing and secretomeanalyses on the Finnish isolate 79. Molecular systematics of the genus Phlebia was inferred by perfoming a four-gene study on North-European Phlebiaspp. isolates including 10 species. Ribosomal RNA-encoding (SSU 18S rDNA; ITS1-5.8S-ITS2; ITS2-28S) regions and two protein-coding genes (gapdh, rpb2)were partially PCR-amplified with fungal or basidiomycete-specific primer pairs. Phylogenetic sequence analyses resulted with no single taxonomic clusterof Phlebia. The genus is obviously polyphyletic, and the various species were scattered together with other genera of Polyporales, like Phanerochaete andRhizochaete, in the families Corticiaceae and Meruliaceae. P. radiata was the most related with Phlebia acerina and P. rufa along with P. tremellosa and P.brevispora, whereas P. subserialis and Phlebiopsis gigantea are more distant. P. radiata transcriptome analysis resulted with 6 590 unique gene transcripts,and similarity searches with blastx (E-value cut-off 1e-6) matched 77% of the unique transcripts to known or predicted protein-coding gene sequences.Functional annotation assigned Gene Ontology term for 54% of the gene transcripts. Most of the genes were annotated with term nucleotide bindingwhereas 7% were oxidoreductases. Among these were several lignin-modifying enzymes (class II heme-including peroxidases and laccases). From thedataset, 16% of the gene transcripts were unknown thus representing proteins potentially unique to P. radiata. Also, the mitochondrial genome of over150 kb in size shows unique features and high degree of genetic flexibility. These results provide an insight into gene content of P. radiata and genomeleveltranscriptional information on fungal genetic machinery for growth on complex liquid media.329. Characterization of molecular mechanisms underlying the multi-drug-resistant phenotypes of Mycosphaerella graminicola field isolates. SelimOmrane 1 , Anne-Sophie Walker 1 , Hind Sghyer 1 , Catherine Lanen 1 , Lamia Aouini 2 , Gert Keema 2 , Sabine Fillinger 1 . 1) BIOGER, INRA, Thiverval-Grignon, France;2) Plant Research International, Wageningen University, Wageningen, The Netherlands.Multidrug resistance (MDR) is a common trait developed by many organisms to counteract chemicals and/or drugs used against them. The basic MDRmechanism is relying on an overexpressed efflux transport system that actively expulses the toxic agent outside the cell. In fungi, MDR (or PDR) has beenextensively studied in Saccharomyces cerevisiae and Candida albicans, but also plant pathogenic fungi, e.g., Botrytis cinerea, Oculimacula yallundae andMycosphaerella graminicola are concerned by this phenomenon. In agriculture, it is currently under investigation if MDR strains may threaten the efficacyof current fungicide treatments. MDR strains were detected in septoria leaf blotch (M. graminicola) field populations since 2008. These strains are slightlymore resistant to DMI (inhibitor of the sterol 14 a-demethylase) fungicides than comparable cyp51 genotypes and cross-resistant to fungicides withdifferent modes of action. The identification of the molecular mechanism explaining the MDR phenotype in two isolated strains (MDR6 and MDR7) wasthe main goal of this study. By the use of C14-prochloraz, a DMI, we demonstrated increased fungicide efflux in both MDR strains in comparison tosensitive strains. RNA-sequencing led to the identification of several overexpressed transporter genes, out of which one MFS (major facilitator family)transporter had particularly abundant mRNA in both MDR strains. Crosses between both MDR strains showed that mdr6 and mdr7 loci are closely linked.We applied bulk-progeny sequencing to progeny of the crosses MDR6 x sensitive and MDR7 x sensitive in order to map the genomic regions co-segregatingwith the MDR phenotypes. SNP frequency analysis in sensitive and resistant bulks showed a clear co-segregation between phenotypes and the left arm ofchromosome 7. This region harbors a gene cluster including the MFS transporter gene mentioned above. After sequencing the mfs promoter, we identifieda 514 bp insertion in both MDR strains. Further studies are needed to validate the role of this insertion leading putatively to mfs overexpression and toclarify its relation to the MDR phenotype in the two studied strains. Financial support: Arvalis Institut du Vegetal, BASF Agro SAS, Bayer SAS, DuPont deNemours SAS, Syngenta Crop Protection AG.330. Candidate pathogenesis gene identification via Ustilago maydis `first gen` genomic analyses. M.E. Donaldson 1 , S. Meng 2 , B.J. Saville 1,3 . 1)Environmental & Life Sciences, Trent University, Peterborough, Ontario, Canada; 2) Lineberge Comprehensive Cancer Center, School of Medicine,University of North Carolina at Chapel Hill, Chapel Hill, N.C., USA; 3) Forensic Science Program, Trent University, Peterborough, ON, Canada.Three different approaches were used to identify candidate pathogenesis genes for the model plant pathogen Ustilago maydis (DC) Corda: 1) suppressivesubtractive hybridization (SSH) cDNA library analysis, 2) a bioinformatics approach, selecting genes unique among pathogenic basidiomycete fungi, and 3)microarray hybridization analyses. The SSH cDNA library was constructed to capture U. maydis genes expressed in planta, as well as Zea mays genes upregulatedduring the infection process. The resulting ESTs represented 23 U. maydis, and 159 Zea mays transcripts, respectively. Analysis of the U. maydistranscripts revealed 14 genes which have been previously characterized, 8 genes of unknown function, and 1 putative non-coding RNA. The fact that notall of the transcripts code secreted proteins is consistent with the observation by others that not all U. maydis effectors have recognizable secretionsignals. RT-PCR results supported in planta transcript levels for a subset these genes. To evaluate the three strategies, one gene from each series ofexperiments was chosen for selective gene deletion experiments. Deletion strains were created for: 1) a hypothetical gene (um03046) that had highrepresentation in the SSH cDNA library, 2) a conserved hypothetical gene (um01632) unique among basidiomycetes, and 3) the calcineurin B regulatorysubunit (cnb, um10226), identified through microarray hybridization as two-fold more highly expressed in the dikaryotic filamentous growth formcompared to the diploid filamentous form. All U. maydis deletion strains were capable of mating on DCM medium containing charcoal. Mutant U. maydisclones disrupted for um03046 and um01632 did not show aberrant growth phenotypes; and the morphology of these cells was indistinguishable fromwild-type strains. In contrast, the cnb mutants did not appear to separate after budding. In pathogenesis assays, the um03046 and um01632 mutants wereslightly more, or less pathogenic than wild-type infections, respectively. Results for the cnb mutants were more striking, with a marked 77% decrease inthe disease index, compared to wild-type infections. Together, these results support SSH cDNA library and microarray hybridization analyses as useful toolsin identifying genes expressed during specific stages of in planta development and those genes involved in pathogenesis.331. A biocontrol agent among pathogens : How Pseudozyma flocculosa genome relates to singular lifestyle. F. Lefebvre 1 , D.L. Joly 2 , G. Bakkeren 2 , F.Belzile 3 , R.R. Bélanger 1 . 1) Centre de recherche en horticulture, Département de phytologie, Université Laval, Quebec, QC, Canada; 2) Pacific Agri-FoodResearch Centre, Agriculture and Agri-Food Canada, Summerland, BC, Canada; 3) Institut de biologie intégrative et des systèmes, Département dephytologie, Université Laval, Québec, QC, Canada.Most fungal species belonging to the Ustilaginales are well known for their pathogenic activity towards a variety of plant species. One interestingexception is Pseudozyma flocculosa that rather acts as a biocontrol agent against powdery mildews. In order to better understand the factors underlyingthese opposed lifestyles among closely related organisms, the genome of P. flocculosa was first sequenced and annotated on the basis of homology to27th Fungal Genetics Conference | 201

FULL POSTER SESSION ABSTRACTSknown protein sequences, RNA-Seq data and ab initio predictors. Then, based on comparative genomics with the pathogenic species Ustilago maydis,Ustilago hordei and Sporisorium reilianum, we identified key features that could explain both the avirulent nature of P. flocculosa toward plants and thevirulent features of the pathogenic Ustilaginales.First, the genome structural annotation showed similarities in total gene number, gene density and average gene length. But these similarities hidesmajor differences with regards to average intron per gene and GC content. In fact, P. flocculosa has about 4 times more introns than U. maydis and has avery high GC content of 65.1%. Moreover, P. flocculosa genome shows a lower level of synteny than pathogenic species compared together.Second, comparison of gene content revealed unexpected results. On one hand, the genome of P. flocculosa harbor many traits usually associated withpathogenic species like plant cell-wall degrading enzymes and genes involved in the synthesis of secondary metabolites that are highly conservedcompared to pathogenic species. On the other hand, genes coding for candidate secreted effector proteins (CSEPs) show a much lower level ofconservation, that seem to explain, in part, the differences in lifestyle.Finally, this work led to the identification of certain of the most interesting genetic features in the study of pathogens and biocontrol agents.332. A tale of two poplar pathogens - Moving from sequence to function. B. Dhillon 1 , N. Feau 1 , P. Tanguay 2 , M. Sakalidis 1 , S. Beausiegle 1 , R. Ohm 3 , A.Aerts 3 , I. Grigoriev 3 , G. H. J. Kema 4 , S. B. Goodwin 5 , R. Hamelin 1 . 1) Forest Sciences, University of British Columbia, Vancouver, BC, Canada; 2) CFSLaurentian Forestry Centre, Succ. Sainte-Foy, Québec, Canada; 3) 3DOE Joint Genome Institute, Walnut Creek, California, USA, DOE, USA; 4) Plant ResearchInternational B.V., Wageningen, The Netherlands; 5) USDA-Agricultural Research Service, Purdue University, West Lafayette, Indiana, USA.Two closely related, morphologically indistinct fungal pathogens of poplars, Mycospharella populorum and M. populicola are prevalent in North America.In natural stands, these two fungal species closely follow the distribution of their host, with M. populorum being found on Aiegeiros botanical section andM. populicola on the Tacamahaca section of poplars. Epidemiologically, M. populorum is considered to be more aggressive, as in addition to leaf-spots, ithas the ability to infect woody tissue and cause cankers, an ability that M. populicola lacks. Moreover, introduction of hybrid plantations has added to M.populorum host range. Availability of genomes will allow us a window into understanding the genetic basis for these observed differences in epidemiologyand host-specificity for these two pathogens. Historical observation of host-specificity was confirmed by comparative sequence analysis the estimated thedivergence time between the two poplar pathogens to be ~6.4 Mya, which agrees with the divergence time estimates for the poplar botanical sections(6.8 - 7.8 Mya). Despite the remarkable macro-synteny exhibited between these two recently diverged pathogens, several genes specific to each pathogenwere identified in genomic regions where synteny broke down. In addition to being candidates for the different physiological and epidemiologicalattributes, these species-specific genes could be utilized for diagnostic and monitoring assays. A consistent expansion of several pathogenicity-relatedgene families was observed in M. populorum, suggesting a role for gene-dosage in determining its ability to cause cankers. Preliminary enzyme assaysshowed significant differences in beta-glucosidase and xylanase activities between these two fungi.333. Defining Open Chromatin Regions in Coprinopsis cinerea Oidia by FAIRE. Virginia K. Hench 1,2 , Patricia J. Pukkila 1,2 . 1) Department of Biology,University of North Carolina at Chapel Hill, NC 27599; 2) Office for Undergraduate Research, University of North Carolina at Chapel Hill, NC 27599.Changes in chromatin organization are principal regulatory mechanisms controlling multiple cellular processes including gene expression and meioticcrossover formation. Here we present FAIRE (formaldehye assisted isolation of regulatory elements) data that reveals regions of open chromatin inCoprinopsis cinerea oidia, the asexual spore stage of the C. cinerea life cycle. A standard FAIRE protocol was developed and optimized for oidia and used toenrich for nucleosome-free stretches of chromatin. FAIRE peaks were identified from single-end read whole genome sequence data using ZINBA (Zero-Inflated Negative Binomial Algorithm), which identified 7,276 peaks covering 6.3% of the genome. FAIRE peaks are predominantly intergenic with 78% ofFAIRE domains overlapping noncoding sequence. The peak widths range from 98-1390 bps, with an average width of 310 bps. Nearly half or 47% ofannotated genes (Broad version 3) contain a FAIRE peak in the proximal promoter region (defined as 500 bps immediately upstream of the gene start).Differential transcription has been characterized throughout the synchronous meiotic process in C. cinerea (Burns, C. et al., PLOS Genetics, vol 6, issue 9,2010), but the extent to which nucleosome occupancy might contribute to gene regulation in this multicellular fungus was not known. We found that aminority of meiotic specific (MS; genes expressed in meiosis and not in vegetative tissue) genes had promoter FAIRE peaks (of 819 genes 37% hadpromoter FAIRE peaks). In contrast, 61% of genes significantly changing during meiosis (SCDM; 2,455 genes) had promoter FAIRE peaks. Out of 295 genesthat were MS and SCDM, 38% had promoter FAIRE peaks in oidia. Genes with known meiotic function including spo11, dmc1, and rec8 were amongst theMS/SCDM genes that did not have promoter FAIRE peaks in oidia. In summary, all examined meiotic gene sets included genes associated with and withoutFAIRE peaks in their promoter regions, indicating that complex gene regulation mechanisms contribute to differential, tissue-specific gene expression in C.cinerea. Supported by the U.S. Department of Energy Joint Genome Institute Community Sequencing Program. The work conducted by the U.S. DOE JGI issupported by the Office of Science of the U.S. Department of Energy under Contract No. DE-AC02-05CH11231.334. Ensembl Fungi - genome-scale data portal from fungal species. Uma Maheswari, Heldér Pedro, Mark McDowall, Daniel M. Staines, Paul Kersey.European Bioinformatics Institute (EMBL-EBI), Cambridge, United kingdom.Ensembl Fungi (http://fungi.ensembl.org) is a portal offering access to genome-scale data from fungal species, using the Ensembl genome analysissystem, through a common set of interfaces shared with non-fungal species also represented in the Ensembl system. These include a web-based genomebrowser, Perl and REST-ful APIs, a public MySQL server and a query-orientated data warehouse (BioMart). The current release (January 2013) providesaccess to 36 fungal genomes across 12 different taxonomic orders, including the model species Saccharomyces cerevisiae and Schizosaccharomycespombe, (for which data are imported from the Saccharomyces Genome Database and PomBase respectively) but focuses mainly on plant pathogenspecies: genomic data from these is being integrated with information about infectious phenotypes (derived from PHI-base (http://www.phibase.org) on aper-gene basis, thro ugh a new targeted resource PhytoPath (http://www.phytopathdb.org).Core data provided for all species includes genome sequence, sequence patterns, annotation of protein and non-coding genes and functional annotationimported from direct curation, UniProt and InterPro. Information about gene regulation, sequence variation, evolution and conservation is also integratedin the system. Protein alignments are used to reconstruct evolutionary trees and infer homology relationships, while pairwise alignments between DNAsequences are performed between closely related species. Genomic polymorphisms are presented in the context of the reference genome sequences ofSaccharomyces cerevisiae and the phytopathogens Gibberella zeae, Puccinia graminis and Fusarium oxysporum.Ensembl Fungi will continue to expand with the increase in genomic data , we seek to work with the communities actively generating and using data, andare participants in a growing range of collaborations involved in the annotation and analysis of genomes.202

FULL POSTER SESSION ABSTRACTS335. A draft genome of the ectomycorrhizal fungus Rhizopogon vesiculosus: Characterization of mating system and heterozygosity within the dikaryon.Alija Mujic, Joseph Spatafora. Botany and Plant Pathology, Oregon State University, Corvallis, OR.Species of Rhizopogon are EM symbionts of trees in family Pinaceae and produce basidiospores within hypogeous false truffles that are dispersed bymycophagous mammals. All known members of R. subgenus Villosuli form obligate EM relationships with Pseudotsuga spp. (Douglas Fir) and are the onlymembers of the genus known to possess this host association. R. vesiculosus, along with its cryptic sister species R. vinicolor, possess a sympatricdistribution where sampled within the range of their host tree, P. menziesii. While the sporocarp and EM morphology of these fungi may be highly similar;they possess striking life history differences with R. vesiculosus producing larger vegetative genets and displaying greater population structure at bothlocal and landscape scales. We have sequenced the genome of R. vesiculosus using dikaryotic tissue and a whole genome shotgun sequencing approach onthe Illumina HiSeq platform. De novo assembly of the genome was performed using VELVET 1.19 and gene predictions were made using AUGUSTUS withLaccaria bicolor as a training model. The draft genome assembled to a total length of 46 Mb in 6700 contigs with an N50 of 26,783, a maximum contig sizeof 446,818 bp, and 12,604 predicted genes. Here we characterize the mating system of R. vesiculosus, which possesses both an A-locus encoding aheterodimer transcription factor, as well a B-locus encoding transmembrane pheromone receptors and pheromone precursor genes. We presentcomparisons of the mating system of R. vinicolor and its similarities to other members of Boletales (e.g., Serpula) and differences with Agaricales (e.g.,Laccaria). Due to the dikaryotic nature of the genome sequence produced for R. vesiculosus, single nucleotide polymorphisms (SNPs) can be observed andused to characterize allelic variation. SNPs observed in protein coding regions of both MAT loci indicate that R. vesisculosus is likely heterothallic. We havealso characterized heterozygosity across the whole genome in order to identify hypervariable regions. This genome will allow for comparative analysis ofgene content, mating type system with other Basidiomycota and, ultimately, for population/species-level genomic studies within Rhizopogon.336. Diverse Lifestyles and Strategies of Plant Pathogenesis Encoded in the Genomes of Eighteen Dothideomycetes Fungi. Robin A Ohm 1 , Nicolas Feau 2 ,Bernard Henrissat 3 , Conrad L Schoch 4 , Benjamin A Horwitz 5 , Rosie E Bradshaw 6 , Lynda Ciuffetti 7 , Richard C Hamelin 2,8 , Gert HJ Kema 9 , ChristopherLawrence 10 , James A Scott 11 , Joseph W Spatafora 7 , B. Gillian Turgeon 12 , Pierre JGM de Wit 13 , Shaobin Zhong 14 , Stephen B Goodwin 15 , Igor V Grigoriev 1 ,Other members of the Dothideomycetes community. 1) United States Department of Energy (DOE) Joint Genome Institute (JGI), Walnut Creek, CA, UnitedStates of America; 2) Faculty of Forestry, Forest Sciences Centre, University of British Columbia, Vancouver, BC, Canada; 3) Architecture et Fonction desMacromolécules Biologiques, Aix-Marseille Université, CNRS, Marseille, France; 4) NIH/NLM/NCBI, Bethesda, MD, United States of America; 5) Departmentof Biology, Technion - IIT, Haifa, Israel; 6) Institute of Molecular BioSciences, Massey University, Palmerston North, New Zealand; 7) Department of Botanyand Plant Pathology, Oregon State University, Corvallis, OR, United States of America; 8) Natural Resources Canada, Ste-Foy, QC, Canada; 9) Plant ResearchInternational, Wageningen, The Netherlands; 10) Virginia Bioinformatics Institute & Department of Biological Sciences, Blacksburg, VA, United States ofAmerica; 11) Division of Occupational & Environmental Health, Dalla Lana School of Public Health, University of Toronto, Toronto, Canada; 12) Departmentof Plant Pathology & Plant-Microbe Biology, Cornell University, Ithaca, NY, United States of America; 13) Laboratory of Phytopathology, WageningenUniversity, Wageningen, The Netherlands; 14) Department of Plant Pathology, North Dakota State University, Fargo, ND, United States of America; 15)United States Department of Agriculture, Agricultural Research Service, Purdue University, West Lafayette, Indiana, United States of America.The class Dothideomycetes is one of the largest groups of fungi with a high level of ecological diversity including many plant pathogens infecting a broadrange of hosts. Here, we compare genome features of 18 members of this class, including 6 necrotrophs, 9 (hemi)biotrophs and 3 saprotrophs, to analyzegenome structure, evolution, and the diverse strategies of pathogenesis. The Dothideomycetes most likely evolved from a common ancestor more than280 million years ago. The 18 genome sequences differ dramatically in size due to variation in repetitive content, but show much less variation in numberof (core) genes. Gene order appears to have been rearranged mostly within chromosomal boundaries by multiple inversions, in extant genomes frequentlydemarcated by adjacent simple repeats. Several Dothideomycetes contain one or more gene-poor, transposable element (TE)-rich putatively dispensablechromosomes of unknown function. The 18 Dothideomycetes offer an extensive catalogue of genes involved in cellulose degradation, proteolysis,secondary metabolism, and cysteine-rich small secreted proteins. Ancestors of the two major orders of plant pathogens in the Dothideomycetes, theCapnodiales and Pleosporales, may have had different modes of pathogenesis, with the former having fewer of these genes than the latter. Many of thesegenes are enriched in proximity to transposable elements, suggesting faster evolution because of the effects of repeat induced point (RIP) mutations. Asyntenic block of genes, including oxidoreductases, is conserved in most Dothideomycetes and upregulated during infection in L. maculans, suggesting apossible function in response to oxidative stress.337. Domains of meiotic DNA recombination and gene conversion in Coprinopsis cinerea (Coprinus cinereus). Patricia J. Pukkila 1 , Wendy Schackwitz 2 . 1)Dept Biol, Univ North Carolina, Chapel Hill, NC, USA; 2) US DOE Joint Genome Institute, Walnut Creek, CA, USA.We have shown previously that rates of meiotic recombination are highly non-uniform along the assembled chromosomes of C. cinerea (Stajich et al.PNAS 107: 11889-11894, 2010). That study revealed an over-representation of paralogous multicopy genes in regions with elevated levels of meioticexchange. In addition, retrotransposon-related sequences were not found in large segments of the genome with low levels of meiotic exchange. However,the study was limited by the available markers, and only 31 Mb of the 36 Mb genome could be mapped. More recently, we have resequenced 45 meioticsegregants and 4 complete tetrads. We developed a simple script to detect crossover and gene conversion events involving over 75,000 SNPs spanning 35Mb. The data were analyzed using MSTmap (Wu et al. PLoS Genetics 4: e1000212, 2008). The new dataset revealed sub-telomeric recombination hotspotsat every chromosome end, and 36% of the crossovers were associated with uninterrupted tracts of gene conversion. The conversion tracts (2-8 SNPs) werequite short (8-219 nt), and the median distance between the flanking SNP markers was also small (500 nt). Since these subtelomeric hotspots correspondto sites of synaptic initiation in C. cinerea (Holm et al. Carlberg Res. Commun. 46: 305-346, 1981), these data may contribute to our understanding of howhomologous chromosome pairing and synapsis are coordinated with meiotic recombination. Supported by the U.S. Department of Energy Joint GenomeInstitute Community Sequencing Program. The work conducted by the U.S. DOE JGI is supported by the Office of Science of the U.S. Department of Energyunder Contract No. DE-AC02-05CH11231.338. FungiDB: An integrated functional genomics database for fungi. Raghuraman Ramamurthy 1 , Edward Liaw 1 , Sucheta Tripathy 7 , John Brestelli 2,3 , SufenHu 3 , Wei Li 3 , Omar Harb 3,4 , Brian Brunk 3,4 , Steve Fischer 2,3 , Deborah Pinney 2,3 , Jessica Kissinger 5,6 , Brett Tyler 8 , David Roos 3,4 , Jason Stajich 1 . 1) Plantpathology and Microbiology, University of California, Riverside, Riverside, CA; 2) Department of Genetics, University of Pennsylvania School of Medicine,Philadelphia, PA; 3) Penn Center for Bioinformatics, University of Pennsylvania, Philadelphia, PA; 4) Department of Biology, University of Pennsylvania,Philadelphia, PA; 5) Center for Tropical & Emerging Global Diseases, University of Georgia, Athens, GA; 6) Department of Genetics and Institute ofBioinformatics, University of Georgia, Athens, GA; 7) Virginia Bioinformatics Institute, Virginia Tech University, Blacksburg, VA; 8) Center for Genome27th Fungal Genetics Conference | 203

FULL POSTER SESSION ABSTRACTSResearch and Biocomputing, Oregon State University, Corvallis, OR.FungiDB (http://FungiDB.org) is a functional genomic database and website tool for fungal genomes to enable data mining and analyses of the panfungalgenomic resources. The resource was developed in partnership with the Eukaryotic Pathogen Bioinformatic Resource Center (http://EuPathDB.org).Using the same infrastructure and user interface as EuPathDB, FungiDB allows for sophisticated and integrated searches to be performed over an intuitivegraphical system. The new 2.2 release contains sequence and annotation for over 50 species spanning the Ascomycota, Basidiomycota, Zygomycota, andChytrid fungi; including pathogenic species from the Cryptococcus, Histoplasma, and Coccidiodes genera. Six Oomycete genomes from Phytophthora andPythium species and RNA-Seq data are also included in the release of the system. Data from Saccharomyces cerevisiae, Candida albicans, Aspergillusnidulans, and Neurospora crassa represent the latest annotation releases of these genomes.Functional genomics data is available for querying including gene expression data from microarray, RNA-Seq, and expressed sequence tags; yeast twohybrid interaction data; and gene ontology from curated and automated sources. New features in the 2.2 release include population genomics data ofSNPs for several ascomycetes including A. fumigatus. A user interface to the precomputed orthology and paralogy of complete gene sets from thesupported fungal genomes along with key metazoan, plant, microbial eukaryotes, and bacteria enable phylogenetic profiling across the tree of life. Thedata-mining interface also permits the ability to make inferences using functional data in one species transformed by orthology into another species,providing a powerful resource for in silico experimentation. Query strategies from the system can be saved and shared as web links to enable reproducibleresults. FungiDB is supported by the Burroughs Wellcome Fund and the Alfred P. Sloan Foundation.339. Letters from the front: The Microbotryum violaceum genome and transcriptome project. Su San Toh 1 , Jared Andrews 1 , Sébastien Duplessis 2 , DavidTreves 3 , Christina Cuomo 4 , David Schultz 1 , Michael Perlin 1 . 1) University of Louisville, Louisville, KY, USA; 2) Centre INRA de Nancy, Champenoux, France; 3)Indiana University Southeast, New Albany, Indiana, USA; 4) Broad Institute, Cambridge, Massachusetts, USA.Microbotryum violaceum is a fungal species complex that includes related smut species primarily infecting members of the Caryophyllaceae (pinks).Individual species of this group are limited to successful infection and reproduction on a specific host species. We have produced a draft sequence at 18xcoverage for a haploid strain derived from meiosis of teliospores isolated from the host Silene latifolia. The draft sequence is currently in the process ofannotation and is publicly available through a website at the Broad Institute. Using Illumina Next Gen sequencing, we are generating deep transcriptomeinformation about a variety of stages in the lifecycle of the fungus, with particular emphasis on the late stages of infection, where teliosporogenesisoccurs. Through the analysis we have performed so far, we were able to identify a suite of secreted proteins (SPs) that are potentially involved in hostpathogeninteractions. Some of these include plant cell degradation enzymes like pectinesterase, laccase, subtilase and glycoside hydrolase. Moreover,some of these SPs are small, unique and cysteine-rich proteins, that might be involved in pathogenicity. Finally, since no reliable transformation system hasbeen adapted for this fungus and, as a consequence, no targeted gene disruption has been demonstrated, we are developing constructs that rely on thenewly completed genome to devise new strategies to allow such functional analyses in the future.340. The Aspergillus and Candida Genome Databases: Recent Developments and Future Plans. Martha B. Arnaud 1 , Gustavo C. Cerqueira 2 , Diane O.Inglis 1 , Marek S. Skrzypek 1 , Jonathan Binkley 1 , Clinton Howarth 2 , Prachi Shah 1 , Farrell Wymore 1 , Gail Binkley 1 , Stuart R. Miyasato 1 , Matt Simison 1 , GavinSherlock 1 , Jennifer Russo Wortman 2 . 1) Dept. of Genetics, Stanford University School of Medicine, Stanford, CA; 2) Broad Institute, Cambridge, MA.The Aspergillus and Candida Genome Databases (AspGD, http://www.aspgd.org and CGD, http://www.candidagenome.org/) are freely available, webbasedresources for researchers studying the molecular biology of these fungi. The interfaces of both web sites and databases now provide streamlined,ortholog-based navigation of the genomic and functional annotation for multiple species concurrently. We have completed manual curation of thepublished literature about multiple Candida and Aspergillus species. As part of our community-oriented mission, we also provide resources to fosterinteraction and dissemination of community information, tools, and data, including collecting, archiving, and providing large-scale datasets for download.AspGD also offers a full-featured genomics viewer to facilitate comparative genomics analysis. We have added new servers to improve web siteperformance and page loading speeds. Areas of future expansion include incorporation and curation of additional species, as well as improvements to thereference genome sequences and gene sets, utilizing high-throughput sequence to correct errors in sequence and gene structure, and display of additionalregulatory elements and gene products, including alternate splice forms. We also plan to develop and incorporate improved tools for query, display andanalysis of data, especially large-scale and comparative data such as gene synteny and the evolution of genes and gene substructure (e.g., intron gain andloss). We welcome, encourage, and appreciate your questions, feedback or suggestions. AspGD and CGD curators can be reached at aspergilluscurator@lists.stanford.eduand candida-curator@lists.stanford.edu, respectively. AspGD is funded by grant R01 AI077599 from the National Institute ofAllergy and Infectious Diseases, and CGD is funded by R01 DE015873 from the National Institute of Dental and Craniofacial Research at the US NationalInstitutes of Health.341. The Trichoderma reesei polyketide synthase gene pks1 is necessary for yellow-green pigmentation of conidia and is involved in the establishmentof environmental fitness. Lea Atanasova 1 , Benjamin P. Knox 2 , Christian P. Kubicek 1 , Scott E. Baker 2 , Irina S. Druzhinina 1 . 1) Microbiology Group, ResearchArea Biotechnology and Microbiology, Institute of Chemical Engineering, Vienna University of Technology, 1060 Vienna, Austria; 2) Chemical and BiologicalProcess Development Group, Pacific Northwest National Laboratory, Richland, WA, USA.The economically important genus Trichoderma (Hypocreales, Ascomycota, Dikarya) is well known for its mycotrophic lifestyle and for the broad range ofbiotrophic interactions with plants and animals. Moreover it contains several cosmopolitan species characterized by their outstanding environmentalopportunism. These properties have given rise to the use of several species in agriculture as biopesticides and biofertilizers while T. reesei is applied forproduction of bioenergy-related enzymes. The molecular basis of the opportunistic success of Trichoderma is not yet well understood. While there is someevidence for a role of secreted enzymes and proteins, less is known about a possible role of secondary metabolites. Recently it was predicted that the PKSencoding gene pks1 from T. reesei and its orthologues are most likely responsible for the characteristic yellow-green pigmentation of conidia. To reveal thefull function of the gene we deleted it from the wild-type strain QM 6a what resulted in complete loss of the green coloration of conidia. Theecophysiological profiling of Dpks1 showed that the gene is also involved in multiple functions at different stages of the T. reesei life cycle. Testing theantagonistic antifungal potential of the T. reesei Dpks1 mutant against several host/prey fungi suggested that the loss of pks1 reduced the ability tocombat them by means of both mechanisms: the pre-contact inhibition and direct overgrowth. However the overall analysis of mycoparasitic interactionssuggests that the gene is most likely involved in protection against other fungi rather than in attacking them. Interestingly, we noticed the increasedproduction of volatile compounds by the Dpks1 strains. The phenotype microarrays showed that PKS1 encoding gene restricts T. reesei from conidiation ona number of the best utilized carbon sources but does not influence the sexual development except the alteration of stromata pigmentation. The data fortranscriptional response of genes putatively involved in above mentioned processes will be presented.204

FULL POSTER SESSION ABSTRACTS342. Functional Analysis of Genes in Regions of Introgression in Coccidioides. Bridget M. Barker. Immunology & Infectious Diseases, Montana State Univ,Bozeman, MT.Coccidioides immitis and C. posadasii are dimorphic fungi endemic to the Americas. Genomic analysis of sequenced strains of C. posadasii and C. immitisreveals insights into the population biology of these organisms. There is strong evidence for hybridization and introgression, such that for many of the C.immitis strains, there are several regions that have a closer match to C. posadasii, but few regions within C. posadasii matching C. immitis. Multiplehybridization regions were located in several genomes analyzed, and at least one region containing ten genes exhibits a pattern consistent withintrogression in C. immitis. This conserved region was further evaluated in a larger collection of isolates. Approximately half of the C. immitis isolatescontain the C. posadasii fragment, and the majority of those are from the southern California and Mexico populations. The region of introgressionrepresents a unique opportunity to functionally assess genes that are likely to be relevant for species-specific virulence and adaptation to mammalianhosts or the environment. This region has a shared recombination point flanking a metalloproteinase, Mep4; genes that are highly expressed in theparasitic phase; and genes of unknown function. Importantly, evolutionary selection has preserved this region in multiple strains of C. immitis furtheremphasizing the possible role in virulence of these genes. Variation among strains for virulence in murine models of coccidioidomycosis has beenobserved, but has not been tested in the context of the newly discovered species or with a targeted underlying genetic mechanism hypothesis to test.Gene deletion mutants are being generated for three genes in the conserved introgression region to determine effects on in vitro growth andmorphological change under host relevant conditions.343. Classification and accurate functional prediction of carbohydrate-active enzymes by recognition of short, conserved peptide motifs. Peter K. Busk,Lene Lange. Biotechnology and Chemistry, Aalborg University, AAU Cph, Copenhagen, Copenhagen, Denmark.Functional prediction of carbohydrate-active enzymes is difficult due to low sequence identity hampering recognition of the functional relationship.However, similar enzymes often share a few short motifs, e.g., around the active site even when the overall sequences are very different. To exploit thisnotion for functional prediction of carbohydrate-active enzymes we developed a simple algorithm, Peptide Pattern Recognition (PPR) that can divideproteins into groups of sequences that share a set of short conserved sequences. When this method was used on 118 functionally characterized GH5proteins with 9 % average pairwise identity and representing four enzymatic functions, 97 % of the GH5 proteins were sorted into groups correlating withtheir enzymatic activity. Furthermore, we analyzed 8138 GH13 proteins including 204 experimentally characterized enzymes with 28 different functions.There was a 91 % correlation between group and enzyme activity. These results indicate that the function of carbohydrate-active enzymes can bepredicted with high precision by finding short conserved motifs in their sequences. The GH61 family is important for fungal biomass conversion but onlyfew GH61s have been functionally characterized. Interestingly, PPR divided 743 GH61 proteins into 16 subfamilies useful for targeted investigation of thefunction of these proteins, and pinpointed three conserved motifs with putative importance for enzyme activity. The conserved sequences were useful fordiscovery and cloning of new, subfamily-specific GH61 proteins from 14 different fungi. In conclusion, identification of conserved sequence motifs is a newapproach to sequence analysis that can predict carbohydrate-active enzyme functions with high precision. Furthermore, these motifs can be used to minegenomes and more complex data such as metagenomes and -transcriptomes for genes encoding proteins with specific, enzymatic activity.344. The mechanism of introner-like element multiplication in fungi. Ate van der Burgt 1 , Edouard Severing 2 , Valeria Ochoa Tufiño 1 , Pierre de Wit 1 , JérômeCollemare 1 . 1) Laboratory of Phytopathology, Wageningen University, Wageningen, Netherlands; 2) Laboratory of Bioinformatics, Wageningen University,6708PB Wageningen, The Netherlands.The recent discovery of introner-like elements (ILEs) in six fungal species shed new light on the origin of regular spliceosomal introns (RSIs). ILEs are novelspliceosomal introns that are found in hundreds of near-identical copies in unrelated genes. They account for the vast majority of intron gains in thesespecies and are not associated with intron losses. Remarkably, ILEs are longer than RSIs and harbor predicted stable secondary structures. However, theyare prone to quickly degenerate in sequence and length to become undistinguishable from RSIs, suggesting that ILEs are predecessors of most RSIs.Further analyses are being performed in order to understand the multiplication mechanism of ILEs, which is hypothesized to resemble the retro-homingmechanism of self-splicing group II introns. The dynamics of ILE’s secondary structures could be predicted and two conserved motifs were identified inalmost all fungal ILEs, which might play an important role in direct insertion into DNA. We also have developed a genetic screen in yeast in order tocapture and characterize ILE insertion events. These ongoing studies should provide hints about the mechanism of ILE multiplication, i.e. how newspliceosomal introns are gained in fungi.345. Fungi use prion folds for signal transduction processes involving STAND proteins. Asen Daskalov, Khalid Salamat, Sven J. Saupe. CNRS, IBGCUMR5095, BORDEAUX, AQUITAINE, France.Prions are proteins embedding genetic information into their structural state. Generally, those proteins exist in a soluble state and sporadically asinfectious amyloid aggregates. Podospora anserina’s [Het-s] is one of the best characterized fungal prions with a remarkably high prevalence in wildpopulations. [Het-s] functions in vegetative incompatibility - a biological process occurring during anastomosis between two genetically incompatiblestrains. The HET-s protein exists in a soluble state - [Het-s*] - or can switch to an aggregated amyloid state - [Het-s] - the prion form. When an [Het-s] prioninfected strain fuses with a strain expressing the alternative allelic variant of the het-s locus - het-S - a cell death reaction of the heterokaryon occurs.Recent studies shed light on the mechanism of [Het-s]/HET-S incompatibility reaction. Differing by 13 amino acids both proteins shares a two domainarchitecture; a globular N-terminal domain called HeLo and a C-terminal Prion Forming Domain (PFD). The latter is able to adopt a b-sheet richconformation with a specific b-solenoid fold. It has been demonstrated that in presence of [Het-s] amyloid fibers HET-S turns into a pore-forming toxin:transconformation of the HET-S PFD by [Het-s] fibers triggers the refolding of the HET-S HeLo domain, inducing the cell death reaction. In an attempt tobetter characterize the conserved features of the [Het-s] b-solenoid fold and identify new distant homologues of HET-S/s, we have generated a minimalconsensus sequence motif of it. Surprisingly, the second best hit in a BLASTp search is in the N-terminal region (3-23) of the product encoded by nwd2, theimmediately adjacent gene to het-S. NWD2 is a STAND protein. STAND proteins form signal transducing hubs through oligomerization upon ligandrecognition. That in mind and several bioinformatics observations led us to propose that HET-S and NWD2 are functional partners in various filamentousfungal species using the amyloid fold in a signal transducing pathway. We will present experimental evidence that NWD2 is able to trigger HET-S toxicity inmuch the same way as [Het-s] does. Further in silico analysis identify a number of these STAND/prion-like gene pairs and suggest that signal transductionthrough an amyloidal prion-like fold is a general widespread mechanism in fungi.346. RNA silencing in poplar anthracnose fungus Colletotrichum gloeosporioides. Simeng Li, Yonglin Wang, Chengming Tian. The Academy of Forestry,Beijing Forestry University, Beijing, China.Poplar anthracnose is one of the most destructive diseases on Poplus sp, whose causal agent is Colletotrichum gloeosporioides. Although the fungus is a27th Fungal Genetics Conference | 205

FULL POSTER SESSION ABSTRACTSbroad-host range plant pathogen, only dozens of genes involved in pathogenesis have been identified and characterized. In order to establish a highthroughputplatform,this study delivered the double-stranded RNA(dsRNA) expression cassette into protoplasts to trigger silencing for functional genomicsresearch in C. gloeosporioides.A new silencing vector pSD-SUR1 based on RNA-silencing vector (pSD1) with a convergent dual promoter was introduced. Inthis silencing system, the target gene was proposed to be transcribed as a chimeric RNA which activates the system. As an indicator of gene silencing, GFPfluorescence is used to evaluate efficiency of this silencing system. The fluorescence observation showed GFP fluorescence significantly decreased in someof the silenced strains, comparing with the recipient strain. The GFP mRNA transcript levels in the strains were analyzed using quantitative RT-PCR. Theresults showed that the reduction range of controls in gfp expression was from 30% to 80%,suggesting an effective gene silencing system and a feasibleapproach to generate detectable phenotypes in C. gloeosporioides. In addition, some genes encoding signal transduction pathways and transcriptionalfactor were inserted respectively into the vector pSD-SUR1 and to be silenced. In conclusion, RNA silencing system opens up new opportunity for exploringgene function in the fungus C. gloeosporioides.347. Comparative Genomics of L and S Morphotypes of Aspergillus flavus. Mana Ohkura, Peter Cotty, Marc Orbach. Division of Plant Pathology,University of AZ, Tucson, AZ.Aspergillus flavus is a widely distributed facultative pathogen of plants and animals and the most common causal agent of crop contamination withaflatoxins. Isolates of A. flavus vary widely in aflatoxin producing ability, ranging from atoxigenic to being capable of producing many mg/g. Variability inaflatoxin production makes specific attribution of etiology very complex. Aspergillus flavus exists in two morphotypes the large (L) and small (S) sclerotialproducing strains. The S strains have consistent high aflatoxin-producing ability while the L strains vary greatly in toxin production with atoxigenic strainscommonly found. Some atoxigenic strains are active ingredients in biocontrol products used commercially to prevent contamination. We are applyingcomparative genomics to L and S strains in an attempt to reveal clues to potential differential adaptations associated with the variation in aflatoxinproductionbetween these morphotypes. In addition to aflatoxin producing potential, several characteristics diverge between the morphotypes includingsclerotial morphology as well as spore and hydrolase production and prevalence. We hypothesize there are genomic differences between the L and Smorphotypes that reflect their divergent evolution leading to differential adaptation. To evaluate this, we have sequenced the genomes of three Lmorphotype and three S morphotype isolates from agricultural fields in Arizona belonging to 6 different vegetative compatibility groups. L and S strainisolates from across Arizona were selected. Strains were sequenced to ~40-45 X coverage on the Illumina HiSeq 2000 platform. The genomes wereassembled de novo using VelvetOptimiser and gaps were filled using GapFiller. Preliminary assessment of the assemblies indicate there is ~90% synteniccoverage with the published genome of A. oryzae RIB40, a close relative of A. flavus. The genomes were annotated transitively with RATT using thegenome of A. oryzae RIB40 as a reference. Comparisons of genome statistics, secondary metabolite clusters, and morphotype specific genes will bepresented.348. Searching for Functional Mobile Elements in Coprinopsis cinerea. Kendra Boyd, Madhura Chitnavis, Marilee A. Ramesh. Dept Biol, Roanoke College,Salem, VA.The genome of Coprinopsis cinerea contains both Class I and Class II repetitive elements, making up about 2.5% of the total genomic DNA. Whilebioinformatics techniques were used to identify and classify these elements based on sequence similarity, it is uncertain whether any of the elements arefunctional. Although the nature of repetitive elements is to expand their numbers within the genome, the genome acts to suppress activity throughmutation and methylation. Detailed analysis to survey functionality and expression was conducted on two families of repetitive elements, the Gypsy-likeretrotransposons (Class I) and the hAT Transposons (Class II). The potential for functionality was determined based on size, structure and flanking repeatsequence integrity. Evidence for expression was determined based on reviewing EST, SAGE and methylation data for these elements. Of the 31 largestgypsy elements analyzed, all appear to be inactive. However, one of the nine hAT elements appears to be structurally intact and shows evidence ofexpression. The dimerization domain of this element is being studied as a potential region to assay for activity.349. Genome-wide analysis of small RNA machineries in fungal kingdom. Jiayao Wu 1 , Jaeyoung Choi 2 , Fred O. Asiegbu 1 , Jari P.T. Valkonen 1 , Yong-HwanLee 1,2 . 1) Forest Science, Helsinki University, Helsinki, Finland; 2) Department of Agricultural Biotechnology, Seoul National University, Seoul, Korea.RNA interference (RNAi) is a phenomenon widely conserved in eukaryotes to regulate gene expression through diverse pathways at transcriptional (TGS)or post-transcriptional level (PTGS). In fungi, the RNAi pathways are found with three major functions: genomeic defence, heterochromatin formation, andgene regulation. The mechanisms of RNAi in fungi seem to be unique and highly differentiated from plant and animal kingdoms, although the coremechanisms are relatively similar. We identified 3 key genes such as Argonaute, Dicer and RNA-dependent RNA Polymerase (RdRP) in the pathway from143 fungal and 66 other genomes. They were found in most genomes with very different gene numbers, while some of fungal genomes appear to be lackof all the components indicating the absent of the whole pathways. In general, fungi have the same domains in Argonauts with plant and animal, butlonger in the length and less in the number. Compared to plant and animal, fungi have more Dicers, but they do not contain PAZ domain, which is essentialfor RNAi in plant and animal. Phylogentic analysis indicates that most fungal Argonuates belong to AGO-like subfamily. However, fungal Dicers could to bedivided into two subfamilies; one is closely related to plant and animal Dicers and the other only exists within fungal kingdom. Further analysis usingcodonW shows RNAi proteins are evolved into different subfamilies under natural selection not due to random mutation. Taken together fungi RNAipathway is likely to be much complex than we expected with multiple functions in diverse regulatory pathways. All information on proteins analyzed isarchived in Fungal Small RNA Machinery Database (http://funrna.riceblast.snu.ac.kr/).350. Comparative Analysis of Malassezia Mating Loci. Jun Xu 1 , Wenjun Li 2 , Anastasia Giotia 3 , Björn Nystedt 4 , Anna Averettec 2 , Charles Saunders 1 , ThomasDawson 1 , Joseph Heitman 2 , Annika Scheyniuse 5 . 1) Procter & Gamble Co., Mason Business Center, USA; 2) Department of Molecular Genetics andMicrobiology, Duke University Medical Center, USA; 3) Science for Life Laboratory, Translational Immunology Unit, Department of Medicine Solna,Karolinska Institutet, Stockholm, Sweden; 4) Science for Life Laboratory, Department of Biochemistry and Biophysics, Stockholm University, Stockholm,Sweden; 5) Translational Immunology Unit, Department of Medicine Solna, Karolinska Institutet, Stockholm, Sweden.Malassezia fungi are naturally found on the skin surfaces of many animals and are associated with skin disorders such as dandruff and atopicdermatitis/eczema. Whole genome sequence analysis showed that M. globosa has a bipolar mating type with two MAT alleles encoding the homeodomain(HD) and pheromone/receptor (P/R) loci separated by 167 kb of intervening sequence. We compared the M. globosa mating locus with the newlysequenced M. sympodialis genome. Our analysis showed that the M. sympodialis MAT region has extensive well-conserved synteny with M. globosa andthe HD and P/R mating subloci are physically linked with a 141 kb separating the two. Interestingly, MAT sequences derived from a population of M.sympodialis isolates suggests that M. sympodialis does not fit a traditional bipolar or tetrapolar system. Instead, it is more similar to a pseudo-bipolarmodel previously reported for Sporidiobolus salmonicolor in which the HD and P/R genes are physically linked similar to bipolar mating type configurations.206

FULL POSTER SESSION ABSTRACTSThis is in contrast to bipolar systems where recombination can still occur, giving rise to more mating types similar to tetrapolar mating systems. We alsoprovide some initial comparative analysis of the MAT region of M. restricta.Education and Professional Development351. The internet effectiveness for gaining students enrolled at college of education The scientific facts and concepts about Biofuel issue according tothe Responsibility Spiral model. Khlood S AlSheikh. Science Education, K.A.U, Jeddah, Saudi Arabia.This study was conducted to ascertain the effectiveness of the internet gaining students enrolled at college of education the scientific facts, concepts anddisagreement about Biofuel issue according to the Responsibility Spiral model. The study used a quasi-experimental design. The study sample consisted of76 students divided into two groups. The experimental group consisted of 37 students, whereas the control group consisted of 39 students. The studyused materials and activities according to the Responsibility Spiral model. Each stages of the waks (1992) model consisted of objectives, content, teachingand learning activities and evaluation tools. The results showed statistically significant differences between the mean score of the experimental groupwhich gained the scientific facts and concepts about Biofuel issue according to the Responsibility Spiral model and the mean score of the control group.The result showed that there is an opposite relationship between the model stages and t-test that if the model expands the value of t-test decreased.Based on the results, the researcher recommends that the internet is not enough tools for advanced stages of the Responsibility Spiral model.352. ComGen Authentic Research Experiences (C-ARE): Fungal genetic analysis. Gita Bangera 1 , Andrea Gargas 2 . 1) Bellevue College, Bellevue, WA, USA; 2)Symbiology LLC, Middleton, WI, USA.ComGen (Community College Genomics Research Initiative) teaches students the skills of self-directed learning, critical thinking, and analysis.Community college students in this program receive a mini-graduate school experience, following a single requisite course in cell biology. Students workon original research projects, learn to troubleshoot their experiments, organize lab meetings and student journal clubs, and network within the scientificcommunity. In one research track students work with DNA from described fungal collections, learning DNA-based techniques including PCR amplification,DNA sequencing and sequence analysis. Student-gathered sequence information is used to advance identification and phylogenetic results for thesecollections. With NSF Award DUE #1225857 ComGen (C-ARE): Dissemination, Enrichment and Expansion Project the project will be expanding tocommunity college partners throughout the Seattle/Tacoma region of Washington State.353. Facilitating an Interdisciplinary Learning Community Amongst Undergraduate Research Fellows By Emphasizing Scientific Inquiry as the UnifyingThread. Virginia K. Hench 1,2 , Patricia J. Pukkila 1,2 . 1) Department of Biology, University of North Carolina at Chapel Hill, NC 27599; 2) Office forUndergraduate Research, University of North Carolina at Chapel Hill, NC 27599.The HHMI-Future Scientists and Clinicians (HHMI-FSC) fellowship is 1 of 3 components of the HHMI Science Learning Communities program at UNCChapel Hill. The HHMI-FSC program was designed to foster an intellectual community that empowers high-ability students from low-incomebackgrounds to engage in biomedical research for 2 summers. Each year, 12 new fellows are matched with mentors in labs spanning a range of biomedicalareas. They work fulltime in labs on their own research project and meet weekly as a group to engage in interactive programming that targets skills criticalfor success in science beyond the bench. One area of emphasis has been the process of inquiry itself. The goal is for students to transition from being apair of hands executing protocols to active learners invested in their own projects and able to speak with authority about why experiments are performedin particular ways and what conclusions can be drawn from data generated. This starts with coaching students to state the questions that they are tryingto answer and think through whether an experimental setup is consistent with what they say they are trying to find out. Assignments and feedback aredesigned to reinforce this principle. One of the most satisfying aspects of doing science is getting to follow one’s own instinctive curiosities and developthe methodologies needed to navigate new terrains. Undergraduates are usually still trying to define their own specific curiosities. Pushing students todescribe what they are curious and passionate about is one feasible strategy that can help students identify pursuits that fit their interests and talents.Another successful strategy has been to require returning second year fellows to share science learning experiences via 15-30 minute long talks for theirpeers. Some took the opportunity to become more immersed in their lab’s focus, while others branched into questions like what motivates scientists towork in foreign countries and what has genomic anthropology told us about human evolution. Project aims were developed through conversationsbetween the fellow and instructor. The one constraint was for fellows to organize their presentations around questions. Feedback indicated thatpresenters benefited from having to give presentations and others enjoyed learning about a broader array of topics.Gene Regulation354. YAB- An Agrobacterium-based vector system for direct cloning of eukaryotic gene constructs via yeast recombination. M Alejandra Mandel, Marc JOrbach. School of Plant Sciences and The Bio5 Institute, University of Arizona, Tucson, AZ.Agrobacterium tumefaciens-mediated transformation (ATMT) has become the preferred method to introduce modified genes in many fungal systems.Therefore, there is a need for a simple and efficient method to create gene constructs in an ATMT vector. Generation of ATMT constructs usually requiresPCR-amplification of DNA fragments and multiple cloning steps into binary Agrobacterium vectors. Advances have been made with attB/attP-basedsystems (e.g. pTroya, Gene-blast, DelsGate) that shorten the number of steps required for generating plasmid constructs, however these constructsrequire a final sub-cloning step into an Agrobacterium vector. One of these systems, OSCAR, combines PCR and attB/attP-based technology in anAgrobacterium platform. All of these methods require the use of commercial enzymes and usually involve one or more PCR amplification steps. There arealso plasmid vectors that use yeast transformation-associated recombination (TAR) which do not require the use of enzymes for cloning, but can only beused to transform fungi via electroporation, protoplast transformation or biolistics. We present here YAB (for Yeast-Agrobacterium-Bacteria), a vector thatcombines the homologous recombination properties of yeast in an Agrobacterium vector backbone that can be used to create any kind of gene construct(e.g. gene deletion mutants, fluorescence-tagged genes, overexpression, RNAi) to directly transform fungi in one step. DNA fragments are cloned into YABvia recombination between 22-nucleotide compatible ends generated by PCR, or by using short oligomers that “bridge” the DNA fragment with the vector,thus avoiding the need for amplification steps. YAB-Hph has a 1.4-kb Hygromycin phosphotransferase cassette flanked by the left and right borders of theTi plasmid. This vector was used to generate whole-gene deletion mutants by TAR in one step, that were directly introduced into fungi.355. Removal of C4-methyl Sterol Accumulation in a SREBP-null Mutant of Aspergillus fumigatus Restores Hypoxia Growth. Sara J. Blosser 1 , Brittney27th Fungal Genetics Conference | 207

FULL POSTER SESSION ABSTRACTSHendrickson 1 , Nora Grahl 2 , Dawoon Chung 2 , Bridget Barker 1 , Robert A. Cramer 2 . 1) Immunology & Infectious Disease, Montana State University, Bozeman,MT; 2) Microbiology & Immunology, Geisel School of Medicine at Dartmouth, Hanover, NH.The human pathogen Aspergillus fumigatus adapts to stress encountered in the mammalian host based on evolutionary mechanisms derived from itsecological niche as a composter or saprobe. SrbA, a member of the helix-loop-helix family of transcription factors, plays a significant role in A. fumigatushypoxia adaptation, antifungal drug responses, and virulence. SrbA is a direct transcriptional regulator of several key enzymes in the ergosterolbiosynthesis pathway, which has been verified by in vivo ChIP-SEQ analyses. The sterol intermediate profile of DsrbA revealed a significant accumulation ofC4-methyl sterols, which correlates with the loss of erg25 (C4-sterol methyl oxidase) mRNA abundance in the SrbA-null mutant. We hypothesized that thisC4-methyl sterol accumulation may contribute to the significant phenotypes observed in DsrbA. We have characterized the two genes predicted to encodeC4-methyl sterol oxidases (Erg25) in A. fumigatus. Genetic deletion of both erg25 genes, A and B, is lethal in A. fumigatus, while single genetic deletions ofthe respective genes are viable. Although loss of both erg25A and erg25B resulted in accumulation of C4-methyl sterols, Derg25A accumulated far moreC4-methyl sterol intermediates than Derg25B, suggesting that Erg25A is the predominant C4-sterol methyl oxidase in A. fumigatus. No dramatic in vitro orin vivo phenotypes under various stress conditions were observed in Derg25A or Derg25B mutants; however, a moderate reduction in hypoxia growth wasobserved in Derg25A. Generation of a strain that constitutively expresses erg25A in the DsrbA background biochemically relieved a majority of the C4-methyl sterol buildup in DsrbA. Significantly, restoration of erg25A mRNA levels in DsrbA with a promoter replacement fully restored the in vitro hypoxiagrowth defect of DsrbA. These results indicate that erg25 transcriptional regulation by SrbA and management of C4-methyl sterol intermediateaccumulation is highly important for hypoxia stress adaptation in A. fumigatus. Future studies will explore the impact of reducing C4-methyl sterol levels inDsrbA on A. fumigatus virulence.356. VeA Regulates Conidiation, Gliotoxin Production and Protease Activity in the Opportunistic Human Pathogen Aspergillus fumigatus. SourabhDhingra 1 , David Andes 2 , Ana M. Calvo 1 . 1) Biological Sciences, Northern Illinois University, DeKalb, IL; 2) Medical Mycology and immunology, University ofWisconsin-Madison, Madison, WI.Aspergillus fumigatus is the causative organism of invasive aspergillosis. Our study shows that normal levels of veA expression are necessary for wildtypemorphological differentiation in this medically important fungus. Specifically, deletion or overexpression of veA reduce conidiation. Parallely, brlAexpression was also affected by alterations in veA transcription levels. In addition, our studies revealed that veA regulates gliotoxin production in A.fumigatus. Gliotoxin production was decreased in the deletion veA and over-expression veA strains, where gliZ and gliP expression was altered.Interestingly, veA also controls hydrolytic enzyme activity in this human pathogen. Deletion of veA resulted in a reduction of protease activity; this is thefirst report of a veA homolog with a role in controlling fungal hydrolytic activity. Although veA affects several cellular processes in A. fumigatus,pathogenicity studies in a neutropenic mouse infection model indicated that veA is dispensable for virulence. More research will be conducted to studythe effect of veA in non-neutropenic mice.357. HapXcess and C-terminal truncation impairs Aspergillus fumigatus' iron homeostasis. Fabio Gsaller 1 , Veronika Klammer 1 , Beatrix E. Lechner 1 , PeterHortschansky 2 , Axel A. Brakhage 2 , Ernst R. Werner 3 , Hubertus Haas 1 . 1) Division of Molecular Biology, Medical University of Innsbruck, Austria; 2)Department of Molecular and Applied Microbiology, Leibniz Institute for Natural Product Research and Infection Biology (HKI), Jena, Germany; 3) Divisionof Biological Chemistry, Medical University of Innsbruck, Austria.The maintenance of iron homeostasis is indispensable as iron is essential for various cellular processes but can be toxic at iron excess. In A. fumigatus thebZIP-like transcription factor HapX is important for adaption to iron starvation and consequently virulence due to its role in repression of iron consumingpathways (i.e. heme biosynthesis, TCA cycle, respiration) and activation of iron uptake (i.e. siderophore biosynthesis and uptake, reductive ironassimilation). In this study we demonstrate that conditional hapX overexpression using the xylose-inducible xylP promoter leads to repression of genesinvolved in iron consumption (i.e. heme biosynthetic hemA and leucine-biosynthetic leuA) and activation of iron acquisition-related genes (i.e.siderophore-biosynthetic sidG and siderophore transporter-encoding mirB) within one hour of induction. In agreement, elevated hapX expressiondecreased the cellular accumulation of protoporphyrin IX, the iron-free precursor of heme, and increased production of the extracellular siderophoreTAFC. HapX-truncation studies revealed that the C-terminal 93 amino acid residues are essential for its activating as well as repressing functions. HapX N-terminally tagged with Venus green fluorescent protein localized to the nucleus during iron starvation but was undetectable after an one hour-shift to ironsufficiency. These data demonstrate tight iron-regulation of hapX expression at the protein level as previously shown at the transcript level. Consistently,HapX-deficiency is detrimental only during iron limitation. Two hapX copies and in particular xylP promoter-mediated overexpression of hapX causedgrowth defects independent of the iron availability, which underscores the importance of a precisely regulated HapX level. This work was supported by theAustrian Science Foundation grant FWF P21643-B11 to HH.358. The CCAAT-Binding-Complex mediates Iron Regulation in Aspergillus fumigatus. Hubertus Haas 1 , Christoph Joechl 1 , Thorsten Heinekamp 2 , Ilse D.Jacobsen 2 , Markus Schrettl 1 , Axel A. Brakhage 2 , Lukas Schafferer 1 . 1) Division of Molecular Biology, Biocenter, Innsbruck Medical University, Austria; 2)Department for Microbial Pathogenicity Mechanisms, Leibniz Institute for Natural Product Research and Infection Biology (HKI), Jena, Germany.Iron is essential for a wide range of cellular processes but its excess is toxic. Therefore, microorganisms evolved fine-tuned mechanisms for uptake andstorage of iron, to sustain iron homeostasis. In the opportunistic fungal pathogen Aspergillus fumigatus, the bZIP-type transcription factor HapX mediatesadaption to iron starvation by activating siderophore biosynthesis and repressing iron-dependent pathways. HapX-deficiency attenuates the virulence of A.fumigatus underlining the importance of adaptation to iron starvation in pathogenicity. The HapX N-terminal amino acid sequence predicts interactionwith the DNA-binding, heterotrimeric CCAAT-binding complex (CBC), which is conserved in all eukaryotes and believed to co-regulate up to 30% of allgenes. Here, we characterized the role of the CBC in iron regulation of A. fumigatus by analysis of the phenotypic consequences of genetic inactivation ofthe CBC subunit HapC. HapC-deficiency was deleterious during both iron starvation as well as iron sufficiency, demonstrating iron-independent regulatoryfunctions of the CBC. In contrast, HapX is important during iron starvation only. As shown previously for HapX-deficiency, HapC-deficiency derepressedgenes involved in iron-consuming pathways during iron starvation but decreased siderophore metabolism at transcriptional and metabolic levels.Inhibition of reductive iron assimilation by ferrous iron chelation blocked colony formation of both HapC-deficient and HapX-deficient conidia. Moreover,inactivation of HapC was epistatic to HapX-deficiency. Taken together, these data indicate that the CBC mediates both the activating and the repressingfunctions of the iron-regulatory transcription factor HapX. The central role of the CBC in environmental adaptation is underlined by HapC-deficiencyrendering A. fumigatus avirulent in a murine model of aspergillosis. This work was supported by the Austrian Science Foundation grant FWF I282-B09 toHH.359. Protein kinase A signaling in Aspergillus fumigatus: Identification of downstream targets. Juliane Macheleidt 1,4 , Wolfgang Schmidt-Heck 2 , Ilse D.208

FULL POSTER SESSION ABSTRACTSJacobsen 3 , Thorsten Heinekamp 1,4 , Axel A. Brakhage 1,4 . 1) Leibniz Institute for Natural Product Research and Infection Biology - Hans-Knöll-Institute,Molecular and Applied Microbiology, Jena, Germany; 2) Leibniz Institute for Natural Product Research and Infection Biology - Hans-Knöll-Institute, SystemsBiology / Bioinformatics, Jena, Germany; 3) Leibniz Institute for Natural Product Research and Infection Biology - Hans-Knöll-Institute, MicrobialPathogenicity Mechanisms, Jena, Germany; 4) Institute of Microbiology, Friedrich Schiller University, Jena, Germany.Aspergillus fumigatus is a saprophytic mould normally inhabiting the soil. The fungus also represents a medically important pathogen causing severesystemic infections in immunocompromised patients. To survive in these entirely different habitats, A. fumigatus needs mechanisms to senseenvironmental signals and transduce them intracellularly. One of these signal transduction pathways is the cAMP dependent protein kinase A (PKA)pathway. For A. fumigatus, components of this signaling cascade have been characterized in detail and its significance for virulence is evident.To identify target genes of PKA, we performed microarray analyses using a mutant strain overproducing the PKA catalytic subunit. Following thisapproach, 23 transcription factors potentially regulated by PKA were identified. From these, 15 were deleted and the mutant phenotypes werecharacterized. A gene encoding a C6 finger domain protein that showed highest upregulation of all identified transcription factors is located in a potentialsecondary metabolite gene cluster. Deletion of the transcription factor gene resulted in reduced growth and sporulation of the mutant strain. Thisphenotype was observed even more drastically for a strain lacking the nonribosomal peptide synthetase of the same cluster. Because genes of this clusterwere shown to be transcribed in infected mouse lungs, a virulence study was performed using an embryonated egg infection model. However, thetranscription factor deletion mutant showed no altered virulence compared to the corresponding wild type.To get deeper insights into the function of the secondary metabolite gene cluster, the gene encoding the C6 finger domain protein was overexpressedusing an inducible promoter. Overproduction of the transcription factor resulted in induced transcription of all cluster genes and furthermore in theformation of a brown substance which is currently under investigation.360. Aspergillus nidulans galactofuranose biosynthesis affects antifungal drug sensitivity. Md. Kausar Alam, Susan Kaminskyj. Biology, University ofSaskatchewan, Saskatoon, SK, Canada.The cell wall is essential for fungal survival in natural environments. Many fungal wall carbohydrates are absent from humans, so they are a promisingsource of antifungal drug targets. Galactofuranose (Gal-f) is a sugar that decorates certain carbohydrates and lipids. It comprises about 5% of theAspergillus fumigatus cell wall, and may play a role in systemic aspergillosis. We are studying Aspergillus wall formation in the tractable model system, A.nidulans. Previously we showed single-gene deletions of three sequential A. nidulans Gal-f biosynthesis proteins each caused similar hyphalmorphogenesis defects and 500-fold reduced colony growth and sporulation. Here, we controlled A. nidulans ugeA, ugmA or ugtA using the alcA(p) orniiA(p) promoter. For repression and expression, alcA(p)-regulated strains were grown on complete medium with glucose or threonine, whereas niiA(p)-regulated strains were grown on minimal medium with ammonium or nitrate. Expression was assessed by qPCR and colony phenotype. The alcA(p) andniiA(p) strains produced similar effects: colonies resembling wild type for gene expression, and resembling deletion strains for gene repression. Gal-fimmunolocalization using the L10 monoclonal antibody showed that ugmA deletion and repression phenotypes correlated with loss of hyphal wall Gal-f.None of the gene manipulations affected itraconazole sensitivity, as expected. Deletion of any of ugmA, ugeA, ugtA, their repression by alcA(p) or niiA(p),OR, ugmA overexpression by alcA(p), increased sensitivity to Caspofungin. Strains with alcA(p)-mediated overexpression of ugeA and ugtA had lowercaspofungin sensitivity. Gal-f appears to play an important role in A. nidulans growth and vigor. We are extending these studies to A. fumigatus UgmA andUgtA to determine which amino acids are critical for function and Gal-f generation. Previously, we showed that wild type AfugmA can restore an A.nidulans ugmA deletion strain to wild type phenotype. Our current results show that certain amino acid residues in A. fumigatus UgmA are critical for Gal-fgeneration: constructs with mutated A. fumigatus sequences failed to rescue the AnugmD phenotype.361. WITHDRAWN362. Sulfur metabolism regulatory mutations induce environmental stress response. J. Brzywczy, M. Sienko, R. Natorff, M. Skoneczny, J. Kruszewska, A.Paszewski. Institute of Biochemistry and Biophysics, 02-106 Warszawa, Poland.Mutations in the cysB, sconB and sconC genes affect sulfur metabolism in Aspergillus nidulans in different ways. The cysB mutation blocks synthesis ofcysteine and leads to a shortage of this amino acid while sconB and sconC mutations lead to elevated levels of cysteine and glutathione. We havecompared transcriptomes of these three mutants to the wild type strain finding that expression of 1263 genes is altered at least twofold. Transcripts of908 genes are elevated and 355 genes exhibit decreased levels of transcripts. Despite opposite effect on sulfur metabolism these mutations influenceexpression of overlapping sets of genes. We have assigned categories of Functional Catalogue to up- and down-regulated genes and have identifiedcategories most enriched with differentially regulated genes. Besides genes involved in sulfur metabolism we find that many up-regulated genes arerelated to stress responses. Two component signal transduction system is a category that is fifteen times enriched with genes up-regulated in the sconCmutant and also highly enriched in the cysB and sconB mutants (eight- and tenfold, respectively). Genes encoding heat shock proteins and enzymes ofglutamate degradation pathway are also up-regulated. The glutamate degradation pathway is also known as a GABA shunt which is induced by anoxidative stress. A large group of up-regulated genes is involved in carbohydrate and energy metabolism including genes coding for enzymes of trehaloseand glycerol synthesis. Genes coding for enzymes of alcohol fermentation, which are induced in response to anaerobic stress, are also up-regulated in thesulfur regulatory mutants. Altered expression of carbohydrate metabolism genes is accompanied by changes in sugar accumulation in mutant mycelia andconidia. Among down-regulated genes there are many encoding membrane proteins and enzymes involved in secondary metabolism including penicillinbiosynthesis cluster. Genes coding for lysozyme are down-regulated too. As secondary metabolites often inhibit growth of other organisms, loweredexpression of genes responsible for their synthesis suggests a decreased response to biotic stress in sulfur metabolism mutants.363. RNA 3' tagging - signalling transcript degradation and translational repression. Mark X. Caddick, Meriel G Jones, Daniel Rigden, Igor Y Morozov. DeptBiological Sci, Univ Liverpool, Liverpool, Merseyside, United Kingdom.A large body of work has elucidated the mechanisms associated with mRNA degradation and translational repression - processes which are fundamentalto biological systems. For the majority of eukaryotic transcripts, deadenylation is known to lead to transcript degradation and translational repression.However, the surveillance mechanism which determines the point at which functional transcripts are effectively switched off is poorly defined. Thistransition is generally associated with the mRNA being tagged at the 3' end with a short run of pyrimidine nucleotides. For example, cell cycle-regulateddecapping and degradation of mammalian histone mRNA is triggered by uridylation. In fission yeast and the filamentous fungus, Aspergillus nidulans,polyadenylated transcripts are decapped and degraded in response to 3' tagging. In A. nidulans and Arabidopsis tagging involves the addition of a C/U richelement when the transcript's poly(A) tail is shortened to ~15 nucleotides. Disruption of the Aspergillus nucleotidyltransferases, CutA and CutB, results inloss of tagging, lower rates of degradation and an accumulation of transcripts with short poly(A) tails. Recently we have shown that 3' tagging is also27th Fungal Genetics Conference | 209

FULL POSTER SESSION ABSTRACTSinduced by nonsense mediated decay (NMD) and is required for efficient dissociation of the resulting termination complex. Upf1, a major NMDcomponent, is implicated not only in this but, surprisingly, in all mRNA tagging to date. This led us to propose that deadenylation of mRNA results in an"NMD-like" event, where dissociation of poly(A) binding protein results in recruitment of Upf1 to the mRNA thereby triggering translational repression,clearance of the termination complex and efficient RNA degradation. We will describe our recent findings which relate to the mechanisms that triggermRNA tagging and the downstream events which ultimately repress transcript expression.364. A novel C2H2 type finger transcription factor, MtfA, regulates mycotoxin biosynthesis and development in Aspergillus nidulans . VellaisamyRamamoorthy, Sourabh Dhingra, Sourabha Shantappa, Ana M. Calvo-Byrd. Biological Sciences, Northern Illinois University, Dekalb, IL.Secondary metabolism in the model fungus Aspergillus nidulans is controlled by the global conserved regulator VeA, which also governs morphologicaldifferentiation. Among the secondary metabolites regulated by VeA is the mycotoxin sterigmatocystin (ST), where the presence of VeA is necessary for thebiosynthesis of this carcinogenic compound. We identified revertant mutants (RM) capable of synthesizing ST in the absence of VeA. The point mutation inthe RM7 mutant occurred at the coding region of a gene encoding a novel putative C2H2 zinc finger domain type transcription factor that we denominatedMtfA. As expected, the A. nidulans mtfA gene product localized at nuclei. Deletion of the mtfA gene restored mycotoxin biosynthesis in the absence of veA,but drastically reduced mycotoxin production when mtfA gene expression was altered, by either deletion or overexpression, in the Aspergillus nidulanswild type strain. Our study revealed that mtfA regulates ST production by controlling the expression of the specific ST gene cluster activator aflR.Importantly, mtfA also controls sexual and asexual development in A. nidulans. Deletion of mtfA results in a reduction of conidiation and sexualdevelopment.365. Histidine 704 of the Aspergillus nidulans GATA factor AreA is required for nuclear export. Damien Downes 1 , Brandon Pfannenstiel 1 , CameronHunter 1 , Kendra Siebert 1 , David Clarke 2 , Meryl Davis 2 , Richard Todd 1 . 1) Department of Plant Pathology, Kansas State University, Manhattan, USA; 2)Department of Genetics, University of Melbourne, Melbourne, AUS.In A. nidulans, the GATA transcriptional activator AreA controls the preferential utilization of nitrogen nutrients as well as the response to nitrogenstarvation. During nitrogen starvation AreA accumulates in the nucleus, and a strong increase in target gene expression is observed. Addition of nitrogennutrients to nitrogen starved cells results in rapid translocation of AreA to the cytoplasm and arrest of elevated AreA-dependent gene expression,indicating that regulated nuclear export is the control mechanism for AreA nuclear accumulation. AreA contains a single conserved CrmA-dependentNuclear Export Sequence (NES). We propose that regulated AreA nuclear export is controlled by post-translational modification of residues within the NES.We show that deletion of the AreA NES confers nuclear accumulation. Substitution of individual amino acids within the AreA NES identified a singlehistidine residue, which when mutated to a non-modifiable alanine residue leads to constitutive nuclear accumulation. This suggests that histidinemodification may promote AreA nuclear export. We show that fusion of the AreA NES to the constitutively nuclear protein PrnA confers nucleocytoplasmicdistribution and a proline utilization loss of function phenotype. We have used this phenotype to select mutants affecting AreA-dependent nuclear export.366. Redundant Nuclear Localization Signals Mediate Nuclear Import of the Aspergillus nidulans Transcription Activator of Nitrogen Metabolic GenesAreA. Cameron C. Hunter 1 , Kendra S. Siebert 1 , Damien J. Downes 1 , Koon Ho Wong 2 , Sara Lewis 2 , James A. Fraser 2 , David F Clarke 2 , Michael J. Hynes 2 , MerylA. Davis 2 , Richard B. Todd 1 . 1) Department of Plant Pathology, Kansas State University, Manhattan, KS; 2) Department of Genetics, The University ofMelbourne, Parkville VIC 3010, Australia.The Aspergillus nidulans GATA transcription factor AreA activates transcription of nitrogen metabolic genes. AreA accumulates in the nucleus duringnitrogen starvation but not in the presence of nitrogen sources. AreA contains five putative classical nuclear localization sequences (NLSs) and oneputative non-canonical bipartite NLS. We used two approaches to identify the functional NLSs. First, we constructed epitope-tagged gene replacementareA mutants affected in individual NLSs or combinations of NLSs to identify sequences required for nuclear localization. Deletion of all five classical NLSsdid not affect utilization of nitrogen sources and did not prevent AreA nuclear localization. Mutation of the bipartite NLS conferred inability to utilizealternative nitrogen sources but did not prevent AreA nuclear localization. Combinations of mutations of the six NLSs indicate redundancy among the AreANLSs. Second, we constructed Green Fluorescent Protein (GFP)-AreA NLS fusion genes and introduced them into A. nidulans. The bipartite NLS stronglydirects GFP to the nucleus, one of the classical NLSs weakly directs GFP to the nucleus and the other four classical NLSs collaborate to direct GFP to thenucleus.367. Conditional expression of the phospho-transmitter gene ypdA and the signaling interaction of YpdA with response regulators; SskA and SrrA inAspergillus nidulans. Mayumi Nakayama 1,2 , Yura Midorikawa 1,2 , Akira Yoshimi 2 , Daisuke Hagiwara 2,3 , Keietsu Abe 1,2 . 1) Tohoku University, Sendai, Miyagi,Japan; 2) ABE-project New Industry Hatchery Center (NICHe) Tohoku University, Sendai, Japan; 3) Present address: MMRC, Univ. of Chiba, Chiba Japan.The histidine-to-aspartate (His-Asp) phosphorelay signaling transduction system has been conserved widely in both prokaryotes and eukaryotes. Thesystems typically consist of three types of common signal transducers: His-kinase (HK), a response regulator (RR), and a histidine-containingphosphotransfer intermediate (HPt). Generally, HPt acts as an intermediate between HK and RR and is indispensable for inducing appropriate responses toenvironmental stresses through His-Asp phosphotransfer signaling. In Aspergillus nidulans, we revealed His-Asp phosphorelay signal transducers: HK(NikA), RR (SskA and SrrA), and HPt (YpdA) were essential for the response of high-osmotic and oxidative stresses. Nevertheless the ypdA is the essentialgene, the molecular mechanism underlying the importance of YpdA remains unclear. To identify the function of the YpdA, we constructed A. nidulansmutant in which expression of the ypdA gene is conditionally regulated under the control of the A. nidulans alcA promoter (CypdA strain) and analyzedtheir phenotype. We constructed mutant strain from CypdA by deleting the response regulator gene srrA (CypdA/DsrrA) and sskA (CypdA/DsskA). WhenypdA was downregulated, CypdA showed remarkable growth retardation and formed abnormal hyphae, and CypdA/DsrrA unexpectedly showed moresevere growth retardation than the parent CypdA, in contrast, the growth retardation of CypdA/DsskA partly recovered. It is suggested that the growthretardation of CypdA was only partly suppressed by switching off the HogA pathway. We further constructed a mutant (CypdA/DsrrADsskA) from CypdA bydeleting the two RR genes. Here, we discuss two-component signaling under the inhibitory conditions of signaling between YpdA and response regulators.368. WITHDRAWN369. FigA, a putative member of low-affinity calcium system, is involved in both asexual and sexual differentiation in Aspergillus nidulans. Shizhu Zhang,Hailin Zheng, Nanbiao Long, Sha Wang, Ling Lu. College of life Sciences, Nanjing Normal University, Nanjing, Jiangsu, China.Calcium-mediated signaling pathways are widely employed in eukaryotes and are implicated in the regulation of diverse biological processes. In baker’syeast Saccharomyces cerevisiae, at least two different carrier systems have been identified—a high-affinity calcium influx system (HACS) and a low-affinity210

FULL POSTER SESSION ABSTRACTScalcium influx system (LACS). In the filamentous fungus Aspergillus nidulans, we identified the homologs of HACS—the voltage gated channel CchA and thestretch activated channel MidA, which formed a complex that played important roles in conidial development, hyphal polarity establishment, and cell wallcomponents in low-calcium environmental condition. In comparison, loss of FigA, a putative member of LACS, showed very severe defects in conidiationand in self-fertility during sexual development under either low or high calcium environmental condition. Interestingly, extracellular Ca2+ was unable toimprove these figA defects substantially. Most importantly, the quantitative PCR results revealed the expression of the major asexual developmentregulator brlA and sexual development regulator nsd, steA had been remarkable regulated in figA mutant. In addition, the localization of Fig1::GFPrevealed that FigA was highly accumulated at the center of septum on the mature hypha , and the sites between vesicle-metulae, between metulaephialide.These data implied that figA likely played important roles in cellular trafficking and communication during in both asexual and sexualdifferentiation in Aspergillus nidulans. *Correspondence to be addressed: linglu@njnu.edu.cn This work was financially supported by the National NaturalScience Foundation of China (NSFC) 31200057 to Z.S, 31070031 to L.L.370. The Saccharomyces cerevisiae FUS3 homologue MAKB in Aspergillus niger is a central regulator connecting differentiation and secondarymetabolite production with nutrient availability and light. Bert-Ewald Priegnitz, Ulrike Brandt, André Fleissner. Institut für Genetik, TU Braunschweig,Braunschweig, Germany.Aspergillus niger is an important cell factory in biotechnology. Productivity of this fungus is strongly influenced by the environmental growth conditionsand the subsequent morphological adaptation. In eukaryotic organism, mitogen activated protein kinase cascades are important signaling pathwaysmediating cellular responses to environmental cues. In order to gain a deeper understanding of the molecular mechanisms of cellular adaptation in A.niger, we characterized the Saccharomyces cerevisiae FUS3 MAP kinase homologue MAKB by using a combination of molecular genetics, biochemicalanalysis and light and fluorescent microscopy. We found that the absence of makb results in specific developmental defects, which strongly depend onenvironmental conditions. For example, the mutant secrets a dark pigment in a nutrient dependent manner in constant darkness, but not while growing ina dark-light-rhythm. In addition, MAKB is also involved in asexual spore formation, indicated by its accumulation in developing conidiophores and areduced sporulation of the makb deletion mutant. Detailed comparison of sporulation in the wild-type and the mutant under different growth conditionslet us to the hypothesis that two independent trigger induce conidiation: nutrient starvation and cell age or density. Interestingly, MAKB appears to beonly involved in the latter one, indicated by suppressed sporulation of the mutant under nutrient sufficient growth conditions. Taken together, theseobservations indicate that in A. niger, MAKB is intimately involved in the adaptation of secondary metabolism and cell differentiation in response toenvironmental influences.371. Engineering and characterizing protein secretion in Aspergillus niger. Y. Zheng, G. Budkewitsch, S. Bourque, J. Burai, N. Geoffrion, J. Richard Albert,E. Munro, S. Sillaots, R. Storms. Biology, Concordia University, Montreal, QC, Canada.Aspergillus niger is widely used commercially and for basic research as a host for native and foreign protein production. This is mainly because of itsability to secrete large amounts of protein into the growth medium, and carry out the eukaryotic post-translational modifications of glycosylation,proteolytic cleavage and disulfide bond formation. Although A. niger can express some native and foreign proteins at high levels, many native proteins andmost foreign proteins are expressed at very low levels. Competition with native proteins for rate limiting steps in the secretory pathway, transcription,mRNA processing and translation are bottlenecks that can limit levels of secreted protein production. Furthermore, proteins expressed using A. niger oftenrequire extensive purification, due to the presence of high levels of many native secreted proteins that can interfere with the downstream characterizationor reduce protein stability. To address these issues we have been developing recombinant promoters that support high transcription rates, engineeringstrains that produce reduced levels of native secreted protein, and identifying secretory pathway bottlenecks that limit secreted protein yields. We havecreated a set of novel expression cassettes that are capable of supporting significantly higher transcription rates than are obtained with the widely usedglucoamylase gene (glaA) promoter. We have also, engineered new "clean" expression strains that combine dramatically reduced levels of"contaminating" native extracellular protein production with increased levels of target protein expression. RNA-seq analysis and conditional geneexpression are being used to identify additional bottlenecks that limit the production of foreign proteins.372. Design of culture conditions for secondary metabolite production of fungi based on large-scale transcriptome data. M. Umemura 1 , H. Koike 2 , M.Sano 3 , N. Yamane 2 , T. Toda 2 , K. Tamano 1 , S. Ohashi 3 , M. Machida 1 . 1) Bioproduction Research Institute, National Institute of Advanced Industrial Scienceand Technology (AIST), Sapporo, Japan; 2) Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST),Tsukuba, Japan; 3) Kanazawa Institute of Technology, Kanazawa, Japan.Fungi produce a wide variety of secondary metabolites, many of which possess bioactivity against bacteria, fungi, and human diseases (ex. penicillin,micafungin, and lovastatin). Inducing the fungal productivity of secondary metabolites is requisite for study and industrial applications of the compounds.Since types and amounts of the secondary metabolites greatly change according to strains and culture conditions, controlling the productivity is difficultand has largely depended on empirical knowledge. More data-based knowledge is required especially when treating the metabolites for which lessempirical knowledge has been accumulated. In this study, we have developed an algorithm to analyze metabolic pathway activities using large-scaletranscriptome data, and applied it to 72 sets of Aspergillus oryzae transcriptome data obtained using different nutritions. Based on the classification ofmetabolic pathways and assignment of A. Oryzae genes to them by Vongsangnak et al. [BMC Genomics, 9,245 (2008)], we evaluated the correlations of thepathways and found some positive and negative correlations between primary and secondary ones. We also found that secondary metabolic pathwayswere divided into two groups depending on the type of nutrition. Combining some culture conditions based on these results, we designed the idealnutrition for inducing each two groups of secondary metabolic pathways. We are now confirming metabolic pathway activity and secondary metaboliteproductivity under the designed culture conditions using DNA microarray and LC-MS, respectively.373. The effect of the clbR overexpression on cellulose degrading enzyme production in Aspergillus aculeatus. S. Tani, A. Kawamura, E. Kunitake, J.Sumitani, T. Kawaguchi. Life & Environmental Sci, Osaka Prefecture Univ, Osaka, Japan.Aspergillus aculeatus has two pathways that control the induction of cellulase and hemicellulase genes in response to cellulose. The expression ofcarboxymethylcellulase 1 (cmc1) and FIb-xylanase (xynIb) genes was controlled by XlnR, in contrast the expression of cellobiohydrolase I (cbhI),carboxymethylcellulase 2 (cmc2), hydrocellulase (cbhIb) and FIa-xylanase (xynIa) genes was controlled by XlnR-independent signaling pathway. We havereported that ClbR, a Zn(II) 2Cys 6 transcription factor, controlled the cellulose-responsive induction of the genes regulated by both XlnR-dependent andXlnR-independent signaling pathways. Therefore, we investigated if their enzyme productions could be improved by the clbR overexpression. The clbRgene was expressed under the promoter of the translation elongation factor 1 alpha gene. Interestingly, xylanase activity in the clbR overexpressing strain(clbRox) increased 6-fold more than that in control strain. Although endoglucanase activity did not increase, its activity was kept at high level for 10 days.Peptide mass fingerprinting revealed that FIa-xylanase production was drastically increased in the clbRox strain, in contrast that hydrocellulase production27th Fungal Genetics Conference | 211

FULL POSTER SESSION ABSTRACTSdecreased. Northern blot analysis revealed that the expression of cbhIb decreased in the clbRox strain under cellulose inducing condition. However, theclbR overexpression increased xynIa expression under wheat bran inducing condition, but not under the avicel inducing condition. The effect of the clbRoverexpression was not equal to the enzyme production and the gene expression even though they were under the control of ClbR. These results suggestthat ClbR coordinately controls their expression with another factors.374. Regulation of the a-tubulin B-encoding gene during early development of the phytopathogenic fungus Botrytis cinerea. Y. Faivre Talmey 1 , I. R.Gonçalves 1 , A. Simon 2 , M. Viaud 2 , C. Bruel 1 . 1) CNRS, University Claude Bernard, LYON, France; 2) INRA, BIOGER, Grignon, France.The fungal plant pathogen Botrytis cinerea is able to infect more than 250 different hosts and is responsible for major economic losses worldwide. Thesuccess of the fungus life cycle relies on distinct developmental stages and specific cellular structures like appressoria, infection cushions, conidiophores,apothecia and sclerotia. Based on the important role that cytoskeleton plays in cellular organization and shape in all eucaryotes, attention was given totubulins and their putative specific role in the virulence of B. cinerea. One b-tubulin and two a-tubulin encoding genes are found in B. cinerea‘s genome.Alpha-tubulin A and B share 69% identity at the protein level and phylogenetic analyses revealed that orthologs to the a-tubulin A gene exist in allascomyceta species whereas the a-tubulin B gene seems to be present in some ascomyceta only. Expression studies showed that the a-tubulin B gene ismore expressed than the a-tubulin A gene. Besides, a peak of expression is observed for the a-tubulin B encoding gene early during conidia-derived fungaldevelopment. In order to understand the regulation of the a-tubulin B encoding gene, combinations of promoter deletions and transcriptional fusionswere used. Putative regulatory regions were identified and the one hybrid yeast system was used to search for putative transcription factors that wouldinteract with these regions and play a role in the regulation of this gene.375. The regulation of D-galacturonic acid utilization in Botrytis cinerea. Lisha Zhang, Joost Stassen, Sayantani Chatterjee, Maxim Cornelissen, Jan vanKan. Laboratory of Phytopathology, Wageningen University, Wageningen, Netherlands.The plant cell wall is the first barrier to pathogen invasion. The fungal plant pathogen Botrytis cinerea produces a spectrum of cell wall degradingenzymes for the decomposition of host cell wall polysaccharides and the consumption of the monosaccharides that are released. Especially pectin is animportant cell wall component, and the decomposition of pectin by B. cinerea has been extensively studied. D-galacturonic acid is the most abundantcomponent of pectin and effective utilization of D-galacturonic acid is important for virulence of B. cinerea. The D-galacturonic acid catabolic pathwaycomprises three enzymatic steps, involving D-galacturonate reductase (encoded by Bcgar1 and Bcgar2), L-galactonate dehydratase (encoded by Bclgd1),and 2-keto-3-deoxy-L-galactonate aldolase (encoded by Bclga1). Therefore, an effective concerted action of the appropriate pectin depolymerisingenzymes, monosaccharide transporters and catabolic enzymes is important for complete pectin utilization by B. cinerea. In this study, RNA sequencing wasperformed to compare genome wide transcriptional profiles in B. cinerea grown in media containing glucose and pectate as sole carbon sources. Weidentified 31 genes that are significantly upregulated in pectate containing culture, including Bcgar2, Bclga1, and a putative monosaccharide transporter.In addition, conserved cis-regulatory elements were predicted in the promoters of genes involved in pectate decomposition and D-galacturonic acidutilization. Functional analysis was carried out of the bidirectional promoter of the Bcgar2-Bclga1 gene cluster to study which of the cis-regulatoryelements is required for induction by D-galacturonic acid. Furthermore, potential regulatory protein(s) were isolated by DNA-protein pull down assaysusing one important cis-regulatory element.376. Epigenetic Regulation of Subtelomeric Gene Noise in Candida albicans. Matthew Z Anderson, Joshua A Baller, Lauren J Wigen, Judith Berman.Genetics, Cell Biology and Development, Univeristy of Minnesota, St Paul, MN.Candida albicans grows within a wide range of fluctuating host niches, and the ability to rapidly adapt enhances its success as a commensal and as apathogen. The recently expanded telomere-associated (TLO) gene family consists of fourteen expressed members in C. albicans. Each TLO gene encodes aparalog of a single Mediator complex component. Thirteen expressed TLOs are located at the chromosome ends as the most telomere-proximal openreading frame. Individual TLO expression at both the transcript and protein level was extremely noisy. Noise originated from single cell variability in TLOexpression due to intrinsic factors. Deletion of chromatin modifying enzymes that function in subtelomeric silencing abolished TLO noise, as did ectopicallyexpressing a TLO from an internal locus. Conversely, transcriptional variation of a low noise gene increased significantly when ectopically expressed in thesubtelomere. Interestingly, deletion of the Mediator component MED3, which inhibits Tlo from incorporating into Mediator, also drastically reduced TLOnoise and supports an autoregulatory mechanism for TLO noise. These data suggest subtelomeric chromatin structure regulates TLO gene noise throughthe action of chromatin modifiers and Mediator. We propose that TLO noise is beneficial to C. albicans by producing heterogeneous cell populations thatincorporate different Tlo proteins in Mediator, producing a range of transcriptional profiles in the population that allows some cells to survive in alteredenvironmental conditions.377. Signaling and Cell Behavior: Pheromone Response in Candida albicans. Ching-Hsuan Lin 1,2 , Kabrawala Shail 2 , Fox Emily 3 , Nobile Clarissa 3 , JohnsonAlexander 3 , Bennett Richard 2 . 1) Dept. Biochem. Sci. and Tech., NTU, Taipei, Taiwan; 2) Dept, MMI, Brown Univ. RI; 3) Dept, Biochem. Biophy. UCSF, CA.Candida albicans is currently the most common human fungal pathogen, and its propensity for causing infection has been closely lined with its ability toform biofilms. Central to understanding its behavior is the white-opaque phenotypic switch, in which cells can undergo an epigenetic transition betweenthe white state and the opaque state. The phenotypic switch regulates multiple properties including biofilm formation, virulence and sexual mating. Inparticular, opaque cells are the mating competent form whereas white cells do not mate but generate biofilms in response to pheromone. In this work, weidentify the master transcriptional regulator involved in pheromone-induced biofilms in white cells as well as sexual mating in opaque cells as C. albicansCph1, ortholog of Saccharomyces cerevisiae Ste12. In contrast, Cph1 is not required for the formation of conventional biofilms (biofilms that can formwithout pheromone treatment). Transcriptional profiling analysis under biofilm conditions during pheromone treatment revealed a number of potentialdownstream targets of Cph1 and Tec1. In total, we observed 23 genes that exhibited decreased induction (>2-fold) by pheromone in both Dcph1 and Dtec1mutants. Of these genes, six candidates were chosen for further analysis due to their dependence on Cph1 and Tec1 for pheromone-induced expression,and because they were also not induced in pheromone-treated opaque cells. Interestingly, one novel gene product shown to influence biofilm formation isHgc1. Similar to the Dtec1, deletion of HGC1 resulted in decreased biofilm formation in white cells responding to pheromone, and also abolished formationof conventional biofilms. It is therefore apparent that Hgc1 is important for cell adhesion and biofilm development in the two distinct models of biofilmformation. Most importantly, these results suggest that both shared and unique components operate in different models of biofilm formation in thisimportant human pathogen.378. Transcriptional regulatory networks controlling the early hypoxic response in Candida albicans. A. Nantel, M. van het Hoog, A. Sellam, C.Beaurepaire, F. Tebbji, M. Whiteway. National Research Council of Canada, Montreal, Quebec, Canada.212

FULL POSTER SESSION ABSTRACTSThe ability of Candida albicans to colonize or invade multiple host environments requires that it rapidly adapts to different conditions. Our group hasbeen exploiting ChIP-chip and transcription profiling technologies, together with computer modeling, to provide a better understanding of selecttranscription factor (TF) networks. We used DNA microarrays to measure the changes in transcriptional profiles that occur immediately following thetransfer of C. albicans to hypoxic growth conditions. The impressive speed of this response is not compatible with current models of fungal adaptation tohypoxia that depend on the inhibition of sterol and heme biosynthesis. Functional interpretation of these profiles was achieved using Gene Set EnrichmentAnalysis, a method that determines whether defined groups of genes exhibit a statistically significant bias in their distribution within a ranked gene list.The Sit4p phosphatase, Ccr4p mRNA deacetylase and Sko1p TF were identified as novel regulators of the early hypoxic response. While cells mutated inthese regulators exhibit a delay in their transcriptional responses to hypoxia their ability to grow in the absence of oxygen is not impeded. Promoteroccupancy data on 26 TFs was combined with the profiles of 375 significantly-modulated target genes in a Network Component Analysis (NCA) to producea model of the dynamic and highly interconnected TF network that controls this process. The NCA also allowed us to observe correlations betweentemporal changes in TF activities and the expression of their respective genes, thus allowing us to identify which TFs are potentially subjected to posttranscriptionalmodifications. The TF network is centered on Tye7p and Upc2p which are associated with many of the genes that exhibit the fastest andstrongest up regulations. While Upc2p only associates with downstream promoters, Tye7p is acting as a hub, its own promoter being bound by itself and 7additional TFs. Rap1 and Ahr1 appear to function as master regulators since they bind to a greater proportion of TF gene promoters, including those ofUpc2p and Tye7p. Finally, Cbf1p, Mrr1p and Rap1p show the greatest numbers of unique gene targets. The high connectivity of these models illustratesthe challenges that lie in determining the individual contributions of specific TFs.379. Gene expression and function during invasive Candida infection. Wenjie Xu 1 , Norma Solis 2 , Carol Woolford 1 , Scott Filler 2 , Aaron Mitchell 1 . 1)Biological Sciences, Carnegie Mellon University, Pittsburgh, PA; 2) Departments of Medicine and Pathology, Harbor-UCLA, Los Angeles, CA.What genes does a pathogen express during infection? What regulatory pathways contribute to expression of those genes in vivo? Which pathogen genefunctions evoke specific host responses? These questions are beginning to be addressed for many plant pathogens, in which a lesion that is enriched forinfected tissue can be isolated readily. There have also been pioneering studies with human and animal pathogens, but they have been limited by theability to isolate infected tissue and by background problems from microarray technology. We implemented a recently developed technology, nanoStringprofiling, to investigate these long-standing questions with the fungal pathogen C. albicans in a murine model of hematogenously disseminatedcandidiasis. We used whole kidneys, a major target organ, for profiling the time course of both pathogen and host gene expression. NanoString technologyis not genome-wide, so we have selected high-priority fungal and host genes for investigation. On the pathogen side, we find that hyphal genes andneutral/alkaline pH response genes are induced early in infection, while oxidative and cell wall stress genes are induced later. These results are consistentwith the idea that the influx of neutrophils causes cell wall and oxidative stress. Among the 222 C. albicans genes that specify transcription factors, thosehighly expressed or highly up-regulated during infection are enriched for previously demonstrated virulence regulators, and also include many genes notpreviously known to govern virulence. We have profiled attenuated mutants despite their limited growth during infection, and we have found thattranscription factor target genes in vivo differ considerably from the target genes identified in vitro. Finally, we find that the host displays progressiveinduction of cytokines, pattern recognition receptors, and innate immune signaling pathways, with expression changes detectable at 12 hours postinfection,when fungal burden is extremely low. An attenuated rim101 mutant causes a muted host response, but also alters the kinetic profile to yieldprecocious induction of late host response genes. Our data allow the most detailed sketch to date of the dynamics and functions at the host-pathogeninterface during disseminated candidiasis.380. Effects of histone H3 point mutations on centromere maintenance. Steven Friedman 1 , Eric Selker 2 , Michael Freitag 1 . 1) Biochemistry & Biophysics,Oregon State University, Corvallis, OR; 2) Institute of Molecular Biology, University of Oregon, Eugene, OR.Post-translational modifications (PTM) of histone amino acid residues are known to play important roles in chromatin structure and function. InNeurospora crassa, trimethylation of histone H3 lysine 9 (H3K9me3) is essential for cytosine DNA methylation [1]. There is also evidence for a role ofH3K9me3 in the maintenance of centromeres [2]. An in-depth study of a larger set of histone H3 point mutations revealed additional recessive anddominant mutations involved in DNA methylation, including some mutations that proved lethal [3]. In this study, hH3 alleles were integrated ectopically orat the his-3 locus in the presence of a mutant hH3 allele at the endogenous hH3 locus. Here we describe a gene replacement system that allows mutanthH3 alleles to be integrated at the endogenous loci by homologous recombination, yielding hH3 replacement strains. The general approach is versatile andapplicable to studying the role of specific point mutations in other genes. We will present results on how hH3 mutations affect the deposition ofcentromere proteins (e.g., CenH3, CEN-T and CEN-C).[1] Tamaru, H. and E.U. Selker, 2001, Nature 414: 277. [2] Smith, K.M. et al. 2011, Mol. Cell Biol. 31: 2528. [3] Adhvaryu, K.K. et al. 2011, PLoS Genetics 7:e1002423..381. Analysis of the transcriptional regulation of genes involved in the synthesis and organization of the cell wall of Ustilago maydis during infection ofan alternatiive host. Angélica Mariana Robledo Briones, José Ruiz Herrera. Centro de Investigaciones y de Estudios Avanzados del IPN, Km 9.6 LibramientoNorte Carretera Irapuato-León.The cell wall is the most external structure of the cell. Its function is to protect it against the difference in osmotic pressure with the environment andprovide the morphology. The wall in fungi is made of microfibrils of structural polysaccharides (chitin and b-1,3-glucans) immersed in a matrix ofglycoproteins . Ustilago maydis is a dimorphic basidiomycota, pathogen of maize, but under axenic conditions it may infect other plants includingArabidopsis thaliana. Considering the role of the wall in the pathogenic process, we made a transcriptomic analysis of the genes involved in its structureand synthesis, and encoding secreted proteins during the infection of A. thaliana. For this study, one channel chips with high density oligonucleotide wereused. Plantlets of Arabidopsis were infected with a haploid or a diploid strain, and at intervals RNA was isolated, complementary cDNA was synthesizedand used to hybridize the microarrays. Data of microarrays allowed to identify genes involved in cell wall synthesis and organization, and encodingproteins from the secretome of U.maydis whose expression was regulated during the transition from saprophytic to pathogenic stages. These accountedto about 60 per cent; of the total of 639 genes existing in U. maydis, a proportion being slightly higher in the haploid strain. Some differences wereobserved in the regulation of these genes in the haploid and diploid strains. We observed that genes involved in N- and O- glycosylation of proteins wereup-regulated during infection. In addition, some CHS and CDA genes, and some genes involved in the synthesis of b-1,6-glucans and GPI proteins, weredifferentially regulated. A great number of genes encoding secreted proteins with a degradative function were up-regulated (more in the haploid). Thisincreased transcription may be related with degradation of the plant cell wall necessary to establish the infection. All these results demonstrate theusefulness of the Ustilago maydis-Arabidopsis thaliana pathosysthem for identification of the pathogenic mechanisms of U. maydis, and in the case of this27th Fungal Genetics Conference | 213

FULL POSTER SESSION ABSTRACTSstudy the role of genes encoding proteins of the wall and secretome, and the differences existing in the behavior of the diploid and haploid, taking intoconsideration that only the first one is pathogenic to the natural host.382. Expression of the Trichophyton rubrum ace2 and pacC genes during degradation of keratinized substrates. Larissa Silva, Nalu Peres, GabrielaPersinoti, Elza Lang, Vanderci Oliveira, Antonio Rossi, Nilce Martinez-Rossi. University of Sao Paulo, Ribeirão Preto/SP/Brazil, Sao Paulo, Brazil.Trichophyton rubrum is a pathogenic, cosmopolitan and anthropophilic fungus that infect keratinized tissues, mainly skin and nails. The genomes ofseveral dermatophytes, including T. rubrum, were sequenced by the Broad Institute/NIH, enabling studies on the regulation of the expression of genesrelated to several cellular processes. The transcription factor (TF) Ace2 participates of a network of genes called RAM (Regulation of Ace2 activity andcellular morphogenesis), involved in the regulation of morphogenesis, cell division, and development of conidiophores. The TF PacC regulates thetranscription of genes in response to extracellular pH and also genes related to the biosynthesis of cell wall, suggesting a crosstalk between these twopathways. Therefore, the aim of this study was to analyze the expression profile of the pacC and ace2 genes in different nutritional sources (nail and skinex vivo infections) to understand the regulation of these TF during pathogenesis and development. In silico analyses of the putative promoter regions ofthe pacC and ace2 genes revealed the presence of the DNA binding motifs of both TFs, suggesting a possible cross- and co-regulation of these TFsexpression in T. rubrum. Gene expression analyses during growth in keratinized tissues suggested an opposite expression profile in nail interaction assays,the ace2 gene was up-regulated and pacC was down-regulated. Moreover, in ex vivo skin infective both genes presented a similar expression level. Theseresults suggest a different gene expression modulation of ace2 and pacC according to the nutrient source and possibly the infection site. Moreover, thisevaluation provides a better comprehension of the involvement of both pathways in regulating a variety of cellular processes that enable cell viabilityduring infection of keratinized tissues.383. Control and Function of Two Fatty Acid Regulators in Neurospora crassa. Erin L. Bredeweg 1 , Fei Yang 2 , Kristina Smith 1 , Rigzin Dekhang 2 , JillianEmerson 3 , Jay Dunlap 3 , Deborah Bell-Pedersen 2 , Matthew Sachs 2 , Michael Freitag 1 . 1) Program for Molecular and Cellular Biology, Department ofBiochemistry and Biophysics, and Center for Genome Research and Biocomputing (CGRB), Oregon State University, Corvallis, OR 97331; 2) Department ofBiology and Program for the Biology of Filamentous Fungi, Texas A&M University, College Station, TX; 3) Department of Genetics, Geisel School ofMedicine at Dartmouth, Hanover, NH.The filamentous saprobe Neurospora crassa is an excellent model for describing the behavior of transcriptional regulators. We describe the genomewidebehavior of two Fatty Acid Regulators (FAR) proteins, transcription factors that modulate the response of N. crassa to the presence of fatty acids. Weused ChIP-seq to find the localization of FAR-1 and FAR-2 under nutrient conditions targeting short and long chain fatty acid carbon sources, with sucroseas a control. Bioinformatic analyses describe variant binding sites for FAR-1 and FAR-2, with overlap in about a third of all target regions. Functions underthe control of ChIP-seq targets were further examined by phenotypic assays for siderophore production, oxidative stress, and linear growth. We foundreduced siderophore production, and increased vulnerability to oxidative stress in far-1 mutants, but not far-2 mutants. Linear growth showed a carbonspecificreduced growth rates for far-2, as well as Tween-20 sensitivity and conidiation defects for far-1. RNA-seq identified numerous differentiallyregulated transcripts under different growth conditions and in the single or double mutants. Many of these transcripts are part of the gene set identifiedby ChIP-seq, and many were affected by the absence of one or both FARs. Our analyses identified groups of co-regulated proteins not previously identifiedas affected by FAR transcription factors, in addition to those involved in the control of the core cellular machinery for energy production by beta-oxidation.384. Characterization of genomic targets for the Neurospora crassa hypothetical transcription factor NCU04390 by ChIP-sequencing. R. Gonçalves 1 , E.Bredeweg 2 , M. Freitag 2 , M. C. Bertolini 1 . 1) Instituto de Química, UNESP, Araraquara, São Paulo, Brazil; 2) Department of Biochemistry and Biophysics, OSU,Corvallis, OR, USA.The mechanisms by which glycogen content is controlled in microorganisms are intricate, involving co-regulation of many proteins. In Neurospora crassa,glycogen reaches maximal levels at the end of the exponential growth phase, however under heat shock, glycogen content and transcription of theglycogen synthase gene (gsn) rapidly decrease. In a previous analysis, the NCU04390 KO strain showed a drastic increase in glycogen levels and upregulationof the gsn gene after heat shock when compared to the wild-type strain, suggesting that the NCU04390 gene product is involved in theregulation of glycogen metabolism. Because the product of this ORF is annotated as a hypothetical transcription factor (TF) with an N-terminal zinc-fingerand a central fungal-specific TF domain, chromatin immunoprecipitation followed by high throughput DNA sequencing (ChIP-seq) was expected to revealgenes that are directly regulated by the NCU04390 gene product. First, GFP tag was fused to the 3'-end of the ORF NCU04390 by gene replacement. ChIPwas performed with NCU04390-GFP at 30ºC and 45ºC with antibodies against the GFP tag. ChIP-libraries were sequenced on a HiSeq2000 (Illumina/Solexa)genome analyzer and data from 45ºC experiment revealed that most of the genes regulated by the transcription factor encode hypothetical proteins.However genes encoding proteins with known functions, such as proteins involved in carbon metabolism and transporters were also identified. Amongthese genes, it is important to mention the glycogen debranching enzyme coding gene (ORF NCU00743), which participates in the glycogen degradation.The His::4390 recombinant protein was produced in E. coli, partially purified by IMAC and used in EMSA experiments to validate the result found in theChIP-Seq assay. The results showed specific binding of the recombinant His::4390 in the NCU00743 promoter, suggesting that the transcription factormight regulate glycogen metabolism under heat stress through the gene encoding the debranching enzyme. Data from EMSA validation analysis for morepeaks found in ChIP-seq will be presented. Supported by FAPESP, CNPq, CAPES and US NIH.385. The KMT6 Histone H3 K27 Methyltransferase Regulates Expression of Secondary Metabolites and Development in Fusarium graminearum. KristinaM. Smith, Lanelle R. Connolly, Michael Freitag. Department of Biochemistry and Biophysics, Center for Genome Research and Biocomputing, Oregon StateUniversity, Corvallis, OR 97331.The cereal pathogen Fusarium graminearum produces secondary metabolites toxic to humans and animals, yet coordinated transcriptional regulation ofsecondary metabolite gene clusters remains largely a mystery. By ChIP-sequencing we found that regions of the F. graminearum genome with secondarymetabolite clusters are enriched for a histone modification, trimethylated histone H3 lysine 27 (H3K27me3), associated with gene silencing. Thismodification was found predominantly in regions that lack synteny with other Fusarium species, generally subtelomeric regions. H3K27me3 and di- ortrimethylated H3K4 (H3K4me2/3), modifications associated with gene activity, are found in mutually exclusive regions of the genome. To betterunderstand the role of H3K27me3, we deleted the gene for the putative H3K27 methyltransferase, KMT6, a homolog of Drosophila Enhancer of zeste, E(z).The kmt6 mutant lacks H3K27me3, as shown by western blot and ChIP-sequencing, displays growth defects, is sterile, and produces mycotoxins underconditions where they are not generated in wildtype (WT) strains. RNA-sequencing showed that genes modified by H3K27me3 are most often silent, asabout 75% of the 4,449 silent genes are enriched for H3K27me3. Surprisingly, we found 22% of the 8,855 expressed genes enriched for H3K27me3. Asubset of genes that were enriched for H3K27me3 in WT gained H3K4me2/3 in kmt6 (1,780 genes), and an overlapping set of genes showed increased214

FULL POSTER SESSION ABSTRACTSexpression. Almost 95% of the remaining 2,720 annotated silent genes showed no enrichment for either H3K27me3 or H3K4me2/3. In these cases absenceof H3K27me3 is insufficient for expression, which suggests a requirement for additional factors for gene expression. Taken together, we show that absenceof H3K27me3 allows expression of 14% of all annotated genes, resulting in derepression of predominantly secondary metabolite pathways and otherspecies-specific functions, including potentially secreted pathogenicity factors. This study provides the framework for novel targeted strategies to controlthe “cryptic genome” and specifically secondary metabolite expression.386. Circadian clock-gated cell division cycles in Neurospora crassa. C. Hong 1 , J. Zamborszky 1 , M. Baek 1 , K. Ju 1 , H. Lee 1 , L. Larrondo 2 , A. Goity 2 , A. Csikasz-Nagy 3 . 1) Molecular and Cellular Physiology, University of Cincinnati, Cincinnati, OH; 2) Departamento de Genética Molecular y Microbiología, Facultad deCiencias Biológicas, Pontificia Universidad Católica de Chile, Casilla 114-D, Santiago, Chile; 3) Randall Division of Cell and Molecular Biophysics, andInstitute for Mathematical and Molecular Biomedicine, King’s College London, London, SE1 UL, UK.Asynchronous nuclear divisions are readily observed in filamentous fungi such as Ashbya gossypii and Neurospora crassa. Our computational simulations,however, predict synchronous circadian clock-gated mitotic divisions if the division cycles of such multinucleated organisms are coupled with circadianrhythms. Based on this hypothesis, we investigate the coupling between the cell cycle and the circadian clock in Neurospora crassa. First, we show WC-1-dependent light-induced expression of stk-29 mRNA (homolog of wee1), which suggests that there exists a conserved coupling between the clock and thecell cycle via STK-29 in Neurospora as in mammals. Second, we demonstrate that G1 and G2 cyclins, CLN-1 and CLB-1, respectively, show circadianoscillations with luciferase bioluminescence reporters. Moreover, clb-1 and stk-29 gene expressions show circadian clock-dependent light-induced phaseshifts, which may alter the timing of divisions. Third, we show circadian clock-dependent synchronized nuclear divisions by tracking nuclear morphologywith histone hH1-GFP reporter. Synchronized divisions occur late in the evening, and they are abolished in the absence of circadian rhythms (frq KO ). Ourfindings demonstrate the importance of circadian rhythms for synchronized mitotic cycles and establish Neurospora crassa as an ideal model system toinvestigate mechanisms that couple the cell cycle and the circadian clock.387. Protein Binding Microarrays and high-throughput real-time reporters studies: Building a four-dimensional understanding of transcriptionalnetworks in Neurospora crassa. A. Montenegro-Montero 1 , A. Goity 1 , C. Olivares-Yañez 1 , A. Stevens-Lagos 1 , M. Weirauch 2 , A. Yang 3 , T. Hughes 3 , L. F.Larrondo 1 . 1) Genética Molecular y Microbiología, Pontificia Universidad Católica de Chile, Santiago, Chile; 2) CAGE, Cincinnati Children`’s Hospital MedicalCenter, University of Cincinnati. U.S.A; 3) Banting and Best Department of Medical Research, University of Toronto, Canada.It has been suggested that ~20% of the Neurospora-transcriptome may be under circadian control. Nevertheless, there is scarce information regardingthe regulators that are involved in the rhythmic expression of clock-controlled genes (ccgs). We are using a high-throughput platform, based on variouscodon-optimized luciferase transcriptional- and translational-reporters, to monitor time-of-day-specific gene expression and to identify key elementsmediating circadian transcriptional control. Thus, we have identified transcription factors -such as SUB-1- that affect the expression of known and novelccgs, among which there are transcriptional regulators that give access to a group of third-tier ccgs. In addition, we are characterizing several rhythmicbZIP-coding genes as potential nodes of circadian regulation. In order to characterize regulatory networks in which these and all Neurospora transcriptionfactors participate, we are using double-stranded DNA microarrays containing all possible 10-base pair sequences to examine their binding specificities andin that way, predict possible targets on a genome-wide manner. Currently, these Protein Binding Microarray studies have provided DNA-bindingspecificities for over 120 Neurospora transcription factors granting an unprecedented and powerful tool for transcriptional network studies. Finally, wehave generated graphic tools to explore the spatial differences observed in the temporal control of gene expression. Funding: Conicyt/Fondecyt/regular1090513.388. Glycogen metabolism is regulated by the circadian clock in Neurospora crassa. S. Virgilio, T. Candido, M. C. Bertolini. Instituto de Química, UNESP,Araraquara, São Paulo, Brazil.The fungus Neurospora crassa has been widely used in studies of circadian rhythms and photobiology. Our research group has been using this modelorganism to study the molecular mechanisms involved in glycogen metabolism regulation and recent findings have revealed that circadian rhythms controla variety of physiological and metabolic functions in different organisms. In a screen of a mutant strains set we identified a number of transcriptionfactors/cofactors likely acting as regulators of glycogen metabolism. Among them, several transcription factors were previously described as controlled byregulators of the circadian clock in N. crassa. The result led us to start to investigate whether glycogen metabolism is under control of circadian clock in N.crassa. Experiments were performed to verify whether glycogen was rhythmically accumulated in a wild-type strain. In circadian clock experiments, theglycogen content varied according to the circadian rhythm, with cyclical periods ranging from 22 to 24 h. The glycogen synthase activity (GSN) wasquantified in the presence and absence of the allosteric activator glucose-6-phosphate (G6P). The -/+ G6P ratio is considered as an index ofphosphorylation, higher levels correlating with lower phosphorylation. The GSN phosphorylation was influenced by the biological clock, showing changesin the GSN phosphorylation status along the experiment. The expression of the gsn and gpn (encoding glycogen phosphorylase) genes was evaluated inthe same experiments and in light-induced experiments as well. In circadian clock analysis, the gsn and gpn transcripts showed rhythmic expressionalthough not as pronounced as the levels of the ccg-1 transcript (positive control). In light-induced experiments, the levels of glycogen were kept constantduring different times after exposure to light, however the expression of the gsn and gpn genes showed to be delayed light-induced. The results suggesteda connection between the energy derived from the glycogen metabolism and the circadian clock in N. crassa. Supported by FAPESP and CNPq.389. Genetic and Molecular Dissection of the Neurospora Circadian Oscillatory System. Qijun Xiang 1 , Bin Wang 1 , Chandru Mallappa 1 , Jennifer Hurley 1 ,Arko Dasgupta 1 , Jennifer Loros 2 , Jay Dunlap 1 . 1) Department of Genetics, Dartmouth Medical School, Hanover, NH03755; 2) partment of Biochemistry,Dartmouth Medical School, Hanover, NH03755.Transcription/ translation feedback loops are central to all eukaryotic circadian clocks. In the circadian oscillator, the negative feedback loop drivesperiodic expression of proteins that feed back to reduce their own expression. A heterodimer of proteins, WC-1 and WC-2, acts as a transcription factor todrive expression of the frq gene. Its product FRQ dimerizes and forms a complex with another protein FRH. This complex inhibits the activity of the WCheterodimer creating the negative feedback loop. While canonical clock proteins such as FRQ work exclusively in timing, all systems utilize additionalproteins performing other functions in the cell. Among these in Neurospora is the essential putative RNA helicase, FRH. A novel, unbiased genetic screenfor circadian negative feedback mutants uncovered a point mutation that completely complements the essential functions of FRH yet is totally arrhythmic,thus genetically separating essential functions from clock-associated roles. In other experiments we used mass spectrometry to look for interactors of FRH,FRQ, and to follow posttranslational modifications of these proteins over the day. Although few modifications are found on FRH, FRQ is extensivelymodified with nearly 100 phosphorylations. By examining the phenotypes of strains bearing mutants that have lost these sites individually and in groups,we begin to see how temporally regulated phosphorylation has opposing effects directly on overt circadian rhythms and FRQ stability. For over 60% of the27th Fungal Genetics Conference | 215

FULL POSTER SESSION ABSTRACTSconfirmed phosphorylation sites, loss of the individual or neighboring sites have no apparent effect on the free running period length, suggesting that sitesmay work in groups as dynamically regulated charged domains. Some domains promote FRQ stability and lengthen period and other promote turnoverand shorten period. Modifications are dynamic such that at near all times of day “FRQ” describes a heterogeneous mix of proteins with the same aminoacid sequence but variable and distinguishable structure and surface chemistry. We have also used luciferase as a reporter to follow the FRQ-WC coreoscillator under conditions where growth rhythms are manifest. Under these conditions the FRQ/WC oscillator cycles with a normal compensatedcircadian period length even when the overt rhythm of growth moves out of the circadian range.390. Non-optimal codon usage determines the expression level, structure and function of the circadian clock protein FREQUENCY. Mian Zhou 1 , JinhuGuo 5 , Joonseok Cha 1 , Michael Chae 1 , She Chen 2 , Jose Barral 3 , Matthew Sachs 4 , Yi Liu 1 . 1) Department of Physiology, UT Southwestern Medical Center,Dallas, TX; 2) National Institute of Biological Sciences, Beijing, China; 3) Departments of Neuroscience and Cell Biology and Biochemistry and MolecularBiology, The University of Texas Medical Branch, Galveston, TX; 4) Departments of Biology, Texas A&M University, College Station, TX; 5) School of LifeSciences, Sun Yat-sen University, Guangzhou, China.Codon usage bias has been observed in the genomes of almost all organisms and is thought to result from selection for efficient and accurate translationof highly expressed genes 1-3. In addition, codon usage is also implicated in the control of transcription, splicing and RNA structure 4-6. Many genes,however, exhibit little codon usage bias. The lack of codon bias for a gene is thought to be due to lack of selection for mRNA translation. Alternatively,however, non-optimal codon usage may also have biological significance. The rhythmic expression and the proper function of the Neurospora FREQUENCY(FRQ) protein are essential for circadian clock function. Here, we show that, unlike most genes in Neurospora, frq exhibits non-optimal codon usage acrossits entire open reading frame. Optimization of frq codon usage results in the abolition of both overt and molecular circadian rhythms. Codon optimizationnot only increases FRQ expression level but surprisingly, also results in conformational changes in FRQ protein, impaired FRQ phosphorylation, andimpaired functions in the circadian feedback loops. These results indicate that non-optimal codon usage of frq is essential for its circadian clock function.Our study provides an example of how non-optimal codon usage is used to regulate protein expression levels and to achieve optimal protein structure andfunction.391. Two putative long non-coding RNAs upstream of transcription factor Znf2 may regulate morphogenesis (or dimorphic transition) in Cryptococcusneoformans. Nadia Chacko, Linqi Wang, Xiaorong Lin. Biology, Texas A&M University, College Station, TX.Cryptococcus neoformans is an opportunistic human pathogen and the causal agent of fungal meningitis, one of the leading causes of death inimmunocompromised patients. The virulence of this dimorphic fungus is closely tied to its morphology as the yeast form is pathogenic while thefilamentous form is non-pathogenic. The morphological switch from yeast to filament occurs typically during unisexual and bisexual mating but can occurunder mating limiting conditions too. Recently we found that transcription factor Znf2 directs morphological transition from yeast-to-filament and itsactivity is reversely correlated with fungal virulence. The means to increase Znf2 activity, either by activation of its activator or inactivation of itsrepressors, could be of great value to alleviate cryptococcosis. In a search to identify upstream regulators of ZNF2, we screened 60,000 insertional mutantsthat mimic znf2D phenotype. Insertions in two of the selected mutants were found to be in a potential long non-coding RNA located in the intergenicregion of ZNF2. This lncRNA was named RZE1. The rze1D mutant phenotype resembles the znf2D mutant phenotype, supporting our hypothesis that RZE1functions upstream of ZNF2. Interestingly, the expression of RZE1 is increased under host relevant conditions but not under mating-inducing conditions,suggesting that RZE1 could be involved in the adaptation to the host during infection. Surprisingly the expression of ZNF2 is only modestly reduced in theRZE1 insertional mutants, indicating the existence of other potential regulators or non-transcriptional regulation of ZNF2 by RZE1. Further analysis of theintergenic region of ZNF2 revealed the presence of another lncRNA, which was named RZE2. Long ncRNAs are known to regulate genes by transcriptionalactivation, repression and epigenetic control. The investigation of extent of regulatory role of RZE1 and RZE2 on ZNF2, the conditions under which theyexert regulation, and the mode of action (transcriptional, translational, or epigenetic control) of these ncRNAs will further clarify the role of ZNF2 inmorphogenesis and virulence in C. neoformans.392. Introns in Cryptococcus neoformans. Carolin Goebels, Sara Gonzalez-Hilarion, Frédérique Moyrand, Guilhem Janbon. Institut Pasteur, Paris, France.Cryptoccus neoformans is a encapsulated basiomycetous yeast responsible for deadly meningoencephalitis in immunocompromised patients. Theanalysis of its genome sequence revealed that nearly all the genes contain introns. These introns are short (67 bp) and each gene contains 5 introns inaverage. RNAseq data analysis showed that alternative splicing is also very common. Moreover, for most of the genes tested introns appeared to beessential for gene expression. We have studied the pathways by which these introns regulate gene expression in C. neoformans. We indentified a polyAbinding protein as a major key factor in this regulation controlling the degradation of the mRNA transcribed from intronless alleles.393. Unravelling of sexual differentiation mediated by Ire1 via Hxl1-independent manners in Cryptococcus neoformans . Kwang-Woo Jung, Yong-SunBahn. Department of Biotechnology, College of Life science and Biotechnology, Yonsei University, Seoul, South Korea.Sexual differentiation is a key biological process for generating genetically diverse offspring, which contributes to the increased fitness of certain speciesin its environmental niches. A human fungal pathogen, Cryptococcus neoformans, undergoes both bisexual and unisexual differentiations. Our previousstudy revealed that UPR (unfolded protein response) components including an evolutionarily conserved ER stress sensor Ire1 and a unique transcriptionfactor Hxl1 modulate ER stress, cell wall integrity, antifungal drug resistance, and virulence in C. neoformans. In this study, we for the first time provideseveral evidences showing that the UPR pathway governs both bisexual and unisexual differentiation of C. neoformans. In the serotype A strainbackgrounds (H99 and KN99a), the ire1D mutants exhibit severe defects in bilateral cross whereas Hxl1 appears to be dispensable for mating in bothunilateral and bilateral crosses. Cell fusion efficiency of unilateral and bilateral crossing with ire1D mutants is significantly decreased when compared toWT crossing, indicating that Ire1 promotes cell-to-cell fusion during mating. Moreover, deletion of the IRE1 gene blocked induction of pheromonemediatedconjugation tubes in crg1D mutants, which lack a RGS protein that negatively regulates pheromone responsive G-protein signaling.Unexpectedly, however, expression of the mating pheromone gene (MFa1) was strongly induced by the mutation of IRE1 gene in serotype A strain,suggesting that Ire1 has both positive and negative roles in mating of serotype A C. neoformans. The ire1D mutants constructed in serotype D (JEC21a andJEC20a) strains also exhibit mating defects similar serotype A ire1D mutant strains, whereas the hxl1D mutants are dispensable for mating. It indicates thatthe role of Ire1 in sexual differentiation is evolutionary conserved in both serotype A and D strains. The unisexual mating, also known as monokaryoticfruiting, is an alternative differentiation process in C. neoformans. The deletion of IRE1 in the XL280 strain, which is used as a tester strain for same-sexmating, causes significant defects in filamentation whereas the hxl1D mutants exhibit levels of filamentation indistinguishable from those of XL280. Inconclusion, the Ire1 regulates both bisexual and unisexual mating of C. neoformans in Hxl1-independent manners.216

FULL POSTER SESSION ABSTRACTS394. Functional analysis of PUF mediated post-transcriptional regulation in Cryptococcus neoformans. Jan Naseer Kaur, John Panepinto. University atBuffalo, Buffalo, NY.The Puf (Pumilio and FBF) family of RNA binding proteins is known to bind to 3’UTRs of mRNAs and is associated with regulatory functions includingtranslation, stabilization and localization of transcripts. Investigation of the C. neoformans genome has revealed that it encodes four PUF proteins. PUFproteins are typically characterized by the presence of 8 consecutive Puf repeats, however variations do occur. Sequence analysis and evolutionary studiesof Puf proteins in fungi has predicted Puf1, Puf2 and Puf3 of C. neoformans to contain 5, 3 and 8 PUM-HD repeats respectively. We hypothesize that Puf1,Puf2 and Puf3 act as RBPs and regulate gene expression. The PUF proteins characterized to date have been reported to bind to 3’ UTR sequenceencompassing a UGUR tetranucleotide in their target RNA. Previous studies have reported that the core motif of UGUA followed by a variable region thatPuf3 binds to is conserved from yeast to humans. Scanning of 3’UTRs of all the annotated genes of C. neoformans revealed Puf3 binding consensussequence in the transcripts involved in pheromone signaling cascade. To test this hypothesis we performed bilateral mating assays for wild type and puf3Dmutants. When equal numbers of opposite mating cells of puf3D mutants were cocultured on V8 agar, we observed that they were defective infilamentation as compared to the wild type cross. To determine the ability of puf3D mutants to produce pheromone, northern blot was done. The RNAobtained from puf3D mutants bilateral cross was probed for mating pheromone MFa in comparison to the wild type cross (H99a x KN99a) and theinduction of MFa was found to be normal in puf3D mutants mating. Also, fusant colony formation assay revealed that filamention defect of puf3D is notdue to impaired cell fusion. Using fluorescent microscopy we have shown that mCherry tagged Puf3 localizes to areas of hyphal growth. Our resultssuggest that mate recognition and fusion do occur when puf3D mutants are crossed. We predict that the defect is in Puf3 mediated post fusion hyphalextension. Future studies will determine the mechanism of Puf3 regulation on potential target transcripts. Also we will identify the target/s of Puf1 andPuf2 and their mechanism of regulation which would enable us to establish a link between the physiology and the Puf regulon of C. neoformans.395. Determining the direct targets of two master regulators of sexual development in Cryptococcus neoformans. Matthew E Mead 1 , Emilia K Kruzel 1 ,Christina M Hull 1,2 . 1) Department of Biomolecular Chemistry, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA, 53706; 2)Department of Medical Microbiology and Immunology, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA, 53706.Cryptococcus neoformans is a major global fungal pathogen that causes disease primarily in immunocompromised individuals. Proposed infectiousparticles include spores, which are produced as a result of sexual development. In crosses between a and a cells this process is in part controlled by thehomeodomain transcription factors Sxi1a and Sxi2a. To understand the molecular events governing development, we set out to identify directtranscriptional targets of the Sxi1a/Sxi2a heterodimer.First, we created a haploid strain in which galactose-inducible promoters control the expression of SXI1a and SXI2a. This SXI-inducible strain allowed usto assess global transcript levels in the presence and absence of SXI expression. At the same time, we compared changes in transcript levels in crossesbetween wild type (a x a) and sxi deletion (sxi1aD x sxi2aD) strains. We discovered that 185 genes exhibited a “Sxi-induced” regulation pattern in bothexperiments.Upstream regions of these highly regulated genes were then analyzed using motif-finding algorithms, and a subset of the Sxi-induced genes was found tocontain a sequence similar to one bound by Sxi1a/Sxi2a in vitro. Individual occurrences of the motif were tested in a Yeast 1-Hybrid system and shown tobe bound by Sxi1a/Sxi2a in a sequence-specific and heterodimer-specific manner. An in vivo reporter assay was then used to show that these binding sitesconfer Sxi-dependent regulation to their downstream targets that is also sequence specific.The list of direct targets studied so far includes numerous uncharacterized genes and putative transcriptional regulators likely important for controllingsubsequent developmental transitions. Future studies will focus on building a complete, Sxi-dependent transcriptional network of development. This workwill help us better understand a process that results in the production of a likely infectious particle in mammalian disease.MEM is funded by the Microbes in Health and Disease Training Grant (NIH T32 AI55397).396. Post-transcriptional gene regulation contributes to host temperature adaptation and virulence in Cryptococcus neoformans. Amanda L. MisenerBloom 1,2 , Kurtis Downey 1 , Nathan K. Wool 1 , John C. Panepinto 1,2 . 1) Microbiology/Immunology, SUNY University at Buffalo, Buffalo, NY; 2) Witebsky Centerfor Microbial Pathogenesis and Immunology, SUNY University at Buffalo, Buffalo, NY.In response to the hostile host environment, pathogens must undergo rapid reprogramming of gene expression to adapt to the stresses they encounter.Upon exposure to host temperature, Ribosomal protein (RP) transcripts are rapidly repressed in C. neoformans. We are interested in investigating specificmechanisms involved in this response, as this repression may be a critical process in host temperature adaptation. Using a mutant null of the majordeadenylase, Ccr4, we have discovered that this repression is in part due to enhanced degradation of RP-transcripts. Ccr4 lacks a nucleic acid bindingdomain and therefore must be recruited to mRNA targets via RNA binding proteins. Using MEME analysis and chromatographic techniques, we haveidentified a shared cis element in the 3’UTR of RP transcripts that is recognized by the zinc knuckle protein, Gis2. We are currently investigating theimportance of this protein-RNA interaction in the expression of RP genes.Host temperature-induced enhanced degradation of RP transcripts is also dependent on the dissociable RNA polymerase II subunit, Rpb4. Specifically, wedemonstrated that in an rpb4D mutant, RP-transcript deadenylation is impaired, suggesting that Rpb4 may be required for Ccr4-targeted degradation. Inaddition, we observed that upon a shift to 37°C, Rpb4 travels from the nucleus to the cytoplasm, supporting a role for Rpb4 in coupling transcription anddegradation. Interestingly, this coupling is not restricted to the RP transcripts, as Rpb4 is also involved in enhanced decay of ER stress transcripts followingtheir peak induction, one hour after a shift to host temperature. We have demonstrated that signaling through PKH enhances the degradation of the RPtranscriptsin response to host temperature, but not the ER stress transcripts, highlighting the complexity of this system. We report that whentranscription and degradation are uncoupled by the loss of Rpb4, growth at host temperature is impaired and virulence in a mouse model of disseminatedcryptococcosis is attenuated. Our data suggests that coupling of transcription and degradation via Rpb4 allows the cell to control the intensity andduration of different responses at specific times following exposure to host temperature, contributing to the ability of C. neoformans to adapt to thisstress.397. Protein arginine methylation in post-transcriptional gene regulation and stress adaptation of Cryptococcus neoformans. J.T. Graham Solomons,Amanda L.M. Bloom, John C. Panepinto. The Department of Microbiology and Immunology, The State University of New York at Buffalo, Buffalo, NY.Cryptococcus neoformans is environmental fungus that opportunistically infects immune compromised individuals, and is widely studied as a modelbasidiomycete. The ability to adapt to host temperature is an essential pathogenic trait of C. neoformans, and the degradation of mRNA initiated by themajor deadenylase Ccr4 appears to play an integral role in the temperature stress response of C. neoformans. Microarray analysis revealed that the largestfunctionally related group of mRNA stabilized in the ccr4D mutant encode ribosomal protein (RP) transcripts. An RNA-binding protein, identified as Gis2,has been shown to interact specifically with a cis element in the 3’UTR of RP transcripts. Gis2 is a small protein (19kDa), predominantly comprised of a27th Fungal Genetics Conference | 217

FULL POSTER SESSION ABSTRACTSseries of a zinc knuckles and a single glycine-arginine rich (GAR) region, which may serve as a target for protein arginine methyltransferases (RMTs).Primary sequence analysis revealed that, Gis2 from C. neoformans (CnGis2) shares significant primary sequence identity with Gis2 from S. cerevisiae(ScGis2) and the human CNBP/ZNF9 protein. However, ScGis2 does not contain a GAR motif; in contrast, the human ortholog contains a GAR element, andhas been shown to undergo methylation by RMT5. An in vitro methylation assay demonstrated that recombinant CnGis2 can be methylated by one (ormore) Cryptococcal RMTs. Investigation of the C. neoformans genome revealed there are 5 putative RMT genes. A C. neoformans rmt5D deletion mutantexhibited a severe growth defect and aberrant cell morphology at 39°C, suggesting that Rmt5 activity impacts temperature adaptation in C. neoformans.Further analysis of the rmt5D mutant and the remaining 4 C. neoformans RMTs will determine to contribution of protein arginine methylation to thevirulence and stress tolerance of C. neoformans.218

FULL POSTER SESSION ABSTRACTS398. UVE1 is a Photo-regulated Gene Required for the Protection of Mitochondrial DNA in Cryptococcus neoformans from UV Induced DNA Damage.Surbhi Verma, Alexander Idnurm. School Of Biological Sciences, University of Missouri-Kansas City, Kansas City, MO 64110.The UVE1 gene is an apurinic/apyrimidinic endonuclease, identified in a T-DNA insertion mutagenesis screen in the pathogenic fungus Cryptococcusneoformans. UVE1 mutation or deletion leads to a UV hypersensitive phenotype. The homologous gene in fission yeast Schizosaccharomyces pombeencodes apurinic/apyrimidinic endonuclease acting in the UVDE-dependent excision repair (UVER) pathway. C. neoformans UVE1 complements a S.pombe uvde knockout strain, hence functionally similar. In Cryptococcus, the Bwc1-Bwc2 photoreceptor complex regulates mating, virulence andultraviolet radiation (UV) stress tolerance. How the Bwc1-Bwc2 complex regulates these functions is not clear. We discovered that UVE1 is photoregulatedin Bwc1-Bwc2 dependent manner in Cryptococcus, and in Neurospora crassa and Phycomyces blakesleeanus that represent two other major lineages inthe fungi. Overexpression of UVE1 in bwc1 mutants rescues their UV sensitivity phenotype and gel mobility shift experiments show binding of Bwc2 to theUVE1 promoter. These experiments indicate that UVE1 is a direct downstream target for the Bwc1-Bwc2 complex, required for UV stress tolerance. Uve1-GFP fusions localize to the mitochondria in C. neoformans. Hence in Cryptococcus, UVE1 is a key photo-regulated gene responsible for tolerance to UVstress for protection of the mitochondrial genome.399. Uve1 endonuclease protects Cryptococcus neoformans from UV damage through regulation by the White collar complex. Surbhi Verma, AlexanderIdnurm. School of Biological Sciences, University of Missouri-Kansas City, Kansas City, MO.In Cryptococcus neoformans the Bwc1-Bwc2 photoreceptor complex regulates mating, virulence and ultraviolet radiation (UV) stress tolerance. Weidentify and characterize a gene, UVE1, whose mutation leads to a UV hypersensitive phenotype. The homologous gene in Schizosaccharomyces pombeencodes a apurinic/apyrimidinic endonuclease acting in the UVDE-dependent excision repair (UVER) pathway. C. neoformans UVE1 complements a S.pombe uvde knockout strain. UVE1 is photoregulated, in a Bwc1-dependent manner in Cryptococcus, as well as in Neurospora crassa and Phycomycesblakesleeanus. Overexpression of UVE1 in bwc1 mutants rescues their UV sensitivity phenotype and gel mobility shift experiments show binding of Bwc2to the UVE1 promoter, indicating that UVE1 is a direct downstream target for the Bwc1-Bwc2 complex. Uve1-GFP fusions localize to the mitochondria. InC. neoformans UVE1 is a key gene regulated in response to light that is responsible for tolerance to UV stress for protection of the mitochondrial genome.400. Multiple laccase genes in Schizophyllum commune. S. Madhavan, K. Krause, E. Kothe. Microbiology Microbial Communication, Friedrich SchillerUniversity, Jena, Germany 07743.The saprophytic white rot fungus S. commune, is involved in the degradation of complex organic molecules including lignin as well as refractory organicmatter from black slate with the help of different exoenzymes. Thus, the genome sequence of S. commune was used to gain an insight into the functionalanalysis of laccases and laccase-like enzymes. Laccases are multi-copper oxidases that catalyze oxidation of a wide spectrum of organic and inorganicsubstances. In most fungi, laccases are found to be multigene families producing isoenzymes with multiple functions. Two laccases and four laccase-likegenes have been identified from the genome of S. commune. Differential regulation of individual genes was analysed at the transcript level by quantitativereal-time PCR. Individual laccase genes showed distinct expression profiles during fungal development, morphogenesis and during substrate utilization.Genes lcc1 and lcc4 seem to play a role during the primordial formation for fruiting bodies phase and lcc2 and lcc6 were found to be related to thedikaryotic phase and vegetative growth. Gene lcc5 is regulated during fruitbody formation. Various stress responsive elements (XRE, STRE) could bedetected in all respective promoters indicating an infuence of aromatic compounds and stress molecules in transcriptional regulation. Characterization oflaccase mutants with respect to organic matter and black slate degradation is linking these data to different functions of single laccase and laccase-likegenes.401. Regulation of DNA repair genes expression by UV stress in Neurospora crassa. Tsukasa Takahashi, Makoto Fujimura, Akihiko Ichiishi. Faculty of LifeScience, Toyo University, Ora-gun, Gunma, Japan.In all organisms, DNA is constantly damaged by endogenous and exogenous factors such as environments and chemicals. In these genotoxins, ultraviolet(UV) irradiation induces DNA damage such as cyclobutane pyrimidine dimers (CPDs) and 6-4 photoproducts (6-4PPs). N.crassa has three mechanisms torepair UV-damaged DNA; nucleotide excision repair (NER), UV dependent repair (UVDR), photoreactivation (PR). Because UV-induced DNA lesions areefficiently removed by these DNA repair systems, N.crassa shows highly resistance to UV as compared to other species. In human and S.cerevisiae, it hasbeen reported that some of DNA repair genes involved in removal of UV-damaged DNA were induced by UV irradiation. Furthermore, some MAP kinasepathways were activated in response to UV irradiation in human. In N.crassa, characterizations of DNA repair gene mutants have been performed in detail,but relationship between expression of these genes and UV stress are not clear yet. Thus, we investigated whether UV stress is involved in regulation ofexpression of DNA repair genes, and whether UV stress activates MAP kinase pathway like a human. We show that some DNA repair genes such as mus-40, mus-43 were up-regulated by UV irradiation. The OS-2 MAP kinase, involved in response to osmotic stress in N.crassa, was activated by UV irradiation,and then expression of mus-40, mus-43 were not induced after UV irradiation in os-2 mutant. In addition, os-2 mutant was more sensitive to UV irradiationthan the wild-type. These data suggest that UV stress upregulates some DNA repair genes and UV signal was partially transmitted by OS MAP kinasecascade, in N.crassa.402. Diverse classes of small RNAs originating from genomic hotspots, tRNA and the mitochondrial genome in Phytophthora infestans. Sultana N.Jahan 1 , Anna K. M. Åsman 1 , Ramesh R. Vetukuri 1 , Anna O. Avrova 2 , Stephen C. Whisson 2 , Christina Dixelius 1 . 1) Department of Plant Biology and ForestGenetics, Uppsala BioCenter, Swedish University of Agricultural Sciences and Linnean Center for Plant Biology, PO-Box 7080, SE-75007, Uppsala, Sweden;2) Cell and Molecular Sciences, The James Hutton Institute, Invergowrie, Dundee DD2 5DA, United Kingdom.Phytophthora infestans is the oomycete pathogen responsible for the devastating late blight disease on potato and tomato. P. infestans is notorious forits ability to evolve to overcome resistant potato varieties. The genome of this pathogen has been sequenced and revealed vast numbers of transposonsequences, and hundreds of disease-promoting effector proteins. We are aiming at understanding gene-silencing mechanisms in P. infestans includingdeciphering roles of small non-coding RNAs. In our previous work we have shown that P. infestans has an active RNA silencing pathway (Vetukuri et al.2011). We also performed deep sequencing of sRNAs from P. infestans and knocked down the genes encoding the RNA silencing components Argonauteand Dicer in order to investigate their roles in sRNA biogenesis (Vetukuri et al. 2012). Here, we describe the distribution of genomic sites from which sRNAsoriginate. Genome-wide analysis of sRNAs revealed diverse classes mapping to genomic sources such as tRNAs, rRNAs, genomic sRNA ‘hotspots’, and themitochondrial genome. Most tRNA-derived RNA fragments (tRFs) mapped to the sense strand of the 5’-halves of mature tRNAs and peaked at 27 and 30 ntlengths. In accordance with reports from other organisms (Franzén et al. 2011), the tRFs mapped to different tRNA isoacceptors with unequal frequencies,the Ile_tRNA_Cluster_0 showing the highest proportion of mapping sRNAs. We are presently using our Dicer knockdown transformants to investigate thetRF biogenesis mechanism. Another interesting group of sRNAs are those that map to transposons that have close-by neighboring RXLR-effector genes.27th Fungal Genetics Conference | 219

FULL POSTER SESSION ABSTRACTSOne such example is PiAvr2, which is located just 231 bp from a class II transposon. The presence of sRNAs mapping to both PiAvr2 and the nearbytransposon indicate that RNA silencing may play a role in regulation of this important effector gene. Over a hundred additional predicted genes werefound to be sRNA hotspots in our data: Crinkler effector genes, arrays of duplicated genes, potentially antisense overlapping transcripts, and genescontaining transposon insertions. Our present task is to reveal the role that sRNAs might play in their regulation.403. Epigenetic control of effector gene expression in the plant pathogen fungus Leptosphaeria maculans. Jessica Soyer, Mennat El Ghalid, Marie-HélèneBalesdent, Thierry Rouxel, Isabelle Fudal. INRA, UR 1290 BIOGER-CPP, Avenue Lucien Brétignière, F-78850 Thiverval-Grignon, France.Plant pathogenic microbes secrete an arsenal of small secreted proteins (SSPs) acting as effectors that modulate host immunity to facilitate infection. InEukaryotic phytopathogens, SSP-encoding genes are often located in particular genomic environments and show waves of concerted expression at diversestages of plant infection. To date, little is known about the regulation of their expression. Leptosphaeria maculans is an ascomycete fungus responsible forthe most devastating disease of oilseed rape (Brassica napus). The sequencing of its genome revealed a bipartite structure alternating gene rich GCisochoresand gene poor AT-isochores made up of mosaics of transposable elements. The AT-isochores encompass one third of the genome and areenriched in putative effector genes that present the same expression pattern (no or a low expression level during in vitro growth and a strong overexpressionduring primary infection). Here, we investigated the involvement of an epigenetic control in the regulation of effector gene expression. For thispurpose, we silenced expression of two key players of heterochromatin remodeling, i.e. HP1 and DIM5, by RNAi and used HP1::GFP as a heterochromatinmarker. Whole genome oligoarrays were done in silenced-HP1 and silenced-DIM5 isolates to analyze the involvement of HP1 and DIM5 on geneexpression according to their function and location. We evaluated the effect of a change of genomic context from AT-isochores to GC-isochores on theexpression of effector genes. Silencing of DIM5 resulted in lack of chromatin condensation. The silencing of HP1 and DIM5 resulted in an over-expressionof pathogenicity-related genes during in vitro growth, with a favored influence on SSP-encoding genes in AT-isochores. The “moving” of effector genescorroborated transcriptomic analysis as it led to a strong overexpression of effector genes during in vitro growth. These data strongly suggest that anepigenetic control represses the expression of effector genes located in AT-isochores during in vitro growth, which is, to our knowledge, the firstdescription of an epigenetic control, relying on HP1 and DIM5, exerted on effector-encoding genes expression. Switch toward pathogenesis lifts thisrepression based on chromatin-structure, rendering promoters of effector genes accessible to specific transcription factors.404. Discovering the link: The NOX-GSA network for sexual development and ascospore germination in Sordaria macrospora. Daniela Dirschnabel,Christian Schäfers, Ines Teichert, Ulrich Kück. General and Molecular Botany, Ruhr-University Bochum, Bochum, Germany.Recently we were successful in establishing a genetic network for sexual development and ascospore germination in the homothallic filamentous fungusSordaria macrospora [1, 2]. Central components of this network are three G-protein alpha subunits (GSA), an adenylat cyclase SAC1, and the transcriptionfactor STE12. The three GSA proteins (GSA1, GSA2 and GSA3) have different roles in developmental processes. GSA1 and GSA2 are important for sexualpropagation and the generation of perithecia, while GSA3 is essential for proper ascospore germination. Interestingly, the phenotypes of mutants lackingfungal NAD(P)H oxidases (NOX) resemble the known Dgsa phenotypes: DnoxA shows an arrest of sexual development and ascospores from a DnoxBmutant fail to germinate. These similarities raised the question, whether the GSA proteins and NOX enzymes are part of identical signaling pathways. Toverify this hypothesis, we generated knockout mutants of both NOX A and B isoforms and their regulator NOXR in S. macrospora. Our hypothesis wasfurther supported by the comparison of these mutants with gsa deletion mutants by measuring hyphal fusion events, quantification of reactive oxygenspecies and ascospore germination. The generation of double mutants and complementation studies with constitutive gsa1 derivatives enabled us topropose an interactive NOX-GSA network for sexual development and ascospore germination. References: 1.Kamerewerd, J., M. Jansson, M. Nowrousian,S. Pöggeler, and U. Kück, Three alpha-subunits of heterotrimeric G proteins and an adenylyl cyclase have distinct roles in fruiting body development in thehomothallic fungus Sordaria macrospora. Genetics, 2008. 180(1): p. 191-206. 2.Engh, I., M. Nowrousian, and U. Kück, Sordaria macrospora, a modelorganism to study fungal cellular development. European journal of cell biology, 2010. 89(12): p. 864-72.405. The bZIP transcription factor Atf1 acts as a global regulator for secondary metabolite production in Fusarium fujikuroi. Sabine E. Albermann,Bettina Tudzynski. IBBP, WWU Muenster, Schlossplatz 8, 48143 Muenster, Germany.The activating transcription factor 1 (Atf1) belongs to the bZIP transcription factor family and is known to have a great impact on stress responsesmediated by the mitogen activated protein kinase (MAPK) cascade in fission yeast. In this pathway, activation of the transcription factor is achieved byphosphorylation via the kinase Sty1. Furthermore, the transcription factor plays a role in sexual and asexual development which was observed for severalfilamentous fungi e.g. in Aspergillus species where it affects conidiospore germination. Atf1 can also act as a virulence factor which was described for itshomologue in the rye pathogen Claviceps purpurea. However, involvement of Atf1 in secondary metabolism was first observed in the grey mould Botrytiscinerea. As Atf1 seems to play a crucial role in different processes, this transcription factor was also investigated in the rice pathogen Fusarium fujikuroi.For this purpose, deletion mutants of atf1 and the Sty1 homologue sak1, the putative kinase for Atf1, were cultivated under varying conditions. HPLCanalysis of the secondary metabolite spectrum revealed a drastic change in the production level of several metabolites. Gibberellic acids, for instance, aredown-regulated up to 50 % in Datf1 compared to the wild-type, whereas the amount of gibberellins in the Dsak mutant is about twice as much as in thewild-type. Furthermore, applied salt stress dramatically enhances mycotoxin production in the Datf1 mutants, while the deletion mutant Dsak1 is not ableto grow at all. Plate assays applying different stressors to the strains revealed involvement of both proteins in the osmotic stress response. However,reactive oxygen species and cell wall damaging agents do not seem to have an impact on their growth. In contrast, reduced protoplast formation wasobserved for Datf1 mutants and even more significantly in Dsak. Therefore, it is very likely, that the cell wall composition and integrity is changed in thesemutants. Summarizing, Atf1 and Sak1 are involved in various processes such as secondary metabolite production, cell wall integrity as well as in stressresponses. The obtained information leads to the conclusion that Sak1 might be the kinase responsible for Atf1 phosphorylation. But there certainly haveto be more factors to be involved in activation of this transcription factor.406. Role of the Vivid ortholog of Fusarium fujikuroi VvdA in carotenoid biosynthesis and development. Marta Castrillo Jimenez, J. Avalos. Genetics,University of Sevilla, Sevilla, Seville, Spain.Fusarium fujikuroi is well known for its ability to produce gibberellins, growth-promoting plant hormones with agricultural applications. Recently, thisspecie has become a model system in the research of other metabolic pathways, including carotenoid biosynthesis. This fungus produces an acidicapocarotenoid, neurosporaxanthin, through the activity of the enzymes encoded by five structural genes, whose expression is induced by light. We areinterested in the molecular basis of this regulation. As usually found in fungi, the F. fujikuroi genome contains genes for WC-1 and WC-2 orthologs. Incontrast to other species with light-induced carotenogenesis, e.g., Neurospora crassa or Phycomyces blakesleeanus, this photoresponse is not impaired innull mutants of the only wc-1-like gene of F. fujikuroi, wcoA. Therefore, we are analyzing the role of other blue-light photoreceptors. Here we described220

FULL POSTER SESSION ABSTRACTSthe identification, regulation and targeted mutation of the gene vvdA, ortholog of the N. crassa vivid (vvd) gene. The predicted F. fujikuroi VvdA protein issimilar to VVD in size (198 aa compared to 186 aa) and sequence (87 identical positions). Deletion of vvdA in F. fujikuroi results in a significant reduction inpigmentation and carotenoid production, a regulatory effect opposite to the enhanced carotenoid accumulation characteristic of the vvd mutants of N.crassa. Additionally, vvdA mutant colonies exhibit a different aspect in the light, apparently due to more compact development or aerial mycelia. As foundfor vvd in N. crassa, expression of vvdA in F. fujikuroi cultures is strongly stimulated by light, an activation which is severely reduced in the wcoA mutants.Accordingly, the alterations exhibited by the vvdA mutants are only apparent under illumination. Our results suggest that VvdA participates in thephotoreceptor machinery responsible for carotenoid photoinduction in F. fujikuroi.407. Gene expression of secondary metabolism gene clusters by different Fusarium species during in planta infection. J. Espino 1 , M. Muensterkoetter 2 ,U. Gueldener 2 , B. Tudzynski 1 . 1) Institut of Plant Biology and Biotechnology, Westf. Wilhelms University,Schlossplatz 8, 48143 Muenster, Germany; 2)2Institute of Bioinformatics and Systems Biology, Helmholtz Zentrum Muenchen (GmbH), Ingolstaedter Landstrasse 1, 85764 Neuherberg, Germany.The Gibberella fujikuroi complex (GFC) comprises about 50 Fusarium species with similar characteristics, which are responsible for an array of plantdiseases, causing devastating losses in agriculture. The majority of their members is able to produce different toxins which can contaminate food and feedworldwide. Despite of their similarities, they differ in their spectrum and amount of secondary metabolites (SM) production, probably due to differentnatural hosts. For example, Fusarium verticillioides is considered as a fumonisin producer and attacks mainly maize, whereas Fusarium fujikuroi causes thebakanae disease in rice, secreting the phytohormone gibberellin beside several other products. Another fungus, Fusarium mangiferae, causes the mangomalformation and is neither able to produce fumonisins nor gibberellins. And Fusarium proliferatum produces a very broad spectrum of mycotoxins andinfects mainly maize. In the present study we compared the in planta expression profiles for different secondary metabolism gene clusters in these fourspecies of the GFC, and also the one of Fusarium oxysporum as an outgroup not belonging to the GFC. So far, gene expression studies have been done forthese fungi mainly in vitro, showing differential regulation mechanisms, e.g. in response to nitrogen availability. But not much is known about the geneexpression during plant infection. We have performed an infection assay in maize and rice and quantified the fungal biomass in the roots by quantitativePCR using genomic DNA to determine the ratio between plant and fungal biomass in infected tissue. The expression of SM genes was followed up in timecourse experiments. The results of this study show differences between the species regarding colonization of the host and expression of SM. Surprisinglythe high expression of some gene clusters, which were never expressed before in vitro, suggests a specific induction by plant signals.408. A cis-acting factor modulating the transcription of FUM1 in Fusarium verticillioides. Valeria Montis 1 , Matias Pasquali 2 , Ivan Visentin 1 , Petr Karlovsky 3 ,Francesca Cardinale 1 . 1) Dipartimento di Scienze Agrarie, Forestali e Alimentari, Università degli Studi di Torino, 10095 Grugliasco (TO), Italy; 2)Environment and Agrobiotechnology Dept, CRP GABRIEL LIPPMANN, Belvaux, Luxembourg; 3) Department of Crop Sciences, Molecular Phytopathologyand Mycotoxin Research, University of Göttingen, D-37077 Göttingen, Germany.Fumonisins-biosynthetic FUM genes are clustered and co-expressed in toxin producers. By overrepresentation analyses, we identified a motif inpromoters of clustered FUM genes in both fumonisins producers F. verticillioides and A. niger. The same motif was not found in various FUM genehomologues of fungi that do not produce fumonisins. Deletion of the main 6-mer in FvFUM1 promoter compromises its gene expression both in plantaand in vitro. We hypothesize that such motif may be important for clustered FUM genes coordinated transcription, being the core of a transcription factorbindingsite for a putative Zn-finger protein.409. Shedding light on secondary metabolite cluster gene expression, sporulation, UV-damage repair and carotenogenesis in the rice pathogenFusarium fujikuroi. Phillipp Wiemann, Bettina Tudzynski. Institut für Biologie und Biotechnologie der Pflanzen Westfälische Wilhelms-Universität MünsterSchlossplatz 8 48143 Münster Germany.The rice pathogen Fusarium fujikuroi produces economically important secondary metabolites like gibberellic acids and carotenoids as well asmycotoxins like bikaverin and fusarin C. Their production is activated in response to environmental stimuli such as light, pH or nutrient availability. In thisstudy, we evaluate the effects of light and different putative light receptors on growth and differentiation as well as secondary metabolism. Bimolecularfluorescence complementation proved that homologs of the Neurospora crassa White Collar proteins in F. fujikuroi (WcoA and WcoB) form a nuclearlocalized complex (WCC) that is needed for full functionality. Deletion and complementation of both genes revealed that the WCC represses bikaverin geneexpression in constant light conditions and induces immediate light-dependent carotenoid gene expression as shown by northern blot analyses.Additionally the WCC represses conidiogenesis in response to light. The effects observed regarding bikaverin and carotenoid gene expressions as well asconidiogenesis are antagonistically to the ones observed in the velvet mutant, making a connection between the WCC and the velvet complex feasible,similarly to the situation in Aspergillus nidulans. Since carotenoid production was maintained in both wcoA and wcoB single as well as in wcoA/B doublemutants in constant light conditions, we focused on characterization of additional putative light receptors in F. fujikuroi. Deletion of the phytochrome-likeencodinggene fph1 did not show any significant phenotype. Deletion of phl1, coding for a cryptochrome/photolyase demonstrated impaired carotenoidbiosynthesis gene expression upon exposure to light. Additionally, gene expression and HPLC analyses of these mutants demonstrated loss of fusarin Cgene expression and concomitant production formation compared to the wild type, suggesting a distinct transcriptional activity for this barelycharacterized class of enzymes. Finally UV mutagenesis experiments and qRT-PCR demonstrate that WcoA, WcoB and Phl1 are involved in UV-damagerepair most likely by transcriptionally activating phr1, encoding a CPD-photolyase. The data presented here allow us to draw a first model of how lightreceptors function in a signaling network in the rice pathogen F. fujikuroi.410. Fgap1-mediated response to oxidative stress in trichothecene-producing Fusarium graminearum. M. Montibus, N. Ponts, E. Zehraoui, F. Richard-Forget, C. Barreau. INRA, UR1264-MycSA, BP81, F-33883 Villenave d’Ornon, France.The filamentous fungus Fusarium graminearum infects cereals and corn. It is one of the main causal agent of “Fusarium Head Blight” and “Maize EarRot”. During infection, it produces mycotoxins belonging to the trichothecenes family that accumulate in the grains. Although the biosynthetic pathwayinvolving specific Tri genes has been elucidated, the global regulation of toxin biosynthesis remains enigmatic. It is now established that oxidative stressmodulates the production of toxins by F. graminearum. H 2O 2 added in liquid cultures of this fungus enhances trichothecenes accumulation and increasesTri genes expression. Our working hypothesis is that a transcription factor regulates redox homeostasis, and is involved in Tri genes regulation. In the yeastSaccharomyces cerevisiae, the transcription factor Yap1p mediates response to oxidative stress via nuclear re-localization and activation of genes codingfor detoxification enzymes. In this study, we investigate the role of Yap1p homolog in F. graminearum, Fgap1, in response to oxidative stress and itseventual role in the regulation of trichothecene production. A deleted mutant and a strain expressing a constitutively activated form of the Fgap1 factor inF. graminearum were constructed. We cultured these mutants in GYEP liquid medium supplemented with H 2O 2 to evaluate their sensitivity to oxidativestress and analyse their toxin production. The nuclear localization of constitutively activated Fgap1p as well as wild-type Fgap1p under oxidative stress by27th Fungal Genetics Conference | 221

FULL POSTER SESSION ABSTRACTSH 2O 2 was analyzed. Expression profiles of genes encoding oxidative stress response enzymes potentially controlled by Fgap1p and of genes involved in thebiosynthesis of type B trichothecenes were analyzed by Q-RT-PCR. Trichothecene accumulation is strongly enhanced in the deleted strain, with an increasein Tri genes expression. On the other hand, Tri genes expression and toxin accumulation are drastically repressed in the mutant in which Fgap1p isconstitutively activated. Moreover, the level of expression of two genes encoding catalases is modulated in both mutants. The involvement of Fgap1 inother types of stress has also been investigated. In particular, cadmium and osmotic stress affect growth in the deleted strain.411. Functional analyses of FgLaeA in Fusarium graminearum. Hee-Kyoung Kim, Seong-mi Jo, Seunghoon Lee, Sung-Hwan Yun. Dept Med Biotech,Soonchunhyang Univ, Asan, Chungnam 336-745, South Korea.Fusarium graminearum (telomorph: Gibberella zeae) is the casual agent of the head blight of cereal crops and produces mycotoxins such astrichothecenes and zearalenone in infected plants. The expression of genes involved in biosyntheses of these mycotoxins are controlled at the differentlevels ranging from by a pathway-specific transcription regulator (encoded by TRI6 or ZEB2) to by a global regulator involved in chromatin remodeling.Here we focused on the function of FgLaeA in F. graminearum, which is an ortholog of the Aspergillus nidulans LaeA, encoding the global regulator forboth secondary metabolism and sexual development. For functional analysis of FgLaeA in mycotoxin production, we used a transgenic F. graminearumstrain expressing a firefly luciferase gene under control of TRI6 or ZEB2 promoter as a reporter system. Targeted deletion of FgLaeA led to a dramaticreduction of luminescence in the reporter strain, indicating that FgLaeA controls the expression of both TRI6 and ZEB2 in F. graminearum; the reducedtoxin accumulation was further confirmed by HPLC analysis. In addition, the FgLaeA deletion strains exhibited not only albino phenotype on CM mediumbut also earlier formation of sexual fruiting bodies (perithecia) on carrot agar than its wild-type progenitor, the latter indicating that FgLaeA seems tonegatively control the perithecial induction. Quantitative real-time PCR revealed that FgLaeA was expressed constitutively under both mycotoxinproduction and sexual development. Overexpression of a GFP-FgLaeA fusion construct in a FgLaeA-deletion strain recovered all the phenotypic changes tothe wild-type levels, and led to constitutive expression of GFP in the entire cells at different developmental stages. A split luciferase assay for in vivoprotein-protein interaction demonstrated that FgLaeA could not interact with FgveA, an ortholog of A. nidulans veA. Taken together, it is likely that FgLaeAcontrols both secondary metabolism and sexual development in F. graminearum, but the regulation pattern operated by FgLaeA is somewhat differentfrom that by LaeA in A. nidulans.412. Molecular cloning and differential expression of two novel Family 1 b-glucosidases genes from the rare fungus Stachybotrys microspora. SalmaAbdeljalil, Houcine Lazzez, Ali Gargouri. Centre of Biotechnology of Sfax, Sfax-Tunisia.The cellulolytic system of the fungus Stacchybotrys microspora is characterized by the existence of several b-glucosidases. From a compilation of fungalb-glucosidases belonging to family GH1, we designed primers to isolate b-glucosidases by PCR. Using different primers combination, three differentfragments genes were firstly obtained. Two of them are overlapping and constitute a novel gene named Smbgl1A while the third one is a part of a secondgene named Smbgl1B. RT-PCR analysis showed the first gene is induced by cellulose and repressed by glucose while Smbgl1B is equally expressed on bothconditions The identification of putative catalytic residues as well as the conserved glycone and aglycone binding sites was performed on SmBgl1Adeduced aminoacid sequence. The predicted secondary structure of Smbgl1 confirmed its appurtenance to GHI family: the presence of a classical (b/a)8barrel and all the characteristic of subsite -1 (glycone site).413. The transcriptional factors XYR1 and CRE1 regulate the expression of Cellulolytic and Xylanolytic genes at carbon source dependent-manner inHypocrea jecorina (Trichoderma reesei). Amanda C.C. Antoniêto, Lílian S. Castro, Wellington R. Pedersoli, Roberto N. Silva. Department of Biochemistryand Immunology, School , University of São Paulo, Ribeirão Preto-SP, São Paulo, Brazil.The ascomycete Hypocrea jecorina (anamorph of Trichderma reesei) is a one of the most well studied cellulolytic fungus and widely used in thebiotechnology industry, such as in the production of second generation ethanol, because it is a strong producer of hydrolytic enzymes such as cellulasesand xylanases. The objective of this study was evaluate the gene expression and enzymatic activity of cellulases and xylanases in the Dxyr1 and Dcre1mutants and compare with the parental T. reesei (QM9414), in three different carbon sources. The strains were grown in Mandels-Andreotti medium,supplemented with cellulose, sophorose or glucose. The expression of 22 set cellulases and xylanases genes were evaluated by real-time PCR (qRT-PCR)and cellulolytic and xylanolytic activities were observed using different substrates. The cel6a, cel3a, cel7b, cel3c, cel3e, xyn2 and swo genes showed asignificantly high expression in the mutant Dcre1 when compared with the parental QM9414 and low expression of the cel1a, cel3d and cel61b genes wasobserved when compared the mutant Dxyr1 with the QM9414 on cellulose, sophorose and glucose. Overall, all of cellulase and xylanase genes showedhigher expression in mutant Dcre1 and low expression in mutant Dxyr1 in all studied conditions, when compared to QM9414. Concerning to enzymaticprofiles, the activity of CMCase, b-glucosidase and Xylanases ranged also for the presence of specific carbon source. These results suggest that the deletionof the genes xyr1 and cre1 affects the formation of cellulases and xylanases directly at transcriptional level and shown to be specific and dependent of thecarbon source.414. Characterization of tannic acid-inducible and hypoviral-regulated CpsHsp1 expression level of the chestnut blight fungus Cryphonectria parasitica.J.-H. Baek 1 , J.-A. Park 1 , J.-M. Kim 2 , S.-M. Park 1 , D.-H. Kim 1 . 1) Institute for Molecular Biology and Genetics, Center for Fungal Pathogenesis, ChonbukNational University, Jeonju, Chonbuk, South Korea; 2) Department of Bio-Environmental Chemistry, Wonkwang University, Iksan, Chonbuk, South Korea.A small heat shock protein gene, CpsHsp1, a ubiquitous chaperone in Cryphonectria parasitica, was characterized. The predicted protein sequence ofCpsHsp1 gene contains a putative conserved domain, which is alpha crystallin domain (ACD) of alpha-crystallin-Hsps_p23-like superfamily. To characterizebiological functions of the CpsHsp1 gene in the C. parasitica, the replacement vector for CpsHsp1-null mutant was designed to favor double crossoverintegration events. Disruption of the CpsHsp1 protein resulted in retarded growth rate, approximately 78.5% of the radial growth observed in the virusfreestrain EP155/2. When the hypovirus CHV1 was transferred to the CpsHsp1-null mutant, all of the virus-containing CpsHsp1-null progeny displayedcharacteristics of invasive feeding hyphae, near absence of the typical mycelial mat on the surface, and sparse aerial hyphae. Northern blot analysisshowed little accumulation of the CpsHsp1 gene transcript under normal growth conditions. However, the accumulation of the CpsHsp1 gene transcriptwas induced in modified Bavendamm’s medium, which is a 0.7% tannic acid-.supplemented malt extract agar. To examine the viral regulation of theinduction, the CpsHsp1 induction pattern in the isogenic hypovirulent strain UEP1 was compared with that in the wild-type strain EP155/2. Northern blotanalysis of RNA from UEP1 cultured under induction conditions with tannic acid showed that hypoviral infection specifically reduced the level of CpsHsp1transcript induced by tannic acid. To determine whether CpsHsp1 is induced by cool or heat stress, we additionally observed difference in the expression,and induction pattern of CpsHsp1 between virus-free EP155/2 and virus-infected hypovirulent UEP1 strains by Northern blot analysis and Western blotanalysis.222

FULL POSTER SESSION ABSTRACTS415. Artificial miRNA constructs for Phytophthora sojae transformation. Stephanie R. Bollmann 1 , Felipe D. Arredondo 1 , Noah Fahlgren 2 , James C.Carrington 2 , Niklaus J. Grünwald 3 , Brett M. Tyler 1 . 1) Center for Genome Research and Biocomputing, and Department of Botany and Plant Pathology,Oregon State University, Corvallis, OR; 2) Donald Danforth Plant Science Center, St. Louis, MO; 3) Horticultural Crops Research Laboratory, USDAAgricultural Research Service, Corvallis, OR.Phytophthora, a genus of fungal-like oomycetes, contains some of the most devastating plant pathogens, causing multi-billion dollar damage to crops,ornamental plants, and natural environments. The genomes of five Phytophthora species, including the soybean pathogen P. sojae, have recently beensequenced, with many more species soon to be completed. Gene regulation by small RNA pathways is highly conserved among eukaryotes, although littleis known about small RNA pathways in the Stramenopile kingdom. Two Dicer homologs, DCL1 and DCL2, and one RDR homolog were cloned andannotated from P. sojae, and gene expression analysis revealed only minor changes in transcript levels among different lifestages and infection timepoints.At this point, the role of the two oomycete Dicer homologs are only speculated. This study aims to down-regulate DCL1 and DCL2 expression in order toanalyze the contribution of each homolog to small RNA biogenesis. Traditional RNAi, such as overexpression of RNA complementary to a target mRNAtranscript, has been used to knockdown gene expression in Phytophthora, although the effect is most often short-lived. Dicer homologs are involved in theRNAi pathway, therefore this method may not be effective, especially for the homolog involved in the siRNA pathway. Artificial miRNAs, designed fromendogenous miRNAs, have recently been used to target transcripts such as these. We designed artificial miRNA constructs based on the conservedPhytophthora miRNA found in P. sojae, targeting both DCL1 and DCL2 as well as the effector Avr1k, the histidine biosynthesis enzyme HISG, and GFP forcontrols. Analysis of transformants is currently underway.416. RNAi-dependent epimutations evolve antifungal drug resistance in the zygomycete fungal pathogen Mucor. Silvia Calo Varela 1 , Cecelia Shertz 1 ,Robert J Bastidas 1 , Soo Chan Lee 1 , Piotr Mieczkowski 2 , Joshua A Garnek 1 , Rosa Ruiz-Vazquez 3 , Santiago Torres-Martinez 3 , Maria E Cardenas 1 , JosephHeitman 1 . 1) Molecular Genetics and Microbiol, DUKE University Medical Center, Durham, NC; 2) High Throughput Sequencing Facility, CCGS, UNC,Chapel Hill, NC; 3) Department of Molecular Genetics and Microbiology, University of Murcia, Murcia, Spain.Microorganisms evolve via a panoply of mechanisms spanning aneuploidy, sexual/parasexual reproduction, mutators, Hsp90, and even prions.Mechanisms that may seem detrimental can be repurposed to generate diversity. The pathogenic fungus Mucor circinelloides grows as a hyphaeaerobically, but as a yeast in anaerobic conditions or in the presence of the immunosuppressive drug FK506. FKBP12 is a protein folding enzyme conservedthroughout eukaryotes that interacts with FK506 and mediates antifungal activity of this drug. The FK506-FKBP12 complex inhibits the proteinphosphatase calcineurin and thereby blocks hyphal growth of M. circinelloides. Continued exposure to FK506 yields resistant isolates, which exhibit hyphalgrowth emerging from the yeast colony. Some isolates harbor a variety of mutations in the fkbA gene that encodes FKBP12. However, other isolatesharbor no mutations in the fkbA gene. These unusual epimutant isolates also revert frequently within several generations of vegetative growth in drugfreemedia and are restored to wild-type (yeast growth in the presence of FK506). Northern and Western analyses revealed a loss of fkbA mRNA andFKBP12 protein in the epimutants. High-throughput sequencing and Northern blot also detected sRNA generated from fkbA in the epimutant strains,revealing a new role for RNAi in the development of transient, reversible resistance to an antifungal drug treatment. RNAi could be triggered via dsRNAproduction from an overlap in the 3’ regions of the mRNA of fkbA and its neighboring gene patA, which encodes a putative polyamine transporter. Ourresults reveal a novel epigenetic RNAi-based epimutation mechanism controlling phenotypic plasticity in fungi.417. Heterochromatic marks are involved in the repression of plant-regulated secondary metabolism in Epichloë festucae and for symbiotic interactionwith the host perennial ryegrass. Tetsuya Chujo, Barry Scott. Institute of Molecular BioSciences, Massey University, Palmerston North, New Zealand.The fungal endophyte Epichloë festucae systemically colonizes perennial ryegrass (Lolium perenne), and produces a range of secondary metabolites toprotect the host plant. The alkaloid peramine provides protection against insect herbivory. Protection against mammalian herbivory is afforded by theproduction of alkaloids, such as ergot alkaloids and lolitrems. Using the E. festucae-perennial ryegrass symbiotic association as a model experimentalsystem we have shown that gene clusters for the synthesis of these bioprotective metabolites are all preferentially and highly expressed in planta, but notexpressed in culture. Recent work showed that disruption of genes encoding either heterochromatin protein-1 (HepA) or the H3K9 methyltransferase(ClrD) in Aspergillus nidulans resulted in enhanced expression of secondary metabolite gene clusters, demonstrating that heterochromatic marks areinvolved in the repression of these clusters. Thus, we hypothesized that plant-regulated E. festucae secondary metabolite gene clusters have a repressivechromatin structure in culture, and chromatin remodeling is an important component for activation of these gene clusters in planta. To test thishypothesis we have deleted the hepA and clrD homologues from E. festucae by targeted gene replacement. Deletion of hepA resulted in a slight reductionin culture radial growth whereas deletion of clrD resulted in a severe reduction. Western blot analysis revealed that the level of H3K9 tri-methylation(H3K9me3) is dramatically decreased in DclrD mutants. Expression levels of ltmG & ltmM (cluster 1) and ltmP & ltmF (cluster 2), as measured by qRT-PCR,increased in both the DhepA and DclrD mutants grown in a defined medium. Introduction of a wild-type allele of either hepA or clrD complemented DhepAor DclrD mutant phenotypes, respectively. In addition, the DhepA mutant has a dramatic host interaction phenotype, inducing severe stunting andpremature senescence of the ryegrass host. On the other hand, DclrD mutant is an infection mutant. These results strongly suggest that heterochromaticmarks regulate both secondary metabolite gene expression and the mutualistic symbiotic interaction of E. festucae with its host perennial ryegrass.418. Cellulose Degradation Regulator 2 Induces Expression of a Conserved Core of Genes for Plant Cell Wall Saccharification in Neurospora crassa andAspergillus nidulans. Samuel T. Coradetti, Yi Xiong, N Louise Glass. Department of Plant and Micorbial Biology, University of California, Berkeley, CA.To better understand mechanisms of cellulase gene regulation and genome-wide gene regulation enabling robust enzyme secretion, we studied theconservation of gene regulation by cellulose degradation regulator 2 (CLR-2) in Neurospora crassa and Aspergillus nidulans. Misexpression of CLR-2 undernormally repressive and non-inducing culture conditions was sufficient for cellulases secretion in N. crassa, but not A. nidulans. We used RNAseq to mapthe trascriptome in wild-type, deletion and mis-expression strains of both species. We identified a cohort of conserved enzymes with conserved sequenceand CLR-2 dependent regulation across evolutionarily divergent ascomycetes, which represent a core of essential enzymes for degradation of complexcellulosic substrates. We also identified non-conserved CLR-2 regulated genes in each species, which may have function specific to a particular substrate orniche. These data suggest that manipulation of CLR-2 has significant potential for improved cellulase production from industrial production strains.419. The transcriptional repressor CRE-1 regulates glycogen metabolism in Neurospora crassa. Fernanda B. Cupertino, Stela Virgilio, Fernanda Z. Freitas,Thiago S. Candido, Maria Célia Bertolini. Instituto de Quimica,UNESP, Araraquara, São Paulo, Brazil.In Neurospora crassa the RCO-1 co-repressor, an orthologue of the yeast Tup1, has been identified as a protein involved in glycogen metabolismregulation in a screening of a transcription factor knocked-out strains set. The Saccharomyces cerevisiae Tup1 protein participates in the Tup-1-Ssn627th Fungal Genetics Conference | 223

FULL POSTER SESSION ABSTRACTScomplex, which represses the expression of genes through its interaction with other protein partners, such as the yeast Mig1 repressor, mediator ofcarbon catabolite repression (CCR). The N. crassa CRE-1 is an orthologue of Mig1p and has been reported to regulate a number of genes by binding to theCreA motif 5’-SYGGRG-3’. In this work we investigated if CRE-1 transcription factor regulates glycogen metabolism and whether depends on the RCO-1/RCM-1 complex, the ortologue of the yeast Tup-1/Ssn6. First, we demonstrated that CRE-1 is involved in catabolic repression in N. crassa. Growth of thewild-type strain in VM medium containing xylose + 1 mM 2-deoxyglucose (2-DG) was drastically reduced, while growth in the cre-1 KO was not affected by 2-DG. The cre-1 KO , rco-1 KO and rcm-1 RIP mutant strains showed impaired glycogen accumulation when compared to the wild-type strain, being the higherlevels observed in the cre-1 KO strain. Glycogen accumulated by this mutant strain was much higher than the wild-type strain under both repressed(glucose) and non-repressed (xylose) carbon source, suggesting that the carbon source has no influence in the glycogen accumulated. The DNA motif 5’-SYGGRG-3’ was identified in the promoters of genes encoding for synthesis (gnn, gsn, and gbn) and degradation enzymes (gpn, gdn). Binding of theGST::CRE-1 recombinant protein to the CRE-1 motifs in the gsn and gpn promoters was confirmed by DNA gel shift, indicating that both genes must beregulated by CRE-1. Gene expression was analyzed by qRT-PCR and all genes were differently expressed in the mutant strains, and for some genes, geneexpression correlated well with the levels of glycogen accumulated. ChIP-PCR assay was performed to confirm in vivo CRE-1 binding to the promoters andthe results suggested that the CRE-1, RCO-1 and RCM-1 proteins likely interacted. All results together indicated that the CRE-1 transcription factor acts as arepressor in glycogen regulation, and might require the interaction with RCO-1 and RCM-1 co-repressors. Supported by FAPESP and CNPq.420. Transcriptional Response to Hypoxia in the Dimorphic Fungus Histoplasma capsulatum . Juwen C. DuBois 1,3 , A. George Smulian 2,3 . 1) Pathobiologyand Molecular Medicine Graduate Program, University of Cincinnati, Cincinnati, OH; 2) Infectious Disease Division, University of Cincinnati, Cincinnati, OH;3) Cincinnati VA Medical Center, Cincinnati OH.Background and purpose: The incidence of life-threatening fungal infections has increased because of a rapidly growing population ofimmunocompromised individuals. Histoplasma capsulatum (Hc) is a thermally dimorphic fungal pathogen which causes pulmonary and systemic disease inboth immunocompetent and immunocompromised individuals. All fungal pathogens encounter microenvironmental stresses as they colonize and infectthe mammalian host and their ability to adapt to environmental changes is critical for pathogenicity. While oxygen is essential for the survival of mosteukaryotic organisms, low oxygen availability or hypoxia is one microenvironmental stress that is known to occur during infection. Therefore, we aim todetermine the mechanism by which Hc is able to survive and adapt to hypoxia by determining the significance of its transcriptional response to hypoxia.Methods: Hc yeast cells were cultured to mid-log phase then transferred to normoxic (~21%) or hypoxic conditions (

FULL POSTER SESSION ABSTRACTSAcuM binding to sites in the 5’ upstream region. Studies with an acuF-lacZ gene fusion indicate positive control by AcuK and AcuM but a loss of the glucoserepression observed in Northerns suggesting negative regulation acting via 3’ sequences in response to growth on glycolytic carbon sources. Support forthis is provided by transcription studies. Modulation of the balance between the opposing activities of these two gene products is proposed to result fromtranscriptional interference involving collision of RNA polymerase molecules.424. ClbR and its paralog, ClbR2, regulate gene expression of cellulase genes in response to cellobiose in Aspergillus aculeatus. E. Kunitake, S. Tani, J.Sumitani, T. Kawaguchi. Life & Environmental Science, Osaka Prefecture University, Sakai, Osaka, Japan.The cellobiose- and cellulose-responsive induction of the cellobiohydrolase I (cbhI) and carboxymethylcellulase 2 (cmc2) genes is not regulated by XlnR, aZn(II) 2Cys 6 transcriptional activator, in Aspergillus aculeatus. We have identified a novel activator containing a Zn(II) 2Cys 6 binuclear cluster motif designatedas cellobiose-response regulator (ClbR), and which is not homologous to Clr-2/ClrB, a transcriptional activator controlling cellobiose-responsive inductionin Neurospora crassa and Aspergillus nidulans. Interestingly ClbR regulates not only the expression of cbhI and cmc2 but also genes regulated by XlnRunder the cellulose-inducing condition. However, the clbR overexpression did not increase the expression of all genes under control of ClbR. Therefore, wepredict that ClbR functions cooperatively with other factor(s). This study reports the ClbR function and the functional relationships among ClbR, ClbRinteractingfactor, and ClrB homolog on cellobiose-responsive induction in A. aculeatus. ClbR-interacting proteins were screened from a prey librarycomposed of ClbR paralogs and transcription factors controlling the expression of glycoside hydrolase genes by yeast two-hybrid method. ClbR2, 42%identity to ClbR, was so far isolated as a ClbR interacting protein. To investigate the correlation between ClbR and ClbR2 function, gene expression profilesunder control of ClbR were assessed in the clbR amd clbR2 single disruption mutants, and clbR/clbR2 double disruption mutant. The expression of cbhI andcmc2 decreased to the same level in all three mutants under the cellobiose-inducing condition. Furthermore, transcripts of the cbhI and cmc2 genesdrastically decreased in the clrB disruption mutant. This result suggests that ClrB regulates gene expression controlled via the XlnR-independent signalingpathway in this fungus. Because the clrB gene is induced by cellulosic compounds, we investigated if clrB expression is regulated by ClbR and ClbR2. In theclbR, clbR2, and clbR/clbR2 mutants, clrB transcripts under the inducing condition reduced to almost the same level as those under the uninducingcondition. Taken together, these data demonstrate that ClbR and ClbR2 regulate the cellulase gene expression in response to cellobiose by regulating theclrB gene expression.425. Expression of a bacterial xylanase in Trichoderma reesei under the egl2 and cbh2 glycosyl hydrolase gene promoters. Helena Nevalainen 1,2 , ShingoMiyauchi 1,2 , Junior Te'o 1,2 , Peter Bergquist 1,2,3 . 1) Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney, NSW, Australia; 2)Biomolecular Frontiers Research Centre, Macquarie University, NSW 2109, Australia; 3) Department of Molecular Medicine & Pathology, University ofAuckland Medical School, Auckland 1142, New Zealand.Our main aim in this study was to isolate a suite of promoters, other than cbh1, to establish a gene expression platform in which they could be usedsynergistically for the expression of recombinant gene products, using a bacterial thermophilic xylanase (XynB) as an example. The egl2 and cbh2promoters were selected on the basis of their relative strength and functionality under the same cultivation conditions that induce cbh1 so that thispromoter, widely used for recombinant expression, could be included in the ‘promoter mix’ when desired in the future. Together, these promoterspossess considerable expression capacity that can be harnessed for the synthesis of recombinant gene products in T. reesei. We also explored the mode ofglycosylation of the recombinant bacterial xylanase in T. reesei to confirm that the higher molecular weight bands seen in activity zymograms were indeedpost-translationally modified by glycosylation and to characterize the sugars attached to the protein. The highest XynB production was achieved from atransformant containing 1-2 copies of the EGL2sigpro vector (egl2 promoter). Best xylanase producers did not show any particular pattern in terms of thenumber of gene copies and their mode of integration into the chromosomal DNA. Transformants produced multiple forms of XynB which were decoratedwith various N- and O-glycans. One of the O-glycans was identified as hexuronic acid, whose presence had not been observed previously in theglycosylation patterns of T. reesei.426. A single argonaute gene participates in exogenous and endogenous RNAi and controls different cellular functions in the basal fungus Mucorcircinelloides. F. E. Nicolas-Molina 1,2 , M. Cervantes 1 , A. Vila 1 , S. Moxon 3 , J. P. De Haro 1 , T. Dalmay 3 , S. Torres-Martínez 1 , R. M. Ruiz-Vázquez 1 . 1) Departmentof Genetics and Microbiology, University of Murcia, Murcia, Spain; 2) Regional Campus of International Excellence "Campus Mare Nostrum", Murcia,Spain; 3) School of Biological Sciences, University of East Anglia, Norwich, UK.Regulation by RNAi of diverse cellular functions in metazoans is largely known. However, although different classes of endogenous small RNAs (esRNAs)have been identified in fungi, their biological roles are poorly described. The Argonaute proteins are the core component of all known RNAi pathways.Here we identified three argonaute genes of the basal fungus Mucor circinelloides and investigated their participation in exogenous and endogenous RNAi.Only ago-1 is required for transgene-induced RNA silencing. Ago-1 is also required for the production of distinct classes of esRNAs derived from exons (exsiRNAs),which differ in the silencing proteins required for their biogenesis. Classes I and II ex-siRNAs bind to Ago-1 to control mRNA accumulation of thetarget protein coding genes. Class III ex-siRNAs do not specifically bind to Ago-1, but require this protein for their production, revealing the complexity ofthe biogenesis pathways of ex-siRNAs. We also show that ago-1 gene is involved in the response to environmental signals, since vegetative developmentand autolysis induced by nutritional stress are affected in ago-1 - mutants. Our results highlight the role of ex-siRNAs in the regulation of endogenous genesin basal fungi and expand the range of biological functions modulated by RNAi silencing. This work was funded by MICINN (BFU2009-07220) and MINECO(BFU2012-32246), Spain.427. The transcription factor, AtrR, regulates the expression of ABC transporter genes and ergosterol biosynthesis genes in aspergilli. Ayumi Ohba 1 ,Kiminori Shimizu 2 , Daisuke Hagiwara 2 , Takahiro Shintani 1 , Susumu Kawamoto 2 , Katsuya Gomi 1 . 1) Div. Biosci. Biotechnol. Future Bioind., Grad Sch. Agric.Sci., Tohoku Univ., Japan; 2) MMRC, Chiba Univ., Japan.We previously demonstrated that a novel Zn(II)2Cys6 transcriptional factor, AoAtrR, regulates gene expression of the ABC transporters that wouldfunction as drug efflux pumps and contributes to the azole resistance in Aspergillus oryzae. Moreover, we showed that a deletion mutant of the AoatrRortholog (AfatrR) in Aspergillus fumigatus was similarly hypersensitive to azole drugs. However, little is known about target genes regulated by AfAtrR. Inthis study, we comprehensively examined the target genes regulated by AfAtrR using next-generation DNA sequencing technology (RNA-seq). RNA-seqanalysis indicated that AfAtrR similarly regulated at least one ABC transporter. In addition, surprisingly, AfAtrR also regulated several ergosterolbiosynthetic pathway genes including erg11(cyp51A). It has been known that the basic helix-loop-helix transcription factor, SrbA, has a critical role inergosterol biosynthesis and resistance to the azole drugs in A. fumigatus. Interestingly, the ergosterol biosynthetic pathway genes regulated by AfSrbAwere nearly identical with those regulated by AfAtrR. Therefore, we investigated difference in function between AtrR and SrbA in A. oryzae. The expressionof ergosterol biosynthetic pathway genes such as erg11, erg24, and erg25 etc. and three ABC transporter genes was significantly down-regulated in the27th Fungal Genetics Conference | 225

FULL POSTER SESSION ABSTRACTSdisruption mutant of AoatrR. Similarly, the expression of same ergosterol biosynthetic pathway genes was also markedly down-regulated in the disruptionmutant of AosrbA, but the expression of ABC transporter genes was not affected. In contrast, the expression of ergosterol biosynthetic pathway genes wasnot up-regulated in an overexpression strain of AoatrR. These results suggest that AtrR and SrbA coordinately regulate ergosterol biosynthetic pathwaygenes in aspergilli. On the other hand, the AoatrR disruptant was more hypersensitive to azole drugs compared to the AosrbA disruptant, suggesting thathypersensitivity of the atrR disruptant to azole drugs is attributed not only to lowered ergosterol levels due to down-regulation of ergosterol biosyntheticpathway genes, but also to reduced efflux transport of the drugs due to down-regulation of ABC transporter genes.428. Effects of intron deletions in production of a heterologous protease in Trichoderma reesei. Marja Paloheimo. Roal Oy, Rajamäki, Finland.A protease gene originating from a filamentous fungus Malbranchea cinnamomea was expressed in Trichoderma reesei using the strong T. reesei cbh1(cel7A) promoter. The heterologous protease was produced at relatively high yields. However, when the protease cDNA was included in the (otherwiseidentical) expression cassette the amount of protease in the culture supernatants of the transformants was very low. The native M. cinnamomea proteasegene contains three introns. To further study the effects of the removal of introns in the production of protease, single-copy transformants wereconstructed that contained expression cassettes in which each of the three introns was separately removed and in which two of the introns (allcombinations) were deleted at a time from the genomic protease gene. The expression cassettes were targeted to the native cbh1 locus. Three single copytransformants from each transformation were chosen for further studies. These strains were cultivated in shake flasks and the amount of protease wasanalysed from the culture supernatants. A clear decrease in the protease activity was detected when any of the introns was removed. However, dependingon the intron removed there were differences in the relative decreases of protease activity. Removal of two introns at a time had a cumulative decreasingeffect on production of protease. The results obtained are shown and discussed.429. Novel core promoter elements in the oomycete Phytophthora infestans and their influence on expression pattern detected by genome-wideanalysis. Laetitia Poidevin, Sourav Roy, Howard S. Judelson. Department of Plant Pathology & Microbiology, University of California Riverside, USA.The core promoter is the region flanking the transcription start site (TSS) that directs pre-initiation complex formation. While core promoters have beenstudied intensively in mammals and yeast, little is known about more diverse eukaryotes including oomycetes. Prior studies of a small collection of clonedoomycete genes proposed that its core promoters contain a 19-nt block bearing both an Initiator-like sequence (INR) and a novel 3' sequence named FPR,but this has not been extended to whole-genome analysis. To learn more about oomycete core promoters, we used expectation maximization to find overrepresentedmotifs near TSSs of Phytophthora infestans, the potato blight pathogen. The motifs corresponded to INR, FPR, and a new element found 25-ntdownstream of the TSS called DPEP. TATA boxes were not detected. Assays of DPEP function by mutagenesis were consistent with its role as a core motif.Genome-wide searches found a well-conserved combined INR+FPR in only 13% of genes after correcting for false discovery, contradicting prior reportsthat INR and FPR are adjacent to each other in most genes. INR or FPR were found alone near TSSs in 18% and 7% of genes, respectively. Promoters lackingthe motifs had pyrimidine-rich regions near the TSS. The combined INR+FPR motif was linked to higher than average mRNA levels, developmentallyregulatedtranscription, and functions related to plant infection, while DPEP and FPR were over-represented in constitutive housekeeping genes. Themotifs were all detected in other oomycetes including Hyaloperonospora arabidopsidis, Phytophthora sojae, Pythium ultimum, and Saprolegnia parasitica,but only INR seemed present in a non-oomycete stramenopile. The absence of a TATA box and presence of novel motifs show that oomycete corepromoters have diverged from that of previously-studied model systems, and likely explains failures in prior heterologous expression studies. Theassociation of the INR+FPR with developmentally-regulated genes shows that oomycete core elements influence stage-specific transcription in addition toregulating pre-initiation complex formation.430. New insights into the phosphate-sensing network in Neurospora crassa. Antonio Rossi 1 , Gabriela F Persinoti 2 , Nalu T A Peres 2 , Diana E Gras 2 , Nilce MMartinez-Rossi 2 . 1) Bioquímica e Imunologia, FMRP-USP, Ribeirão Preto, SP, Brazil; 2) Genética, FMRP-USP, Ribeirão Preto, SP, Brazil.The filamentous fungus Neurospora crassa is an excellent model system for examining molecular responses to ambient signals in eukaryoticmicroorganisms. Inorganic phosphate is an essential growth-limiting nutrient in nature and is crucial for the synthesis of nucleic acids and the flow ofgenetic information. Numerous ambient signals activate the recruitment of mitogen-activated protein kinase (MAPK) cascades. Thus, to identify genesinvolved in metabolic responses to exogenous phosphate sensing and the functioning of an MAPK, MAK-2, we performed microarray experiments using amak-2 knockout strain (Dmak-2) grown under phosphate-shortage conditions by comparing its transcription profile to that of a control strain grown in lowandhigh-phosphate cultures. These experiments revealed 912 unique differentially expressed genes involved in a number of physiological processesrelated to phosphate transport, metabolism, and regulation as well as posttranslational modification of proteins, and MAPK signaling pathways.Quantitative real-time-PCR gene expression analysis using independent RNA samples of 10 arbitrarily chosen genes validated our microarray results. A highPearson correlation between microarray and quantitative real-time-PCR data was observed. The analysis of these differentially expressed genes in theDmak-2 strain provide evidence that the mak-2 gene is a component of the hierarchical phosphate-signaling pathway in N. crassa in addition to itsindependent involvement in other metabolic routes such as the isoprenylation pathway. In this extended model, MAK-2 is functional regardingtranscription of Pi-repressible phosphatases when N. crassa is cultured under Pi shortage and is non-functional under abundant Pi conditions, thusrevealing novel aspects of the N. crassa phosphorus-sensing network. Financial support: FAPESP, CNPq, CAPES, FAEPA.431. Carbon source and light dependent regulation of gene clusters in Trichoderma reesei (Hypocrea jecorina). Doris Tisch 2 , Monika Schmoll 1 . 1) Healthand Environment, Bioresources, Austrian Institute of Technology AIT, Tulln, Austria; 2) Vienna University of Technology, Institute of Chemical Engineering,Vienna, Austria.Trichoderma reesei (anamorph of Hypocrea jecorina) is one of the most prolific producers of plant cell wall degrading enzymes. Regulation of the genesencoding these enzymes occurs in response to the nutrient sources available in the environment and many of them are responsive to light as well.Cellulose as the natural substrate induces the most complete enzyme set, while induction of cellulases also occurs on sophorose and lactose. In contrast,no cellulases are induced on glycerol and the respective genes are repressed on glucose. We therefore investigated the transcriptome on these five carbonsources in light and darkness and aimed to identify genes specifically expressed under cellulase inducing conditions. These conditions are characterized bya significant enrichment of genes involved in C-compound and carbohydrate degradation and transport among the upregulated gene set. Genes downregulatedunder inducing conditions show a significant enrichment in amino acid metabolism and energy metabolism. We were further interested whetherlight dependent regulation is clustered in the genome and if the carbon source is relevant for activation of light dependent clusters. We found that lightdependent clustering predominantly occurs upon growth on cellulose, with the most significant regulation in a gene cluster comprising env1. This clusterappears on glucose as well, but is not down regulated in mutants of blr1 or blr2. Also cbh2, the arabinofuranosidase gene abf2 and the histoneacetyltransferase gene gcn5 are part of light dependent clusters. Hierarchical clustering of gene expression patterns was performed to reveal functional226

FULL POSTER SESSION ABSTRACTSdivergence of gene regulation with respect to light response or carbon specific regulation. Glycoside hydrolase genes follow the whole transcriptomepattern with carbon source being superior to light in terms of regulation. ENV1 in in part the G-protein beta subunit GNB1 were found to be crucial forcarbon source specific regulation of G-protein coupled receptors, genes involved in secretion, sulphur metabolism and oxidative processes as well astransporters. We conclude that clustered regulation of light responsive genes preferentially occurs upon growth cellulose and that ENV1 and to a lesserextent GNB1 play a role in carbon source dependent regulation of specific gene groups in light.432. Identification of regulators for enzyme production in Trichoderma reesei using genome-wide approaches. M. Häkkinen, M. Valkonen, N. Aro, M.Vitikainen, A. Westerholm-Parvinen, M. Penttilä, M. Saloheimo, T. Pakula. VTT, Espoo, Finland.Trichoderma reesei (anamorph Hypocrea jecorina) is an efficient producer of enzymes degrading cellulosic and hemicellulosic biomass. The cellulases andhemicellulases produced by the fungus are widely employed in industry, and also in biorefinery applications. Various environmental and metabolic factorstogether with the physiological state of the cell affect the enzyme production of T. reesei. Thus, a complex signalling cascade and regulatory network isneeded for the accurate timing of hydrolytic enzyme production and to control the pattern of enzyme activities produced. In previous studies, bothpositively and negatively acting regulatory factors for cellulase and hemicellulase genes have been characterised in T.reesei. In this study, an expressionmicroarray data on T. reesei cultivated in the presence of different carbon sources was analysed in order to identify additional regulatory genes forcellulase and hemicellulase production. In total, 28 putative regulatory factors for T. reesei cellulases and hemicellulases were identified and selected forfurther studies. The genes were overexpressed in T. reesei QM9414. Cultivated modified strains were tested for their ability to produce cellulases,xylanases and total secreted protein. Over-expression of seven of the genes led to increased production of cellulases and/or xylanases.433. The central core of the response to light and injury and their regulation by RNAi machinery in the filamentous fungus Trichoderma atriviride. J.M.Villalobos-Escobedo, N. Carreras-Villaseñor, A. Herrera-Estrella. LANGEBIO-CINVESTAV, Irapuato, Guanajuato, Mexico.All living organisms must sense and respond appropriately to different environmental stimuli in order to survive. Trichoderma atroviride is a filamentousfungus with wide adaptability to different environmental conditions and is considered a good morphogenetic model because it respond to light and injuryproducing asexual reproductive structures (conidia). The mechanisms used by this fungus to respond to these stimuli have been studied independentlyand models of perception and signal transduction for each stimulus have been proposed. In our research group, we found that the Ddcr2 mutant, involvedin small RNA biogenesis, is affected in conidiation in response to light and injury, indicating that conidiation is regulated by small RNAs. In this work wediscovered that when the Ddcr2 mutant receives simultaneously light and injury, it is able to conidiate. Based on this observation, we propose that there isa central core of genes needed to coordinate the response to both stimuli and in the Ddcr2 the signaling pathways act synergistically to achieve the correctexpression of these genes in order to conidiate. To test the above hypothesis we analyzed the transcriptome in response to light and injury of the wild type(WT), and identified 38 genes that have the same expression profile in response to the two stimuli, 19 of them are induced and 19 are repressed, calledthe central core genes, most of the up-regulated genes are involved in RNA processing, ribosome biogenesis, chromatin remodeling and other cellularprocesses indicating that this core regulates gene expression to respond to the stimulus. While genes that are repressed are mainly involved in lipid,carbohydrate and protein metabolic processes suggesting that a metabolic arrest is necessary to respond to both stresses. The expression analysis of thiscore of genes in the Ddcr2 revealed some of them are deregulated, but other genes respond similarly to WT in either light or injury in Ddcr2, so this groupof genes is regulated by the RNAi machinery.434. RNA-mediated Gene Silencing in Candida albicans: Reduction of Fungal Pathogenesis by Use of RNAi Technology. M. Moazeni 1 , MR.Khorramizadeh 2 , P. Kordbacheh 1 , H. Zeraati 3 , F. Noorbakhsh 4 , L. Teimoori-Toolabi 5 , S. Rezaie 1,2 . 1) Dept. of Medical Mycology & Parasitology, TehranUniversity of Medical Sciences, Tehran, Iran; 2) Dept. of Medical Biotechnology, School of Advanced Technologies in Medicine, Tehran University ofMedical Sciences, Tehran, Iran; 3) Dep. of Epidemiology and Biostatistics, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran; 4)Dept of Biology, Islamic Azad University, Varamin-Pishva Branch, Varamin, Iran; 5) Dept of Molecular Medicine, Biotechnology Research Center, PasteurInstitute of Iran, Iran.Background: The introduction of RNA silencing machinery in fungi has led to the promising application of RNAi methodology to knock down essentialvital factor or virulence factor genes in the microorganisms. Efg1p is required for development of a true hyphal growth form which is known to be essentialfor interactions with human host cells and for the yeast’s pathogenesis. In addition, it is responsible for positive regulation of the expression of severalhyphal-specific genes: SAP5, which encodes secreted aspartic proteinase, and ALS3, which encodes a multi-functional adhesive polypeptide. In this paper,we describe the development of a system for presenting and studying the RNAi function on the EFG1 gene in Candida albicans. Materials and Methods:The 19-nucleotide siRNA was designed on the basis of the cDNA sequence of the EFG1 gene in Candida albicans and transfection was performed by use ofa modified-PEG/LiAc method. To investigate EFG1 gene silencing in siRNA-treated cells, the yeasts were grown in human serum; to induce germ tubes asolid medium was used with the serum. Quantitative changes in expression of the EFG1 gene as well as two Efg1-associated genes, ALS3 and SAP5, wereanalyzed by measuring the cognate EFG1, SAP3 and ALS3 mRNA levels by use of a quantitative real-time RT-PCR assay. Results: Images taken byfluorescent microscopy method indicated the effectiveness of transfection. Compared with the positive control, true hyphae formation was significantlyreduced by siRNA at concentrations of 1 mM, 500 nM, and 100 nM (P227.05). According to REST® software data analysis, a considerable decrease in EFG1gene expression was observed when applying both 500 nM and 1mM of siRNA (P

FULL POSTER SESSION ABSTRACTSparticipation of the RNAi machinery in these processes. In addition, different classes of sRNAs are differentially expressed between the WT and Ddcr2strains. An in depth analysis of features of these small RNAs, e.g. size, type of sRNAs and 5'nucleotide bias, show that those present in WT are lost in thesRNAs of the Ddcr2 strain. All together these data show that in Trichoderma atroviride, the RNAi machinery has a central role in endogenous processessuch as the development and fitness, beyond controlling the protection of genome against invasive nucleic acids as reported for other fungi.436. Genome-wide analysis of light responses in Mucor circinelloides. Victoriano Garre, Sergio López-García, Eusebio Navarro, Santiago Torres-Martínez.Departamento de Genética y Microbiología, Universidad de Murcia, 30100 Murcia, Spain.Light regulates developmental and physiological processes in a wide range of fungi. Particularly, Zygomycete fungi have developed complex mechanismsto control the responses to light that await detailed characterization at molecular level. The zygomycete Mucor circinelloides is a good model for thispurpose because its genome has been sequenced and several molecular tools are available. Mucor, like other Zygomycetes, has three white collar-1 genes(mcwc-1a, mcwc-1b and mcwc-1c) that code for proteins which present characteristics of photoreceptors. Each mcwc-1 gene controls a specific responseto light. Thus, mcwc-1a and mcwc-1c control phototropism and photocarotenogenesis, respectively, whereas the mcwc-1b function in regulation by lighthas not been proved. In order to deepen in the regulation by light in Mucor, a systematic approach using microarrays was followed to characterize whitelight-inducible transcriptional changes in wild-type and knockout mutants for each mcwc-1 gene. Analysis of microarray data revealed that light is mainly apositive signal for transcription in Mucor, as in other fungi, since 123 genes were up-regulated in the wild-type strain in response to light, whereas only 26were down-regulated, considering a threshold of threefold change. Genes strongly induced by light included genes known to be up-regulated by light, likethe carotenogenic gene carB (74-fold), cryptochrome (45-fold) and mcwc-1c (22-fold), supporting reliability of the microarray data. Although many of upregulatedgenes code for proteins implicated in protection against light-induced damage, several of them code for protein involved in signal transductionthat could be involved in light responses like phototropism. Transcriptomic analysis of mcwc-1 mutants showed that induction of around 60% of the genesis mediated by mcwc-1a, whereas only 1% is mediated by mcwc-1c and none is mediated by mcwc-1b, suggesting that mcwc-1a is the main photoreceptor.Searching for cis-acting regulatory motifs upstream of genes regulated by mcwc-1a identified consensus sequences similar to those found in lightregulated genes of Neurospora crassa. Moreover, the identification of a small group of genes regulated by the three mcwc-1 genes points out that thethree proteins form complexes to regulate gene expression. Funded by MINECO (BFU2012-32246), Spain.437. Gene expression profiling of the basidiomycetous fungus Lentinula edodes after light stimulation. H. Sano 1 , Y. Sakamoto 2 , M. Abe 3 , S. Kaneko 4 , M.Nakamura 1 , Y. Miyazaki 1 . 1) Department of Applied Microbiology, Forestry and Forest Products Research Institute, Tsukuba, Ibaraki, Japan; 2) Departmentof Biological Resources Research, Iwate Biological Research Center, 22-174-4 Narita, Kitakami, Iwate 024-0003,Japan; 3) Forest and Forestry ResearchInstitute, Tokushima Agriculture, Forestry and Fisheries Technology Support Center, 5-69 Nanshocho, Tokushima, Tokushima 770-0045, Japan; 4)Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsutacho, Midori-ku, Yokohama,Kanagawa 226-8503, Japan.Light is an important environmental signal for various organisms and is known to regulate their physiological and developmental processes. In fungi, lightinfluences sexual and asexual reproductions, mycotoxin productions and pigmentation. Morphological differentiations of mushroom-forminghomobasidiomycetes are also affected by light and stimulation of light is one of the important factors regulating fruiting body formation. The molecularmechanisms regulated by light are well studied in ascomycetes, and a number of light-regulated genes are identified and are characterized. Inbasidiomycetes, the photoreceptor-encoding genes homologous to Neurospora crassa wc-1 have been isolated from several species and those geneticalevidences revealed the involvement in fruiting body formation. However, the details of light-sensing systems and light-dependent regulation of genes areunclear. In this study, we performed super serial analysis of gene expression (SuperSAGE) using Illumina/Solexa genetic analyzer and analyzed the changeof gene expressions stimulated by light during fruiting body formation in Lentinula edodes. The samples for high-throughput SuperSAGE were preparedfrom mycelia cultivated under darkness and after exposure to light with low temperature treatment. The mycelium which had been exposed to lightformed a number of primordium and developed into normal fruiting bodies, whereas the mycelium cultivated under darkness could produce noprimordium. The obtained transcriptome data showed that there were many kinds of genes expressed in L. edodes after light irradiation (5251 genes),compared with the data under darkness (2876 genes). The comparison analysis revealed that the expressions of 2500 genes were different between lightand dark condition, and that over 2000 genes were strongly transcribed in L. edodes after exposure to light. Light irradiation also caused the decrease inexpression levels of 500 genes. The up- and down-regulated genes were categorized by Gene Ontology and were assigned by the KEGG pathway mapping.These analyses suggested that several genes encoding putative fungal-specific proteins were regulated by light. The cataloged data of the expressed genesprovide valuable information on understanding of light-sensing system in L. edodes.438. Further Characterization of Surface Recognition Mechanisms in Magnaporthe oryzae. Guanghui Wang 1 , Xiaoying Zhou 2 , Guotian Li 2 , Jin-Rong Xu 1,2 .1) College of Plant protection, Northwest A & F University, Yangling, Shaanxi, China; 2) Department of Botany and Plant Pathology, Purdue Universtiy,West Lafayette, USA.Surface recognition and appressorium penetration are critical infection processes in the rice blast fungus Magnaporthe oryzae and many other plantpathogenic fungi. Various chemical and physical surface signals are known to be recognized by germ tubes to activate the Pmk1 MAP kinase that isconserved in fungal pathogens for regulating appressorium formation and penetration. Recently, the Msb2 mucin gene was found to function as a surfacesensor upstream from the Pmk1 pathway. However, it is not clear how is Msb2 activated and what is its relationship with other surface sensors. In thisstudy, we found that the cleavage domain and transmembrane helics are essential for Msb2 functions. Site-directly mutagenesis was used to verify twocandidate cleavage amino acid sites. In addition, we conducted deletion analysis with the cytoplasmic tail of Msb2 that likely plays a role in intracellularsignaling. We also assayed the effects of over-expressing the C-terminal region of Msb2 and identified proteins co-precipitated with it by affinitypurification. Because CBP1 and PTH11 are two other putative surface sensor genes, we also generated the msb2 cbp1 and msb2 pth11 double mutantsand triple mutants with sho1. The msb2 cbp1 mutant rarely formed appressoria and was non-pathogenic, indicating that Msb2 and Cbp1, the only twomucins in M. oryzae, may have overlapping functions in surface recognition. Detailed phenotype characterization of the msb2 pth11 and triple mutants areunder the way. A model of Msb2 activation and relationship among different receptors will be presented.439. Evidence of Microbial Epigenetics; Loss-of-function mutant of the Bck1 Homolog, CpBCK1, from the chestnut blight fungus Cryphonectria parasiticaresulted in the sectoring accompanied with the changes in DNA methylation. J.-M. Kim 1 , S.-H. Yun 2 , K.-Y. Jahng 2 , D.-H. Kim 2 . 1) Department of Bio-Environmental Chemistry, Wonkwang University, Iksan, Jeonbuk, South Korea; 2) Institute for Molecular Biology and Genetics, Center for Fungalpathogenesis, Chonbuk National University, Jeonju, Jeonbuk, South Korea.Cpbck1, encoding a mitogen-activated protein (MAP) kinase kinase kinase from the chestnut blight fungus Cryphonectria parasitica, is an ortholog of228

FULL POSTER SESSION ABSTRACTSBck1 from Saccharomyces cerevisiae. Colony morphology of the Cpbck1-null mutants differed dramatically from the wild type that mutants showed theinvasive growth pattern characterized by slower growth rate, absence of distinctive aerial hyphae resulting in almost absence of conidia-bearing structureand conidia, sparse mycelial growth on the surface of agar plate with abnormal pigmentation, and irregular mycelial mat within the restricted area.Feeding hyphae growing under the plate showed less branched and relatively slower growth pattern. Interestingly, the Cpbck1-null mutant producedsectors appeared as thick rubbery patches of matted growth without pigmentation and sporulation. Complementation of the Cpbck1-null mutant with awild-type allele rescued mutant phenotypes indicating that the mutant phenotypes were due to the absence of the Cpbck1 gene. Intracellular structureobserved by electron microscope revealed both invasive growth-type and sectored-type showed the occurrence of hypertrophy of cell wall, multiple nucleiwithin swollen cells and intrahyphal hyphae. DNA methylation, an indicative of epigenetic marker, examined by Southern blot analysis and bisulfite DNAmodification of putative target genes revealed that there was difference in the DNA methylation pattern between original Cpbck1-null mutant andsectored isolate. This study suggests that epigenetic changes are predisposed by the loss of function mutation of a specific gene Cpbck1 and it will be ofinterest to determine what decide the transition of the mycelia growth pattern from the invasive and very-sick hyphal growth type to compact-mat type.The Cpbck1-null mutant showed the sectored phenotype accompanied with the changes in DNA methylation demonstrated that the fungal signalingpathway implicated in the control of epigenetic processes, without which abnormal degeneration such as sectoring occurred.440. NUP-6 (Importin a) is required for DNA methylation in Neurospora crassa. Andrew D. Klocko 1 , Michael R. Rountree 1 , Paula L. Grisafi 1 , Shan M. Hays 2 ,Eric U. Selker 1 . 1) Institute of Molecular Biology, University of Oregon, Eugene, OR 97448; 2) Department of Natural and Environmental Sciences, WesternState College of Colorado, Gunnison, CO 81231.Heterochromatic regions on chromosomes are essential for numerous cellular processes, including centromere function and gene silencing. Therepetitive DNA found in heterochromatin is highly compacted, frequently A:T rich, and in some species such as Neurospora crassa, methylated atcytosines. This DNA methylation can effectively silence genes. Interestingly, unlike the situation in some eukaryotes, loss of DNA methylation is notrequired for viability in Neurospora. The dispensability of DNA methylation in Neurospora allows for the identification of defective in methylation (dim)genes that have critical roles in the establishment, maintenance, and/or regulation of DNA methylation. This approach established that, at least inNeurospora, DNA methylation is initiated by the histone methyltransferase activity of a multi-subunit complex, DCDC (DIM-5/-7/-9 CUL-4 DDB-1 dim-8Complex), which catalyzes tri-methylation of lysine 9 on histone H3 (H3K9me3). While the identification of the components of the DCDC was an importantstep to understanding heterochromatin formation, much is still unknown about the DCDC, including its detailed function, regulation, and assembly. Here,we characterize the action of a previously unidentified dim mutant, dim-3. We found that dim-3 corresponds to the nup-6 gene, which encodes theImportin a subunit (NUP-6) for classical nuclear transport. NUP-6 dim-3 virtually abolishes H3K9me3 and significantly reduces DNA methylation, and causesDIM-5 and DIM-7 to be mislocalized from heterochromatin, suggesting DCDC activity is impacted in a dim-3 strain. Curiously, nuclear transport of DCDCcomponents in a dim-3 strain background appears to be equal to or greater than in a wild type background. The possibility exists that NUP-6 could beimportant in directing the DCDC to heterochromatin or in assembly of the DCDC, and we will address these hypotheses. In addition, the mutations found indim-3 could prevent its gene product, NUP-6, from facilitating DCDC action.441. Identification and characterization of a Blastomyces dermatitidis mutant with a bidirectional defect in the phase transition. Amber J. Marty,Gregory M. Gauthier. University of Wisconsin - Madison, 1550 Linden Drive, Microbial Sciences Building, Madison, WI, 53706.Collectively, the dimorphic fungi are the most common cause of invasive fungal disease worldwide. The ability of these fungi to undergo a shift betweenmold and yeast is critical for pathogenesis. In the soil (22°C), these fungi grow as mold, which produce infectious conidia. Following soil disruption,aerosolized conidia inhaled into the lungs of a host (37°C) convert into yeast to cause pneumonia. Knowledge of the mechanisms used to regulate thisphase transition is limited. To uncover genes that control the phase transition, Agrobacterium tumefaciens-mediated DNA transfer was used tomutagenize Blastomyces dermatitidis conidia. We generated and screened 22,000 insertional mutants for defects in the phase transition. We identified amutant, 11-9-75, with a single site of T-DNA insertion that grew as pseudohyphae at 37°C and 22°C, rather than yeast or mold. Adaptor PCR, DNAsequencing, and reverse transcription PCR (RT-PCR) revealed the T-DNA was located in the 5’ UTR of an uncharacterized gene (referred to as BKY1) thatwas not annotated in the B. dermatitidis genome. Analysis of cDNA indicated BKY1 was 1546 bp in length, lacked introns, and the ORF was predicted toencode a 156 amino acid protein. BLAST analyses against the NCBI database failed to reveal homologs of BKY1 in other fungi. The T-DNA insert alteredtranscription of BKY1 in mutant 11-9-75. BKY1 transcript in the mutant was 35-fold higher versus wild type (WT) by quantitative real-time PCR (qPCR) andtruncated at the 5’ UTR when analyzed by rapid amplification of cDNA ends (RACE). RT-PCR supported the qPCR and RACE analyses, and indicated theGAPDH promoter, which is upstream of a hygromycin resistance cassette in the T-DNA was driving increased transcription of truncated BKY1. The T-DNAinsert also altered alternative splicing of a gene with unknown function, Bd594, which was less than 1.2 kb downstream of BKY1. Although transcriptabundance of Bd594 in mutant 11-9-75 was similar to WT, the frequency of intron excision was reduced. In conclusion, we have identified an insertionalmutant with a bidirectional defect on the phase transition; it grows as pseudohyphae instead of yeast at 37°C or mold at 22°C. The T-DNA insert alterstranscription of adjacent genes, BKY1 and Bd594, in a poorly characterized region in the B. dermatitidis genome.27th Fungal Genetics Conference | 229

FULL POSTER SESSION ABSTRACTS442. Transcriptional regulation of peptidases and nitrogen transporters during the assimilation of organic nitrogen by the ectomycorrhizal fungi Paxillusinvolutus. Firoz Shah 1 , Francois Rineau 2 , Tomas Johansson 1 , Anders Tunlid 1 . 1) Microbial Ecology Group, Department of Biology, Lund University, SE-22362,Lund, Sweden; 2) Centre for Environmental Sciences, Hasselt University, Building D, Agoralaan, 3590 Diepenbeek, Limburg, Belgium.Proteins and amino acids form a major part of the organic nitrogen (N) sources in soils. Though a poorly characterized process, this N is mobilized andbecomes available to plants due to the activity of ectomycorrhizal (ECM) fungi. We have examined the role of ectomycorrhizal extracellular peptidases andamino acid transporters in the degradation, uptake and transfer of various protein sources (BSA, Gliadin and pollen) as well as of plant litter material usingthe ECM model fungus Paxillus involutus. During N-deprived conditions, all substrates induced secretion of peptidase activities. The activity had acidic pHoptimum (2.3-3.0), and it was mainly due to aspartic peptidases and with minor contribution of metallo and serine peptidases. The activity was partly andtemporarily repressed by low concentrations of ammonium (1mg/L). Transcriptional analysis showed that P. involutus expressed a large array of proteinsand enzymes involved in the assimilation of organic N including peptidases, N-transporters and enzymes of the N-metabolism. Extensive in-silico analysisrevealed the presence of genes encoding 312 peptidases, 129 N transporters and 284 enzymes involved in amino acid metabolism. Out of these, 89peptidases and 37 N-transporters and 109 amino acid metabolism enzymes encoding genes were significantly upregulated during organic N assimilation.The genes were encoding a variety of secreted (23) and non-secreted (20) peptidases which were differentially expressed depending on the medium withthe highest expression of the aspartic and metallo peptidases. Apart from the YAAH/ATO family, upregulated genes were found in all the other families oftransporters for amino acids, oligopeptides, ammonium, urea and allantoate/allantoin. The results shows that the expression levels of peptidases andtransporters in P. involutus are coordinately regulated during the assimilation of organic N sources.443. Characterization of genome maintenance components in Neurospora crassa using whole-genome high-throughput approach. Evelina Y. Basenko,Zack Lewis. Department of Microbiology, University of Georgia, Athens, GA.Eukaryotic DNA is packaged into a higher order DNA-protein structure also known as chromatin, which can regulate and impact an array of nuclearprocesses including DNA repair and genome maintenance. Disruptions in genome integrity can lead to serious ailments in humans and also contribute tocancer emergence. DNA repair and genome maintenance have been extensively studied in yeast. We, however, have chosen to investigate genomemaintenance in the filamentous fungus Neurospora crassa. N. crassa possesses a much larger genome than budding yeasts, and it also containsapproximately 1400 genes which are conserved in higher eukaryotes including humans and are absent in yeasts. We utilized the whole-genomeNeurospora knockout library to search for novel regulators of genome maintenance. We screened the knockout library for mutants sensitive to the DNAdamage agent methyl methane sulfonate (MMS). We have further confirmed and investigated additional mutagen sensitivities of confirmed MMSsensitivestrains. For our further studies, we have decided to focus on one of the MMS-sensitive mutants, which contains a deletion of a SNF2-like protein.The whole-genome high-throughput approach is a powerful method to identify novel players of genome maintenance. We have identified several hundredmutants sensitive to DNA damage, that fall within various categories of cellular processes, including but not limited to DNA repair, RNA metabolism, andchromatin maintenance. Our findings and current progress will be reported.444. Circadian regulation and carbon catabolite repression in Neurospora crassa: Two integrated regulatory systems? Rodrigo Díaz-Choque, Luis FLarrondo. Dept Molecular Genetics & Microbiology, Pontificia Universidad Católica de Chile, Santiago, Chile.Circadian clocks are autonomous timers composed of interconnected transcriptional/transcriptional feedback loops. They are thought to confer aselective advantage to individuals by temporally coordinating several processes and contributing to cellular homeostasis. In Neurospora crassa, a modelorganism in circadian studies, ~20%; of its genes are under circadian control and interestingly; many of them are related to metabolism. Indeed, the ideaof a crosstalk between metabolism and the circadian clock has become stronger in the last years, and several examples have been obtained in mammaliansystems. However, we still don’t know in Neurospora the actual influence of circadian regulation in its physiology and its real impact in the “real-world”.Moreover, the different transcriptional regulators linking time-of-day information and the expression of genes involved in metabolically relevantprocesses, like Carbon-Catabolite Repression (CCR) or cellulose degradation remain largely unknown. Thus, we are analyzing glucose repression in acircadian context, using N. crassa as a model. We hypothesize that there is an intimate crosstalk between both regulatory systems over the expression ofseveral rhythmic CCR-controlled genes. We are using gene expression assays and a codon-optimized luciferase transcriptional reporter, to evaluate therole of the transcription factor CRE-1 (carbon catabolite regulation-1), a conserved metabolic regulator, in this potential cross-talk. Also, CRE-1 is a crucialtranscription factor involved in several important cellular processes as cellulose degradation and catabolic repression. Further, we are studying how bothinputs are integrated to control the expression of target genes. Our results suggest CRE-1 as a link between both pathways, as it appears to be importantfor both CCR and circadian control of target genes. In addition, we describe for the first time In Neurospora the presence of a functional clock in cellulose(Avicel) -containing media. This observation strengthens the hypothesis that a circadian clock may regulate the expression of several cellulase-encodinggenes, having a real impact in such a physiologically relevant process.445. Opposing activities of the HCHC and DMM complexes maintain proper DNA methylation in Neurospora crassa. Shinji Honda 1,2 , Eun Yu 1 , Eric Selker 1 .1) University of Oregon, Institute of Molecular Biology, Eugene, OR; 2) University of Fukui, Life Science Unite, Fukui, Japan.Proper regulation of heterochromatin and DNA methylation is critical for the normal function of cells. We show that heterochromatin and DNAmethylation are faithfully controlled in Neurospora by opposing activities of the silencing complex HCHC and the anti-silencing complex DMM. Theworkings of these two complexes were investigated. HCHC consists of four proteins, the two chromo domain proteins HP1 and CDP-2, the histonedeacetylase HDA-1 and the AT-hook motif protein CHAP. We found that histone deacetylase activity is critical for HCHC function but the H3K9me3 bindingactivity of the CDP-2 chromo domain is not. Instead, CDP-2 serves as an essential bridge between HP1 and HDA-1. CHAP interacts directly with HDA-1,binds in a methylation-independent way to the A:T-rich DNA that forms the cores of methylated regions and is important for stable association of HDA-1with chromatin. HCHC is involved in the spreading of DNA methylation in dmm mutants. The DMM complex consists of a presumed histone demethylase,DMM-1, plus DMM-2, which is characterized by a fungal-specific Zn(II) 2Cys 6 DNA-binding domain (“Zn-Cys”). We found that DMM-2 strongly binds to DNAfrom euchromatin/heterochromatin junctions, thereby promoting the stable association of DMM-1 at the edge of heterochromatin domains to preventaberrant spreading of DNA methylation.446. The transcription factor FL is phosphorylated and interacts with a trehalose related protein in Neurospora crassa. Carmen Ruger-Herreros 1 , GencerSancar 2 , Michael Brunner 2 , Luis M. Corrochano 1 . 1) Departamento de Genetica, Universidad de Sevilla, Spain; 2) BZH, Universität Heidelberg, Germany.Several environmental cues, including light, promote a developmental transition in Neurospora crassa that leads to the formation of conidia. Conidiationis controlled by FLUFFY (FL), a zinc finger transcription factor. Light activates the transcription of fl through the transient binding of the WC complex to thefl promoter. Light also activates the transcription of several conidiation genes in Aspergillus nidulans, and their Neurospora homologs have been identified230

FULL POSTER SESSION ABSTRACTSin the Neurospora genome. We have assayed the activation by light of the Neurospora homologs of A. nidulans conidiation genes (flbA, flbC, flbD, medAand stuA), and the Neurospora conidiation gene con-10 as a control. Unlike con-10, none of the Neurospora homologs of the A. nidulans conidiation geneswere induced by light in vegetative mycelia. However, we found that deletion fl resulted in light-dependent mRNA accumulation for all the conidiationgenes. This result indicated that the absence of FL allows the binding of the WC complex to the promoter of these genes to activate transcription in a lightdependentmanner. We have assayed the amount of WC proteins in the Dfl and wild type strains but we did not find any difference between the twostrains. We expect to identify additional genes deregulated by the absence of FL after massive sequencing of total RNA (RNAseq) using a Dfl strain andwild-type strain in dark and light conditions. We have investigated the role of FL during conidiation in Neurospora using a tagged version of FL. FL ispresent in vegetative mycelia but the amount increses after light exposure. We observed several forms of FL due to phosphorylation, and and we havedetermined by mass spectrometry that FL is phosphorylated in several residues. We have immunoprecipitated FL to identify proteins that may interactwith FL. We have found a protein that interacts with FL in different growth conditions. This protein has been described in other organisms and plays a rolein the ability to grow in the presence of trehalose. Since FL is a transcription factor, we have use FL::3XFLAG strain to do ChIPseq in order to identify theputative binding sites of FL to the DNA. We expect that the results from these experiments will help us to understand in more detail the role of FL in theactivation of gene transcription during development.447. Transcriptomic profiling of fumonisin B biosynthesis by Fusarium verticillioides. N. Ponts, E. Zehraoui, L. Pinson-Gadais, F. Richard-Forget, C.Barreau. INRA, UR1264-MycSA, 71 avenue Edouard Bourlaux, BP81, F-33883 Villenave d’Ornon, France.The plant fungal pathogen Fusarium verticillioides can infect various plants worldwide, including maize, and contaminate kernels with mycotoxins of thefumonisin family. Fumonisins B are stable polyketides that resist agrifood processing and are classified as potentially carcinogenic. As such, contaminationof food and feeds with these toxic secondary metabolites must be avoided. Numerous factors influence fumonisins B accumulation on maize, including thecomposition of the grains on which Fusarium develops. In particular, several phenolic compounds were shown to inhibit fumonisin B biosynthesis.Preliminary analyses showed that free phenolic acids are particularly abundant in immature grains, i.e., at the onset of toxin production, from cerealcultivars on which mycotoxins tend to accumulate less. We tested in vitro the effect of chlorogenic, caffeic, and ferulic acid on fumonisin B production in F.verticillioides. All three compounds inhibit fumonisin B accumulation, caffeic acid being the most efficient with that regard. We investigated themechanisms by which these phenolic acids may exert their inhibitory properties and analyzed whole genome expression levels by RNA-seq. Sequencedreads were mapped to the reference genome of F. verticillioides and results were analyzed according to the current annotation available at the FusariumComparative Database. Doing so, we identified 175 and 1133 potential new genes and transcripts, respectively. We also found that the genes involved inthe fumonisins biosynthetic pathway are all inhibited in the presence of any of the three tested phenolic acids. Finally, we identified sets of genes that areregulated specifically by a given phenolic acid, and others that follow similar patterns in all tested conditions. As a whole, our results show a large reorganizationof Fusarium’s transcriptome upon phenolic acid treatment.448. Differential transcriptome analysis of Zymoseptoria tritici infecting wheat reveals novel effectors. Stefano F.F. Torriani 1 , Marcello Zala 1 , DanielCroll 1 , Patrick C. Brunner 1 , Eva H. Stukenbrock 2 , Dee Carter 3 , Bruce A. McDonald 1 . 1) Integrative Biology, ETHZ, Zurich, Switzerland; 2) Max Planck Institutefor Terrestrial Microbiology, Marburg, Germany; 3) University of Sydney, Sydney, Australia.Zymoseptoria tritici (formerly called Mycosphaerella graminicola) is a hemibiotrophic fungus belonging to the Dothideomycetes, the largest class ofascomycetes that includes many plant pathogens. Like other hemibiotrophic pathogens Z. tritici uses different strategies for obtaining nutrition during itslife cycle. For the first 10 days post inoculation (dpi) the pathogen colonize the host as a biotroph without causing visible symptoms. The necrotrophicphase lasts until the affected plant cells have died. Depending on the strain-cultivar interaction, plant cell death occurs from 18 to 20 days afterpenetration. Z. tritici concludes its life cycle by surviving as a saprotroph on dead leaves for several months. Thus Z. tritici presents a powerful system tostudy host-pathogen interactions during different stages of disease development. Next generation sequencing technology was used to analyze changes intranscription during the complete infection cycle of Z. tritici on wheat. The total RNA was extracted from inoculated plants at six time points (3-, 7-, 11-,14-, 21- and 56- dpi). RNA-Seq analyses allowed us to trace the expression profile of 10,251 genes and identify genes that differed in expression betweenthe biotrophic, necrotrophic and saprotrophic stages of infection. About 14% and 34% of the genes showed statistically significant differences inexpression from the biotrophic to necrotrophic and from the necrotrophic to saprotrophic stages of infection, respectively. Putative effector genes werepreferentially transcribed at 11 dpi during the transition between biotrophy and necrotrophy. Through this screen we identified five putative effectorgenes for further characterization, using Agrobacterium-mediated transformation to determine their role in pathogenicity. Although recent experimentalefforts focused mainly on proteinaceous effectors, we investigated the role of non-proteinaceous metabolites as they can also manipulate host cells. Twodifferent clusters of genes (PKS4 and PKS5-related genes) involved in the biosynthetic pathways of different secondary metabolites showed expressionpatterns similar to the putative effectors. Confirmation of function of the putative virulence genes will be based on gain or loss of virulence in planta usinggene knock-outs and knock-ins.449. Role of the xprG gene in autolysis, secondary metabolism and asexual development in Aspergillus nidulans . Margaret E. Katz, KatharynBraunberger, Sarah Cooper. Dept Molec & Cellular Biol, Univ New England, Armidale, Australia.The Aspergillus nidulans xprG gene encodes a transcriptional activator that is a member of the Ndt80 family in the p53-like superfamily of proteins.Previous studies have shown that XprG controls the production of extracellular proteases in response to starvation. We undertook transcriptional profilingto investigate whether XprG has a wider role as a global regulator of the carbon nutrient stress response. Our microarray data showed that the expressionof a large number of genes, including genes involved in secondary metabolism, development, and autolysis, were altered in an xprGD null mutant. Many ofthese genes are known to be regulated in response to carbon starvation. We confirmed that sterigmatocystin and penicillin production is reduced in xprGmutants.The loss of fungal mass and secretion of pigments that accompanies fungal autolysis in response to nutrient depletion was accelerated in anxprG1 gain-of-function mutant and decreased or absent in an xprG- mutant. We found that conidophore development occurred in carbon-starvedsubmerged cultures of both the xprGD1 loss- and xprG1 gain-of-function mutants, though the number of metulae appeared to be reduced. Thus, thereduction of brlA expression observed in the xprGD1 mutant is not sufficient to block conidiophore development in response to carbon starvation. .However, the xprG1 gain-of-function mutation partially suppresses VeA-mediated repression of conidiophore development and the conidiophoredevelopment defect in the fluG701 mutant. These results support the hypothesis that XprG plays a major role in the response to carbon limitation and thatnutrient sensing may represent one of the ancestral roles for the p53-like superfamily.450. Fugal-specific sirtuin HstD coordinates the secondary metabolism and development via the LaeA. M. Kawauchi 1,2 , K. Iwashita 1,2 . 1) Dept. Mol.Biotech., Grad. Sch. Adv. Sci. Mat., Hiroshima Univ., Hiroshima, Japan; 2) Natl. Res. Inst. Brewing, Hiroshima, Japan.27th Fungal Genetics Conference | 231

FULL POSTER SESSION ABSTRACTSThe sirtuins are members of the NAD + -dependent histone deacetylase family that contribute to various cellular functions which are affected aging,disease and cancer development. However, physiological roles of the fungal-specific sirtuin family are still poorly understood, especially with regard totheir participation in the genomic stability of yeast. Here, we determined the novel function of the fungal-specific sirtuin HstD, which is homolog of yeastHst4 in Aspergillus oryzae. The deletion of HstD indicated that both conidial development and secondary metabolism were regulated by HstD in A. oryzae.Furthermore, the gene expression of LaeA, which is the most studied coordinator for the regulation of secondary metabolism and development, wasinduced in the DHstD strain, and we found a significant genetic interaction between HstD and LaeA using double-disrupted or overexpression strains. Thus,we concluded the fungal-specific sirtuin HstD coordinates the fungal development and secondary metabolism via the regulation of LaeA gene expression infilamentous fungi. The HstD is fungal-specific, but it is conserved in the vast family of filamentous fungi. Therefore, HstD has great potential as a drugtarget for mycosis or plant disease, because the fungal development and secondary metabolism are virulence determinants of pathogenic fungi. Inaddition, our findings are also important for improving the productivity of useful secondary metabolites and developing an attractive host for theproduction of several heterogeneous secondary metabolites.451. Improved flavor production by manipulation of the Ehrlich pathway in ascomycetes. D. Ravasio, A. Walther, J. Wendland. Carlsberg Laboratory,Copenhagen V, Denmark.The Ehrlich pathway utilizes amino acids to generate higher alcohols with distinctive flavor in three enzymatic steps including a transaminase, adecarboxylase and an aldehyde dehydrogenase. Comparative genomics revealed the absence of key genes of the Ehrlich pathway in Eremotheciumcymbalariae whereas these genes were found to be present in the closely related species Ashbya gossypii. A. gossypii produces a very fruity flavor both inliquid culture and on solid media. The biological significance of this is unknown. Here, we present the functional analysis of A. gossypii key genes of theEhrlich pathway, ARO8a, ARO8b, ARO10, and ARO80. Deletion of any one component resulted in a noticeable reduction of flavor production asdetermined by GC/MS. In Saccharomyces cerevisiae ARO80 has been described as the main transcription factor regulating other genes of the Ehrlichpathway. Therefore, we analyzed the effect of deletion and overexpression of this gene on flavor production in yeast. As expected, overexpressionresulted in a marked increase in flavor production, particularly in isoamyl alcohol, a banana-like flavor. Next to chemical analyses we generated a lacZbasedreporter gene assay using ARO-gene promoters. With such a tool we can determine the status of flavor production under various conditions and in avariety of yeast strains. Initial results will be presented.452. Suppressor mutagenesis of a DlaeA mutant reveals novel regulators of secondary metabolism in Aspergillus nidulans. Alexandra Soukup, Jerry Luo,Jin Woo Bok, Nancy P. Keller. UW-Madison, Madison, WI.Aspergillus nidulans is a filamentous fungus known to produce a variety of complex natural products known as secondary metabolites (SM). Regulation ofthese bioactive SM can occur through cluster specific transcription factors, or through global regulators such as LaeA. Deletion of laeA results in drasticallydecreased amounts of multiple secondary metabolites. A multi-copy suppressor screen for genes capable of phenotypically returning norsolorinic acid(NOR) production to the DlaeA mutant resulted in identification of 17 plasmids containing inserts ranging from one to four genes. Further analysis of thesuppressor plasmids confirmed of a subset to increase SM production both in the original laeA deletion strain and in wild type backgrounds.453. A network of HMG-box transcription factors regulates sexual cycle in the fungus Podospora anserina. J. Aït-Benkhali 1,2 , E. Coppin 1,2 , S. Brun 1,2,3 , T.Martin 4 , C. Dixelius 4 , R. Debuchy 1,2 . 1) Univ Paris-Sud, Institut de Génétique et Microbiologie, Orsay, France; 2) CNRS, Institut de Génétique etMicrobiologie, Orsay, France; 3) UFR des Sciences du Vivant, Université Paris-7 Diderot, Paris, France; 4) Department of Plant Biology and Forest Genetics,Uppsala BioCenter, Swedish University of Agricultural Sciences and Linnean Center for Plant Biology, Uppsala, Sweden.High-mobility group B proteins are eukaryotic DNA-binding proteins characterized by the HMG-box functional motif. These transcription factors play apivotal role in global genomic functions and in the control of genes involved in specific developmental or metabolic pathways. The filamentous ascomycetePodospora anserina contains 12 HMG-box genes. Of these, four have been previously characterized; three are mating-type genes that control fertilizationand development of the fruiting-body, whereas the last one encodes a factor involved in mitochondrial DNA stability. Systematic deletion analysis of theeight remaining uncharacterized HMG-box genes indicated that none were essential for viability, but that seven were involved in the sexual cycle. TwoHMG-box transcription factors display striking features. Pa_1_13940, an ortholog of SpSte11 from Schizosaccharomyces pombe, is a pivotal activator ofmating-type genes in P. anserina, whereas Pa_7_7190 is a repressor of several phenomena specific to the stationary phase, most notably hyphalanastomoses. Constitutive expression of mating-type genes in a DPa_1_13940 strain did not restore fertility, indicating that Pa_1_13940 has additionalfunctions related to sexual reproduction besides activating mating-type genes. RT-qPCR analyses of HMG-box genes in different HMG-box deletion strainsindicated that Pa_1_13940 is at the hub of a network of several HMG-box factors that regulate the sexual cycle. Complementation experiments with astrain deleted for mating-type genes revealed that this network control fertility genes in addition to mating-type target genes. This study points to thecritical role of the HMG-box members in sexual reproduction in fungi, as 11 out of 12 members were involved in the sexual cycle in P. anserina.Pa_1_13940 and SpSte11 are conserved transcriptional regulators of mating-type genes, although P. anserina and S. pombe have diverged 1.1 billion yearsago. Two HMG-box genes, SOX9 and its upstream regulator SRY, play also an important role in sex determination in mammals. The mating-type genes andtheir upstream regulatory factor form a module of HMG-box genes similar to the SRY/SOX9 module, suggesting it may be ancestral in Opisthokonta.454. Sclerotinia sclerotiorum MAT genes function in fertility and apothecial morphogenesis. Benjamin Doughan, Jeffrey Rollins. Plant Pathology,University of Florida, Gainesville, FL.Sclerotinia sclerotiorum (Lib.) de Bary is an omnivorous, polyphagus, phytopathogenic fungus that relies on the completion of the sexual cycle to initiatemost new disease cycles. The sexual cycle is characterized by the development of apothecia that forcibly discharge ascospores for local and, under suitableconditions, long distance dissemination. A strategy for understanding the regulation of apothecial multicellular development is being pursued throughfunctional characterization of the mating type genes in S. sclerotiorum. These genes are hypothesized to encode master regulatory proteins required foraspects of sexual development ranging from fertilization through fertile fruiting body development. Experimentally, gene deletion strategies wereperformed to create loss-of-function mutants in the two conserved “core” mating type genes common to most ascomycete fungi as well for two lineagespecificgenes found only in S. sclerotiorum and closely related fungi. mat 1-1-1 and mat 1-2-1 mutants are able to form ascogonia but are blocked in allaspects of apothecia development. These mutants also exhibit defects in secondary sexual characters including the production of smaller sclerotia andlower numbers of spermatia. mat 1-2-4 mutants are delayed in apothecia production and form apothecia with aberrant disc morphogenesis and ascosporeproduction. They too produce lower numbers of spermatia and smaller sclerotia and additionally, exhibit a slower hyphal growth rate. Phenotypes of themat 1-1-5 gene knockouts are under evaluation and will be reported. Our findings demonstrate that mat genes are involved in both sexual fertility anddevelopment in S. sclerotiorum.232

FULL POSTER SESSION ABSTRACTS455. The Sclerotinia sclerotiorum mating type locus (MAT) contains a 3.6-kb region that is inverted in every generation. Patrik Inderbitzin 1 , PeriasamyChitrampalam 2 , Karunakaran Maruthachalam 1 , Bo-Ming Wu 3 , Krishna Subbarao 1 . 1) Department of Plant Pathology, University of California-Davis, Davis,CA, USA; 2) Department of Plant Sciences, University of Arizona, Tucson, AZ, USA; 3) Department of Plant Pathology, China Agricultural University, 2 WestYuanmingyuan Rd., Haidian District, Beijing, China.Sclerotinia sclerotiorum is a filamentous ascomycete in the Sclerotiniaceae (Pezizomycotina) and a necrotrophic pathogen of more than 400 hostsworldwide, including many important agricultural crops. In California, the biggest lettuce producer in the United States, S. sclerotiorum is a causal agent oflettuce drop that reduces overall annual lettuce yield by 15%. Little is known about the details of sexual reproduction in S. sclerotiorum, but the structureof the S. sclerotiorum mating type locus MAT, the master regulator of sexual reproduction in ascomycetes, has previously been reported. As in otherhomothallic (self-fertile) ascomycetes, S. sclerotiorum MAT contains both idiomorphs (divergent alleles) fused end-to-end at a single locus. Using 283isolates from lettuce in California and from other states and hosts, we investigated the diversity of S. sclerotiorum MAT, and identified a novel version ofMAT that differed by a 3.6-kb inversion and was designated Inv+, as opposed to the previously known S. sclerotiorum MAT that lacked the inversion andwas Inv-. The inversion affected three of the four MAT genes: MAT1-2-1 and MAT1-2-4 were inverted and MAT1-1-1 was truncated at the 3’-end.Expression of MAT genes differed between Inv+ and Inv- isolates. In Inv+ isolates, only one of the three MAT1-2-1 transcript variants of Inv- isolates wasdetected, and the alpha1 domain of Inv+ MAT1-1-1 transcripts was truncated. Both Inv- and Inv+ isolates were self-fertile, and the inversion segregated ina 1:1 ratio regardless of whether the parent was Inv- or Inv+. This suggested the involvement of a highly regulated process in maintaining equalproportions of Inv- and Inv+, likely associated with the sexual state. The MAT inversion region, defined as the 3.6-kb MAT inversion in Inv+ isolates and thehomologous region of Inv- isolates, was flanked by a 250-bp inverted repeat on either side. The 250-bp inverted repeat was a partial MAT1-1-1 thatthrough mediation of loop formation and crossing over, may be involved in the inversion process. Inv+ isolates were widespread, and in California andNebraska constituted half of the isolates examined. We speculate that a similar inversion region may be involved in mating type switching in thefilamentous ascomycetes Chromocrea spinulosa, Sclerotinia trifoliorum and in certain Ceratocystis species.456. Repression of the phosphor-transmitter gene ypdA resulting in growth defect in Aspergillus fumigatus. Daisuke Hagiwara 1 , Hiroki Takahashi 1 ,Mayumi Nakayama 2 , Keietsu Abe 2 , Tohru Gonoi 1 , Susumu Kawamoto 1 . 1) Medical Mycology Research Center, Chiba university, Chiba, Japan; 2) NICHe,Tohoku university, Sendai, Japan.Two-component system (TCS) is a conserved signal transduction system implicated in cellular responses to a variety of environmental stimuli in fungi.Aspergillus fumigatus has 13 histidine kinases, single HPt (histidine-containing phosphor transmitter), and 3 response regulators, which together constitutea TCS signaling. According to studies of HPt in several fungi, ypdA encoding a single HPt of A. fumigatus has been thought to be an essential gene. In thisstudy, we tested if absence of YpdA leads to cell lethality in A. fumigatus, and investigated what the molecular mechanisms underlying the lethality is. Toaddress these questions, we constructed a conditional ypdA-expressing strain by replacing a native promoter of ypdA with thiA promoter (PthiA). PthiA isstrictly repressed in the presence of thiamine, while moderately expressed in the absence of thiamine. The conditional PthiA-ypdA strain showed severegrowth defect on a plate containing thiamine, while the strain grew normally on a plate without thiamine as the wild-type stain. We, then, investigated theexpression profiles of catA, dprA, and dprB genes, which are regulated under the control of SakA MAPK cascade, since the SakA MAPK cascade isdownstream of A. fumigatus TCS. In a liquid culture, expressions of catA, dprA, and dprB were gradually increased after addition of thiamine, suggestingthat inactivation of ypdA expression leads to the activation of SakA MAPK cascade. To get more insight into a response to ypdA-repression, thetranscriptome profiles were obtained by RNA-seq. Expression levels of each gene at 3h, 6h, and 9h after addition of thiamine were compared to that at 0h.More than 2-fold or less than 1/2-fold of expression changes were regarded as up- or down-regulated, respectively. Through statistical analysis oncategory of gene ontology, the groups concerning ribosome biogenesis or RNA metabolism were found to be significantly down-regulated after theinactivation of ypdA. Based on these results, we will discuss the cellular responses to YpdA deprivation and try to find out the molecular mechanismsattributed to the lethality.457. Unravelling the GTPase polarity complex in Claviceps purpurea. Andrea Herrmann 1 , Janine Schürmann 1 , Britta Tillmann 2 , Michael Bölker 2 , PaulTudzynski 1 . 1) IBBP, WWU Muenster, Schlossplatz 8, 48143 Muenster, Germany; 2) Philipps-Universität, Karl-von-Frisch-Strasse 8, 35032 Marburg,Germany.Claviceps purpurea is a plant pathogen infamous for its production of toxic alkaloids on infected host plants like barley. Consumption of infected grainsleads to severe symptoms up to the death of the patient. Infection patterns are complex and the topic of intensive research. One interesting aspect is thestrict polarity of the hyphal growth during the first infection stage which seems to be crucial for the non-recognition of C. purpurea as a pathogen by thehost. To address the question of the importance of polarity the structure and dynamics of the polarity complex are the focus of this work. The guaninenucleotide exchange factors (GEFs) Cdc24 and Dock180 belong to different families, Cdc24 being a member of the Dbl GEF family and Dock180 a CZH GEF.Cdc24-GFP localises cytosolically and to hyphal tips whereas Dock180-GFP is present in small vesicles in the hypha, though concentrated at the tip region,too. Cdc24 DHPH domains are able to activate the small GTPases Rac and Cdc42 of C. purpurea and U. maydis In vitro, whereas the catalytic domain ofDock180 only activates Rac in both organisms. Despite the proven activation Cdc24 does not interact with any GTPase in yeast two hybrid assays. Dock180shows a weak interaction with Rac and the two p21-activated kinases (PAKs) Ste20 and Cla4. Thus, both GEFs do not share many characteristics apart fromtheir GEF activity. The PAKs Ste20 and Cla4 and the scaffold protein Bem1 are involved in the polarity complex, too. Ste20 localises to hyphal tips andinteracts with Cdc42 in a loading status dependent manner, whereas Cla4 is the main partner of Rac. Other interactions of Ste20 with Dock180 and Cla4could also be shown. Bem1 is present in the cytosol - concentrated at the hyphal tip - and links most of the proteins of the polarity complex as interactionswith Cdc24, Cla4, Ste20 and Dock180 have been detected. Taken together we postulate at least two different polarity complexes, the Rac complex and theCdc42 complex. Both are gathered by Bem1, but Cla4 is the main partner of Rac, whereas Ste20 plays a similar role for Cdc42. Dock180 is mainly linked toRac, Cdc24 can be active in both complexes. We are interested in the spatial and temporal formation and regulation of these complexes and its influenceon polarity and virulence which will be the subject of further studies.458. Atypical Gb and RACK homolog Gib2 is a signal transducing adaptor protein affecting growth and virulence of Cryptococcus neoformans. YanliWang 1 , Gui Shen 1 , Jinjun Gong 1 , Amy Whittington 2 , Ping Wang 1,2,3 . 1) Res Inst for Children, Children's Hospital, New Orleans, LA USA; 2) Dept Microbiology,Immunology and Parasitology, LSUHSC, New Orleans, LA USA; 3) Dept Pediatrics, LSUHSC, New Orleans, LA USA.Virulence in Cryptococcus neoformans is a multifaceted trait underpinned by complex signaling pathways. The atypical G-protein b subunit Gib2 displaysversatility in interactions with signaling molecules such as Ga Gpa1 that governs cAMP signaling and intersectin Cin1 that regulates intracellular trafficking.This and the conserved seven-bladed b-propeller motif are highly suggestive that Gib2 functions as an adaptor protein. We here show that Gib2 binds to27th Fungal Genetics Conference | 233

FULL POSTER SESSION ABSTRACTSGpa1 and Gpg1/Gpg2 in a direct manner, and Gib2 promotes cAMP levels through novel interactions with phosphodiesterase Pde2 and RAS proteins Ras1and Ras2. Using TAP technology, we have identified additional 42 proteins whose putative function range from signal transduction, energy generation,metabolism, and stress response to ribosomal function. Finally, through establishing a protein-protein interactive network, we illustrate that Gib2 adapts ascaffold role to mediate specific protein-protein interactions that drive the formation of various protein complexes. This includes fostering aheterotrimeric complex with Gpa1 and Gpg1/Gpg2, targeting Pde2 (direct) and adenylyl cyclase Cac1 (indirect) to regulate cAMP levels, and likely servingas a conserved ribosomal core protein facilitating fundamental cellular processes to underlie growth and virulence. Our studies reveal the complexity ofthe regulatory network in fungi and advocate Gib2 as a novel target for antifungal therapy.459. Consequences of the loss of transcription factors SreA and HapX on siderophore biosynthesis and iron homeostasis in the perennial ryegrassendophyte, Epichloë festucae. Natasha T. Forester 1,2 , Geoffrey A. Lane 1 , Iain L. Lamont 2 , Linda J. Johnson 1 . 1) Plant Fungal Interactions Team, AgResearchLtd, Palmerston North, New Zealand; 2) Biochemistry Dept, University of Otago, Dunedin, NZ.Siderophores are low molecular weight ferric iron chelators that are made by microorganisms to compete for and to sequester iron, an essentialmicronutrient. Epichloë festucae, a fungal endosymbiont of perennial ryegrass, synthesises two siderophores, epichloënin A and ferricrocin to harvest andutilise iron from its host grass. Work by our group has implicated epichloënin A in the maintenance of symbiosis and is described in another abstract. Toexplore the regulation of siderophore biosynthesis and iron homeostasis processes, we have characterised mutants of two major iron-responsivetranscription factors, SreA and HapX that coordinate cellular responses to iron availability. To evaluate the effect of loss of SreA and HapX on siderophorebiosynthesis, we measured the production of epichloënin A and ferricrocin by LC-MS/MS. Relative to wild type; both siderophores were over-produced inDsreA mycelia grown in the presence of iron, while ferricrocin was produced in excess in DhapX mycelia grown under iron deprived conditions. Irondependentphenotypic deviations from wild type fungal growth were also observed in culture and in planta but neither mutant disrupted the E. festucae -L. perenne association under standard soil conditions. However, under iron-limiting conditions through hydroponic control of symbiotic iron supply, wedemonstrated that DsreA mutants can induce chlorosis in their hosts, indicating that DsreA mutants compete for host iron. In planta, the DsreA fungalhyphae are also markedly increased in girth and lack growth in vascular bundles. In DhapX infected plants grown hydroponically in iron deprivedconditions, we observed inappropriate fungal growth such as highly convoluted and compressed hyphae and elongated fungal structures which hinted atreduced resource conservation by DhapX under growth limiting conditions. Collectively, these results suggest that Epichloë fungi have a tightly regulatediron management system for niche adaptation and actively set limits on iron withdrawal from the host, presumably to prevent competition with its host topromote mutualistic interactions. Mutations that interfere with fungal iron management, either by deregulating siderophore synthesis, can destabilise thefungal-plant association.460. Who is to blame: defining the host responses that lead to ToxA-induced susceptibility. Iovanna Pandelova, Viola Manning, Ashley Chu, LyndaCiuffetti. Botany and Plant Pathology, Oregon State Univ, Corvallis, OR.Pathogenicity by the necrotrophic pathogen of wheat, Pyrenophora tritici-repentis (Ptr), is attributed to the production of host-selective toxins (HSTs).Understanding the mode-of-action of HSTs is essential for a complete characterization of how these pathogenicity factors condition plant diseasesusceptibility. One of the proteinaceous HSTs produced by Ptr, PtrToxA (ToxA), induces necrosis in sensitive cultivars. Several studies suggest that ToxAinteracts with a high affinity receptor, enters mesophyll cells and localizes to chloroplasts. Additionally, ToxA acts as an elicitor of defense responses byincreasing production of phenolic compounds and by the up-regulation of genes involved in jasmonic acid and ethylene production pathways. After ToxAtreatment and incubation in constant light, there is a decrease in photosystem (PS) I and II transcripts observable already at 9 and 14 hours post infiltration(hpi), which is followed by the drastic reduction in levels of both PSI- and PSII-complex proteins. It is proposed that photosystem dysfunction leads to lightdependentaccumulation of reactive oxygen species (ROS) and the development of necrosis. To better understand the role of ROS, photosynthesis anddefense responses in necrosis development induced by ToxA, plants were incubated in light (presence of ROS) or in dark (absence of ROS). In order todetermine the impact of ToxA on gene regulation and to establish when early changes in protein content of PS complexes occur, both biochemical andmicroarray analyses were performed. Some defense-related genes are up-regulated in ToxA-treated leaves incubated in the dark (ToxA/dark), althoughthe number of probesets was considerably less compared to ToxA-treated leaves incubated in light (ToxA/light). Furthermore, ethylene biosynthesis genes,that play a role in symptom development in ToxA/light treatrment are not significantly up-regulated in ToxA/dark-treated leaves. Finally, only a smallfraction of PSI- and II-related and chlorophyll a/b-binding genes are down-regulated in ToxA/dark compared to ToxA/light treatment. These data suggestthat only certain defense-related pathways are involved in ToxA-induced necrosis development, and help to identify those genes whose differentialregulation by ToxA is light and/or ROS-dependent.461. RNA silencing of pacC increases aflR transcript levels under alkaline pH conditions in Aspergillus flavus. Benesh M Somai, Kyle W van der Holst, EssaSuleman. Department of Biochemistry and Microbiology, Nelson Mandela Metropolitan University, Port Elizabeth, 6031, Eastern Cape, South Africa.Aspergillus flavus produces aflatoxin B1 which is an important hepatocarcinogen, especially amongst the developing third world countries which have alarge number of poor, rural, subsistence communities with little access to fungicides. The master regulator of aflatoxin production is aflR which, in turn,appears to be negatively regulated by pacC. However, until now, there were never any direct measurements of the relative aflR/pacC transcript ratiosproduced under aflatoxin conducive and non-conducive conditions. In the current study, pacC was down-regulated in two transformants by a syntheticpacCRNAi construct under the control of a thiamine inducible promoter. Expression of pacC and aflR transcripts was then measured via RT-qPCR incultures grown under alkaline or acid conditions. At pH 4, between pacCRNAi inducing and repressing conditions, an aflR/pacC transcript ratio of 1.09relative to the reference gene was obtained indicating the production of an equal abundance of aflR and pacC mRNA. It is generally accepted that at acidicpH the majority of pacC mRNA is unprocessed, remains untranslated and non-functional thereby being incapable of repressing AFLR protein production.This stimulates aflatoxin production at acidic pH. Between pH 8 and pH 4, when pacCRNAi was suppressed, the aflR/pacC ratio was 0.2 indicating that pacCproduction was higher than that of aflR. aflR transcript levels were reduced between 76% and 80% therefore explaining the normal lack of aflatoxindetection at pH 8. Between pH 8 and pH 4, when pacCRNAi was induced, the aflR/pacC ratio was between 1.77 and 13.21 indicating that at alkaline pH,suppression of pacC allowed a large increase in AFLR which stimulated aflatoxin production. It is concluded that pacC is produced at acidic pH, but remainslargely non-functional. Furthermore, at pH 8, aflR production decreases only by about 80% and therefore it is possible that the remaining 20% oftranscripts still stimulates aflatoxin production. Finally, via RNAi silencing it is conclusively proved that pacC negatively regulates aflR production at pH 8.462. High-throughput prediction and functional validation of promoter motifs regulating gene expression in spore and infection stages of Phytophthorainfestans. H. Judelson, S. Roy, M Kagda. Dept of Plant Pathology and Microbiology, University of California, Riverside, CA.Most filamentous pathogens have complex life cycles in which gene expression networks orchestrate the formation of cells specialized for dissemination234

FULL POSTER SESSION ABSTRACTSor host colonization. In the oomycete Phytophthora infestans, the potato late blight pathogen, we identified major shifts in mRNA profiles duringdevelopmental transitions using microarrays. We then used those data with three search algorithms to discover more than 100 motifs that are overrepresentedin promoters of genes up-regulated in hyphae, sporangia, sporangia undergoing zoosporogenesis, swimming zoospores, or germinated cystsforming appressoria. Most of the putative stage-specific transcription factor binding sites (TFBSs) thus identified had features typical of TFBSs such asposition or orientation bias, palindromy, and conservation in related species. Each of six motifs tested in P. infestans transformants using the GUS reportergene conferred the expected stage-specific expression pattern, and were shown to bind nuclear proteins in gel-shift assays. Several motifs linked to theappressoria-forming stage were over-represented in promoters of genes encoding effectors and other pathogenesis-related proteins. To understand howpromoter and genome architecture influence expression, we also mapped transcription patterns to the P. infestans genome assembly. Adjacent geneswere not typically induced in the same stage, including genes transcribed from a small shared promoter region. Analyses of global expression, however,demonstrated that co-regulated gene pairs occurred more than expected by random chance. These data help illuminate the processes regulatingdevelopment and pathogenesis, and will enable future attempts to purify the cognate transcription factors. Our approach should be applicable to bothoomycetes and fungi.463. Cooperative regulation of Aspergillus nidulans cellulase genes by transcription factors McmA and ManR/ClrB. Tetsuo Kobayashi 1 , Nuo Li 1 , MikiAoyama 1 , Yohei Yamakawa 1 , Masahiro Ogawa 2 , Yasuji Koyama 2 . 1) Grad Sch of Bioagricultural Sci, Nagoya Univ, Nagoya, Aichi, Japan; 2) Kikkoman Corp,Noda, Chiba, Japan.Expression of the endoglucanase A gene (eglA) in A. nidulans is inducible by cellobiose. Previously, the cis-element responsible for the inductiveexpression, designated CeRE (Cellulose Responsive Element), was identified based on mutational analysis of the eglA promoter. CeRE contained thebinding consensus of SRF-MADS proteins, suggesting involvement of McmA, the sole SRF-MADS protein in A. nidulans. While two Zn 2Cys 6 transcriptionfactors in Aspergillus were recently reported to be essential to cellulase induction. One is ManR in A. oryzae, which regulates both mannanase andcellulase genes, and the other is ClrB in A. nidulans, a homolog of the cellulase regulator CLR-2 in N. crassa. Since these factors were orthologous sharing63% identity, we use the name ManR/ClrB. In this presentation, we provide evidences that McmA and ManR/ClrB cooperatively regulate induction ofcellulase genes. Effects of mcmA mutation and manR/clrB deletion on expression of cellulase genes were examined by qRT-PCR. Expression of eglA, eglB,and cbhA was highly induced by cellobiose in the wild type strain. The induction was significantly impaired by the mcmA mutation and abolished by themanR/clrB deletion, indicating that the cellulase genes under control of McmA and ManR/ClrB are overlapped. Binding of His-tagged McmA and FlagtaggedManR/ClrB-DBD (DNA Binding Domain), which were produced in E. coli and purified, to the CeRE containing region of the eglA promoter wasexamined by EMSA. McmA gave two shifted bands corresponding to single and double occupation of the binding sites, which lay within and just upstreamof CeRE. ManR/ClrB-DBD alone showed very weak binding to the region. When both McmA and ManR/ClrB-DBD were added, a slower-migrating and moreabundant shifted band was appeared, which suggested cooperative binding of McmA and ManR/ClrB-DBD. Presence of McmA and ManR/ClrB-DBD in theshifted band was confirmed by supershift assay with the anti-his-tag and anti-flag-tag antibodies. These results indicated that inductive expression of eglAis regulated by cooperative binding of McmA and ManR/ClrB to its promoter, and suggests that regulation of eglB and cbhA would be similar. This workwas supported by the Program for Promotion of Basic and Applied Researches for Innovations in Bio-oriented Industry.464. Transcription factor shuttling during cellulase induction in Trichoderma reesei. Alex Lichius, Christian P. Kubicek, Verena Seidl-Seiboth. Institute ofChemical Engineering, Vienna University of Technology, Vienna, Austria.For economically feasible production of liquid fuels and other value-added compounds from lignocellulosic plant material, strategies are required toboost cellulolytic and hemicellulolytic enzyme production by industrially relevant fungi. One promising approach is to modulate the transcriptional controlmediating release from carbon catabolite repression (CCR) and induction of cellulase, hemicellulase and xylanase gene expression. To better understandthe underlying molecular dynamics during induction, we characterized nucleo-cytoplamic shuttling of the two transcription factors carbon cataboliterepressor 1 (CRE1) and xylanase regulator 1 (XYR1) of Trichoderma reesei by means of live-cell imaging. In submerged cultures, nuclear import and exportof CRE1 upon repression and induction, respectively, occurred within minutes and therefore was generally faster than shuttling of XYR1. Under CCRconditions XYR1 expression levels were very low, and its nuclear signal required up to one hour to significantly increase upon replacement into an inducingcarbon source. Cultured directly under inducing conditions, nuclear accumulation of XYR1 was detectable after about 20h post inoculation, and stronglyincreased within the following 24 hours. CRE1 under the same conditions was localized exclusively to the cytoplasm. In plate cultures, nuclear recruitmentof CRE1 and XYR1 differed within the central area, the subperiphery and the periphery of the colony depending on the provided carbon source. Mostinterestingly, under inducing conditions we found evidence for increased nuclear recruitment of CRE1 in the central area, correlating with strong nuclearimport of XYR1 in the same region. Notably, the cytoplasmic signal of CRE1 was usually elevated in leading hyphae, whereas XYR1 was never significantlyrecruited to the colony periphery. Taken together our data provide the first temporal resolution of transcription factor shuttling during the induction ofcellulase gene expression in Trichoderma reesei, and reveal some interesting differences between the subcellular localization of CRE1 and XYR1 insubmerged and plate cultures, respectively. These differences indicate that the mycelial organization during fungal growth might be another importantregulatory element to consider for the industrial scale production of cellulolytic enzymes.465. Trichophyton rubrum ap-1 gene expression in response to environmental challenges. Nalu TA Peres, Gabriela F Persinoti, Larissa G Silva, Tiago RJacob, Antonio Rossi, Nilce M Martinez-Rossi. School of Medicine, Univiversity of Sao Paulo, Ribeirao Preto, Brazil.Several families of transcription factors (TF) are found in fungal cells, which contribute for the broad range of cellular responses triggered byenvironmental changes in these organisms. These TF regulate the expression of genes involved in different cellular processes allowing cell survival understressful as well as physiological conditions. The AP-1 TF belongs to the bZIP family (basic leucine zipper), and is involved in the conidiation process,response to oxidative stress, multidrug resistance, and pathogenicity of some fungi. In dermatophytes, a group of keratinophilic fungi, little is known aboutthis TF and the processes in which it is required. Trichophyton rubrum is the major etiologic agent isolated from clinical cases of cutaneous mycoses inhumans, and studies of the responses of this fungus to several environmental conditions allow a better understanding of its physiology and pathogenicity,thus providing information of how to decrease its growth and to establish more efficient therapeutic measures. Here, we evaluated the ap-1 geneexpression profile during growth of T. rubrum in several nutrient sources (keratin, ex vivo skin and nail), exposure to antifungal drugs, andoxidative/osmotic stresses. An up-regulation of ap-1 gene expression was observed during ex vivo infection and keratin utilization, compared to growth ina glucose containing medium. In response to different antifungal agents, ap-1 was up-regulated, while osmotic and oxidative stress did not alter itsexpression level. These results provide insights into the regulation of the ap-1 TF gene expression, suggesting its involvement in T. rubrum pathogenicity,possibly regulating several genes that allow the utilization of proteins from the host tissues as nutrient sources, and also protecting the cell against thedamages caused by antifungal drugs. Financial Support: FAPESP, CNPq, CAPES, and FAEPA.27th Fungal Genetics Conference | 235

FULL POSTER SESSION ABSTRACTS466. Vib-1 is required for cellulose utilization in Neurospora crassa. Yi Xiong, Jianping Sun, N. Louise Glass. Department of Plant and Microbial Biology,Univ California, Berkeley, Berkeley, CA.Vib-1 (vegetative incompatibility blocked) encodes a transcriptional regulator that is required for nonself recognition and heterokaryon incompatibility inNeurospora crassa. It is also required for the production of extracellular proteases upon carbon and nitrogen starvation. We found that vib-1 null mutantwas severely defective in utilizing cellulose as a carbon source and this growth defect could be rescued by constitutively expressing clr-2, an essentialtranscription factor for cellulase gene expression. Our transcriptional profiling with RNA-seq showed that most genes that were induced in wild type oncellulose were not induced in the vib-1 null mutant, but were expressed in vib-1 null mutant under non-inducing conditions when clr-2 transcription factorwas constitutively expressed. Our data suggests that vib-1 functions in a signaling pathway upstream of clr-2 for cellulose utilization. By comparing thetranscriptomes of vib-1 and clr1/clr-2 mutants versus wild type in response to different carbon sources including sucrose and cellulosic materials as well ascarbon and nitrogen starvations, we differentiated vib-1 dependent gene expression into several categories. We propose that vib-1 may play an importantrole in carbon starvation signaling ,thus positively regulating preparation for utilization of cellulosic materials.467. The stringency of start codon selection in the filamentous fungus Neurospora crassa. Jiajie Wei 1 , Ying Zhang 1 , Ivaylo P. Ivanov 2 , Matthew S. Sachs 1 .1) Dept Biol, Texas A&M Univ, College Station, TX; 2) BioSciences Institute, University College Cork, Ireland.In eukaryotic cells, initiation may occur from near-cognate codons that differ from AUG by a single nucleotide. The stringency of start codon selectionimpacts the efficiency of initiation at near-cognate codons and the efficiency of initiation at AUG codons in different contexts. We used a codon-optimizedfirefly luciferase reporter initiated with AUG or each of the nine near-cognate codons in preferred context to examine the stringency of start codonselection in the model filamentous fungus Neurospora crassa. In vivo results indicated that the hierarchy of initiation at start codons in N. crassa (AUG >>CUG > GUG > ACG > AUA » UUG > AUU > AUC) is similar to that in human cells. Similar results were obtained by translating mRNAs in a homologous N.crassa in vitro translation system or in rabbit reticulocyte lysate. We next examined the efficiency of initiation at AUG, CUG and UUG codons in differentcontexts in vitro. The preferred context was more important for efficient initiation from near-cognate codons than from AUG. These studies demonstratedthat near-cognate codons are used for initiation in N. crassa. Such events could provide additional coding capacity or have regulatory functions. Analysesof the 5’-leader regions in the N. crassa transcriptome revealed examples of highly conserved near-cognate codons in preferred contexts that could extendthe N-termini of the predicted polypeptides.468. A temperature-dependent complex transcriptional network controls cell shape and virulence in Histoplasma capsulatum. Sinem Beyhan 1 , MatiasGutierrez 1 , Mark Voorhies 1 , Anita Sil 1,2 . 1) Microbiology and Immunology, University of California, San Francisco, San Francisco, CA; 2) Howard HughesMedical Institute, Chevy Chase, MD.Histoplasma capsulatum, which is a respiratory fungal pathogen of humans, is endemic in the United States. Depending on the exposure dose and theimmune status of the host, the infection can lead to mild-respiratory or life-threatening and systemic disease. H. capsulatum has a dimorphic life cycle,switching from an infectious filamentous form in the soil to a pathogenic yeast form in mammalian hosts. This morphological switch, which requires adramatic shift in the gene expression profile of the cells, can be easily recapitulated in the laboratory simply by changing the temperature from roomtemperature to 37°C. We previously identified three regulators, Ryp1, Ryp2 and Ryp3, which are required for the yeast-phase growth. ryp1, ryp2 and ryp3mutants are unable to respond to change in temperature and grow constitutively in the filamentous form even at 37°C. Ryp1 belongs to a conservedfamily of fungal proteins that regulate cellular differentiation in response to environmental signals. The best-studied member of this family of proteins isWor1, which is a master regulator of white-to-opaque switching in Candida albicans. Ryp2 and Ryp3 are orthologous to VosA and VelB, respectively, whichare developmental regulators in Aspergillus nidulans. In this study, using transcriptional profiling and chromatin immunoprecipitation (ChIP) experiments,we explored complementary and unique roles of Ryp1, Ryp2, and Ryp3 in regulating yeast-phase growth. Our results reveal that Ryp1, Ryp2 and Ryp3physically interact and associate with DNA throughout the genome. Additionally, we identified a fourth transcription factor, Ryp4, which is a direct targetof Ryp1, Ryp2 and Ryp3, as a novel regulator of yeast-phase growth in H. capsulatum. Further transcriptional profiling and ChIP experiments show thatRyp4 regulates and associates with the upstream regions of a subset of Ryp1, Ryp2, and Ryp3 targets, which are involved in morphology and virulence in H.capsulatum. Finally, we identified two distinct cis-regulatory elements that are utilized by Ryp1 or the Ryp2/Ryp3 complex to facilitate gene expression.Our results reveal a tightly regulated and interwoven transcriptional network that controls the ability of a pathogenic fungus to cause disease in responseto host temperature.469. Evolutionary analysis of Dicer proteins: a preliminary analysis to study of microRNAs in the mushroom, Coprinopsis cinerea. Xuanjin Cheng, HoiShan Kwan. Life Sciences, The Chinese University of Hong Kong, New Territory, Hong Kong.Coprinopsis cinerea is a mushroom of limited edible value and is extensively used as a model organism to study the development of homobasidiomycetefungi. Unraveling the molecular basis of the fungus developmental processes would contribute to evolutionary studies and lead to improvement in thebreeding and cultivation of edible or medical homobasidiomycete mushrooms. MicroRNA (miRNA) is a group of endogenous non-coding regulatory RNAsof ~22 nt that regulate gene expression in various biological processes such as cell differentiation, development regulation and heterochromatinformation. Dicer is a key enzyme involved in the biogenesis of miRNAs and is highly conserved through eukaryotes. A miRNA-like RNA cannot be defined asa miRNA unless a Dicer (or Dicer-like) protein is found participating in its biogenesis. There are three Dicer homologs (CC1G_00230, CC1G_03181,CC1G_13988) identified in the C. cinerea genome. In order to gain an insight into the roles of Dicer proteins in C. cinerea and to investigate whether Diceris involved in miRNA biogenesis, we employed a comprehensive phylogenetic analysis of the Dicer protein family in all of the three kingdoms underEukaryota - animal, plant and fungus - and highlighted the results of Dicer homologs in C. cinerea. We showed that Dicer genes duplicated and diversifiedindependently in early animal, plant and fungus evolution, coincident with the origins of multicellularity. Besides, identified a group of Dicer homologs thatare specific to mushroom-forming fungi. We also showed that changes in one of the Dicer domains, the double-stranded RNA binding domain (dsRBD),alone may lead to diversification of Dicer proteins. As a whole, we revealed a dynamic picture in which the evolution of Dicer proteins has drivenelaboration of parallel RNAi functional pathways in the animal, plant and fungus kingdoms.470. Effect of the trp1 gene on transformation frequencies in Coprinopsis cinerea. Bastian Doernte, Ursula Kües. Molecular Wood Biotechnology andTechnical Mycology, University of Goettingen, Germany.Genetic transformation of the basidiomycete Coprinopsis cinerea has first been described by Binninger et al. in 1987 (1). For the transfer of geneticmaterial, chromosomal integrative vectors are used, which contain a selectable marker gene and/or a gene of interest. During transformation the geneticmaterial integrates at ectopic sites into the host chromosomes. Binninger et al. (1987) created the vector pCc1001, by cloning a 6.5 kb PstI genomic236

FULL POSTER SESSION ABSTRACTSfragment, with the tryptophan synthetase gene (trp1) of C. cinerea, into the ColE1 vector pUC9. The inserted gene allows the complementation of trp1auxotrophies and can be used as a selection marker. Several transformation experiments using this vector reveal a surprising phenomenon. Singletransformationwith solely pCc1001 gives only low numbers of transformants, whereas co-transformation with an additional plasmid yields about 2x moretransformants. To explore this phenomenon, the length of the trp1 harboring fragment was changed and the existing replicon was replaced by a modifiedColE1 replicon. All single-transformations resulted in the same observations. An alternative selection marker (pab1 encoding) and different relative vectorvectorconcentrations were tested in several co-transformation experiments. The obtained results lead to the conclusion that tryptophan feedbackinhibition might be responsible for the reduced transformation efficiencies in single-transformations of trp1 vectors. (1) Binninger et al. (1987). DNAmediatedtransformation of the basidiomycete Coprinus cinereus. EMBO J 6:835-840.471. The first promoter for conditional gene expression in Acremonium chrysogenum: iron starvation-inducible mir1 P . Fabio Gsaller 1 , Michael Blatzer 1 ,Beate Abt 1 , Markus Schrettl 2 , Herbert Lindner 3 , Hubertus Haas 1 . 1) Christian Doppler Laboratory for Fungal Biotechnology, Division of Molecular Biology,Medical University of Innsbruck, Austria; 2) Sandoz GmbH, Kundl, Austria; 3) Division of Clinical Biochemistry, Medical University of Innsbruck, Austria.The filamentous fungus Acremonium chrysogenum is of enormous biotechnological importance as it represents the natural producer of the beta-lactamantibiotic cephalosporin C. However, a limitation in genetic tools, e.g. promoters for conditional gene expression, impedes genetic engineering of thisfungus. Here we demonstrate that in A. chrysogenum iron starvation induces the production of the extracellular siderophores dimerumic acid, coprogen B,2-N-methylcoprogen B and dimethylcoprogen as well as expression of the putative siderophore transporter gene, mir1. Moreover, we show that thepromoter of mir1, mir1 P , is suitable for conditional expression of target genes in A. chrysogenum as shown by mir1 P -driven and iron starvation-inducedexpression of genes encoding green fluorescence protein and phleomycin resistance. The obtained iron-starvation dependent phleomycin resistanceindicates the potential use of this promoter for selection marker recycling. Together with easy scorable siderophore production, the co-regulation of mir1expression and siderophore production facilitates the optimization of the inducing conditions of this expression system. This work was funded by SandozGmbH (Kundl, Austria) and the Christian Doppler Society (Vienna, Austria).472. Mutagenic effect of high-LET ion beam irradiation in Neurospora crassa. Liqiu Ma 1* , Yusuke Kazama 2 , Tomoko Abe 2 , Shuuitsu Tanaka 1 , ShinHatakeyama 1 . 1) Regulation Biol, Saitama Univ, SAITAMA, Japan; 2) Radiation Biology Team, RIKEN, SAITAMA, Japan.Heavy ion beams cause great damages to cellular components particularly generating severe DNA damages, DNA double strand breaks (DSBs). Weexamined the biological effect and mutagenesis of irradiation of high-LET ion beam (Fe-ion) to DSB repair defect mutants in filamentous fungus Neuosporacrassa. Fe-ion beam ( 56 Fe 24+ : 90 MeV/u, LET=641 keV/mm) was irradiated to two DSB repair deficient mutants and wild-type strain. By lower doses (100 Gy), sensitivity to irradiation of the mus-52 strain (non-homologousend-joining deficient) is higher than that of the wild type, whilst lower than that of the mei-3 strain (homologous recombination deficient). Frequency offorward mutation occurred in the ad-3 loci was similar to previously examined C-ion beam irradiation, i.e. mei-3 > wild type > mus-52 strains. However,characteristic difference of mutation was observed as the scale of deletions; large deletions were frequently in the Fe-ion beam irradiated wild type strain,comparing to that 1 bp-deletions were mainly observed in the C-ion irradiation. Differences of mutagenesis and killing effect between the irradiation oftwo heavy ions, Fe-ion and C-ion, were discussed based on types of DNA damages.473. The Mad complex binds to light-regulated promoters in Phycomyces blakesleeanus. Alejandro Miralles-Duran, LM Corrochano. Genetica, Facultadde Biologia, University of Sevilla, Sevilla, Spain.The zygomycete Phycomyces blakesleeanus responses to light include phototropism of the fruiting body, activation of beta-carotene biosynthesis, andregulation of fruit body development. These photoresponses require the Mad complex, a protein complex composed of proteins MadA and MadB. Theseproteins are homologous of WC-1 and WC-2 from Neurospora crassa and presumably play a similar role in the regulation by light of gene expression.MadA and MadB have a zinc finger domain at the carboxyl end, and MadA has a LOV domain that should serve as the binding site for a flavinchromophore. In Phycomyces, the Mad complex should operate as a photoreceptor and transcription factor complex. The Phycomyces genome containstwo additional wc-1 homologs, wcoA and wcoB, and three additional wc-2 homologs, wctB, wctC, and wctD, but their function is unknown. We haveexpressed MadA and MadB in E. coli, and we have shown that these proteins bind the promoter of the light-regulated gene hspA by electrophoresismobility shift assays (EMSA). Protein binding to the hspA promoter was observed with each isolated protein or with the two proteins associated in theMad complex. The binding site to the hspA promoter will be identified by DNA footprinting analysis. We are performing similar assays with the otherPhycomyces Wc proteins and we hope that the results will help us to understand the role of the multiple Wc proteins in light-dependent gene regulation inPhycomyces.474. Down Regulation of sidB Gene by Use of RNA interference Technology in the Filamentous Fungi Aspergillus nidulans. S, Rezaie 1,2 , H, Eslami 1 , M.R.Khorramizadeh 1 , M.R. Pourmand 1 , M. Moazeni 2 . 1) Medical Biotechnology Dept, Tehran University of Medical Sciences, PhD; 2) Div. of Molecular Biology,Dept. of Medical Mycology and Parasitology, Tehran University of Medical Sciences, PhD.Background: RNA interference (RNAi) is a natural process by which short double-stranded RNA (siRNA) silences the expression of complementary targetRNAs by inducing RNA cleavage and subsequent reduction in protein expression levels. Introduction of the RNA interference machinery has guided theresearchers to discover novel methodologies for knocking down essential vital factor or virulence factor genes in the microorganisms such as fungi. Infilamentous fungi, Aspergillus nidulans, the gene sidB plays essential role in septation, conidiation and vegetative hyphal growth. In the present study, webenefited from the RNA interference strategy for down-regulating of a vital gene in the fungus Aspergillus nidulans. Materials and Methods: The 21-nucleotide siRNA was designed on the basis of the cDNA sequence of the sidB gene of A. nidulans. Transfection was performed via uptaking siRNAs frommedium by germinated spores. After 18 hours of incubation, total RNA was extracted and quantitative changes in expression of the sidB gene wereanalyzed by measuring the cognate sidB mRNA level by use of a quantitative real-time RT-PCR assay. Results: In the presence of 25 nM of siRNA, asignificant inhibition in germ tube elongation was observed compared with positive control samples (21 VS 42 mM). In addition, at the concentration of 25nM , a considerable decrease in sidB gene expression was revealed. Conclusion: Usage of RNA interference as a kind of post-transcriptional gene silencingmethods is a promising approach for designing new antifungal agents and discovering new drug delivery systems.475. SmallRNA mediated meiotic silencing of a transposable element in Neurospora crassa. Yizhou Wang, Jason E. Stajich. Plant Pathology &Microbiology, Univ. of CA, Riverside, Riverside, CA.Meiotic silencing of unpaired DNA plays an important role in protecting the genome integrity of Neurospora crassa. It is thought to fight against theinvasion of virus and endogenous transposable elements. Our previous work has shown that a 10 KB MULE (mutator-like element)-related DNA27th Fungal Genetics Conference | 237

FULL POSTER SESSION ABSTRACTStransposable element, named sly-1, uniquely exists in the wild type strain OR74A (FGSC#2489) of N. crassa. Here we show that in the cross betweenOR74A and D60 (FGSC#8820), a strain lacking sly-1, the unpaired sly-1 induced the production of small RNAs 4 days after fertilization. The small RNAs weregenerated from both strands of the sly-1 region and demonstrated typical Dicer-processed smallRNA features in Neurospora crassa: 25bp long with astrong preference for uridine at the 5’ end. An RNA-dependent RNA polymerase (SAD-1) was found to be required for such small RNA production (1). Wegenerated draft genome sequencing of D60 with Illumina HiSeq and compared it to the OR74A genome to identify additional unique regions where meioticsilencing of unpaired DNA may have occurred. These unique regions were also found to produce smallRNA with the same features as those from sly-1.These results provide strong support for the endogenous silencing role of meiotic silencing against a natural intact transposable element and describe theRNA interference pathway-involved silencing pattern of meiotic silencing. 1) Shiu PK, Raju NB, Zickler D, Metzenberg RL. Cell 2001; 107(7):905-16.Pathogenic and Mutalistic Interactions476. Functional Characterization of Small, Cysteine-Rich Secreted Effectors from the Filamentous Fungus Magnaporthe oryzae. William C. Sharpee,Yeonyee Oh, Bill Franck, Ralph A. Dean. Plant Pathology, NC State University, Raleigh, NC.The filamentous fungus Magnaporthe oryzae is the most destructive pathogen of rice worldwide. It is described as having two distinct lifestyles withinthe host plant: a biotrophic phase during the early stages of infection followed by a necrotrophic phase characterized by host cell death and lesionformation. To identify candidate effector proteins that may contribute to pathogenesis, the genome of M. oryzae strain 70-15 was mined for predictedproteins that contain a signal peptide, have greater than 3% cysteine content, and are less than 250 amino acids in length. These criteria were selectedbased upon the characteristics of known effectors from other plant-pathogenic fungi and oomycetes. To investigate the roles of these candidates in thebiotrophic or necrotrophic phases of infection, they were transiently expressed in Nicotiana benthamiana leaves via agroinfiltration. When expressedwithin plant cells, candidate effectors that induce necrosis in the N. benthamiana leaves could potentially act as inducers of host cell death during thenecrotrophic phase of infection. Conversely, candidate effectors that prevent necrosis when co-infiltrated with known inducers of host cell death arepotentially involved in suppressing host plant defenses and therefore may contribute to the biotrophic phase of infection. Of 70 candidate effectors testedto date, 10 were found to induce necrosis when transiently expressed in N. benthamiana. In addition, to test for suppression of host cell death, candidateeffectors are currently being co-agroinfiltrated with the BAX gene, a known inducer of host cell death in both plant and mammalian cells, or a knownnecrosis inducer from M. oryzae. Those candidates that show an interesting phenotype will be selected for further characterization as potential effectorsby analyzing their expression in planta and activity when expressed within rice protoplasts.477. Penetration-specific effectors from Phytophthora parasitica favour plant infection. Edouard Evangelisti 1* , Benjamin Govetto 2 , Naima Minet-Kebdani 1 , Marie-Line Kuhn 1 , Agnes Attard 1 , Franck Panabieres 1 , Mathieu Gourgues 1 . 1) UMR Institut Sophia Agrobiotech, INRA/CNRS/Université de Nice,Sophia Antipolis, France; 2) Institut Méditerranéen de Biodiversité et d'Écologie marine et continentale (IMBE), CNRS-INEE - IRD -Aix Marseille Université -Université d'Avignon - Institut Pytheas.Oomycetes are major crop pests which cause million dollars losses every year. To date only a few efficient chemicals are available against thesefilamentous microorganisms. A better understanding of the molecular events occuring during plant-oomycete interactions will help to propose newstrategies for crop protection. We performed a transcriptional analysis in order to identify oomycete penetration-specific genes and identified a set ofpenetration-specific effectors (PSE) bearing a RXLR motif. This motif was previously shown to promote effector import into plant cells during the biotrophicstage in feeding structures called haustoria. Here we report the functional analysis of three candidate genes, referred to as PSE1, PSE2 and PSE3. The threeeffectors were able to abolish plant defense responses when transiently expressed in Nicotiana plants. Moreover, constitutive expression of PSE1 andPSE3 in A. thaliana led to an enhanced susceptibility to P. parasitica infection suggesting a role for these proteins in P. parasitica pathogenicity. TransgenicArabidopsis lines accumulating PSE1 protein showed several developmental perturbations that were associated with altered auxin physiology. Rootgrowth inhibition assays showed that auxin signaling pathway is not altered by PSE1 accumulation. Nevertheless, the coiled-root phenotype and theenhanced susceptibility of PSE1-expressing lines to P. parasitica were reverted by synthetic auxin 2,4-D supply, or treatment with the auxin efflux inhibitorTIBA suggesting that a reduced auxin accumulation is responsible for these phenotypes. This hypothesis was confirmed by a reduced activity of the pDR5auxin sensitive promoter at the root apex. The alteration of the expression pattern observed for two auxin efflux carriers, PIN4 and PIN7 suggests that aperturbation of auxin efflux could be responsible for the PSE1 associated defects. We proposed that PSE1 could favour P. parasitica virulence byinterfering with auxin content. Our results show that penetration specific effectors can modulate general plant functions to facilitate plant infection.Perturbation of hormone physiology was previously reported for other plant pathogens, including nematodes and bacteria, supporting the hypothesis thatinfection strategies from distant pathogens species could converge onto a limited set of plant targets.478. Transcriptional regulatory circuits necessary for appressorium-mediated plant infection by Magnaporthe oryzae. Miriam Oses- Ruiz, Darren M.Soanes, Nicholas J. Talbot. University of Exeter, Exeter, United Kingdom.Rice blast disease is caused by the fungus Magnaporthe oryzae and is the most destructive disease of cultivated rice. The pathogen elaborates aspecialized infection structure called the appressorium. The morphological and physiological transitions that lead to appressorium formation of M. oryzaeduring plant infection are stimulated through perception of environmental signals including surface hydrophobicity and hardness, and the presence ofcutin monomers and leaf surface waxes. The fungus perceives and internalizes these stimuli by a variety of intracellular MAP kinase signaling pathways.The homeobox and C2/H2 Zn finger domain transcription factor, MST12 (ScSte12 homogue) is part of the PMK1 MAP kinase signalling pathway, which isrequired for appressorium formation and invasion. The Mst12 null mutant is able to form completely normal melanised appressoria but it is nonpathogenic. The Mst12 null mutant is unable to form a penetration peg and therefore to cause disease in the rice plant. To understand the mechanism ofthe penetration peg formation, we have recently carried out genome-wide comparative transcriptional profiling analysis for mst12 null mutant using RNAseqand HiSeq 2000 sequencing. In this way, we will show the transcriptional signature associated with penetration peg differentiation in the rice blastfungus. Moreover we will show the set of genes that are likely to be MST12 regulated and therefore help define the regulatory circuits necessary forappressorium-mediated plant infection by plant pathogenic fungi.479. Differential activation of ammonium transporters during the accumulation of ammonia by Colletotrichum gloeosporioides and its effect onappressoria formation and pathogenicity. Dov B. Prusky 1 , Chen Shnaiderman 1 , Itay Miyara 1 , Ilana Kobiler 1 , Sherman Amir 2 . 1) Post Harvest Sci, AgriculturalRes Org, Bet Dagan, Israel; 2) Genomic Unit, Plant Sciences Institute, ARO, Bet Dagan, Israel.Ammonium secreted by the post-harvest pathogen Colletotrichum gloeosporioides during host colonization accumulates in the host environment due to238

FULL POSTER SESSION ABSTRACTSenhanced fungal nitrogen metabolism. Two types of ammonium transporter encoding genes, AMET and MEP, are expressed during pathogenicity. Genedisruption of AMET- a gene modulating ammonia secretion, showed twofold reduced ammonia secretion and 45% less colonization on avocado fruits,suggesting a contribution to pathogenicity. MEPB a gene modulating ammonium transport is expressed by C. gloeosporioides during pathogenicity andstarvation conditions in culture. Gene disruption of MEPB, the most highly expressed gene of the MEP family, resulted in twofold overexpression of MEPAand MEPC but reduced colonization, suggesting MEPB expression's contribution to pathogenicity. Analysis of internal and external ammonia accumulationby DmepB strains in mycelia and germinated spores showed rapid uptake and accumulation, and reduced secretion of ammonia in the mutant vs. WTstrains. Ammonia uptake by the WT germinating spores, but not by the DmepB strain with compromised ammonium transport, activated cAMP andtranscription of PKA subunits PKAR and PKA2. DmepB mutants showed 75% less appressorium formation and colonization than the WT, which waspartially restored by 10 mM exogenous ammonia. Thus while both AMET and MEPB genes modulate ammonia secretion, only MEPB contribute toammonia accumulation by mycelia and germinating spores that activates the cAMP pathways, inducing the morphogenetic processes contributing to C.gloeosporioides pathogenicity.480. Functional analysis of Nbs1 of Magnaporthe oryzae. K. Sasaki 1 , K. Amano 1 , T. Sone 2 , M. Narukawa 1 , T. Kamakura 1 . 1) Applied Biological Science,Tokyo Univ. of Science, Noda, Chiba, Japan; 2) Research Faculty of Agriculture, Hokkaido University, Sapporo, Japan.The filamentous fungus Magnaporthe oryzae causes rice blast, the most serious disease that affects global rice production. On the surface of host plant,a specialized infection structure called appressorium is formed on tip of germ tube. Induction of the development of appressorium requires severalexternal stimulants and a complete cycle of cell division. Although many studies have revealed some of process of appressorium formation in M. oryzae,the complete mechanism is still obscure. We selected Nbs1 from germ tube expressing cDNA library and made Nbs1 disruptants. The cDNA library mainlycontains the genes that express in the period of germ tube development and/or appressorium formation. Nbs1 is presumed to have forkhead associated(FHA) domain, which is contained in many proteins that are involved in DNA repair and cell cycle. In our previous study, Nbs1 disruptants showed growthdelay, abnormality of conidia formation and nuclear division, reduction of germination rate and appressorium formation rate, abnormal pigmentation andhigh sensitivity to DNA-damaging agents. Although Neurospora crassa knock-out mutants of rcaA, which share sequence similarities with Nbs1, showedsimilar phenotypes to Nbs1 disruptants, rcaA did not seem to contain FHA domain. Toward further study of the function of Nbs1, we induced a plasmidcarrying an rcaA (pNB51) or FHA domain-deleted Nbs1 (pCB51dF) into Nbs1 disruptants. Consequently, pNB51 and pCB51dF were able to partiallycomplement phenotypes of Nbs1 disruptants. This result suggested that rcaA has at least partial similar functions of Nbs1 in N. crassa and anotherfunctional domain exists in Nbs1.481. Influence of hypoxia on antifungal susceptibility, sterole pattern and biomarker release of Aspergillus spp. Ulrike Binder 1 , Elisabeth Maurer 1 ,Christoph Müller 2 , Franz Bracher 2 , Cornelia Lass-Flörl 1 . 1) Division of Hygiene and Medical Microbiology, Medical University Innsbruck, Innsbruck, Tirol,Austria; 2) Department of Pharmacy, Ludwig Maximilians University Munich, Germany.Invasive aspergillosis (IA) is a major life-threatening disease in immunocompromised patients, with mortality rates from 40% up to 90% in high-riskpopulations. The most common species causing aspergillosis is Aspergillus (A.) fumigatus, accounting for approximately 90% of infections. Depending onregional distinctions, A. flavus and A. terreus are frequently reported. During infection, fungal pathogens must adapt to microenvironmental stresses,including hypoxia as well as high CO2 levels. Such oxystress conditions are usually not taken into account in current in vitro models of infection, theassessment of antifungal sensitivities or the release of biomarkers used for diagnosis. Therefore, we compared the in vitro activity of amphotericin B(amB), different azoles and echinocandins in hypoxic conditions (1% O2, 5% CO2) to their activity in normoxic conditions against isolates of A. fumigatusand A. terreus and other aspergilli. Using Etest strips, we found a reduction of the minimal inhibitory concentration (MIC) for amB for all aspergilli inhypoxic conditions. Similarly, a significant reduction in the MIC for all tested azoles was demonstrated for A. terreus isolates, while for A. fumigatusisolates differences were less pronounced. For echinocandins, little or no change in the MEC (minimal effective concentration) was detected betweenhypoxic and normoxic conditions for all aspergilli. Most interestingly, A. terreus strains, that are resistant to amB in normoxia, exhibited sensitivity to amBin hypoxic conditions, defining a breakpoint of > 2 mg/ml. Notably, for none of the strains tested, MIC/MEC values increased in hypoxia. Currently we areinvestigating if changes in the sterole pattern or the amount of ergosterol contribute to these changes in antifungal susceptibility in hypoxia. The detectionof circulating fungal antigens in serum for Aspergillus galactomannan or b-D-glucan has become an accepted diagnostic strategy. However, sensitivity andspecificity vary extremely and the reasons are only partially clear; therefore, we are currently checking whether hypoxia influences the physiologicalkinetics of GM and b-glucan release.482. Sit and wait: Special features of Aspergillus terreus in macrophage interactions and virulence. M. Brock 1 , I.D. Jacobsen 2 . 1) MicrobialBiochemistry/Physiology, Friedrich Schiller University and Hans Knoell Institute, Jena, Germany; 2) Molecular Pathogenicity Mechanisms, Hans KnoellInstitute Jena, Germany.While Aspergillus fumigatus is known as the main cause of invasive pulmonary aspergillosis in immunocompromised patients, Aspergillus terreus is anemerging pathogen prevalent in some local hot spots. When tested in embryonated egg or murine infection models A. terreus required substantiallyhigher infectious doses compared to A. fumigatus to cause high mortality rates. Furthermore, when A. fumigatus and A. terreus infections were followedby in vivo imaging using bioluminescent reporter strains, germination and tissue invasion of A. terreus was significantly delayed. To elucidate differences inmore detail, the interaction of A. terreus and A. fumigatus with macrophages was compared. A. terreus was phagocytosed significantly faster, whichappears mainly due to higher exposure of galactomannan and glucans on the surface of conidia. Additionally, although phagocytosis of both speciesresulted in phagolysosome maturation, A. fumigatus efficiently inhibited acidification, which was not the case for A. terreus. However, within this acidicenvironment of phagolysosomes A. terreus showed long-term persistence without significant inactivation of conidia. Further analyses revealed thatinefficient blocking of acidification by A. terreus was due to differences in the spore colour pigment of both species. Recombinant production of anaphthopyrone synthase from Aspergillus nidulans enabled A. terreus to inhibit the acidification to a similar extent as observed for A. fumigatus. Thisalteration of the phagolysosomal environment resulted in an increased escape from macrophages and was accompanied by increased virulence in amurine infection model. We speculate that the long-term persistence of A. terreus wild-type strains in acidified phagolysosomes might be responsible forhigh dissemination rates observed in infected human patients, because A. terreus might hitchhike inside immune effector cells to reach secondary sites ofinfection.483. Identification and characterization of an RXLR-like effector family from medically relevant fungi. Shiv D. Kale 1* , Kelly C. Drews 1,2 , Helen R. Clark 1,3 ,Hua Wise 1,4 , Vincenzo Antignani 1 , Tristan A. Hayes 1,2 , Christopher B. Lawrence 1,2 , Brett M. Tyler 4,5 . 1) Virginia Bioinformatics Institute, Virginia Tech.,Blacksburg, VA; 2) Department of Biological Sciences, Virginia Tech., Blacksburg, VA; 3) Department of Biochemistry, Virginia Tech., Blacksburg, VA; 4)27th Fungal Genetics Conference | 239

FULL POSTER SESSION ABSTRACTSCenter for Genome Research and Biocomputing, Oregon State University, Corvallis, OR; 5) Department of Botany and Plant Pathology, Oregon StateUniversity, Corvallis, OR.Fungal infections have become an increasingly significant problem for immunocompromised individuals, transplant recipients, the elderly, several casesinvolving healthy individuals. There is a significant growth in incidences of morbidity and mortality associated with medically important fungi, specificallyAspergillus species. Aspergillus fumigatus virulence has been attributed to production of pigments, adhesins on the surface of the cell wall, secretedproteases, and mycotoxins. Current treatments consist of oral corticosteroids, antifungal medications, and/or surgery to remove aspergillomas. Many ofthese treatments have substantial shortcomings. Detection and diagnosis is also weighty problem as most clinical tests take weeks for results allowing theinfection to proceed. Appropriately, the paradigm for human fungal interactions has been focused on the host deficiencies mediating virulence ofopportunistic pathogenic fungi. There has been substantial progress in identifying and characterizing secreted proteins (effectors) from bacterial,oomycete, and fungal plant pathogens. A subset of these effector proteins are able to enter host cells and modulate host intracellular functions. Using ourbioinformatics pipeline we have been able to identify a family of secreted proteins from A. fumigatus sharing a conserved N-terminal RXLR-like motif. Wefound this family is expanded amongst primary fungal pathogens. The RXLR and RXLR-like motifs from known intracellular effectors of plant pathogenicand mutualistic oomycetes and fungi have been shown to facilitate effector entry into plant cells via binding external phosphatidylinositol-3-phosphate(PI3P). Here we describe AF2, a candidate effector from A. fumigatus that contains a N-terminal RxLR-like motif. Through the use of confocal microscopyand flow cytometry we show AF2 is rapidly able to enter several primary and immortalized mammalian cell lines. Through the use of isothermal titrationcalorimetry and liposome binding assays we show AF2 has nanomolar binding affinity for PI3P, and does not bind other mono or poly-PIPs that we havetested thus far. Based on our bioinformatics and biochemical analysis we postulate AF2 is a secreted effector protein capable of rapidly translocating intomammalian cells. We will present our latest findings on the physiological relevance of AF2.484. A role for PalH-mediated signal transduction in A. fumigatus virulence and cell wall integrity: An exploitable target for combination therapy? M.Bertuzzi 1 , C.M. Grice 1 , L. Alcazar-Fuoli 2 , A.M. Calcagno-Pizarelli 1 , J. Kalchschmidt 1 , S. Gill 1 , K. Fox 1 , A. Cheverton 1 , Hong Liu 3 , V. Valiante 4 , E.A. Espeso 5 , S.GFiller 3 , A. Brakhage 4 , E.M. Bignell 1 . 1) Centre for Molecular Bacteriology & Infection , Imperial College London , London (UK); 2) Mycology Referencelaboratory, National Centre for Microbiology, Instituto de Salud Carlos III, Madrid (Spain); 3) David Geffen School of Medicine at UCLA, Division ofInfectious Diseases, Los Angeles Biomedical Research Institute at Harbor-UCLA Medical Center (USA); 4) Leibniz-Institute for Natural Product Research andInfection Biology Hans Knöll Institute, Molecular and Applied Microbiology, Jena (Germany); 5) Dept. of Cellular and Molecular Biology, AspergillusMolecular Genetics Unit, Centro de Investigaciones Biológicas (C.S.I.C.), Madrid (Spain).Adaptation to host-imposed stress is a crucial requirement for persistence of Aspergillus fumigatus in the mammalian lung. In Aspergillus species, PacCsignalling promotes tolerance of alkaline environments via signal-dependent proteolytic processing of the transcription factor PacC. The aim of this studywas to test the requirement for A. fumigatus PalH during infection and to decipher its role in PacC-mediated signalling. The role of PalH in alkalinemediatedPacC processing was tested using electrophoretic mobility shift assay, and A. fumigatus virulence was examined in a murine neutropenic modelof pulmonary aspergillosis. To probe the mechanistic basis of PalH-mediated signalling, we utilised a split-ubiquitin Membrane Yeast Two-Hybrid (MYTH)assay to assess protein interactions amongst candidate A. fumigatus signalling proteins of this pathway. A. fumigatus isolates expressing epitope-taggedPalH protein were constructed to assess the relevance of PalH oligomerisation. Analysis of PacC processing identified the requirement for PalH to initiatealkaline-mediated PacC signalling. A DpalH mutant is somewhat sensitive to alkaline pH, and attenuated for virulence in a murine model of pulmonaryaspergillosis. The mutant is also sensitive to cell wall-perturbing agents, and in the presence of the cell wall-active antifungal caspofungin undergoesextensive hyphal branching and ballooning compared to the parental and reconstituted strains. In the absence of PalH A. fumigatus-mediated damage ofepithelial cells is abrogated in vitro. By using a MYTH assay a significant interaction between A. fumigatus PalH and PalF was detected in Saccharomycescerevisiae. In A. fumigatus PalH-mediated PacC signalling, likely implemented in a (PalF) arrestin-like manner, commands a central role in the expression ofvirulence-determining functions. The impairment of PacC signalling exerts a synergistically inhibitory effect upon fungal viability in the presence of cellwall-active antifungal drugs and therefore represents an attractive target for the development of novel antifungal mono- and combination therapies. Ourresults support a scenario whereby PalH is an oligomerising receptor, responsive to extracellular pH, and required for virulence and echinocandintolerance. Future studies will focus upon the mechanism of PalH-mediated pH sensing.485. Aspergillus fumigatus trehalose-6-phosphate regulates innate immune responses and virulence through modulation of fungal cell wallcomposition. Srisombat Puttikamonkul 2 , Vishu K. Amanianda 3 , Jean-Paul Latge 3 , Kelly M. Shepardson 2 , John R. Perfect 4 , Nora Grahl 2 , Bridget M. Barker 2 ,Robert A. Cramer 1 . 1) Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, NH; 2) Immunology and Infectious Diseases,Montana State University; 3) Unite des Aspergillus, Institut Pasteur; 4) Medicine, Division of Infectious Diseases, Duke University Medical Center.Mechanism(s) behind the attenuated fungal virulence of trehalose biosynthesis pathway mutants are not fully understood. We observed previously thatTPS2/OrlA, a key enzyme in TPS1/TPS2 trehalose biosynthesis is required for cell wall integrity and fungal virulence in A. fumigatus. In this study, we testedthe hypothesis that the significant in vivo attenuated virulence and in vitro impaired cell wall integrity of DorlA is due to accumulation of Trehalose-6-Phosphate (T6P). Our data suggest that the mechanism behind the attenuated virulence of the A. fumigatus TPS2 null mutant, DorlA, in a murine model ofX-linked chronic granulomatous disease (X-CGD) is mediated by an increased susceptibility of DorlA to polymorphonuclear leukocyte (PMN) killing. In theabsence of PMNs in the xCGD murine model, DorlA exhibited restored fungal burden and virulence similar to wild-type inoculated animals. Null mutationsin putative trehalose biosynthesis proteins TslA and TslB in the DorlA background were able to ameliorate T6P accumulation and restore cell wall integrityand virulence strongly suggesting that accumulation of T6P is the key factor associated with DorlA virulence. Our results identify a previously unknownmechanism of immune modulation by the fungal carbohydrate metabolite T6P that has significant implications for targeting trehalose biosynthesis as anantifungal drug target.486. Fungal lipoxygenases: a novel instigator of asthma? Gregory J. Fischer 1 , Katharyn Affeldt 3 , Erwin Berthier 2 , Nancy P. Keller 1,2,3 . 1) Department ofGenetics, University of Wisconsin-Madison, Madison, WI; 2) Department of Medical Microbiology and Immunology, University of Wisconsin-Madison,Madison, WI; 3) Department of Bacteriology, University of Wisconsin-Madison, Madison, WI.Statement of Purpose: Fungi have long been associated with asthmatic diseases, yet the exact mechanism(s) by which fungi induce asthma is unknown.We propose that fungal lipoxygenase enzymes and their eicosanoid products are involved in asthmatic diseases. Human 5-lipoxygenase derivedleukotrienes induce inflammation, mucus secretion, vasodilation, and bronchial constriction. We hypothesize that the fungal pathogen Aspergillusfumigatus is capable of secreting a 5-lipoxygenase homolog, LoxB, that participates in eicosanoid production, including leukotrienes. This secretedhomolog is translocated into lung epithelial cells, participates in the production of leukotriene and other eicosanoids, and exacerbates asthmaticresponses, such as bronchoconstriction. Together, this work will help delineate the role fungal products play in asthmatic diseases. Methods: We are240

FULL POSTER SESSION ABSTRACTSassessing fungal interactions with lung epithelial cells using a microfluidic in-vitro platform followed by murine asthma model research. To assess theeffects of LoxB overexpression, mass spectrometry was used to identify eicosanoid oxylipins within culture supernatants. Results: We have identified anAspergillus fumigatus lipoxygenase, LoxB, with high identity to human 5-lipoxygenase. Moreover, we have identified a motif in LoxB that may mediateentry into lung epithelial cells. To fully understand the impact of LoxB in asthma, we have developed an Aspergillus fumigatus strain that overexpressesLoxB. Overexpression of LoxB results in increased levels of various eicosanoids that are known to cause airway hyperresponsiveness and increased mucusproduction. Future work will focus on characterizing the effect these eicosanoid products have on the airway and whether fungal effector translocationresult in increased leukotriene levels.487. F-box protein 15 (Fbx15) links virulence of Aspergillus fumigatus to protein degradation and stress response. Bastian Jöhnk 1 , Özgür Bayram 1 , OliverValerius 1 , Thorsten Heinekamp 2 , Ilse D. Jacobsen 3 , Axel A. Brakhage 2 , Gerhard H. Braus 1 . 1) Institute for Microbiology and Genetics, Department ofMolecular Microbiology and Genetics, Georg August University, Göttingen, Germany; 2) Department of Molecular and Applied Microbiology, LeibnizInstitute for Natural Product Research and Infection Biology (HKI) and Friedrich Schiller University, Jena, Germany; 3) Department for MicrobialPathogenicity Mechanisms, Leibniz Institute for Natural Product Research and Infection Biology (HKI) and Friedrich Schiller University, Jena, Germany.Rapid adaptation to a versatile host represents a challenge for the opportunistic human pathogen Aspergillus fumigatus for successful infection. F-boxproteins are the adaptor subunits of E3 SCF (Skp1 cullin-1 F-box protein) ubiquitin ligases. They recognize target proteins, which are marked by the SCFcomplex for degradation in the 26S proteasome. Here we have identified Fbx15 as an F-box protein, which links A. fumigatus virulence to proteindegradation. A. fumigatus deletion strains which have lost fbx15 are unable to infect immunocompromised mice in a murine model of invasiveaspergillosis. Fbx15 is required for growth during stress including increased temperature, oxidative stress and amino acid starvation. Fbx15 is also requiredfor controlling the synthesis of the antiphagocytic gliotoxin. Fbx15 interacts in the nucleus with the linker protein Skp1/SkpA suggesting that SCF Fbx15primarily targets nuclear proteins. Four nuclear subunits of the COP9 sigalosome are putative Fbx15 interaction partners. We propose an interdependentstabilization of Fbx15 and the COP9 sigalosome, which is required to link protein degradation and stress response to virulence.488. The sfp-type phosphopantetheinyl transferase, PPTA, is critical for the virulence of Aspergillus fumigatus. A. E. Johns, P. A. Warn, P. Bowyer, M. J.Bromley. Inflammation and repair, Univ. of Manchester, Manchester, United Kingdom.Aspergillus fumigatus is the leading cause of invasive aspergillosis (IA), a fungal disease which is increasing annually on a global scale. IA poses as acommon threat to patients with a weakened immune response due to disorders such as leukaemia, HIV, AIDS and also persons undergoing chemotherapytreatments. The ability of A. fumigatus to produce a wide array of secondary metabolites is thought to contribute to the pathogenicity of this organism.We have identified an enzyme, PPTA that plays a key role in secondary metabolism in A. fumigatus. PPTA is a sfp-type phosphopantetheinyl transferaseand is required to activate non-ribosomal peptide synthases, polyketide synthases and a protein required for lysine biosynthesis aminoadipate reductase(AARA). Disruption of pptA prevents the production of most secondary metabolites and renders the fungus avirulent in both insect and murine infectionmodels. To investigate which aspects of pptA activity are essential to virulence a series of knock out mutant strains were generated; DaarA, DpksP andDsidA. These genes play a vital role in lysine, melanin and siderophore biosynthesis pathways respectively. The sidA gene proved vital to virulence in theinsect model whereas the DaarA and DpksP mutants were unaffected. The pathogenicity of both the pptA and sidA knock out strains was restored by coinjectinglarvae with iron. We postulate that, at least in the larval model, it is PPTAs role in siderophore biosynthesis and not the activation of othersecondary metabolism pathways that is critical for the virulence of A. fumigatus.489. Characterization of effectors of the barley pathogen Rhynchosporium commune. Daniel Penselin, Wolfgang Knogge. Stress and DevelopmentalBiology, Institute of Plant Biochemistry, Halle, Germany.R. commune is the causal agent of barley leaf scald. This disease is a persistent threat and widespread in particular in cool and moist barley-growingareas of the world. Yield losses as high as 35-40% have been reported, but a yield loss of only 5% may already lead 2012 to an economic loss of >700 Mio €in Europe. R. commune colonizes the leaves of its host plants by growing beneath the cuticle, mainly in the pectic layer of the outer epidermis cell walls,without directly contacting the plant plasma membrane. Therefore, the fungus needs to secrete effectors to manipulate the host physiology. Previousstudies have shown that three secreted necrosis-inducing proteins (NIP1, NIP2, NIP3) affect fungal virulence in a quantitative manner depending on thehost genotype. NIP1 was also identified as the avirulence factor that is recognized by barley resistance gene Rrs1.After obtaining the genome sequence of R. commune it turned out that NIP1 and NIP3 are encoded by single genes. In contrast, a small family of highlyhomologous NIP2 genes was identified, precluding a simple targeted deletion strategy for further functional analysis of NIP2. In addition, deletion of oneNIP2 homolog affected the expression of the others. For further investigations an approach to simultaneously silence all members of the NIP2 family isbeing followed using a recombination-based cloning strategy. To this end, a plasmid expressing an intron-containing hairpin RNA (ihpRNA) wasconstructed. Transfection of R. commune with the ihpRNA plasmid and qRT-PCR-based assessment of the transcriptional down-regulation of NIP2homologues are in progress. Establishing a gene silencing system will be of great value for future functional studies of fungal effectors involved in plantpathogeninteractions.490. Molecular and genetic basis guiding the establishment of a mutualistic relationship between Epichloë festucae and perennial ryegrass. SladanaBec, JinGe Liu, Christopher L. Schardl. Dept Plant Pathology, Univ Kentucky, Lexington, KY.The relationship established between Epichloë festucae and perennial ryegrass (Lolium perenne) is a model system for studying mutualism betweenendophytes and cool season grasses. E. festucae colonizes all above-ground plant organs, growing by intercalary hyphal extension in elongating grassleaves. During the reproductive phase of growth, the fungus exhibits a dual nature: retaining its benign endophytic growth and seed transmission, orforming external stromata and suppressing seed production on affected tillers. From our previous work regarding the genes involved in the switchbetween benign plant colonization and formation of stromata, we have identified a number of genes encoding small secreted proteins (ssp) that are highlyup-regulated in benign infected inflorescences. Two of those genes, sspB, and sspX, are located in a subtelomeric region, and preliminary evidencesuggests that they may play a role in host specificity. Although E. festucae is reported to be compatible with two related host species, L. perenne andLolium pratense (meadow fescue), strains generated from a series of crosses and backcrosses showed a range of compatibility with L. perenne, butconsistently were compatible with L. pratense. One such strain, E2368, had low compatibility with L. perenne, whereas a subculture (variant E4844)showed improved compatibility with this host. Genomes of E4844 and E2368 were compared, revealing that the variant had lost the subtelomeric regioncontaining sspB and sspX. The possible roles of sspB and sspX, and of other gene losses and genomic changes in the variant, are under investigation. Also,the parents and full siblings of strain E2368 are being tested for SNPs segregating for phenotypes related to the establishment of stable mutualisticsymbioses with L. perenne. The set of progeny strains has been screened for the establishment of host specificity with perennial ryegrass, and is slated for27th Fungal Genetics Conference | 241

FULL POSTER SESSION ABSTRACTSIllumina genome sequencing and subsequent bulk segregant analysis to identify SNP markers correlated with host-specificity phenotypes.491. Puccinia graminis and Brachypodium distachyon: contrasting profiles of host-pathogen incompatibility. Melania Figueroa 1 , Sergei Filichkin 1 , SeanGordon 2 , Henry Priest 3 , John Vogel 2 , David Garvin 4 , Todd Mockler 3 , William Pfender 1 . 1) Oregon State University, Corvallis, OR; 2) USDA-ARS, WRRC,Albany, CA; 3) Donald Danforth Plant Science Center, St. Louis, MO; 4) USDA-ARS, PSRU, St. Paul, MN.The causal agent of stem rust, Puccinia graminis, is a devastating pathogen that affects the production of cereals and temperate-zone grasses. Someimportant examples of P. graminis and their typical hosts are P. graminis f. sp. tritici (Pg-tr) on wheat and barley, P. graminis f. sp. lolii (Pg-lo) on perennialryegrass and tall fescue, and P. graminis f. sp. phlei-pratensis (Pg-pp) on timothy grass. The biological and evolutionary attributes of Brachypodiumdistachyon have led to its development as a model to study cereals and grasses. To assess the applicability of Brachypodium to investigate non-hostresistance to stem rust, disease severity caused by Pg-tr, Pg-pp and Pg-lo was evaluated across a collection of Brachypodium inbred lines. The differentfungal isolate/Brachypodium line combinations demonstrated significant variation in stem rust resistance and revealed the contrasting pathogenic/virulentcharacteristics among these stem rust isolates. Given the distinct phenotypes obtained when inoculating with Pg-tr, Pg-lo or Pg-pp, Brachypodium lineBd1-1 was selected for further analyses. Histological analysis of the early infection events (first 68 h of infection) indicated that Pg-lo and Pg-pp are moreefficient than Pg-tr in establishing a biotrophic interaction, and that Bd1-1 exhibits pre-haustorial resistance to Pg-tr and post-haustorial resistance to Pg-loand Pg-pp. A comparative transcriptome analysis (RNA-Seq) of the early responses of Bd1-1 to Pg-lo, Pg-pp and Pg-tr was performed. Gene expressionprofiles were determined to capture the transcriptional events in response to 1) appressorium formation (12 hpi, hours post-inoculation), and 2) fungalpenetration (18 hpi) and initial stages of fungal growth in the plant mesophyll for each fungal isolate. The data show distinctive profiles for each fungalisolate/Brachypodium combination. Our results demonstrate a significant transcriptional re-programming that leads to the activation of early plantdefenses associated with quantitative resistance (i.e., phenylpropanoid pathway, cytochrome P450s, and different types of transcription factors).Additionally, several receptor-like proteins and uncharacterized proteins were identified as putative players in pathogen recognition.492. Magnaporthe oryzae has evolved two distinct mechanisms of effector secretion for biotrophic invasion of rice. Martha C. Giraldo 1 , Yasin F. Dagdas 2 ,Yogesh K. Gupta 2 , Thomas A. Mentlak 2,4 , Mihwa Yi 1 , Hiromasa Saitoh 3 , Ryohei Terauchi 3 , Nicholas J. Talbot 2 , Barbara Valent 1 . 1) Plant Pathology, KansasState University, Manhattan, KS. USA; 2) School of Biosciences, University of Exeter, EX4 4QD, UK; 3) Iwate Biotechnology Research Center, Kitakami,Iwate, 024-0003 Japan; 4) Cambridge Consultants Ltd, Cambridge, CB4 0DW, U.K.Pathogens secrete effector proteins into host tissue to suppress immunity and cause disease. Pathogenic bacteria have evolved several distinct secretionsystems to target specific effector proteins during pathogenesis, but it was not previously known if fungal pathogens require different secretorymechanisms. We present evidence that the blast fungus Magnaporthe oryzae possesses distinct secretion systems for delivering effector proteins duringbiotrophic invasion of rice cells. M. oryzae secretes cytoplasmic effectors targeted for delivery inside rice cells and apoplastic effectors targeted to theextracellular space. Cytoplasmic effectors preferentially accumulate in the biotrophic interfacial complex (BIC), a novel in planta structure located besidethe tip of the initially filamentous invasive hypha and then remaining next to the first differentiated bulbous invasive hypha cell. In contrast, apoplasticeffectors remain in the extracellular compartment uniformly surrounding the invasive hypha inside the invaded cell. Disruption of the conventional ER-Golgi secretion pathway by Brefeldin A (BFA) treatment blocked secretion of apoplastic effectors, which were retained in the ER, but not secretion ofcytoplasmic effectors. Fluorescence Recovery After Photobleaching experiments confirmed that cytoplasmic effectors continued to accumulate in BICs inthe presence of BFA. Analysis of mutants showed that the BIC is associated with a novel form of secretion involving exocyst components, Exo70 and Sec5,and the t-SNARE Sso1, which are required for efficient delivery of effectors into plant cells and are critical for pathogenicity. By contrast, effectors whichfunction between the fungal cell wall and plant plasma membrane are secreted from invasive hyphae to the apoplast by the ER-Golgi secretory pathwayconserved in eukaryotes. We propose a model for the distinct secretion systems that the rice blast fungus has evolved to achieve tissue invasion.493. Trichoderma rhizosphere’s competency, endophytism and plant communication: A molecular approach. Artemio Mendoza 1 , Johanna Steyaert 1 ,Natalia Guazzone 1 , Maria Fernanda Nieto-Jacobo 1 , Mark Braithwaite 1 , Robert Lawry 1 , Damian Bienkowski 1 , Christopher Brown 2 , Kirstin MacLean 1 , RobertHill 1 , Alison Stewart 1 . 1) Bio-Protection Research Centre, Lincoln University, Lincoln, New Zealand; 2) Biochemistry Department and Genetics Otago,University of Otago, New Zealand.Establishment of root symbiosis is one the key drivers of biocontrol success for members of the fungal genus Trichoderma. This root symbiosis isdescribed as a two-step process, whereby Trichoderma species colonise the soil surrounding the root (rhizosphere) and then penetrate the root tissue andestablish an endophytic relationship. The ability to colonise and then proliferate over time within the rhizosphere is termed rhizosphere competence (RC).There have been numerous reports of Trichoderma biocontrol strains which persist within the rhizosphere for the growing season of the crop plant. Ourresults strongly suggest that RC is widespread among members of the genus Trichoderma and that RC interactions are strain and host plant specific. Forendophytes and their host plants to maintain a mutualistic relationship requires a constant molecular dialogue between the organisms involved. Forexample, the fungal-derived phytohormone, indole acetic acid (IAA), plays an important role in signalling between Trichoderma and the model plantArabidopsis thaliana. There are however, additional, currently unknown, chemical signals which may be even more important for a positive interactionbetween Trichoderma and plants. By using a soil-maize-Trichoderma as a model system in in situ sterile conditions we are currently analysing the RC andendophytism transcriptomes of two Trichoderma species: T. virens and T. atroviride. Using a combination of bioinformatics, quantitative RT-PCR (for stagespecific genetic markers from Trichoderma) and fluoro-labelled Trichoderma strains we are currently identifying and analysing promising Trichodermacandidates involved in endophytism and RC. A comprehensive panorama of the Trichoderma-soil-plant interaction will be discussed in this conference.494. Ustilago bromivora - Brachypodium distachyon: a novel pathosystem. Franziska Rabe, Regine Kahmann, Armin Djamei. Organismic Interactions, MaxPlanck Institute for Terrestrial Microbiology, Marburg, Germany.The Ustilago maydis - Zea mays pathosystem is a well established model system to investigate basic principles of biotrophic plant-pathogen interactions.However, due to the long generation time, space requirements, and difficulties in transformation of maize studying the plant side is demanding. Recently,it has been shown that the yet uncharacterized smut fungus Ustilago bromivora infects Brachypodium distachyon, a model grass species. Short generationtime, small size, sequenced diploid genome, and accessible reverse genetics make this monocot highly suitable for the analysis of biotrophic interactionswith focus on the pathogen as well as the plant side. The primary goal of this study is therefore the characterization of U. bromivora and the interactionwith its host in order to evaluate the suitability of the U. bromivora - B. distachyon interaction as a new host-pathogen model system.We could show that haploid U. bromivora strains obtained after spore germination can be cultivated and transformed with self-replicating plasmids usedfor U. maydis transformation. A hallmark of smut fungi is that the pathogenic form is the dikaryon which arises after mating of compatible haploids.Haploid cells are produced when diploid spores germinate and undergo meiosis. Usually after germination of a single teliospore, cells with compatible242

FULL POSTER SESSION ABSTRACTSmating types can be isolated. After germination of U. bromivora spores (kindly provided by T. Marcel, INRA-AgroParisTech, France) all haploid progenieswere of the a1 mating type. This suggests a mating type bias where the a2 mating type might be linked to a deleterious recessive allele making theisolation of strains harboring this mating type under laboratory conditions impossible. Hence, the genomes of several strains harboring the a1 mating typeand of diploid spore material containing both mating types were sequenced via an Illumina-Next-Generation Sequencing approach. Based on thesesequences we plan to reconstruct the a2 mating type and to generate a strain containing this mating type as well as a solopathogenic haploid strain.495. T-DNA mediated insertional mutagenesis: evidence of a new gene implied in the early phase of pathogenic development of Botrytis cinerea.Nathalie Poussereau 1* , Eytham Souibguy 1 , Marie-Pascale Latorse 2 , Geneviève Billon-Grand 1 , Cindy Dieryck 1 , Vincent Girard 1 , Adeline Simon 3 , Muriel Viaud 3 ,Julia Schumacher 4 , Paul Tudzinsky 4 . 1) Unité Mixte CNRS BayerSAS, Université LYON I, 14 impasse Pierre Baizet, BP 99163, 69263, Lyon cedex09, France; 2)Centre de Recherche Bayer SAS 14 impasse Pierre Baizet, BP 99163, 69263, Lyon cedex09, France; 3) BIOGER, INRA Versailles, route de Saint Cyr, 78026,Versailles France; 4) Institut fûr Biologie and Biotechnolgie der Pflanzen, West.Wihelms-universitât, Hindenburgplatz 55, 48143 Mûnster, Germany.A collection of mutants of the grey mould fungus Botrytis cinerea has been constructed in order to provide the support for the identification of bothfungal functions that are essential for the pathogenic development and/or plant defence traits raised in the host plant. A random insertional mutagenesisstrategy based on the Agrobacterium tumefaciens-mediated transformation (ATMT) is used to enlarge an existing mutant library (2367 lines, Giesbert etal. 2011). 2144 additional T-DNA integrated transformants have been generated. The insertion sites of the T-DNA are being determined using TAIL-PCR andcapacity to infect the host plant is assayed. These data are organized into a genome-orientated database of tagged genes and will be soon available for thescientific community. One exploitation of this mutant library focuses on the characterization of mutants whose parasitic development in planta ishampered. We present here an example of the study of a new gene encoding a DnaJ domain protein. The T-DNA mutant exhibited a drastic alteration ofthe infectious process on bean leaves. Deletion of the studied gene confirmed this phenotype and revealed that colonization process was also altered ondifferent host plants. A defect in penetration and an abnormal infection cushion formation were registered. A dramatic reduced conidiation and anabnormal hyphal morphology were also observed. Resistance/sensitivity to ROS, formation of ROS, organic acids and cell wall degrading enzymes secretionwere investigated. Finally, proteomic analyses are currently developed in order to attribute a function to this gene.496. The NADPH Oxidase Complexes in Botrytis cinerea. Ulrike Siegmund, Jens Heller, Sabine Giesbert, Paul Tudzynski. IBBP, WWU Muenster, Muenster,Germany.Reactive oxygen species (ROS) are generated in all aerobic environments and therefore play a major role for many organisms depending on oxygen. Forexample they act as messenger molecules for intercellular signaling or play a role during defense mechanisms against pathogens (Takemoto et al., 2007).One good example is the oxidative burst; plants rapidly produce large amounts of ROS as the first defense reaction towards pathogen attacks. NADPHoxidases (Nox) are the most common enzymatic system to produce these ROS. Nox are enzyme complexes, which transport electrons through biologicalmembranes and therewith reduce oxygen to superoxide. In fungi they are shown to be involved in differentiation processes and pathogenicity and aretherewith in our focus to gain insights into plant - fungi interactions. In the phytopathogenic fungus Botrytis cinerea two NADPH oxidases (BcNoxA andBcNoxB) as well as their putative regulator (BcNoxR) were previously identified (Segmueller et al., 2008). Besides their involvement in pathogenicity andsclerotia production, deletion studies have revealed that BcNoxA and BcNoxR are also involved in hyphal germling fusions (Roca and Weichert et al., 2011).Recent analyses show a localization of the catalytical subunits BcNoxA and BcNoxB to the ER and partly to the plasma membrane of hyphae, while theregulator BcNoxR is localized in vesicles and at the hyphal tips. Nox are multi-enzyme complexes, whose regulatory process and the participating proteinsare well described in mammals. However, in fungi not all components have been identified, yet. For B. cinerea interaction studies with potentialcandidates identified the small GTPase Rac, the GEF BcCdc24, the scaffold protein BcBem1 and the PAKs BcCla4 and BcSte20 as interacting proteins withinthe BcNox complex. Roca M.G. and Weichert M. et al., (2012) Fungal Biol 116(3): 379-387. Segmueller N. et al., (2008) Mol Plant Microbe Interact 21: 808-808-819. Takemoto D. et al., (2007) Fungal Genet Biol 44(11): 1065-1076.497. A putative function of small RNAs in the plant pathogen Botrytis cinerea. Arne Weiberg, Ming Wang, Hailing Jin. Plat Pathology & Microbiology, UCRiverside, Riverside, CA.Small RNAs (sRNAs) are a class of non-coding transcripts that regulate gene expression. sRNA-directed gene regulation is a common phenomenon ineukaryotes, and in fungal systems function in differentiation, genome defense, and heterochromatin formation has been described. However, it isunknown in any systems whether sRNAs play an important role in fungal pathogenicity. To study sRNAs in the plant pathogen Botrytis cinerea we arecurrently undertaken a dual approach: I) sRNA deep sequencing was performed analyzing RNA profiles of fungal in vitro culture samples and Botrytisinfectedplant tissues using two host systems, the model plant Arabidopsis thaliana and tomato (Solanum lycopersicum). The goal is to identify infectionspecificB. cinerea-derived sRNAs (Bc-sRNAs). II) Genetic analysis of important sRNA biogenesis factors in B. cinerea is currently piloted. B. cinereapossesses all relevant RNAi components including two Dicer-like genes (Bc-DCL1 and Bc-DCL2) and two Argonaute-like genes (Bc-AGO1 and Bc-AGO2).Targeted gene disruption by homologous recombination of Bc-DCL1 and Bc-DCL2 led to growth retardation on artificial media and to delay of massiveconidiospore production. In planta, no reduction in virulence was observed. However, a dcl1dcl2 double mutant was strongly impaired in virulence andwas unable to produce a set of Bc-sRNAs. Taken our observations together, it is proposed that B. cinerea expresses Bc-sRNAs during infection in order toregulate important processes to facilitate pathogenesis.498. The Role of Quorum-sensing Molecules in Interactions between Candida albicans and its Host. Jessica C. Hargarten 1 , Thomas M. Petro 2 , Kenneth W.Nickerson 1 , Audrey L. Atkin 1 . 1) School of Biological Sciences, University of Nebraska, Lincoln, Lincoln, NE; 2) Department of Oral Biology, University ofNebraska Medical Center, Lincoln, NE.Candida albicans is a polymorphic fungus that is capable of causing the life threatening disease Candidiasis once it reaches the bloodstream of asusceptible host. The capability to switch between morphologies, and its ability to synthesize and secrete the quorum sensing molecule (QSM) farnesol areknown virulence factor. Previously, we showed that C. albicans mutants that produced less farnesol are less pathogenic to mice than their parental strainin a tail vein assay. Also, oral administration of farnesol to the mice prior to infection increased mortality. In contrast, farnesol blocks the yeast to myceliatransition in vitro, which should have a protective effect. These observations pose the dilemma of finding a mechanism whereby a molecule which blocksthe yeast to mycelia transition can also act as a virulence factor. We hypothesize that farnesol functions as a virulence factor by modulating the hostinnate immune response. Distinct Candida morphologies elicit different host immune responses. Both white and opaque cells stimulate leukocytemovement, but only white cells secrete a small molecular weight chemoattractant that draws the leukocyte directly towards the white cell and stimulatesengulfment by mouse macrophages. The white cells are also less susceptible to killing by human macrophages and neutrophils than opaque cells, possiblydue to their increased capabilities of escape once phagocytosed. The chemical identity of this chemoattractant is currently unknown, but the reason27th Fungal Genetics Conference | 243

FULL POSTER SESSION ABSTRACTSbehind its continued secretion by white cells is intriguing. One likely candidate is farnesol because opaque cells, unlike white cells, do not accumulatedetectable levels of farnesol. Macrophages are capable of detecting and responding to exogenous farnesol. Earlier our group reported that farnesolstimulates the expression of both pro-inflammatory and regulatory cytokines by mouse macrophage. The production of these warning signals is animportant indicator of how the body ultimately hopes to clear the infection. Others have shown that farnesol suppresses the anti-Candida activity ofmacrophages through its cytotoxic effects, thus making it all the more difficult to eliminate the fungus early in infection. Here we report the in vitro role offarnesol and other known QSM in macrophage chemotaxis and relative phagocytosis of C. albicans.499. The Role of ISW2 for in vitro and in vivo Chlamydospore Production in Candida albicans. Ruvini U. Pathirana 1 , Dhammika H. M. L. P. Navarathna 2 ,David D. Roberts 2 , Kenneth W. Nickerson 1 . 1) School of Biological Sciences, University of Nebraska - Lincoln, Lincoln, NE; 2) Laboratory of Pathology, Centerfor Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD.The production of chlamydospores is an unusual feature in the medically important opportunistic pathogen Candida albicans which is commonly used asan in vitro diagnostic tool. These thick walled spherical structures arise from a filament tip which is termed a suspensor cell. In the process of evolution, itis hard to believe that C.albicans makes a spore that does not contribute to its biology and thus the function of chlamydospores is of interest. Upon carefulobservation of the chronic stage of C.albicans colonization in mouse kidneys, we often find large cells similar in appearance to chlamydospores. Wecharacterized these large cells using sucrose density gradients and compared them with in vitro induced chlamydospores. The in vivo cells had the samebuoyancy and were physiologically similar to in vitro chlamydospores. So we hypothesized that chlamydospores may promote the persistence of thesepathogens during pathogenesis, particularly in kidneys. To test the role of chlamydospores during host infection, we used the wild type strain SC5314 andcreated a ISW2 knock out mutant. An ISW2 knock out had been reported to be completely abolish chlamydospore formation. We found that the ISW2mutant had significantly reduced virulence in mouse model of disseminated candidiasis and also failed to induce chlamydospores in mouse kidneys duringpathogenesis . In vitro studies confirm the ability of these mutants for normal filamentous growth, but they failed to produce typical chlamydospores fromsuspensor cells. However, after three weeks they produced chlamydospore-like structures that differed from normal chlamydospore production by thecomplete absence of suspensor cells. As an essential ATP dependent chromatin remodeling factor in yeasts, ISW2 affects the regulation of transcription,recombination, and DNA repair. Our findings suggest that ISW2 may also down regulate the genes for suspensor cell formation but not the genes forchlamydospore formation indicating that these are two independent processes. Further, our investigation into in vivo role of chlamydospores andsuspensor cells suggest that ISW2 could be a future drug target. Further studies on gene regulation by ISW2 in C.albicans will be paramount to ourunderstanding of development and regulatory steps for chlamydospore formation and their contribution to host infection.500. Nutrient immunity and systemic readjustment of metal homeostasis modulate fungal iron availability during the development of renal infections.Joanna Potrykus 1 , David Stead 2 , Dagmar S Urgast 3 , Donna MacCallum 1 , Andrea Raab 3 , Jörg Feldmann 3 , Alistair JP Brown 1 . 1) Aberdeen Fungal Group,University of Aberdeen, Aberdeen, United Kingdom; 2) Aberdeen Proteomics, University of Aberdeen, Aberdeen, United Kingdom; 3) Trace ElementSpeciation Laboratory, University of Aberdeen, Aberdeen, United Kingdom.Iron is a vital micronutrient that can limit the growth and virulence of many microbial pathogens. Here we show, that in the murine model ofdisseminated candidiasis, the dynamics of iron availability are driven by a complex interplay of localized and systemic events. As the infection progresses inthe kidney, Candida albicans responds by broadening its repertoire of iron acquisition strategies from non-heme iron (FTR1-dependent) to heme-ironacquisition (HMX1-dependent), as demonstrated in situ by laser capture microdissection, RNA amplification and qRT-PCR. This suggested changes in ironavailability in the vicinity of fungus. This was confirmed by 56 Fe iron distribution mapping in infected tissues via laser ablation-ICP-MS, which revealeddistinct iron exclusion zones around the lesions. These exclusion zones correlated with the immune infiltrates encircling the fungal mass, and wereassociated with elevated concentrations of murine heme oxygenase (HO-1) circumventing the lesions. Also, MALDI Imaging revealed an increase in hemeand hemoglobin alpha levels in the infected tissue, with their distribution roughly corresponding to that of 56 Fe. Paradoxically, whilst iron was excludedfrom lesions, there was a significant increase in the levels of iron in the kidneys of infected animals. This iron appeared tissue bound, was concentratedaway from the fungal exclusion zones, and was accompanied by increased levels of ferritin and HO-2. This iron accumulation in the kidney correlated withdefects in red pulp macrophage function and red blood cell recycling in the spleen, brought about by the fungal infection. Significantly, this effect could bereplicated by selective chemical ablation of splenic red pulp macrophages by clodronate. Collectively, our data indicate that systemic events shapemicronutrient availability within local tissue environments during fungal infection. The infection attenuates the functionality of splenic red pulpmacrophages leading to elevated renal involvement in systemic iron homeostasis and increased renal iron loading. Simultaneously, localized nutrientimmunity limits iron availability around foci of fungal infection in the kidney. In response, the fungus modulates its iron assimilation strategies.501. Identification of the gut fungi in humans with nonconventional diets. Mallory Suhr, Heather Hallen-Adams. Food Science and Technology,University of Nebraska-Lincoln, Lincoln, NE.Identification of the microorganisms that establish themselves inside and outside the human body is crucial to explore how the microbiome impactshuman health. The recent Human Microbiome Project provides an initial compilation and identification of the gut microbiome ecosystem. It is wellresearched and understood that a large part of the gastrointestinal microbiota spans across the prokaryotic domain, but few studies have investigated thecontribution of fungi to the human gut microbiome. Factors such as diet, genetics, and environment can play an influential role in explaining whydifferences in microbiota exist between human hosts. Expanding on work from our lab, this study examines the effect of nonconventional diets (e.g.vegetarians, vegans, gluten-free and lactose-free) on the GI tract fungi. DNA from fecal samples of healthy human subjects was isolated and fungal-specificITS primers were used to target fungal DNA to obtain a baseline of data for gut fungi. Candida tropicalis and C. albicans were both detected, with C.tropicalis more prevalent. This relative abundance of C. tropicalis is in keeping with our earlier studies in people with conventional diets, and may be aregional phenomenon.502. The mutational landscape of gradual acquisition of drug resistance in clinical isolates of Candida albicans. Jason Funt 1 , Darren Abbey 7 , Luca Issi 5 ,Brian Oliver 3 , Theodore White 4 , Reeta Rao 5 , Judith Berman 6 , Dawn Thompson 1 , Aviv Regev 1,2 . 1) Broad Institute of MIT and Harvard, 7 Cambridge Center,Cambridge, MA 02142; 2) Howard Hughes Medical Institute, Department of Biology, Massachusetts Institute of Technology, 77 Masscahusetts Ave,Camridge, MA 02140; 3) Seattle Biomedical Research Institute, Seattle, WA; 4) School of Biological Sciences, University of Missouri at Kansas City, MS; 5)Worcester Polytechnic Institute, Department of Biology and Biotechnology, 100 Institute Road, Worcester MA 01609; 6) Tel Aviv University, Ramat Aviv,69978 Israel; 7) University of Minnesota, Minneapolis MN 55455 USA.Candida albicans is both a member of the healthy human microbiome and a major pathogen in immunocompromised individuals1. Infections are most244

FULL POSTER SESSION ABSTRACTScommonly treated with azole inhibitors of ergosterol biosynthesis. Prophylactic treatment in immuncompromised patients2,3 often leads to thedevelopment of drug resistance. Since C. albicans is diploid and lacks a complete sexual cycle, conventional genetic analysis is challenging. An alternativeapproach is to study the mutations that arise naturally during the evolution of drug resistance in vivo, using isolates sampled consecutively from the samepatient. Studies in evolved isolates have implicated multiple mechanisms in drug resistance, but have focused on large-scale aberrations or candidategenes, and do not comprehensively chart the genetic basis of adaptation5. Here, we leveraged next-generation sequencing to systematically analyze 43isolates from 11 oral candidiasis patients, collected sequentially at two to 16 time points per patient. Because most isolates from an individual patientwere clonal, we could detect newly acquired mutations, including single-nucleotide polymorphisms (SNPs), copy-number variations and loss ofheterozygosity (LOH) events. Focusing on new mutations that were both persistent within a patient and recurrent across patients, we found that LOHevents were commonly associated with acquired resistance, and that persistent and recurrent point mutations in over 150 genes may be related to thecomplex process of adaptation to the host. Conversely, most aneuploidies were transient and did not directly correlate with changes in drug resistance.Our work sheds new light on the molecular mechanisms underlying the evolution of drug resistance and host adaptation.503. Yeast-Hypha transition and immune recognition of Candida albicans influenced by defects in cell signal transduction pathways. Pankaj Mehrotra,Rebecca A Hall, Jeanette Wagener, Neil A.R. Gow. Aberdeen Fungal Group, Aberdeen.During the infection process C. albicans has to respond to various stresses imposed by the host environment including oxidative and osmolarity stressgenerated by phagocytic cells such as macrophages and neutrophils, and also the cell wall stress agents such as exposure to caspofungin and otherantifungal antibiotics. These stress responses area orchestrated through the activation of multiple stress pathways including the cAMP-PKA, several MAPKpathways and the Ca 2+ -calcineurin pathway influence the cell wall shape and composition. We are investigating the effect of the activation or inhibition ofthese pathways on immune recognition mechanisms. We therefore determined the importance of the MAPK, cAMP-PKA and Ca 2+ -calcineurin pathways onthe fungal innate immune response by examining uptake, phagocytosis, and cytokine profile induced by mononuclear and polynuclear lymphocytes inresponse to a library of mutants in each of the above pathways under stressed and non-stressed conditions. We find that the activation and inhibition ofthese pathways plays a important role in remodeling of cell wall and hence the immunological profile. For example, deletion of TPK1 and CNA1 resulted inlower pro-inflammatory cytokine production. Immune- recognition was also affected by the exposure of C. albicans signaling mutants with Calcofluorwhite,caspofungin , oxidative and osmotic stress and changes in temperature. These results suggest that stress signaling pathways act in a co-ordinatedfashion to regulate yeast-hypha morphogenesis and the changes in the cell wall which in turn affects the immunological signature of the cell. Thusexposure to different microenvironments significantly modifies the immunological response to fungal cells, suggesting that responses to local stressesmakes the fungal cell surface is a moving target for immunological surveillance.504. GPI PbPga1 of Paracoccidioides brasiliensis is a surface antigen that activates macrophages and mast cells through the NFkB signaling pathway. C.X. R. Valim, L. K. Arruda, P. S. R. Coelho, C. Oliver, M. C. Jamur. Faculdade de Medicina de Ribeirão Preto, USP, Ribeirão Preto, São Paulo, Brazil.Paracoccidioides brasiliensis is the etiologic agent of paracoccidioidomycosis (PCM), one of the most prevalent mycosis in Latin America. P. brasiliensiscell wall components interact with host cells and influence the pathogenesis of PCM. PbPga1 is a GPI anchored protein that is up-regulated in the yeastpathogenic form. GPI anchored proteins are involved in cell-cell and cell-tissue adhesion and have a key role in the interaction between fungal and hostcells. PbPga1 is an O-glycosylated protein that is localized on the yeast cell surface. Recombinant PbPga1 (rPbPga1) induces nitric oxide (NO) productionand TNF-a release in murine peritoneal macrophages (Valim et al.Plos One, 2012). In the present study, rPbPga1 was able to activate NFkB in macrophagelikeRaw cells that had been transfected with NFkB luciferase as well as in a reporter cell line for NFkB activation derived from RBL-2H3 mast cells. Theresults show that like macrophages, rPbPga1 also activates the transcription factor NFkB in mast cells. However, rPbPga1 does not activate NFAT nor is itable to induce liberation of beta hexosaminidase . The lack of beta hexosaminidase release suggests the PbPga1 is not able to activate RBL-2H3 mast cellsvia the high affinity IgE receptor. Mast cell activation by rPbPga1 does result in activation of the transcription factor NFkB suggesting stimulation ofcytokine production. Taken together these results indicate that the surface antigen PbPga1 may play an important role in PCM pathogenesis by activatingmacrophages and mast cells.505. Cladosporium fulvum effector Ecp6 outcompetes host immune receptor for chitin binding through intrachain LysM dimerization. Andrea Sánchez-Vallet 1 , Raspudin Saleem-Batcha 2 , Anja Kombrink 1 , Guido Hansen 2 , Dirk-Jan Valkenburg 1 , Jeroen R. Mesters 2 , Bart P.H.J. Thomma 1 . 1) Laboratory ofPhytopathology, Wageningen University, Wageningen, Netherlands; 2) Institute of Biochemistry, Center for Structural and Cell Biology in Medicine,University of Lübeck, Lübeck, Germany.Successful pathogens secrete effector proteins to deregulate host immunity which is triggered upon detection of pathogen-associated molecularpatterns (PAMPs). Several fungal pathogens employ LysM effectors, such as Ecp6 from Cladosporium fulvum, to sequester fungal cell wall-derived chitinoligomers which act as PAMP and would otherwise be recognized by host immune receptors and trigger defense responses. The mechanism by whichLysM effectors scavenge chitin molecules remained unknown thus far. Based on crystal structure analysis of Ecp6, we reveal a novel mechanism for chitinbinding by intrachain LysM dimerization, leading to a binding groove in which chitin is deeply buried in the effector protein. Isothermal titrationcalorimetry experiments show that the concerted action of two LysM domains mediates a single chitin binding event with ultra-high (pM) affinity.506. Genotypic and phenotypic characterization of Setosphaeria turcica reveals population diversity and a candidate virulence gene location. SantiagoMideros 1 , Chia-Lin Chung 1,3 , Jesse Poland 2,4 , Gillian Turgeon 1 , Rebecca Nelson 1,2 . 1) Cornell University, Dept. of Plant Pathology and Plant-Microbe Biology,Ithaca, NY, USA; 2) Cornell University, Dept. of Plant Breeding and Genetics, Ithaca, NY, USA; 3) National Taiwan University, Dept. of Plant Pathology andMicrobiology, Taipei, Taiwan; 4) USDA-ARS, Hard Winter Wheat Genetics Research Unit, Kansas State University, Manhattan, KS, USA.The dothideomycete maize pathogen Setosphaeria turcica (anamorph Exserohilum turcicum) causes Northern Leaf Blight, one of the most commonfungal diseases of maize worldwide. Little is known about the genetic basis of virulence and aggressiveness in this pathogen, although several races havebeen described based on their compatibility with maize resistance genes Ht1, Ht2, Ht3 and HtN. To study the genetic basis of virulence and aggressiveness,we generated a F1 population consisting of 221 monosporic progeny of a cross between a race 1 strain and a race 23N strain. Genotyping-by-sequencing(GBS) was conducted on the population and an additional 13 diverse isolates that included the parental lines. We obtained between 341,000 and 428,000sequence tags for each of the 234 isolates. Alignment to the S. turcica Et28A v1.0 genomic sequence(http://genome.jgi.doe.gov/Settu1/Settu1.home.html) yielded 27,174 single nucleotide polymorphisms (SNPs) at a density of 0.63 SNPs per kb. In the 13isolates, using 9,526 filtered SNPs, we found an average nucleotide diversity (p) of 0.297. Using 564 polymorphic markers with less than 35% missing calls,we created a high-density genetic map that resulted in 23 linkage groups and a total length of 1,686 cM. The Et28A sequence has 407 scaffolds, fourscaffolds formed a single linkage group in our genetic map. The rest of the genome remains fragmented. To identify genomic regions controlling virulence27th Fungal Genetics Conference | 245

FULL POSTER SESSION ABSTRACTSand aggressiveness, isolates were characterized for in vitro and in planta phenotypes. For the former, mycelial abundance, colony diameter, pigmentation,and sporulation were rated in replicated trials. For the latter, incubation period, primary diseased leaf area and a qualitative differential score were ratedon maize near isogenic lines with and without Ht2. Linkage mapping identified a 54.3 kb sequence of Et28A as a robust candidate region carrying S. turcicaavrHt2. In order to manage the vast amount of genotypic and phenotypic data a MySQL database was created.507. The secretome is linked to virulence in the yeast pathogen Cryptococcus. Leona Campbell 1 , Anna Simonin 1 , Janna Ferdous 1 , Matthew Padula 1 ,Elizabeth Harry 2 , Ben Herbert 3 , Dee Carter 1 . 1) School of Molecular Bioscience, University of Sydney, Sydney, N.S.W. Australia; 2) iThree Institute,University of Technology, Sydney, NSW, Australia; 3) Department of Chemistry & Biomolecular Sciences, Macquarie University, Sydney, NSW, Australia.The disease caused by pathogenic Cryptococcus spp. begins after inhalation of infectious propagules leading to infection of the lung. In some cases thepathogen disseminates to the central nervous system, resulting in meningoencephalitis, which can be fatal if left untreated. However, closely relatedstrains of Cryptococcus gattii and its sibling species C. neoformans can exhibit significantly different degrees of pathogenesis in the mammalian host. Asfungi utilize absorptive nutrition producing a range of secreted degradative enzymes, and as these may invoke a host response, the fungal secretome islikely to be very important in modulating host-pathogen interactions. To investigate this the secreteome was determined for a hypovirulent and ahypervirulent strain of C. gattii, R272 and R265 respectively, and a virulent strain of the opportunistic pathogen C. neoformans, KN99a. All strains weregrown under conditions designed to mimic those encountered in vivo. Secreted proteins were analysed using two different mass spectrometry-basedtechniques: 1D nanoLC-MS/MS and Imaging Mass Spectrometry (IMS). The three strains secreted significantly different protein cohorts. A total of 70proteins were identified with 47, 13 and 22 identified from R272, R265 and KN99a respectively. Only one protein was shared by all strains, a putativeglycosyl hydrolase. The secretomes of R265 and KN99a primarily included uncharacterized proteins, and bioinformatic analysis suggested these proteinscontained catalytic regions with roles in carbohydrate degradation. In contrast the less virulent R272 strain secreted a more diverse set of proteinsincluding canonical cytosolic proteins such as enolase and transaldolase. These proteins have been described as fungal allergens that bind IgE. Thesefindings indicate that virulence and the secretome are linked in Cryptococcus. The secretion of proteins with a putative role in nutrient scavenging byvirulent strains R265 and KN99a suggest they can source nutrients from a range of available substrates, which may allow them to exploit a wider range ofecological niches including the mammalian host. In contrast, the potentially allergenic proteins secreted by strain R272 suggest this strain triggers a moreeffective immune response, leading to clearance of the pathogen.508. Post-Transcriptional Regulation of the ER Stress Response in Cryptococcus neoformans. Virginia E. Havel, John C. Panepinto. Microbiology andImmunology, University at Buffalo, Buffalo, NY.Cryptococcus neoformans is one of a small number of fungi able to make the transition from ambient environmental temperatures to human core bodytemperature. We have previously reported that the ER stress response plays an important role during host temperature adaptation. Deletion of the RNAbinding protein, Puf4, results in temperature sensitivity and increased resistance to the ER stress inducing drug tunicamycin, leading us to hypothesize thatPuf4 post-transcriptionally regulates the ER stress response during host temperature adaptation. The ER stress response is initiated by the transcriptionfactor Hac1 (Hxl1 in C. neoformans). Hac1 translation requires unconventional splicing of the pre-spliced HAC1 mRNA to the translated HAC1 mRNA at theER surface by Ire1. Time courses measuring HXL1 mRNA splicing during a shift to 37°C demonstrate a delay in the splicing of HXL1 in puf4D when comparedto wild type. The delay in HXL1 splicing in puf4D results in a delayed and persistent induction of ER stress response transcripts in puf4D compared to wildtype as measured by northern blot. We hypothesize that Puf4 is required for localization of the HXL1 transcript to the ER outer surface where it is cleavedby Ire1, thereby promoting the induction of the ER stress response. We have also shown through EMSA analysis and RNA-immunoprecipitationexperiments that Puf4 is able to bind to ALG7 mRNA. Alg7 is involved in protein glycosylation at the ER surface and is the target of tunicamycin. Based onthe observation that puf4D has increased resistance to tunicamycin, and results demonstrating that Puf4 is able to bind ALG7 mRNA we hypothesize thatPuf4 may regulate ALG7 mRNA by repressing translation. In our model Puf4 has a bi-modal mechanism of regulating the ER stress response. ER stressresponse initiation requires Puf4-mediated localization of pre-spliced HXL1 mRNA to the ER surface. During the attenuation phase of the ER stressresponse, we hypothesize that ER transcripts are translationally repressed by Puf4, resulting in attenuation of the ER stress response and allowing the cellto return to homeostasis. Despite the well-studied mechanism of unconventional splicing by Hac1 and Hac1 homologs in yeast and other model systems,this study is the first to identify a RNA binding protein potentially involved its activation.509. A morphogenesis regulator controls cryptococcal neurotropism. Xiaorong Lin 1 , Bing Zhai 1 , Karen Wozniak 2 , Srijana Upadhyay 1 , Linqi Wang 1 , ShupingZhang 3 , Floyd Wormley 2 . 1) Biology, Texas A&M University, TAMU-3258, TX; 2) Biology, the University of Texas at San Antonio, San Antonio, Texas, USA; 3)Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, Texas, USA.Cryptococcus neoformans is the major causative agent of cryptococcal meningitis, a disease that is responsible for more than 600,000 deaths each year.This ubiquitous environmental pathogen enters host lungs through inhalation and typically establishes asymptomatic latent infections. However,extrapulmonary dissemination often occurs in individuals with weakened immunity and Cryptococcus has a predilection to infect the brain. Braininfections are fatal and formidable to treat due to the poor penetration of most antifungals to the brain. Unfortunately, little is known about cryptococcalfactors that control its neurotropism. Here we report that a morphogenesis regulator Znf2 controls the tissue tropism of cryptococcal infection. Inparticular, activation of Znf2 abolishes Cryptococcus extrapulmonary dissemination and consequently leads to the absence of fatal brain infections in theinhalation infection model. Although Znf2 overexpression strains are avirulent in this animal model, these strains are capable of proliferating in the animallungs during the early stages of infections. Histological examinations and cytokine profiling revealed that the Znf2 overexpression strain causes enhancedmonocyte infiltration in the animal lungs. Consistently, the Znf2 overexpression strain stimulates pro-inflammatory host responses while suppressesdeleterious Th2 host responses during early stage of infection in the pulmonary infection model. Such protective host defense responses might haveprevented the extrapulmonary dissemination of Cryptococcus. In the intravenous infection model where the lung infection was bypassed and there wasuniform hematogenous dissemination, the Znf2 overexpression strain showed a specific defect in the brain infection. Taken together, our data indicatethat Znf2 helps polarize the host immunity towards protection and that it mediates cryptococcal tissue tropism during infection.510. Extracellular and intracellular signaling orchestrates morphotype-transition and virulence in human pathogen Cryptococcus neoformans. LinqiWang, Xiuyun Tian, Rachana Gyawali, Xiaorong Lin. Biology, College Station, TX.Interactions with the environment and divergent species drive the evolution of microbes. To sense and rapidly respond to these dynamic interactions,“simple” microbes developed bet-hedging social behaviors, including the construction of heterogeneous biofilm communities and transition betweendifferent morphotypes. The human fungal pathogen Cryptococcus neoformans can undergo morphotype transition between the yeast and the filamentousform. Most recently, we demonstrated that the zinc-finger regulator Znf2 bridges the bi-direction yeast-hypha transition and virulence in this pathogen.246

FULL POSTER SESSION ABSTRACTSOne of Znf2 downstream targets is extracellular protein Cfl1. Cfl1 is a cell-wall bound adhesin and a signaling molecule when it is released. This matrixprotein Cfl1 plays a similar but less prominent role than Znf2 in orchestrating morphogenesis and virulence in C. neoformans. Through transcriptomeanalyses and screening Znf2 downstream targets by overexpression, we identified an additional player in the control of morphogenesis and biofilmformation. This factor is an intracellular RNA-binding protein Pum1. As expected, Pum1 affects filamentation in a Znf2 dependent manner. However, theeffect of Pum1 on morphogenesis is independent of Cfl1. The pum1D cfl1D double mutant shows a more severe defect in filamentation than either of thesingle mutant, indicating that Pum1 and Cfl1 act in two parallel pathways. Two of Pum1’s targets, Fad1 and Fad2, form a Cryptococcus-specific adhesinfamily. Like Cfl1, these two extracellular adhesins show regulatory roles in conducting morphogenesis and virulence in C. neoformans and thus may beinvolved in extracellular signaling transduction. Our results indicate that complex regulatory cascades composed of extracellular and intracellular circuitsmay be responsible for mediating morphological transition in response to the cues in the environments and the host.511. Evidence for alkaloid diversity and independent hybridization events of Elymus endophytes. Nikki D. Charlton 1 , Juan Pan 2 , Daniel G. Panaccione 3 ,Christopher L. Schardl 2 , Carolyn A. Young 1 . 1) Forage Improvement Division, The Samuel Roberts Noble Foundation, Ardmore, OK; 2) Department of PlantPathology, University of Kentucky, Lexington, KY; 3) Division of Plant & Soil Sciences, West Virginia University, Morgantown, WV.The epichloae form mutualistic symbioses with cool-season grasses and have been shown to impart biotic and abiotic fitness benefits to their hosts.Endophyte-infected plants often have greater resistance to biotic stresses such as mammalian and insect herbivory due to the presence of fungalsynthesized alkaloids. Four classes of bioprotective alkaloids have been described, which include ergot alkaloids, indole-diterpenes, a pyrrolopyrazine(peramine), and saturated aminopyrrolizidines (lolines). Elymus species, such as Elymus canadensis and E. virginicus, are cool-season bunchgrasses nativeto much of North America and are known to harbor Epichloë endophytes. Three species are able to associate with Elymus: the non-hybrids Epichloëamarillans and Epichloë elymi and Epichloë canadensis, a hybrid with E. elymi and E. amarillans ancestral progenitors. The distribution and alkaloidgenotypic variation of these fungi was examined to determine endophyte variation that may provide ecological benefits to the host. Endophyte infectionfrequencies from natural populations and germplasm resources ranged from uninfected to highly infected. Analyses of microsatellite loci and mating typegenes characterized the prevalence and distribution of hybrid and non-hybrid endophytes among and between Elymus populations. Overall, non-hybridswere more prevalent than hybrids in the northern region of the U.S., whereas hybrids were more ubiquitous in the southern region. Genotypic analysisbased on presence and absence of key alkaloid biosynthesis genes provided information about the potential alkaloid diversity within these populations.Thirteen unique alkaloid genotypes were identified that showed variation within the EAS (ergot alkaloid), LOL (loline) and PER (peramine) loci thatindicates some genotypes are likely to accumulate pathway intermediates. Evaluation of the mating-type idiomorphs from the hybrid E. canadensisindicates this species has resulted more than once through independent hybridization events thus explaining variation found among the alkaloid genes.Chemical analyses of representative endophyte-infected plants are being conducted to correlate alkaloid predictions with actual alkaloid production.Chemotype diversity will be evaluated to determine how this translates into differences in fitness and persistence of the host.512. The functional characterization of candidate genes involved in host specialization of Zymoseptoria grass pathogens. Stephan Poppe, Petra Happel,Eva Stukenbrock. Fungal Biodiversity, Max Planck Institute Marburg, Marburg, Germany.The ascomycete fungus Zymoseptoria tritici (synonym: Mycosphaerella graminicola) emerged as a new pathogen of cultivated wheat during cropdomestication about 11.000 years ago. To understand the molecular basis of host specialization in this pathogen we have sequenced complete genomes ofZ. tritici and closely related species infecting wild grasses. Evolutionary genomic analyses allowed us to identify 17 genes that show strong evidence ofpositive selection between Z. tritici and the closely related sister species Zymoseptoria pseudotritici. We hypothesize these evolved in a co-evolutionaryarms race with different hosts. None of the genes encode proteins with known function. In this study we focus on three candidate genes Mgr80707, Mgr89160 & Mgr 103264 and investigate their role in Z. tritici and its two closest relatives Z. pseudotritici and Z. ardabiliae during host infection. QuantitativeReal time PCR experiments from the three fungal species infecting four different grass species show that the three genes are strongly up-regulated inplanta and that candidate gene expression differs over a time course of 28 days supporting a role in host pathogen interaction. In addition, we show thatthree different host species (wheat, Elymus repens and Lolium multiflorum) differentially induce gene expression in the fungi. Confocal Laser ScanningMicroscopy conducted at different time points reveals clear differences between Zymoseptoria species during infection and within host development inwheat and Brachypodium distachyon. Deletion strains for each candidate gene have been created by Agrobacterium tumefaciens mediatedtransformation. The single deletion of two candidate genes Mgr80707 & Mgr103264 led to a reduced virulence of Z. tritici on wheat. The deletion of thethird gene Mgr89160 led to a hyper-virulence phenotype suggesting an avirulence function of the gene product. Our study confirms that genes involved inhost specialization can be identified based on footprints of natural selection.513. Diversity and Phylogeny of genus Suillus (Suillaceae, Boletales) from Pakistan (Asia). Samina Sarwar, Abdul Nasir Khalid. Botany, University of thePunjab, Lahore, Punjab, Pakistan.Coniferous forests of Pakistan are rich in mycodiversity. However, only a few scientific researches have been conducted in these forests. This paper aimsto document diversity of Suillus in these forests. During a survey conducted during 2008-2010, a total of thirty two (32) basidiomata were collected. Mostof them were found associated with Pinus wallichiana and Abies pindrow. Only a few were found with Cedrus deodara, Populus ciliata and Quercus spp.These basidiomata were characterized morphologically as well as by molecular analysis by amplifying rDNA. Fungal specific primers ITS3 & ITS6R and ITS2& ITS8F were used to amplify the ITS1 and ITS2 along with partially 5.8S gene. Out of these, twelve (13) different Suillus species were found. Among themtwo (2) species seem undescribed and three (3) as new records for Pakistan. Their Phylogenetic relationships have also been discussed.514. Saprolegnia species can switch hosts to cause infection: a new insight into host pathogen interaction. Mohammad N. Sarowar 1* , A. Herbert van denBerg 1 , Debbie McLaggan 1 , Mark Young 2 , Pieter van West 1 . 1) Institute of Medical Sciences, Foresterhill, Aberdeen, AB25 2ZD, United Kingdom; 2)Department of Zoology, University of Aberdeen, Aberdeen, AB24 2TZ, United Kingdom.Saprolegnia species are destructive oomycete pathogens of many aquatic organisms and are found in all parts of the world. Phylogenetic analysis hasshown that Saprolegnia strains isolated from different aquatic organisms have a close relationship to fish pathogenic Saprolegnia species. We have nowdemonstrated, for the first time, that Saprolegnia spp. can actually switch hosts. Saprolegnia australis, Saprolegnia hypogyna and 2 strains of Saprolegniadiclina were isolated from insects. We also collected other oomycete species, including S. australis, S. ferax, Pythium pachycaule and Pythium sp., in waterof a medium to fast running river. The ITS region of all these isolates was sequenced. Four isolates collected from the aquatic insects together with isolatesof S. parasitica (collected from salmon), S. diclina (collected from trout eggs) and S. ferax (collected from an amphibian) were tested for pathogenicity onnymphs of a stonefly (Perla bipunctata), Atlantic salmon eggs and frog (Xenopus leavis) embryos. Most of the isolates were highly pathogenic on all testedaquatic animals. These results suggest that Saprolegnia spp. are capable of switching host, which may be related to seasonal variation of host availability in27th Fungal Genetics Conference | 247

FULL POSTER SESSION ABSTRACTSfresh water environments.515. The Plant-Microbe Interfaces project: defining and understanding relationships between Populus and the rhizosphere microbiome. ChristopherSchadt 1 , Dale Pelletier 1 , Timothy Tschaplinski 1 , Edward Uberbacher 1 , Hurst Gregory 2 , E. Peter Greenberg 3 , Caroline Harwood 3 , Amy Schaefer 3 , RytasVilgalys 4 , Francis Martin 5 , Mitchel Doktycz 1 , Gerald Tuskan 1 , and other PMI researchers (http://pmi.ornl.gov). 1) Bioscience Division, Oak Ridge NationalLaboratory, Oak Ridge, TN, USA; 2) Chemical Sciences Division, Oak Ridge National Laboratory, Oak Ridge, TN, USA; 3) Department of Microbiology,University of Washington, Seattle, WA, USA; 4) Department of Biology, Duke University, Durham, NC, USA; 5) Institut National de la RechercheAgronomique, Nancy, FRANCE.Microbial interactions benefit plant health by affecting nutrient uptake, hormone signaling, water and element cycling in the rhizosphere and/orconferring resistance to pathogens. The model tree species Populus provides an opportunity for microbiome research relevant to bioenergy, carbon-cycleresearch, and other ecosystem processes. In an effort to define Populus’ microbiome, root and rhizosphere samples from P. deltoides in the eastern USand P. trichocarpa in western US were subjected rRNA pyrosequencing and an isolate collection of over 5000 bacteria and 500 fungi obtained. We showthat the rhizo- and endo-sphere environments feature highly developed, diverse and to a large degree often exclusive communities of bacteria and fungi.Endophytic bacterial diversity was found to be highly variable, but on average tenfold lower than the rhizosphere, suggesting root tissues provide a distinctenvironment supporting relatively few species. Fungal endophytic species were more numerous, but also less than rhizosphere spp. Both fungal andbacterial rhizosphere samples showed distinct phylogenetic composition patterns compared to the more variable endophyte samples. Contrary to initialexpectations, both Populus spp. have low natural levels of colonization by ectomycorrhizal (ECM) and arbuscular mycorrhizal fungi, but high levels ofpresumed fungal endophytic taxa such as Nectria, Mortierrella, and members of the Atractiellales. Select isolates are being studied at a whole-genomelevel to enable comparative work on on the basis for observed symbioses. Thus far ~43 bacterial and 5 fungal isolates have been sequenced. Initialcomparative genomics of these isolates suggest highly divergent physiological and molecular mechanisms of the interactions, even within rather closelyrelated species. Efforts to understand ECM interactions have shown that host defense networks and the ability to bypass such networks through smallprotein and phytohormone signals has a large effect on the ability of Populus to form ECM relationships. Laccaria bicolor is able to modulate host defenseresponse in P. trichocarpa, yet unable to do so in P. deltoides. Mycorrhizal Helper Bacteria from the genus Pseudomonas partially alleviate this colonizationweakness through yet unknown molecular mechanisms, illustrating the value of integrated microbiome wide studies.516. Do the fungal homologs of Verticillium dahliae effector Ave1 act as virulence factors? Jordi C. Boshoven 1 , Melvin D. Bolton 2 , Bart P.H.J. Thomma 1 . 1)Laboratory of Phytopathology, Wageningen University, 6708 PB Wageningen, Netherlands; 2) Agricultural Research Service, Northern Crop ScienceLaboratory, US Department of Agriculture, Fargo, ND 58102.Verticillium species cause vascular wilt disease in over 200 plant hosts, including economically important crops. In tomato, the Ve1 immune receptorconfers resistance to race 1 strains of V. dahliae, but not to race 2. By population genome sequencing of race 1 and race 2 strains, the effector that isrecognized by Ve1 was recently identified as Ave1 (Avirulence on Ve1 tomato). Ave1 has homology to plant natriuretic peptides that are regulators ofhomeostasis, and acts as a virulence factor on tomato plants that lack Ve1 as well as on Arabidopsis. In addition to plants, Ave1 homologs were also foundin a few fungal pathogens, including Fusarium oxysporum, Cercospora beticola and Colletotrichum higginsianum, as well as in the bacterial plant pathogenXanthomonas axonopodis. Co-expression of V. dahliae Ave1 and tomato Ve1 in Nicotiana tabacum resulted in the activation of a hypersensitive response.Remarkably, also co-expression of some of the Ave1 homologs with Ve1 activated a hypersensitive response. Here, we evaluate whether the variouspathogen-derived Ave1 homologs are virulence factors. Expression of the Ave1 homologs of Fusarium, Cercospora and Colletotrichum during infection ontomato, sugarbeet, and Arabidopsis, respectively, was analysed. To investigate the potential role of the Ave1 homologs in virulence, a V. dahliae Ave1deletion mutant was complemented with the Ave1 homologs of Fusarium, Cercospora and Colletotrichum, and tested for full aggressiveness on tomato.Finally, targeted gene deletion was pursued in Fusarium, Cercospora and Colletotrichum and the corresponding deletion strains were inoculated ontomato, sugarbeet, and Arabidopsis, respectively.517. The candidate effector repertoire of closely related Venturia pathogens of the Maloideae revealed by whole genome sequence and RNAsequencing analyses. Cecilia Deng 1 , Daniel Jones 2 , Bruno Le Cam 3 , Kim Plummer 2 , Carl Mesarich 4 , Matthew Templeton 1 , Joanna Bowen 1 . 1) Plant & FoodResearch, Auckland, New Zealand; 2) La Trobe University, Melbourne, Australia; 3) IRHS, INRA Angers, France; 4) Wageningen University, The Netherlands.The genus Venturia includes pathogens that infect members of the Rosaceae. The most widely researched of these is V. inaequalis that causes thedisease apple scab. Related species cause disease of other woody hosts closely related to Malus; eg V. pirina infects European pear. Certain isolates thatare classified as V. inaequalis are unable to infect Malus but instead infect different hosts belonging to the subfamily Maloideae, such as loquat. Hostcultivarspecificity is also demonstrated by isolates of V. inaequalis that infect Malus; 17 gene-for-gene pairings between effectors (pathogen proteins thatenhance disease) and resistance gene products have been identified to date. Thus the effector repertoire of isolates of V. inaequalis determines theircultivar specificity and most probably host specificity. Effectors have yet to be cloned from V. inaequalis. Draft whole genome sequences (WGS) of three V.inaequalis isolates (two from apple, one from loquat) and an isolate of V. pirina have been assembled and candidates that share the characteristics offungal effectors (small, secreted proteins) have been identified. Of the 13333 predicted genes in the WGS of V. inaequalis isolated from apple, 1088encode putatively secreted proteins identified utilising algorithms to detect secretion signals and putative cellular location. The expression (measured bytranscriptome analysis) of 119 of these is up-regulated, with a false discovery rate less than 0.05 and a log-fold change greater than 2, in planta comparedwith in vitro at both 2 and 7 days post inoculation (dpi), 73 at 2 dpi, and 54 at 7 dpi. Of these 246, only 43 have similarities (

FULL POSTER SESSION ABSTRACTScells via type three section systems, a syringe like apparatus that directly penetrates the host cell plasma membrane. Currently, there is no evidence forsuch translocation machinery being present or utilized by fungal and oomycete symbionts to deliver effector proteins. Several bacterial and fungal toxinsutilize external glycosphingolipids to mediate translocation into cells. Upon internalization a subset of these toxins escape endosomal compartments viaretrograde transport to the Golgi-endoplasmic reticulum trans network, while others are capable of flipping across the endosomal membrane. The genusPhytophthora contains a number of highly destructive plant pathogens. Comparative genomics and the sequence availability of known intracellulareffectors resulted in the discovery of a highly conserved N-terminal motif RXLR-dEER that defines an expansive super family of secreted proteins. TheRXLR-dEER motif of PsAvr1b, PsAvr1k, and PsAvh5 has been shown to facilitate effector entry into a plant and animal cells in the absence of any pathogenencoded machinery. Entry is believed to be mediated by binding cell surface phospholipid, phosphatidylinositiol-3-phosphate. Using isothermal titrationcalorimetry we have characterized the phospholipid binding properties of the RXLR effectors PsAvr1b, PiAvr3a, and several RXLR-like effectors to monoand poly phosphatidylinositol-phosphates in a variety of experimental conditions. As controls we have characterized the phospholipid binding propertiesof 2xFYVE and Vam7pX, two known PI3P binding domains.519. Investigating virulence effectors in the poplar-poplar rust pathosystem. Sebastien Duplessis 1 , Benjamin Petre 1 , Hugo Germain 2,3 , Arnaud Hecker 1 ,Stéphane Hacquard 1 , David L Joly 4 , Armand Séguin 2 , Nicolas Rouhier 1 . 1) UMR1136 IAM, INRA, Champenoux, France; 2) Natural Resources Canada,Canadian Forest Service, Laurentian Forestry Centre, Quebec, QC, Canada; 3) Université du Québec à Trois-Rivières, Trois-Rivières, QC, Canada; 4)Agriculture and Agri-Food Canada, Pacific Agri-Food Research Centre, Summerland, BC, Canada.Foliar rust caused by Melampsora larici-populina is a major disease affecting poplar plantations throughout the world. The obligate biotrophic status ofthe fungus and the perennial status of the host plant, make molecular investigation of this interaction a real challenge. However, availability of bothPopulus trichocarpa and Melampsora larici- populina sequenced genomes has allowed for setting an emerging model pathosystem to decipher themolecular bases of disease resistance in trees and of biotrophic growth in rust fungi. Comprehensive analyses of rust transcripts encoding small secretedproteins expressed in planta identified putative virulence factors and we focused our attention on a few candidates that show evidence of purifyingselection between paralogs. In particular, two gene families encode modular SSP with a conserved N-terminal part and a C-terminal part evolving underpositive selection except highly-conserved cysteine residues. Gene family 5464 contains 13 members, homologs of Melampsora lini AvrP4 avirulencefactors whereas H1 family gather 31 genes specific to M. larici-populina. Some of these genes present a transient peak of expression during the biotrophicgrowth of the fungus in poplar leaves, and the corresponding proteins are able to enhance bacterial growth when delivered in Arabidopsis thaliana fromPseudomonas syringae pv tomato DC3000. Currently, we combine a range of approaches including biochemical and structural characterization ofrecombinant proteins, yeast two-hybrid and pull-down assays to characterize these candidate effectors.520. Functional analysis of Aphanomyces euteiches effectors, a legume root pathogen. D. Ramirez-Garces, L. Camborde, H. San Clemente, A. Cerutti, B.Dumas, E. Gaulin. LRSV UMR5546 CNRS/UPS, Castanet-Tolosan, France.Aphanomyces euteiches is an oomycete infecting roots of various legumes species such as pea, alfalfa and the model legume Medicago truncatula. Thegenus Aphanomyces, which belongs to the group of Saprolegniales, is phylogenically distant from the well known Phytophthora genus and comprises bothanimal pathogen and plant pathogen species. The first genome draft of Aphanomyces euteiches (ATCC201684, 57 Mb) will be soon released and genomesequencing of zoo- and phytopathogen species are under progress. A. euteiches genome miming revealed the expansion of CRNs (Crinkling and Necrosis)genes, initially identified in Phytophthora infestans. These modular proteins contain a conserved N-terminal characterized by the presence of a LFLAKamino acid motif implicated in the protein translocation from the pathogen to the host cell, whereas the modular C-terminal effector domain is highlydiverse. The proposed role of the CRNs effectors is to suppress plant defense or to modulate other host cell processes that increase susceptibility andenhance pathogen virulence. In A. euteiches, the active translocation LYLAK motif was detected, and conserved, as well as original effector subdomains,were identified. Functional studies conducted on two types of A. euteiches CRNS, AeCRN5 and AeCRN13, showed that both proteins are highly inducedduring infection of M. truncatula roots. In planta expression of both proteins revealed host nucleus localization and cell-death induction or alteration ofroots architecture when expressed in plant cells. Such observations suggest that A. euteiches CRNs are virulence proteins exerting their function throughthe interaction with nuclear compounds. Latest results regarding the putative function of AeCRNs will be presented.521. Participation of effector proteins from Trichoderma spp. in interaction with Arabidopsis thaliana. P. Guzman-Guzman 1 , M.I. Aleman-Duarte 2,3 , L.J.Delaye-Arredondo 3 , A. Herrera-Estrella 2 , V. Olmedo-Monfil 1 . 1) Biology Dept,University of Guanajuato, Guanajuato, Guanajuato, Mexico; 2) CINVESTAV,Langebio Unit, Irapuato Unit, Irapuato, Guanajuato, Mexico; 3) CINVESTAV, Irapuato Unit, Irapuato, Guanajuato, Mexico.When a plant-pathogen interaction is established, plant activates its immune response system, recognizing pathogen’s virulence factors. In plantresponse are involved several phytohormones. Studies on plant pathogenic processes triggered by different organisms have shown the involvement ofsome pathogen proteins that are capable of altering the function and structure of the host cell, facilitating their entry and affecting overall host physiology,these proteins are known as "effectors". The effectors activity inside the host takes place through conserved mechanisms of molecular interaction, such aseffectors that have the RXLR translocation motif, which is highly conserved among different pathogens, directing its entry into the host cells. In plantpathogeninteractions, effectors are recognized, activating the plant response system. However, some of these molecules have been involved in theestablishment of plant interactions with non-pathogenic organisms; little is known about the mechanisms controlling the establishment of this beneficialinteraction. Fungi of the genus Trichoderma establish beneficial interactions with plants, promoting their growth and defense systems. To betterunderstand this biological process, it is important to identify effector-like molecules in species like Trichoderma, and to establish their role in promotingplant development and defense systems activation. To achieve this goal, we will test the interaction of Trichoderma species with Arabidopsis thaliana,focusing on the selection and identification of effector-like molecules that are expressed during this interaction process. Additionally, we will generate nullTrichoderma mutants on these effector-like candidates to evaluate their participation in the plant-fungus interaction. Until now, we have establishedinteraction conditions, confirming the effect of the fungus presence over plant biomass production and root growth, showing interesting morphologicchanges in the root system, such as a significant increase in lateral root formation. Based on conserved characteristics and bioinformatics tools, weselected 16 encoding sequences for effector-like molecules among T. virens and T. atroviride genomes, and we confirmed the differential expression of 7of them in interaction with A. thaliana, by RT-PCR analysis.522. The role of LysM effectors in fungal fitness. Anja Kombrink 1 , Jason Rudd 2 , Dirk-Jan Valkenburg 1 , Bart Thomma 1 . 1) Phytopathology, WageningenUniversity, Wageningen, Netherlands; 2) Department of Plant Pathology and Microbiology, Rothamsted Research, Harpenden, Hertfordshire, UnitedKingdom.LysM effector genes are found in the genomes of a wide range of fungal species. The encoded LysM effectors are secreted proteins that contain a varying27th Fungal Genetics Conference | 249

FULL POSTER SESSION ABSTRACTSnumber of LysM domains, which are carbohydrate-binding modules. Ecp6, secreted by tomato leaf mould fungus Cladosporium fulvum, is the firstcharacterized LysM effector. We demonstrated that Ecp6 specifically binds chitin, the major constituent of fungal cell walls that acts as a microbialassociatedmolecular pattern (MAMP) and triggers immune responses upon recognition by the host. Ecp6 outcompetes plant receptors for chitin binding,and thus prevents the activation of immune responses. Many fungal genomes, including saprophytes, carry multiple LysM effector genes that share onlylow sequence conservation and encode a varying number of LysM domains. We speculate that fungal LysM effectors might bind different carbohydratesand exert various functions in fungal fitness. In the fungal wheat pathogen Mycosphaerella graminicola, two LysM effectors were identified. Mg3LysM, butnot Mg1LysM, suppresses chitin-induced immune responses in a similar fashion as Ecp6. Interestingly, unlike Ecp6, both Mg1LysM and Mg3LysM inhibitdegradation of fungal hyphae by plant chitinases, revealing an additional function for LysM effectors in pathogen virulence. We recently observed thatMg1LysM binds to the bacterial cell wall constituent peptidoglycan. Similarly, a LysM effector from the saprophytic fungus Neurospora crassa showedpeptidoglycan binding. We hypothesize that peptidoglycan binding by LysM effectors plays a role in the interaction of fungal species with bacterialcompetitors. The soil-borne fungal pathogen Verticillium dahliae contains seven LysM effectors genes of which one (Vd2LysM) is induced during tomatoinfection. Inoculation with two independent knock-out mutants revealed that Vd2LysM is required for full virulence of V. dahliae. However, Vd2LysM doesnot specifically bind chitin and does not function in a similar fashion as previous characterized LysM effectors. Thus, its function in virulence remainsunclear.523. WITHDRAWN524. Functional analysis and localization of SnTox1, a necrotrophic effector produced by the wheat pathogen Stagonospora nodorum. Zhaohui Liu 1 ,Weilin Shelver 2 , Justin Faris 3 , Timothy Friesen 1,3 . 1) Department of Plant Pathology, North Dakota State University, Fargo, ND; 2) USDA-ARS, BiosciencesResearch Laboratory, Fargo, ND; 3) USDA-ARS, Northern Crop Science Laboratory, Fargo, ND.SnTox1 is one of the necrotrophic effectors produced by the fungus Stagonospora nodorum, the causal agent of wheat Stagonospora nodorum blotch. Itinteracts, directly or indirectly, with the product of the wheat gene Snn1 to induce host cell death and promote disease. Previously, we showed thatSnTox1, a cysteine-rich protein, triggers programmed cell death-like responses in the host and plays an important role in fungal penetration. In the presentwork, we are investigating the biochemical and molecular function of SnTox1 as well as its mode of action. Based on a Prosite motif search of SnTox1,multiple predicted sites including a putative chitin binding domain were targeted for site-directed mutagenesis. SnTox1 activity was significantly reducedwhen mutations were produced at a casein kinase II phosphorylation site and a predicted helical region where lysine residues are abundant.Using a fungalstrain expressing an SnTox1-GFP fusion protein, we examined the location of the SnTox1 protein during fungal growth and infection. SnTox1 was observedin higher concentration on several fungal structures, including the surface of conidia and mycelium, hyphal septa, and hyphal tips. The accumulation ofSnTox1-GFP is particularly obvious at hyphal regions where new hyphae are arising. This observation suggests a protection mechanism of SnTox1 that issimilar to that of chitin binding proteins in other fungal pathogens. In planta, SnTox1 is highly expressed in the hyphopodia where the penetration isinitiated, providing further evidence that SnTox1 plays a role in penetration. The cellular localization of SnTox1 was also investigated using fluorescinelabeled SnTox1 in combination with cytological methods and preliminary data has indicated that SnTox1is likely not internalized into mesophyll cells butremains in the apoplast. Interestingly, SnTox1 is able to induce host cell death by directly spraying onto the leaf surface of sensitive lines. We are currentlyinvestigating if SnTox1 is transported through epidermal cell layer.525. Host-targeting protein 3 (SpHtp3) from the oomycete Saprolegnia parasitica translocates specifically into fish cells in a pH and tyrosine O-sulfatedependentmanner. Lars Löbach 1* , Stephan Wawra 2 , Irene de Bruijn 2 , Aleksandra Toloczko 2 , Tim Rasmussen 3 , Christopher Secombes 1 , Pieter van West 2 . 1)Scottish Fish Immunology Research Centre, University of Aberdeen, School of Biological Sciences, Aberdeen, Scotland, UK; 2) Aberdeen OomyceteLaboratory, University of Aberdeen, School of Medical Sciences, Foresterhill, Aberdeen, Scotland, UK; 3) University of Aberdeen, School of MedicalSciences, Foresterhill, Aberdeen, Scotland, UK.The success of eukaryotic oomycete pathogens depends largely on effector proteins, molecules which manipulate or interfere with host defencemechanisms in the extracellular space or inside their host cells. One economical important oomycete parasite is the fish pathogen Saprolegnia parasitica,which is the causal agent of the disease Saprolegniosis. S. parasitica is responsible for devastating losses in the aquaculture industry worldwide. In order toeffectively fight any pathogen it is crucial to understand the key molecular mechanisms that lead to the disease. With the focus on putative effectorproteins we screened the genome of S. parasitica in the present study for potential effector candidates. Analysis identified a novel putative secreted S.parasitica effector protein, which we named host-targeting protein 3 (SpHtp3). Gene expression analyses showed that mRNA levels of SpHtp3 are highestin mycelium, sporulating mycelium and during the later stages of infection. Recombinant SpHtp3 was able to translocate specifically into fish cells in atyrosine O-sulfate and pH dependent manner. SpHtp3 was found in vesicular structures inside fish cells and was released from these upon infection of thecells with S. parasitica. Interestingly, SpHtp3 possesses an N-terminal RTLR tetra-peptide sequence at a similar location as found in RxLR-effectors fromplant pathogenic oomycetes. However, this RTLR-sequence was not required for the fish cell translocation property of SpHtp3. These findings suggest thatSpHtp3 from S. parasitica is a novel intracellular protein that might play an important role in Saprolegniosis.526. Ave1-like orthologs in Venturia: another expanded effector family emerges. Adam Taranto 1 , Daniel Jones 1 , Jason Shiller 1 , Shakira Johnson 1 , NathanHall 1 , Ira Cooke 1 , Gert Talbo 1 , Carl Mesarich 2 , Bart Thomma 2 , Jordi Boshoven 2 , Joanna Bowen 3 , Cecilia Deng 3 , Matthew Templeton 3 , Kim M. Plummer 1 . 1)Dept Botany, La Trobe Univ, Melbourne, Victoria, Australia; 2) Laboratory of Phytopathology, Wageningen University, The Netherlands; 3) Plant & FoodResearch, Auckland, New Zealand.Effectors are secreted by pathogens to modify plant physiology and establish disease. Plant immune receptors have evolved to recognise effectors andcounter attack with defence responses. Most fungal effectors are lineage-specific, i.e. they are unique to a species, or to physiological races within aspecies. The availability of many whole genome sequences has revealed that some effectors are found in a discontinuous distribution within the fungalkingdom; a few phytopathogenic fungi (Colletotrichum higginsianum, Cercospora beticola, Fusarium oxysporum) possess an ortholog of Ave1 fromVerticillium dahliae, an effector that activates Ve1-mediated resistance in tomato. A subset of these orthologs were shown to activate Ve1-mediatedresistance in tomato. Unusually, Ave1 also shares similarity to an ortholog in the phytopathogenic bacterium Xanthomonas axonopodis, as well as to awidespread family of plant natriuretic peptides and expansins, involved in plant homeostasis and plant cell wall modification (de Jonge & van Esse et al.2012). We have identified an expanded Ave1-like gene family in apple and pear scab fungi, Venturia inaequalis and V. pirina. These species also haveexpanded gene families with similarity to the Leptosphaeria maculans effector AvrLm6. V. pirina has 14 unique hits (best,1.43e -18 ) to VdAve1. V. inaequalishas 17 unique hits (best,1.07e -22 ) to VdAve1. The distribution of Ave1 orthologs is suggestive of one or more cross-kingdom gene transfer events. We arecharacterising Venturia Ave1-like genes to investigate the mode of gene multiplication; seek evidence of horizontal gene transfer; and determine the role250

FULL POSTER SESSION ABSTRACTSof Ave1-like genes in pathogenicity. Ave1-like genes from non-Venturia fungi (and the bacterial gene) do not contain predicted introns, however, several(not all) V. inaequalis Ave1-like genes are predicted to contain introns. Codon usage bias among fungal, plant, and bacterial Ave1 orthologs, are beingcompared with the aim of inferring the kingdom of origin of the Venturia Ave1 orthologs. At least two ViAve1 orthologs are up-regulated during infectionof apple. To determine whether the Venturia Ave1 proteins also activate a Ve1-mediated hypersensitive response, each has been co-expressed withtomato Ve1 in Nicotiana tabacum, using an Agrobacterium tumefaciens-mediated transient transformation assay.527. Identification of targets of mycorrhizal effector proteins in planta. Natalia Requena, Carolin Heck, Ruben Betz. Molecular Phytopathology, KarlsruheInstitute of Technology, Karlsruhe, Germany.Plant roots are constantly approached by a myriad of microorganisms and are thus challenged to recognize friends from foes. Most plant roots engage ina mutualistic association with fungi from the Glomeromycota Phylum forming the arbuscular mycorrhiza (AM) symbiosis. The establishment of thisbeneficial association requires an intensive signal exchange including the down-regulation of PAMP triggered responses. We have shown that secretionand delivery of the effector proteins contribute to the manipulation of the plant defense response. In a previous work we showed that one of this proteinstravels to the plant nucleus and interacts with a plant transcription factor to down-regulate plant defenses. In order to identify further mechanisms of howsymbiotic effectors function we are investigating new plant targets of mycorrhizal effector proteins and how do they differ from targets from pathogenicfungi. Progress in this area will be presented.528. Structural basis for interactions of the Phytophthora sojae RXLR effector Avh5 with phosphatidylinositol 3-phosphate and for host cell entry.Furong Sun 2,3 , Shiv Kale 4 , Hugo Azurmendi 3 , Dan Li 2 , Brett M. Tyler 1,4 , Daniel Capelluto 2 . 1) Center for Genome Research and Biocomputing, Oregon StateUniversity, Corvallis, OR; 2) Dept of Biological Sciences, Virginia Tech, Blacksburg, VA 24061; 3) Dept of Chemistry, Virginia Tech, Blacksburg, VA 24061; 4)Virginia Bioinformatics Institute, Virginia Tech, Blacksburg, VA 24061.Oomycetes, such as Phytophthora sojae, employ protein effectors that enter host cells to facilitate infection. Entry of some effector proteins into plantcells is mediated by conserved RxLR motifs in the effectors and phosphoinositides (PIPs) resident in the host plasma membrane such asphosphatidylinositol 3-phosphate (PtdIns(3)P). Recent reports differ regarding the regions on RxLR effector proteins involved in PIP recognition. To clarifythese differences, we have structurally and functionally characterized the P. sojae effector, avirulence homolog-5 (Avh5). Using NMR spectroscopy, wedemonstrate that Avh5 is helical in nature with a long N-terminal disordered region. Heteronuclear single quantum coherence titrations of Avh5 with thePtdIns(3)P head group, inositol 1,3-bisphosphate (Ins(1,3)P2), allowed us to identify a C-terminal lysine-rich helical region (helix 2) as the principal lipidbindingsite in the protein, with the N-terminal RxLR (RFLR) motif playing a more minor role. Mutations in the RFLR motif slightly affected PtdIns(3)Pbinding, while mutations in the basic helix almost abolished it. Avh5 exhibited moderate affinity for PtdIns(3)P, which increased the thermal stability of theprotein. Mutations in the RFLR motif or in the basic region of Avh5 both significantly reduced protein entry into plant and human cells. Both regionsindependently mediated cell entry via a PtdIns(3)P-dependent mechanism. Our findings support a model in which Avh5 transiently interacts withPtdIns(3)P by specific electrostatic interactions mainly through its positively charged helix 2 region, enabling the RFLR domain to promote PI3P-mediatedhost entry. This study, including the identification of the PtdIns(3)P-binding site, provides an improved and updated model for how RxLR effector proteinsrecognize phosphoinositides and for the contributions of the RxLR motif and basic-rich C-terminal regions to the internalization process.529. Dose-dependent induction of plant immunity by application of the Fusarium mycotoxin deoxynivalenol. Antje Blümke, Christian A. Voigt. MolecularPhytopathology, Biocenter Klein Flottbek, Hamburg, Germany.The mycotoxin deoxynivalenol (DON) is associated with Fusarium head blight (FHB). This disease causes vast losses by reducing grain quantity andquality. Our study was aimed at analyzing cell wall changes in the host model grass Brachypodium distachyon due to infection with different F.graminearum mutants. The mutants vary in their ability to infect spikelets and to produce DON. As a reference, we used a F. graminearum wild-type strainwith the full capacity to produce DON and to cause FHB. The results were compared to the infection with the Dtri5, the Dfgl1, and the Dgpmk1 mutant.The Dtri5 mutant cannot produce DON due to disruption of the DON biosynthetic pathway. This mutant infects the directly inoculated spikelet withoutfurther propagation into the head tissue. A similar disease phenotype is described for the lipase-disruption mutant Dfgl1, which is able to produce DON.The MAP kinase disruption mutant Dgpmk1 is apathogenic but still able to produce DON. We observed similar disease phenotypes and amounts of DONfor all F. graminearum mutants during B. distachyon infection as described for wheat. 7 days post-inoculation (dpi), we analyzed the non-cellulosicmonocarbohydrate cell wall composition of B. distachyon spikelets by high-performance anion exchange chromatography with pulsed amperometricdetection (HPAEC-PAD). Only the infection with F. graminearum mutants that showed reduced virulence but still produce DON, namely the Dfgl1 andDgpmk1 mutants, resulted in compositional changes of the cell wall, an increase in the amount of glucose. Next, we wanted to know to what extend themycotoxin DON itself can induce cell wall changes. We applied DON solutions at different concentrations to B. distachyon spikelets. 7 dpi, the HPAEC-PADanalysis revealed an increase in the glucose amount only at relatively low DON concentrations of 1, 10, 100, and 1000 ppb whereas higher DONconcentrations of 50 and 100 ppm did not change the cell wall composition. However, only these high DON concentrations caused necrosis of florets.Interestingly, F. graminearum wild-type infection was significantly reduced on spikelets sprayed with a DON solution at a concentration of 1000 ppb 7 daysprior fungal inoculation. This suggests that the mycotoxin DON can induce an effector-triggered-like immunity in a dose-dependent manner.530. How Oomycete Pathogens Exploit PI3P to Target Secreted RxLR Effectors into Host Cells. Q. Wang 1 , S. Ferrer 1 , J. Carlough 1 , F. Arredondo 1 , S. Kale 2 ,B. Tyler 1 . 1) Botany and Plant Pathology, OREGON STATE UNIVERSITY, Corvallis, OR. 97330; 2) Virginia Bioinformatics Institute, Virginia Tech, Blacksburg,VA 24060.Effector proteins from diverse oomycetes and fungi can enter plant cells to facilitate infection. Recent research suggests that phosphoinositides (PIPs)resident in the host plasma membrane such as phosphatidylinositol 3-phosphate (PI3P) mediate the entry of some oomycete RxLR effectors. The PIPrecognition domain of these effectors is still controversial. Current evidence shows that either the RxLR domain or positive residues in the C-terminaldomains (Ct) of some effectors such as the P. sojae effectors Avr1b and Avh5 can bind PI3P, it has been unresolved which of these domains, if either, orboth, are involved in cell entry during natural pathogen infection. Here we have used heterologous PI3P-binding proteins, such as the yeast VAM7p PXdomain to replace the RxLR or Ct domains of Avr1b in P.sojae transformants. Our results reveal that the VAM7p PX domain can replace the RxLR domain ofAvr1b in carrying the C-terminal domain of Avr1b into soybean cells, conferring an avirulent phenotype on the transformants. Mutations that abolish thebinding of VAM7p to PI3P substantially reduce but do abolish avirulence conferred by the construct. Mutations in the PI3P-binding residues of the Avr1b Ctalso substantially reduce avirulence, while the double mutant cannot confer avirulence at all. These results strongly support the hypothesis that PI3Pbindingis essential for Avr1b cell entry during natural infection, and further suggest that efficient entry by Avr1b may require two PI3P binding sites.27th Fungal Genetics Conference | 251

FULL POSTER SESSION ABSTRACTS531. Identification and functional assay of Phytophthora sojae avirulence effectors. Yuanchao Wang, Suomeng Dong, Weixiao Yin. Plant Pathology Dept,Nanjing Agri Univ, Nanjing, China.Phytophthora sojae is a notorious oomycete pathogen producing a great loss on global soybean production annually. The disease outcome betweensoybean and P. sojae depends on whether hosts could recognize pathogen avirulence effectors. Recently identified oomycete avirulence effectors arecharacterized by N-terminal host entry motif (RxLR motif), sequence and transcriptional polymorphisms between virulent and avirulent strains. Benefitfrom 454 genome sequencing and solexa transcriptome sequencing of P. sojae strains, eight RxLR effectors are bioinformatically identified, geneticmapping suggested that two of them perfectly matched Avr3b and Avr1d phenotype respectively. Transient expression of the ORF from avirulence strainon soybean specifically triggered Rps3b and Rps1d mediated program cell death, respectively. confirming that they encodes avirulence effector Avr3b andAvr1d. Transient expression of Avr3b and Avr1d on Nicotiana benthamiana could promote the infection of Phytophthora capasici, suggesting bothavirulence effectors could suppress plant immunity and contribute to pathogen infection. Silencing of Avr3b impaired the virulence of Phytophthora sojae.Our progress in elucidating the mechanism under the inhibiting plant immunity by these effectors will be presented.532. Evaluating the translocation- and phospholipid binding abilities of the Phytophthora infestans AVR3a and Phytophthora sojae Avr1b RxLR-leaders.Stephan Wawra 1 , Armin Djamei 2 , Isabell Küfner 3 , Thorsten Nürnberger 3 , Justin A. Boddey 4 , Stephen C. Whisson 5 , Paul R.J. Birch 5 , Regine Kahmann 2 , Pietervan West 1 . 1) Sch Med Sci, Univ Aberdeen, Aberdeen, United Kingdom; 2) Department of Organismic Interactions, Max Planck Institute for TerrestrialMicrobiology, Germany; 3) Department of Plant Biochemistry, University Tübingen, Germany; 4) Department of Medical Biology, University of Melbourne,Australia; 5) Cell and Molecular Sciences, James Hutton Institute, Dundee, UK.Plant pathogenic oomycetes have a large set of secreted effectors that are directed into their host cells during infection. One group of these effectors arethe RxLR-effectors found in plant pathogenic oomycetes. These RxLR-effectors are defined as putative secreted proteins that contained a conservedtetrameric amino acid sequence motif, Arg-Xaa-Leu-Arg. This motif has to be within 40 amino acids C-terminal of the predicted cleavage sites of canonicalsignal peptides. Often this sequence is followed by a Glu-Glu-Arg (EER) motif. It has been shown, in a few cases, that the RxLR-motif is important for thedelivery of these proteins into host cells. However, how these proteins translocate into the cytoplasm of their host is currently the object of intenseresearch activity and debate. One model suggests that the RxLR-leader sequences of these effectors are sufficient to translocate the respective effectorsinto eukaryotic cells through binding to surface exposed phosphoinositol-3-phosphate. However, analysing the translocation behaviour of the RxLR-leadersfrom Phytophthora infestans avirulence protein 3a (AVR3a) and Phytophthora sojae avirulence protein 1b (Avr1b) we were unable to obtain conclusiveevidence for specific RxLR-mediated translocation. Importantly, we confirm that the reported phospholipid binding properties of AVR3a and Avr1b are notmediated by their RxLR-leaders. In addition, we will present data showing that the observed phospholipid interaction of the AVR3a effector domain isattributable to a weak association with denatured protein molecules, and is therefore most likely physiologically irrelevant.533. Identifying essential effectors from the soybean pathogen Phytophthora sojae. Hua Z. Wise 1,2 , Ryan G. Anderson 3 , John M. McDowell 3 , Brett M.Tyler 1,2 . 1) Center for Genome Research and Biocomputing and Department of Botany and Plant Pathology, Oregon State University; 2) VirginiaBioinformatics Institute, Virginia Tech; 3) Department of Plant Pathology, Physiology and Weed Science, Virginia Tech.Breeding for resistance to plant pathogens is one of the most effective means of disease control. However, the ability of plant pathogens evolve newpathogenicity factors and evade host defense mechanisms drives the continual necessity to identify new resistance genes. We are exploiting genomictechnologies in an effector-directed breeding approach that augments traditional breeding efforts against Phytophthora sojae, the causal agent of soybeanroot and seedling rot. This approach is founded on identifying monomorphic P. sojae effector genes that are essential for virulence, and using these genesas probes to identify new sources of resistance in soybean and related legumes. These essential effectors will make excellent candidates for screening fornew, durable resistance to P. sojae, as these genes presumably cannot be mutated or deleted without a significant fitness penalty. The majority ofpredicted P. sojae RXLR effector genes are polymorphic amongst sequenced isolates of P. sojae, however, a subset of P. sojae RXLR effectors displays littleor no allelic diversity. We have established a workflow for transient gene silencing and quantitative virulence assays. To date, we have silenced andassessed the virulence contribution of 17 PsAvh genes. Silencing of 13 of these effectors produced reduced virulence. Among these effectors, Avh16,Avh180 and Avh240 showed substantially reduced pathogen growth at early stages of host colonization and reduced disease symptoms at later stages ofinfection. We are currently using these three effectors as candidates in a high throughput screen system utilizing Pseudomonas Type III secretion system toscreen for new resistance genes against P. sojae.534. The LysM effector, Ecp6, is a virulence factor in the interaction of the hemibiotroph, Setosphaeria turcica, but not the necrotroph, Cochliobolusheterostrophus, with their common host, maize. Dongliang Wu 1 , Qing Bi 2 , Gillian Turgeon 1 . 1) Department of Plant Pathology & Plant-Microbe Biology,Cornell University, Ithaca, NY 14853, USA; 2) State Key Program of Microbiology and Department of Microbiology, College of Life Sciences, NankaiUniversity, Tianjin, China, 300071.Fungal phytopathogens are characterized as biotrophs, which derive nutrients from living cells, and necrotrophs, which kill host cells and retrievenutrients from dead tissue. Hemibiotrophs are intermediate in that they initially establish themselves in living host tissue, then undergo rapid killing ofplant cells later on. Hemi- and bio-trophic fungi utilize specialized effectors to prevent host recognition triggered by pathogen associated molecularpatterns (PAMPs), whereas necrotrophs often produce toxins that induce programmed cell death. Chitin, a signature component of fungal cell walls, is onetype of PAMPs known to trigger the plant resistance response. Several fungal chitin-binding LysM effectors have been identified that broker counterdefenseagainst chitin-triggered immunity, including the first characterized one, Ecp6, from the tomato biotroph, Cladosporium fulvum, Mg3LysM, fromthe wheat hemibiotroph, Mycosphaerella graminicola, and Slp1, from the rice hemibiotroph, Magnaporthe oryzae. In this study, Ecp6 homologs wereidentified and deleted from the genomes of two maize pathogens which differ in pathogenic lifestyle, Setosphaeria turcica (hemibiotroph), causal agent ofNorthern Leaf Blight and Cochliobolus heterostrophus (necrotroph), causal agent of Southern Corn Leaf Blight. Deletion of StECP6 caused reducedvirulence, whereas absence of ChECP6 did not alter virulence to the host. Real time RT-PCR demonstrated that expression of pathogenesis related maizegenes, PR1 gene and a chitinase gene was increased in Stecp6 mutants compared to wild type at 4 days post inoculation. Additional in planta geneexpression analyses are underway to compare host responses to these two fungi differing in pathogenic lifestyle on the same host.535. Nematode-trapping fungi eavesdrop on nematode pheromones. Yen-Ping Hsueh 1 , Parag Mahanti 2 , Frank Schroeder 2 , Paul Sternberg 1 . 1) HowardHughes Medical Institute and Division of Biology, California Inst of Technology, Pasadena, CA; 2) Boyce Thompson Institute and Department of Chemistryand Chemical Biology, Cornell University, Ithaca, NY.The recognition of molecular patterns associated with specific pathogens or food sources is fundamental to ecology and plays a major role in theevolution of predator-prey relationships. Recent studies showed that nematodes produce an evolutionarily highly conserved family of small molecules, the252

FULL POSTER SESSION ABSTRACTSascarosides, which serve essential functions in regulating nematode development and behavior. Here we show that nematophagous fungi, naturalpredators of soil-dwelling nematodes, can detect and respond to ascarosides. Nematophagous fungi use specialized trapping devices to catch andconsume nematodes, and previous studies demonstrated that most fungal species do not produce traps constitutively but rather initiate trap-formation inresponse to their prey. We found that ascarosides, which are constitutively secreted by many species of soil-dwelling nematodes, represent a conservedmolecular pattern used by nematophagous fungi to detect prey and trigger trap formation. Ascaroside-induced morphogenesis is conserved in severalclosely related species of nematophagous fungi and occurs only under nutrient-deprived condition. Our results demonstrate that microbial predatorseavesdrop on chemical communication among their metazoan prey to regulate morphogenesis, providing a striking example of predator-prey coevolution.We anticipate that these findings will have broader implications for understanding other inter-kingdom interactions involving nematodes, whichare found in almost any ecological niche on Earth.536. Molecular diagnosis to discriminate pathogen and apathogen species of the hybrid Verticillium longisporum on the oilseed crop Brassica napus.Van Tuan Tran, Susanna Braus-Stromeyer, Christian Timpner, Gerhard Braus. Institut für Mikrobiologie und Genetik, Georg-August-Universität Göttingen,Grisebachstr. 8, 37077 Göttingen, Germany.The cruciferous fungal pathogen Verticillium longisporum represents an allodiploid hybrid with long spores and almost double the amount of nuclearDNA compared to other Verticillium species. V. longisporum evolved at least three times by hybridization. In Europe, virulent A1xD1 and avirulent A1xD3hybrids were isolated from the oilseed crop Brassica napus. Parental A1 or D1 species are yet unknown whereas the D3 lineage represents Verticilliumdahliae. The V. longisporum isolates from Europe or California corresponding to hybrids A1xD1 or A1xD3 were analyzed. Only one single characteristic typeof ribosomal DNA (rDNA) could be assigned to each hybrid lineage. The avirulent A1xD3 isolates carried exclusively D3 rDNA, which corresponds to V.dahliae, whereas the rDNA of the virulent A1xD1 isolates originates from A1. Both hybrid lineages carry distinct isogene pairs of conserved regulatorygenes corresponding to either A1 or D1/D3. D1 and D3 paralogues show high identities but differ in several single nucleotide polymorphisms. Distinctsignatures of the VTA2 regulatory isogene pair allow the identification of V. longisporum hybrids by a single PCR and the separation from haploid speciesas A1 or D1/D3. The combination between the VTA2 marker as a barcode marker and differentiation of the rDNA type represents an attractive diagnostictool to discriminate allodiploid from haploid Verticillia and to distinguish between A1xD1 and A1xD3 hybrids, which differ in their virulence towards B.napus. Furthermore, the VTA2 gene was demonstrated to be a virulence factor that is required for fungal morphogenesis and plant infection.537. Investigating the Pathogenicity of Armillaria. Kathryn Ford 1 , Beatrice Henricot 3 , Kendra Baumgartner 2 , Gary D. Foster 1 , Andy M. Bailey 1 . 1) MolecularPlant Pathology, University of Bristol, Bristol, United Kingdom; 2) USDA-ARS, Plant Pathology, University of California, Davis, CA; 3) Royal HorticulturalSociety, Plant Pathology, Surrey, United Kingdom.Armillaria sp., or 'honey mushroom', is a generalist pathogen of fruit, nut and timber trees in gardens, forests and agricultural systems worldwide,causing Armillaria root disease and resulting in significant yield losses and millions of dollars worth of damage annually. Several questions regarding theinfection mechanisms used by basidiospores, hyphae and rhizomorphs and their subsequent colonisation processes remain unanswered. We established areproducible method of producing fruiting bodies in culture in order to generate basidiospores for use in Agrobacterium-mediated transformation tofacilitate further exploration of Armillaria’s pathogenicity. Results will be presented on the construction and utilisation of various plasmids conferringhygromycin resistance and fluorescent protein expression that have been used to transform A. mellea in order to study the infection mechanisms inherbaceous plants.538. Detoxification of nitric oxide by flavohemoglobin and the denitrification pathway in the maize pathogen Fusarium verticillioides. ThomasBaldwin 1,2 , Anthony Glenn 2 . 1) Plant Pathology Department, Univ of Georgia, Athens, GA; 2) USDA, ARS, R.B. Russell Research Center, Toxicology andMycotoxin Research Unit, , Athens, GA.The ephemeral nitric oxide (NO) is a free radical, highly reactive, environmentally rare, and a potent signaling molecule in organisms across kingdoms oflife. This gaseous small molecule can freely transverse membranes and has been implicated in aspects of pathogenicity both in animal and plant hosts.Fusarium verticillioides is a mycotoxigenic pathogen of maize, notable for its ability to persist as an asymptomatic endophyte. One potential determinantof this lifestyle conversion between overt pathogen and symptomless endophyte may be the regulation of NO. Detoxification of NO is a knownpathogenicity factor for the fungal human pathogen Candida albicans and the bacterial plant pathogen Erwinia chrysanthemi. Both mediate detoxificationby a flavohemoglobin protein (CaYHB1 and HmpX, respectively). BLASTP search of the F. verticillioides genome revealed two putative flavohemoglobinhomologs, denoted FHB1 and FHB2. Microarray analysis revealed a significant induction of FHB2 (13-fold) when the fungus was exposed to exogenous NO.FHB1 had a 2-fold increase. Also noteworthy from the microarray data is the distinct induction of genes within the denitrification pathway, includingdissimilatory nitrate reductase (dNaR, 16-fold increase), dissimilatory nitrite reductase (dNiR, 226-fold), and P450 nitric oxide reductase (P450nor, 27-fold).Flavohemoglobin has been noted as a component of the denitrification pathway, having a role in converting NO to nitrate. Thus, FHB2 is postulated to bethe paralog involved in the F. verticillioides denitrification pathway. Deletion mutants are being created in dNiR, P450nor, FHB1, and FHB2 to furtherevaluate functions of these genes in F. verticillioides. Mutants will be assayed for their endogenous production and regulation of NO, response toexogenous NO, virulence against maize, and mycotoxin production. Elucidating the function of these genes will give insight into the role of NO in F.verticillioides development, maize-fungal interactions, and denitrification, which has previously only been assessed in relation to anaerobic growth.539. Family disintegration: One Fusarium verticillioides beta-lactamase gene at a time. Scott E. Gold, Xiu Lin, Nicole J. Crenshaw, Anthony E. Glenn.Toxicology & Mycotoxin Research, USDA-ARS, Athens, GA.Fusarium verticillioides is a mycotoxigenic fungus found commonly on maize, where it primarily exhibits asymptomatic endophytic growth. The F.verticillioides genome possesses approximately 30 regions that potentially encode beta-lactamase enzymatic domains. These enzymes are classicallyinvolved in bacterial resistance to beta-lactam antibiotics, for example penicillinase. Our attention was drawn to this enzymatic function by the recentfinding that the gene FVEG_08291 is essential for resistance to maize phytoanticipins such as 2-benzoxazolinone (BOA), which possesses a gamma-lactammoiety, the presumed enzymatic target (see poster by Glenn et al.). FVEG_08291 belongs to a subset of these enzymes known as metallo-beta-lactamases.Beta-lactamase enzyme function is not well studied in the fungi, so, in order to further evaluate the roles of these enzymes in F. verticillioides, we are inthe process of deleting the members of their encoding gene family. We assigned directed-research undergraduates each a specific gene, for which theyproduced deletion constructs by DelsGate and/or OSCAR methodology and generated fungal transformants for analysis. Deletion mutants in one of theother metallo-beta-lactamase encoding genes (FVEG_12159) showed a dramatic defective growth phenotype. This observation raises the interestinghypothesis that perhaps this mutant is no longer resistant to a lactam moiety containing compound produced by F. verticillioides itself. Data will bepresented on initial progress with this project.27th Fungal Genetics Conference | 253

FULL POSTER SESSION ABSTRACTS540. A Fungal Metallo-Beta-Lactamase Necessary for Biotransformation of Maize Phytoprotectant Compounds. Anthony E. Glenn, C. Britton Davis,Maurice E. Snook, Scott E. Gold. Toxicology & Mycotoxin Research, USDA-ARS, Athens, GA.Xenobiotic compounds such as phytochemicals, microbial metabolites, and agrochemicals can impact the diversity and frequency of fungal speciesoccurring in agricultural environments. Resistance to xenobiotics may allow plant pathogenic fungi to dominate the overall fungal community, withpotential negative impacts on crop yield and value. The mycotoxigenic Fusarium verticillioides is such a fungus commonly associated with maizeworldwide, often contaminating maize kernels with the fumonisin mycotoxins. The dominance of F. verticillioides as an endophyte may be due in part toits ability to metabolize phytoprotectants produced by maize. The benzoxazinoids and benzoxazolinones are broad spectrum allelopathic, antimicrobial,and anti-herbivory compounds from maize, yet F. verticillioides can rapidly biotransform these phytochemicals into non-toxic metabolites. We haveidentified the genes responsible for the biotransformation process. Two gene clusters were identified that correspond to the previously characterizedFDB1 and FDB2 loci, with both loci being necessary for metabolic tolerance to 2-benzoxazolinone (BOA), one of the maize phytoprotectants. Analysis of thenine ORFs (FVEG_08287 to FVEG_08295) at the FDB1 locus indicated that one of the genes (FVEG_08291) encodes a protein having a metallo-betalactamasedomain, and deletion of the gene in wild-type strain M3125 resulted in the fungus being unable to grow on BOA-amended agar due to aninability to metabolize the compound. Deletion mutants were complemented to wild-type phenotype by transformation with the native allele. Other ORFswere not found to be essential when deleted in M3125. Microarray analysis indicated the metallo-beta-lactamase (FVEG_08291) had a 13-fold induction inresponse to BOA (2-hr incubation), with other genes in the cluster ranging from 3-fold (FVEG_08287) to 42-fold induction (FVEG_08292). Beta-lactamasesare well-known for conferring bacterial resistance to lactam-type antibiotics, but to our knowledge this is the first report of fungal enzymes of this typemetabolizing lactam-like xenobiotics. We are investigating other beta-lactamase encoding genes in F. verticillioides (see poster by Gold et al.) to furtherevaluate their possible role in tolerance to both exogenous as well as endogenous metabolites having lactam-type moieties.541. Nectria haematococca DNase: role in dynamics and localization of pea root infection. D. Huskey, G. Curlango-Rivera, Z. Xiong, H. Van Etten, M.Hawes. University of Arizona, Tucson, AZ.Root tips of pea (Pisum sativum L.) are protected from N. haematococca infection by an extracellular DNA (exDNA)-based trapping process similar to thatoccurring in mammalian defense responses to bacterial and fungal pathogens. N. haematococca spores germinate rapidly in response to root border cellpopulations programmed to export exDNA and antimicrobial proteins as they detach from the root cap. Within 24 h, hyphae and border cells togetherform a mantle which ensheaths the root tip and separates, leaving the root apex uninvaded: >98% of root tips escape infection. When the exDNA isdegraded with DNase added to the root at the time of inoculation, resistance is abolished: 100% of root tips are invaded by proliferating hyphae and rootgrowth ceases within 24 hours. In N. haematococca isolates harboring a conditionally dispensable (CD) chromosome, the process occurs more rapidly thanin CD-minus isolates; a ten-fold increase in spores from CD-minus isolates results in comparable dynamics. Putative DNase-encoding sequences have beendetected on two different CD chromosomes, and direct tests have revealed increased extracellular DNase activity from CD-plus isolates compared with CDminusisolates. The goal of this study is to examine predictions of the hypothesis that CD chromosome encoded DNase activity plays a role in pea rootinfection.542. FvSNF1, a protein kinase of Fusarium virguliforme that affects SDS development in Soybean. K.T. Islam, Ahmad Fakhoury. Plant, Soil and AgSystems, Southern Illinois University, Carbondale, IL.Fusarium virguliforme is a soil-borne pathogen that causes Sudden Death Syndrome (SDS). SDS is one of the top four yield-robbing fungal diseases insoybean resulting in significant economic losses to producers. Despite the importance of SDS, a clear understanding of fungal genetic factors that affectthe development of the disease is still lacking. The aggressiveness of F. virguliforme on infected soybean plants is believed to require the activity of cellwall-degrading enzymes (CWDE). The production of these CWDEs in phytopathogenic fungi is under catabolic repression. In Saccharomyces cerevisiae,catabolic repression is regulated by SNF1 (sucrose non-fermenting 1). To investigate the role of cell wall-degrading enzymes as determinants of F.virguliforme aggressiveness, the F. virguliforme SNF1 homologue FvSNF1 was targeted for disruption. The resulting FvDsnf1 transformant failed to grow ongalactose and grew poorly when arabinose or sucrose where the main carbon source. The mutation did not seem to affect the ability of the fungus to growwith glucose, fructose, maltose, or xylose as the main source of carbon. More importantly, in greenhouse experiments, the FvDsnf1 transformant wasseverely impaired in its ability to cause SDS on challenged soybean plants.543. Functional and molecular analysis of AstA sulfate transporter in pathogenic Fusarium sambucinum with respect to its virulence and ability to infectpotato. Sebastian Pilsyk 1 , Hanna Gawinska-Urbanowicz 2 , Renata Natorff 3 , Marzena Sienko 3 , Joanna S. Kruszewska 1 . 1) Laboratory of Fungal Glycobiology,Institute of Biochemistry and Biophysics, Warsaw, Poland; 2) The Plant Breeding and Acclimatization Institute (IHAR), Bonin, Poland; 3) Department ofGenetics, Institute of Biochemistry and Biophysics, Warsaw, Poland.AstA protein (alternative sulfate transporter) represents a little known type of sulfate trasporter, belonging to an extensive and poorly characterizedfamily of allantoate permeases Dal5. In Aspergillus nidulans the astA gene is under the control of Sulfur Metabolite Repression (SMR). The closesthomologs of astA are frequent in evolutionarily distant fungi belonging to the Pezizomycotina subphylum which exhibit similar plant pathogenicity.Fusarium sp. fungi, like F. sambucinum, contribute to serious devastation of potato crops and increase the cost of cultivation due to the application ofpesticides. Due to the similarity on the metabolic level between pathogenic fungi and the host, there is a problem with efficient plant protection.The aim of this project is elucidation of AstA function upon infection and colonization of potato tubers by the fungal pathogen, Fusarium sambucinumand by its astA deletion mutant. We have observed a high expression level of astA in infected potato tubers and its regulation by SMR as in A. nidulans.The study also involves the identification of amino acid residues crucial for sulfate binding and transportation by the generation of point mutations anduptake analysis. Elucidation of the biological function of AstA will help understanding of fungal pathogenic adaptations upon changes in plant hostmetabolism and definition of a new promising target for a potential fungicide.544. WITHDRAWN545. The adenylate cyclase of the cereal pathogen Fusarium graminearum controls infection structure development, mycotoxin production andvirulence to wheat. Jörg Bormann, Marike Johanne Boenisch, Elena Brückner, Demet Firat, Cathrin Kröger, Birgit Hadeler, Wilhelm Schäfer. MolecularPhytopathology, University Hamburg, Hamburg, Germany.Fusarium graminearum is one of the most devastating pathogens of cereals. Mycotoxins accumulating in infected grains are a serious threat to food andfeed worldwide. Knowledge about the molecular basis of infection and mycotoxin production is still limited. Cyclic 3’,5’-adenosine monophosphate (cAMP)254

FULL POSTER SESSION ABSTRACTSis a nucleotide derived from adenosine triphosphate that acts as a second messenger throughout all kingdoms. Intracellular cAMP levels are subject to alarge membrane-bound protein, the adenylate cyclase. In order to analyze the function of this gene and the importance of cAMP in the life cycle of F.graminearum, the adenylate cyclase gene (FGSG_01234) was deleted from the genome (DFgac1). DFgac1 displayed a drastically reduced growth oncomplete medium. This reduction in growth could partially be complemented by addition of a cAMP analog. Furthermore, the mutant was unable toproduce perithecia on detached wheat nodes but more artificial conditions like carrot agar allowed perithecia development. Possibly, this points to asensing problem of DFgac1. Although growth on agar was reduced, conidia production was increased. Pathogenicity towards wheat was drasticallyreduced in DFgac1 compared to the wild type. Point-inoculated spikelets showed only small lesions even after 21 days post inoculation. No deeperinfection occurred and mycelial growth was never detectable near the rachis. Thus, fungal hyphae never grew from the inoculated spikelet to the adjacentone. Fluorescence microscopy using a DFgac1-strain expressing dsRed constitutively in the cytosol revealed that FgAC1 controls the development ofinfection structures like lobate appressoria and infection cushions. Removal of hyphae superficially colonizing flower leaves and subsequent analysis byscanning electron microscopy demonstrated the lack of any fungal penetration holes. Instead, hyphae on flower leaves produced massively new conidia,thereby circumventing the infection cycle, something never observed in the wild type. DFgac1-strains are unable to produce the mycotoxin deoxynivalenolboth in vitro and during wheat infection. In this study, for the first time, we implicate the cAMP signaling pathway to important processes in F.graminearum like development of infection structures, pathogenicity, secondary metabolite production and sexual reproduction.546. The ATF/CREB transcription factor Atf1 is essential for full virulence, deoxynivalenol production and stress tolerance in the plant pathogenFusarium graminearum. Thuat Van Nguyen, Birgit Hadeler, Cathrin Kröger, Wilhelm Schäfer, Jörg Bormann. Molecular Phytopathology, UniversityHamburg, Hamburg, Germany.The filamentous ascomycete Fusarium graminearum is a highly organ specific pathogen that resides on small grain cereals like rice, wheat, barley, andmaize. Grains infected with F. graminearum accumulate high amounts of mycotoxins, most prominent of which are deoxynivalenol (DON) and zearalenone(ZEA). The stress-activated MAP-kinase FgOS-2 (Saccharomyces cerevisiae HOG1) is a central regulator in the life cycle of F. graminearum (Nguyen et al.,2012. MPMI 25:1142-1156). FgOS-2 regulates, among others, virulence to wheat and maize, and DON- and ZEA-production. Here, we present data on thefunctional characterization of a putative downstream regulator, the ATF/CREB activating transcription factor FgAtf1. We created deletion and overexpressionmutants of Fgatf1, the latter one also in an FgOS-2 deletion mutant. Like FgOS-2, FgAtf1 is mainly involved in osmotic stress response.Bimolecular fluorescence complementation demonstrates an interaction of both proteins under osmotic stress conditions. Deletion mutants in Fgatf1(DFgatf1) are more sensitive to osmotic stress (e.g. mediated by NaCl) and less sensitive to oxidative stress mediated by H 2O 2 compared to the wild type.Furthermore, sexual reproduction is delayed: perithecia develop much slower and some remain immature even after prolonged incubation. DFgatf1strains show an increased DON-production under in-vitro induction conditions compared to the wild type. However, during wheat infection, DONproductionis strongly reduced. Expression of genes encoding for key enzymes in the DON-biosynthesis pathway is regulated accordingly. In infectionassays on wheat and maize, the DFgatf1 strains show a reduced virulence compared to the wild type. Interestingly, constitutive expression of Fgatf1 leadsto hypervirulence on wheat, maize and Brachypodium distachyon. Moreover, constitutive expression of Fgatf1 in a DFgOS-2 mutant background partiallycomplements DFgOS-2-phenotypes regarding growth on osmotic-stress medium, sexual reproduction, and virulence towards wheat and maize.Furthermore, FgAtf1 is involved in the regulation of light-responsive genes. Taken together, these results provide new insights in the stress responsesignaling cascades of F. graminearum and assign the transcription factor FgAtf1 a central role in pathogenic development and secondary metabolism.547. The stress-activated protein kinase FgOS-2 is a key regulator in the life cycle of the cereal pathogen Fusarium graminearum. Thuat Van Nguyen,Birgit Hadeler, Cathrin Kröger, Wilhelm Schäfer, Jörg Bormann. Molecular Phytopathology, University Hamburg, Hamburg, Germany.Fusarium graminearum is one of the most destructive pathogens of cereals and a threat to food and feed production worldwide. It is an ascomycetousplant pathogen and the causal agent of Fusarium head blight disease in small grain cereals and of cob rot disease in maize. Infection with F. graminearumleads to yield losses and mycotoxin contamination. Zearalenone (ZEA) and deoxynivalenol (DON) are hazardous mycotoxins; the latter is necessary forvirulence towards wheat. Deletion mutants of the F. graminearum orthologue of the Saccharomyces cerevisiae Hog1 stress-activated protein kinase, FgOS-2 (DFgOS-2), showed drastically reduced in planta DON and ZEA production. However, DFgOS-2 produce even more DON than the wild type under in vitroconditions, whereas ZEA production is similar to that of the wild type. These deletion strains are dramatically reduced in pathogenicity towards maize andwheat. We constitutively expressed the fluorescent protein dsRed in the deletion strains and the wild type. Microscopic analysis revealed that DFgOS-2 isunable to reach the rachis node at the base of wheat spikelets. During vegetative growth, DFgOS-2 strains exhibit increased resistance against thephenylpyrrole fludioxonil. Growth of mutant colonies on agar plates supplemented with NaCl is reduced but conidia formation remained unchanged.However, germination of mutant conidia on osmotic media is severely impaired. Germ tubes are swollen and contain multiple nuclei. The deletion mutantscompletely fail to produce perithecia and ascospores. Furthermore, FgOS-2 also plays a role in reactive oxygen species (ROS)-related signalling: FgOS-2deletion mutants are more resistant against oxidative stress mediated by H 2O 2. We found that the transcription and activity of fungal catalases ismodulated by FgOS-2. Among the genes regulated by FgOS-2 we found a putative calcium-dependent NADPH-oxidase (noxC) and the transcriptionalregulator of ROS metabolism, Fgatf1. The present study describes new aspects of stress-activated protein kinase signalling in F. graminearum.548. Innate Immunity in Fusarium graminearum. Vong shian Simon Ip Cho 1,2 , Gitte Erbs 3 , Thomas Sundelin 3 , Peter Busk 4 , Mari-Anne Newman 3 , StefanOlsson 1 . 1) Genetics and Microbiology, University of Copenhagen, Copenhagen, Denmark; 2) USDA-ARS Cereal Disease Laboratory, University ofMinnesota, Saint Paul, MN, USA; 3) Transport Biology, University of Copenhagen, Copenhagen, Denmark; 4) Dept. Biotechnology, Aalborg University,Copenhagen, Denmark.Fungi are often mostly recognized as plant pathogens that cause harm to important economical plants. In nature however, fungi are frequently victims ofbacterial parasitism but little is known about fungal defense mechanisms. The potential existence of fungal innate immunity was studied using Fusariumgraminearum as model organism and bacterial flagellin to mimic the presence of bacteria in an in vitro environment. The presence of flagellin triggered aninitial mitochondrial and cell membrane hyperpolarization which was detected using the florescent dye DiOC 7(3). This was followed by the production ofthe secondary signalling molecule Nitric Oxide (NO), common to innate immunity signalling in other eukaryotes. NO was monitored using the fluorescentdye DAF-FM. NO appears to be produced by an inducible enzyme that is regulated by complex mechanisms but centrally modulated byCalcium/Calmodulin. Inhibition studies suggest the presence of a Nitric Oxide Synthase (NOS), but no typical arginine utilizing NOS was identified withinthe F. graminearum’s genome by homology search. Various genes bearing resemblance to the archetypal NOS, as well as argininosuccinate lyase weredeleted. However, the mutants still produced NO. The presence of alternative pathways contributing towards the production of NO was investigated byadding a variety of potential substrates to challenged cultures. Various reactions were observed suggesting that several pathways are present. Inconclusion, F. graminearum reacts strongly to the presence of the bacterial Microbial Associated Molecular Pattern (MAMP) flagellin with an up-regulation27th Fungal Genetics Conference | 255

FULL POSTER SESSION ABSTRACTSof NO production showing the presence of innate immunity-like responses also in fungi.549. Balanced posttranslational activation of eukaryotic translation initiation factor 5A is required for pathogenesis in Fusarium graminearum. Ana LiliaMartinez-Rocha 1 , Mayada Woriedh 1,2 , Jan Chemnitz 3 , Peter Willingmann 1 , Joachim Hauber 3 , Wilhelm Schäfer 1 . 1) Molecular Phytopathology, University ofHamburg, Hamburg, Hamburg, Germany; 2) Cell Biology and Plant Biochemistry, University of Regensburg, Germany; 3) 3Heinrich-Pette-Institute forExperimental Virology and Immunology, Martinistrasse 52, D- 20251 Hamburg, Germany.Activation of the eukaryotic translation initiation factor 5A (EIF5A) requires a posttranslational modification, changing a lysine into the unique amino acidhypusine. This activation is a two steps reaction mediated by deoxyhypusine synthase (DHS) and deoxyhypusine hydroxylase (DOHH). First DHS cleavageand transfers the 4-aminobutyl moiety from the spermidine to an specific lysine in EIF5A to form the deoxyhypusine intermediate, during the second stepDOHH hydroxylate the 4-amonibutyl moiety to create the active form of eIF5A containing hypusine. The activated protein transports mRNAs from thenucleus to the ribosomes, where initiates protein biosynthesis. This system is conserved from Archea to humans and is involved in diseases as diverse asHIV infection, malaria, cancer, and diabetes. Until now, inhibition or silencing of DHS or DOHH has been tested to modify hypusination. For the first time,we evaluate its importance by over-expressing the enzymes that control hypusination of EIF5A. Over-expression of DOHH (oexDOHH) prevents virulence ofFusarium graminearum to wheat and maize. In contrast, over-expression of DHS (oexDHS) leads to an increase of virulence to wheat. Simultaneous overexpressionof both enzymes results in virulence comparable to the wild type strain. GFP assisted histology revealed that oexDOHH mutant is unable toform infection structures on wheat flower leaves and barely grows in point inoculated wheat spikelets. OexDHS results in an increase of infectionstructures and accordingly in an increase of virulence. We determined general hypusine formation by incorporating radiolabeled spermidine in EIF5Aduring in culture growth. Wild type and oexDHS showed similar hypusination intensity, whereas oexDOHH showed an increased incorporation. Thedifferential hypusination state of ElF5A was determined by 2D gels and western blot due to the difference in isoelectric point of the three states; inactive(lysine), intermediate (deoxyhypusine) or active (hypusine). Preliminary results show the wild type strain with all three hypusination states, oexDHS anddouble over-expressing mutants with an increased inactive and intermediate forms, and oexDOHH mutant lacked the inactive and intermediate form, onlythe activated form was detectable. We conclude that a balanced hypusination is required for proper function of EIF5A.550. The Con7 transcription factor, essential for pathogenicity, regulates the expression of genes involved in glycolysis and virulence in Fusariumoxysporum. Carmen Ruiz-Roldán 1,2 , Yolanda Pareja-Jaime 1,2 , M. Isabel G. Roncero 1,2 . 1) Department of Genetics, University of Cordoba, Spain; 2) Campusde Excelencia Agroalimentario (ceiA3).Transcription factors (TF) regulating the different stages of infection like adhesion to the host surface, differentiation of infection structures andpenetration represent potential molecular targets for fungicides with specific modes of action. Our studies on Fusarium oxysporum have demonstrated theessential role of morphogenetic regulation in pathogenesis, including processes such as cell-wall biogenesis, cell division and differentiation of infectionstructures. We identified the Con7 TF whose inactivation produces non-pathogenic mutants with altered morphogenesis, including abnormal polar growthand hyphal branching. To identify genes dependent on Con7 protein profiles of wild type and Dcon7 mutant were compared by 2D electrophoresis.Expression of 126 proteins varied quantitatively between both strains with statistical significance. Among the 80 proteins identified by MALDI-TOF/TOF-MS15, 9, and 4 were associated with secondary metabolism, glycolysis/gluconeogenesis (Gly/Glu) and pentose and glucuronate interconversions,respectively. Additionally, 6 proteins were known virulence factors, including citochrome P450 monooxygenase, O-methyltransferase, peptidyl-prolyl cistransisomerase D, peroxidase/catalase, phospholipase C and superoxide dismutase. Expression of the responsible genes was confirmed by qRT-PCR. Toverify the role of Con7 in Gly/Glu pathways, the intracellular ATP levels and alcohol dehydrogenase (ADH) activity in Dcon7 were compared with wt. Wedetected 2.4 fold increased ATP and 27.5% reduced ADH activity in Dcon7. Additionally, Dcon7 showed a dramatic growth reduction in the presence ofglucose, glycin or polygalacturonic acid, indicating nutrient assimilation defects. No significant differences were detected in intracellular glucoseaccumulated by both strains, while extracellular glucose levels in Dcon7 were significantly higher, suggesting defective sugar transport. RT-PCR analyses inthe wt revealed the existence of four con7 transcripts that differ in size and abundance, indicating alternative intron splicing. To determine thefunctionality of the different deduced proteins, Dcon7 was complemented with cDNA fragments from each version of the mature Con7 protein.Phenotypic and pathotypic characterization of the transformants should reveal their role in the different phenotypes observed in the Dcon7.551. Fusarium oxysporum produces volatile organic compounds that enhance the growth and stress resistance of Arabidopsis thaliana. Vasileios Bitas 1 ,Michael Axtell 2 , James Tumlinson 3 , Seogchan Kang 1 . 1) Department Plant Pathology and Environmental Microbiology, Pennsylvania State Univ, UniversityPark, PA; 2) Department of Biology, Pennsylvania State Univ, University Park, PA; 3) Department of Entomology, The Pennsylvania State University,University Park, PA.Production of volatile organic compounds (VOCs) as signal molecules is a widespread and efficient mode of communication utilized by all organisms.Microbial VOCs promoting plant growth and stress resistance present an environmentally and economically attractive alternative to fertilizers andpesticides. Fungi are some of the most predominant and yet, under-investigated organisms that employ VOCs in order to regulate and affect surroundingenvironments including neighboring organisms. Certain isolates of F. oxysporum, a well-known soilborne fungus, produce VOCs that promote plantgrowth, alter morphological and physiological properties, and enhance biotic and abiotic stress resistance in the model plant Arabidopis thaliana. In orderto investigate the mode of action by which those volatiles function, we have employed genetic and molecular resources and tools including A. thalianamutants that are defective hormonal signaling pathways and gene expression analysis. We are also trying to identify those compounds that affect A.thaliana through the use of GC-MS. Identification of the fungal VOCs triggering these plant responses and elucidation of the physiological and molecularalterations occurring in plants will help us shed light into the mechanism underpinning complex VOC-mediated interactions.552. Lipolytic system of the tomato pathogen Fusarium oxysporum f.sp. lycopersici. G. A. Bravo Ruiz 1,2 , C. Ruiz Roldán 1,2 , M. I. González Roncero 1,2 . 1)Departamento de Genética, Universidad de Córdoba; 2) Campus de Excelencia Agroalimentario (ceiA3), E-14071 Cordoba, Spain.The lipolytic profile of Fusarium oxysporum f.sp lycopersici has been determined on the basis of in silico analyses search and validated by biochemicalenzyme activity determination of Wheat Germ Oil (WGO) induced cultures. Twenty five identified structural secreted lipases show the conservedpentapeptide -Gly-X-Ser-X-Gly- characteristic of fungal lipases and the signal sequence for extracellular secretion. On the other hand, two transcriptionalregulatory predicted lipase genes have been identified showing nuclear localization signals (NLS) and the Zn2Cys6 zinc finger DNA binding motifs. Thetranscription profile of twelve structural genes, during tomato plant colonization in the wild type strain, shows variable expression levels (100 fold-range)being lip1, lip3, and lip22 the highest induced (20% relative to the actin gene). The maximal level of expression is observed in roots at 21-96 hours postinoculation.Targeted replacement of four structural (lip1, lip2, lip3 and lip5), and two regulatory ctf1 (orthologue to Fusarium solani ctf1 and to Aspergillusnidulans farA) and ctf2 (orthologue to F. solani ctf2 and to A. nidulans farB), lipase predicted genes originated the corresponding single deletion mutants256

FULL POSTER SESSION ABSTRACTSand the double deletion mutant Dctf1Dctf2. In vitro qRT-PCR expression analyses of twelve structural lipase genes in the regulatory mutants Dctf1, Dctf2and Dctf1Dctf2, in comparison with the wild type strain, demonstrate the existence of a complex lipase regulation network in F. oxysporum. The reductionof total lipase activity (10-30%), besides the severe altered pathogenic behaviour on tomato plants shown by the single Dctf1, Dctf2, and the doubleDctf1Dctf2 mutants suggest an important role of the lipolytic system of this fungus in pathogenicity.553. Role of glycogen metabolism in the pathotypic behavior of Fusarium oxysporum f.sp. lycopersici on tomato plants. C. Corral Ramos 1,2 , C. RuizRoldan 1,2 , M. I. González Roncero 1,2 . 1) Departamento de Genética, Universidad de Córdoba; 2) Campus de Excelencia Agroalimentario (ceiA3), E-14071Córdoba, Spain.Glycogen can play different roles in biological systems, such as storage carbohydrate for energy and/or carbon or control of glucose metabolism. In mosteukaryotes, glycogen is built on a self-glucosylating initiator protein core, glycogenin (gnn), which acts as a primer for glycogen synthase (gls). Thebranches are created by a specific enzyme (gbe) which transfers a block of 6-7 residues from the end of a linear chain of glucose units to an internalglucose residue by an a-1,6 linkage. De-branching, which is essential for the degradation of glycogen, is catalyzed by a distinct enzyme (gdb) which acts onbranches containing only four residues, transferring three of them to the end of a linear chain and then hydrolyzing the final residue. The level of glycogenfound in a particular situation results from the balance between glycogen synthase and glycogen de-branching activities, resulting in the synthesis anddegradation of this compound, respectively. Additionally, both activities are regulated by the action of a glycogen phosphorilase (gph). In order to studythe role of glycogen metabolism in Fusarium oxysporum pathotypic behavior we generated single deletion mutants of the genes Dgnn, Dgls, Dgbe, Dgdband Dgph, and the double mutant DgphDgdb, by direct targeted replacement. Quantification of glycogen reserves during in vitro growth indicated anincrease along the time period (72 h) up 700mg glucose equivalents per mg protein in a pH independent manner. As expected, no detectable glycogen wasaccumulated in any of three single structural deletion mutants Dgnn5, Dgls10 or Dgbe17. By contrast, glycogen levels were 10% higher in the single Dgdb2and Dgph8, and the double Dgph8Dgdb2 mutants in comparison to wild type. Similar hyphal agglutination patterns were observed in the three singlemutants Dgnn5, Dgls10 and Dgbe17 compared to the wild type, whereas those strains affected in glycogen catabolism, Dgdb2, Dgph8, and the doublemutant Dgph8Dgdb2, showed a dramatic reduction in hyphal agglutination. This phenotype did not exactly correlate with conidial anastomosis tube (CAT)formation since all the mutants, except Dgls10, showed a 40-50% reduction in hyphal fusions. We are currently performing tomato plant infection assayswhat will help us to gain insight into the role of glycogen metabolism in the virulence of F. oxysporum.554. Identification of chemoattractant compounds from tomato root exudate that trigger chemotropism in Fusarium oxysporum. El Ghalid Mennat,David Turra, Antonio Di Pietro. Departamento de Genética, Universidad de Córdoba, 14071 Córdoba, Spain.Fusarium oxysporum is a soilborne pathogen that causes vascular wilt disease on a wide range of plant species, including tomato (Solanum lycopersicum).The host signals that trigger fungal infection are currently unknown. A chemotropic response of F. oxysporum towards tomato root exudate was observedusing a plate assay that measures directed growth of fungal germ tubes towards chemoattractants. To purifiy the chemoattractant coumpound(s) fromtomato root exudate, we applied a series of purification methods including extraction with organic and inorganic solvents, fractionation by size exclusionand ion exchange chromatography. The compound(s) showing chemoattractant activity were found in the hydophilic fraction, had a molecular weightbetween 30 and 50 kDa and were sensitive to boiling and treatment with proteinase K, suggesting that they correspond to one or several secreted tomatoproteins. Polyacrylamide gel electrophoresis of the active fraction revealed multiple protein bands of the expected size, two of which displayedchemoattractant activity when eluted from the gel. Identification of the active protein(s) by LC-ESI-MS is currently ongoing. Identification of the secretedchemoattractant(s) from tomato roots will advance our understanding of the molecular events that trigger fungus-root interactions.555. TOR-mediated control of virulence functions in the trans-kingdom pathogen Fusarium oxysporum. Gesabel Y. Navarro Velasco, Antonio Di Pietro.Departamento de Genética, Universidad de Córdoba, 14071 Córdoba, Spain.Infectious growth of fungal pathogens is controlled by environmental cues, including nutrient status. The soilborne fungus Fusarium oxysporum producesvascular wilt disease in more than a hundred different crop species and can cause lethal systemic infections in immunodepressed humans. Previous workshowed that the preferred nitrogen source ammonium causes repression of infection-related processes in F. oxysporum that could be reversed byrapamycin, a specific inhibitor of the conserved protein kinase TOR. Here we generated mutations in upstream components that should result inconstitutive activation of TOR, including null mutants in tuberous sclerosis complex 2 (TSC2), a small GTPase that represses TOR activity, as well as strainsexpressing a dominant activating allele of the small GTPase Rag (ragA Q86L ), an activator of TOR. The Dtsc2 mutants and, to a minor extent, the ragA Q86Lstrains showed defects in hyphal growth and colony morphology on several amino acids, as well as decreased efficiency in cellophane penetration andvegetative hyphal fusion. These phenotypes were exacerbated in Dtsc2ragA Q86L double mutants and could be reversed by rapamycin, suggesting that theyare caused by hyperactivation of TOR. The mutants caused significantly lower mortality on tomato plants and on larvae of the animal model host Galleriamellonella. These results suggest that TOR functions as a negative regulator of fungal virulence on plant and animal hosts.556. Components of the urease complex govern virulence of Fusarium oxysporum on plant and animal hosts. Katja Schaefer, Elena Pérez-Nadales,Antonio Di Pietro. Departamento de Genética, Universidad de Córdoba, 14071 Cordoba, Spain.In the soilborne pathogen Fusarium oxysporum, a mitogen-activated protein kinase (MAPK) cascade homologous to the yeast filamentous growthpathway controls invasive growth and virulence on tomato plants. Full phosphorylation of Fmk1 requires the transmembrane protein Msb2, a member ofthe family of signalling mucins that have emerged as novel virulence factors in fungal plant pathogens. A yeast two-hybrid screen for proteins interactingwith the Msb2 cytoplasmic tail identified UreG, a component of the urease enzymatic complex. UreG belongs to a set of accessory proteins needed toactivate Apo- urease, which converts urea to yield ammonia and carbon dioxide. The F. oxysporum genome contains two structural urease genes, ure1 andure2. Mutants in ureG or ure1 showed reduced growth on urea as the sole carbon and nitrogen source. Lack of urease activity in the mutants resulted infailure to secrete ammonia and to increase the extracellular pH. The DureG mutants caused significantly reduced mortality on tomato plants and on theanimal model host Galleria melonella, while Dure1 mutants only showed reduced virulence on tomato plants. Real-time qPCR analysis of key genesinvolved in nitrogen uptake and assimilation, as well as in the urea cycle, during infectious growth of F. oxysporum in G. melonella revealed increasedtranscript levels of arginase, which converts arginine to urea. Our results suggest a role for the urease accessory protein UreG in fungal virulence on plantand animal hosts.557. Knock-out of the Fusarium oxysporum f.sp. lycopersici homologs of the DNA-methylation genes DIM2 and HP1 does not affect effector geneexpression. Charlotte van der Does, Jerom van Gemert, Karlijn Klei, Ido Vlaardingerbroek, Martijn Rep. Molecular plant pathology, SILS, University ofAmsterdam, Amsterdam, Netherlands.27th Fungal Genetics Conference | 257

FULL POSTER SESSION ABSTRACTSIn the tomato pathogen Fusarium oxysporum f. sp. lycopersici, most known effector genes reside on a pathogenicity chromosome that can be exchangedbetween strains through horizontal transfer. Expression of these effector genes is induced upon infection, but the mechanism by which this is regulated isunknown. We noticed that targeted deletion of the effector genes on the pathogenicity chromosome has a particular low rate of succes, when comparedto genes on the core genome. Possibly, the pathogenicity chromosome has a more compact (less accessible) chromatin structure. It has been shown thatrelease of chromatin condensation can be a way to regulate gene expression, for example of secondary metabolite gene clusters in Fusarium [Reyes-Dominguez et al, FGB 2012]. To test whether DNA methylation in F. oxysporum can influence effector expression, knock-outs of HP1 (heterochromatinprotein) and DIM2 (DNA methylase) were tested for expression of the effector gene SIX1. No differences compared to wild-type were observed. Previouslyit was shown that expression of SIX1 requires Sge1, a conserved transcription factor encoded in the core genome. Loss of DNA methylation did, however,also not bypass the requirement of Sge1 for SIX1 expression (in Dhp1Dsge1 and Ddim2Dsge1 double mutants). Both DIM2 and HP1 are not required forpathogenicity of F. oxysporum f.sp. lycopersici, and DNA methylation in this strain in general seems to be very low. To obtain more insight in the regulationof effector gene expression we are currently focussing on the potential targets of the transcription factors encoded on the pathogenicity chromosomeitself.558. Mechanistic investigation of Trichoderma cf. harzianum SQR-T037 mycoparasitism against Fusarium oxysporum f. sp. cubense 4, (banana wiltdisease). Jian Zhang 1,2 , Ruifu Zhang 1,2 , Irina S. Druzhinina 3,4 , Qirong Shen 1,2 . 1) Key Laboratory of Plant Nutrition and Fertilization in Low-Middle Reaches ofthe Yangtze River, Ministry of Agriculture. 210095, Nanjing, China; 2) Jiangsu Key Lab and Engineering Center for Solid Organic Waste Utilization, NanjingAgricultural University, 210095, Nanjing, China; 3) Microbiology Group, Institute of Chemical Engineering, Vienna University of Technology Getreidemarkt9/1665, A-1060 Vienna, Austria; 4) ACIB - Austrian Centre of Industrial Biotechnology, Petersgasse 14, 8010 Graz, Austria.Besides the effective stimulation of banana growth, the wild strain of Trichoderma cf. harzianum SQR-T037 (SQR-T037) is capable to control the soil-bornpathogen Fusarium oxysporum f. sp. cubense 4 (Foc4), the causative agent of banana wild disease . In this work we focused on mechanisms involved in themycoparasitic attack of SQR-T037 on Foc4. In vitro, in dual confrontation assays, SQR-T037 was able to cover (overgrow) the hyphae of Foc4 what resultedin damage of the cell wall of the pray fungus and its death. At early stages of the interactions, when SQR-T037 hyphae started to combat the colony ofFoc4, the droplets of the yellowish exudate putatively secreted by SQR-T037 were observed. The GC-MS analysis identified that the exudate containedalmitic and stearic acids, several hydrolytic enzymes (mainly chitinases and proteases) and essential amount of H 2O 2. It allowed us to assume that thesecompounds play the major role in killing the Foc4. We have detected that hyphae of SQR-T037 indeed accumulated H 2O 2 when it physically made contactwith Foc4 and moreover that H 2O 2 suppressed Foc4 growth. The results of transcriptomics analysis of genes putatively involved in the mycoparasitic attackthrough H 2O 2 or its regulation will be presented.559. Epichloënin A, a unique siderophore of Epichloae endophytes and its role in restricting fungal growth in planta. Natasha Forester 1 , Geoffrey A.Lane 1 , Iain Lamont 2 , Linda J. Johnson 1 . 1) Forage Improvement, AgResearch Limited, Palmerston North, Manawatu, New Zealand; 2) University of Otago,Dunedin, New Zealand.We have previously shown, through characterisation of a non-ribosomal peptide synthetase gene (NRPS), sidN, that the biosynthesis of a novelextracellular ferric iron-chelating siderophore designated as epichloënin A is required for maintaining mutualism of E. festucae with its grass host,perennial ryegrass (Lolium perenne). We have extended our investigation of the role of fungal siderophores in iron homeostasis through thecharacterization of other siderophore biosynthetic genes, including sidA which encodes a putative L-ornithine N5-oxygenase that catalyses the firstenzymatic step in siderophore biosynthesis and sidC, encoding a NRPS siderophore synthetase. Using DsidA, DsidC and DsidN mutants we have discoveredthat E. festucae synthesises two siderophores, epichloënin A that requires SidN and an intracellular siderophore, ferricrocin (FC) that requires SidC;production of both siderophores is totally abolished in DsidA mutants. In contrast to DsidN mutants, DsidC-infected plants showed very little phenotypicconsequences due to loss of FC, and this is likely to be due to functional redundancy of epichloënin A. The levels of iron-bound epichloënin A inside the cellare significantly higher than FC, suggesting that epichloënin A acts both as an extracellular and intracellular siderophore. While investigating the influenceof iron on the siderophore mutants in planta with hydroponically supplied nutrients, we unexpectedly demonstrated that iron exacerbates rather thancomplements the DsidN mutant phenotype. We observed increases in fungal proliferation, production of dense mycelial mats, and increased plantstunting. In infected plants, compared to wild-type (WT), there was also an increased uptake of iron in DsidN cells by reductive iron assimilation. Analysesin culture indicated that in the DsidN mutant more iron was stored (in FC and in vacuoles) than in WT fungi, suggesting that DsidN has increased access toiron relative to WT. However, ultra-structural studies of DsidN hyphae in stunted plants (that were not associated with the host vasculature) suggestedthat DsidN hyphae were often non-viable, implicating resource exhaustion from inappropriate growth and misuse of available iron. We therefore proposethat epichloënin A is a multitasking siderophore specialising in iron sequestration to moderate cellular iron supply.560. Interaction between the saprotrophic fungus Serpula lacrymans and living pine roots. Nils OS Högberg 1 , Anna Rosling 1 , Annegret Kohler 2 , MartinFrancis 2 , Stenlid Jan 1 . 1) Department of Forest Mycology, BioCenter, SLU, Uppsala, Sweden; 2) INRA, Nancy, France.Recently it has been shown, with a Comparative genomic perspective, that brown rot and mycorrhiza fungi have evolved from white rot ancestors. Woodis a composite material composed of lignin, cellulose and hemicellulose. White rot fungi are able to degrade all of these components with a combination ofcarbohydrate active and oxidative enzymes. During the course of evolution brown rot and mycorrhiza have lost most of the genes in these gene families.Nevertheless, brown rot fungi are efficient wood decomposers that degrade cellulose and hemicellulose by means of hydroxyl radical production andremaining carbohydrate active enzymes. The family Boletales includes both brown rot fungi and mycorrhiza and it is tentative to speculate that there hasbeen a parallel evolution of these ecological strategies. Here we test the effect of infecting pine roots with the brown rot fungus Serpula lacrymans. Theinteraction was neutral since plant growth was not stimulated but not reduced either. The fungus formed a mantle around the pine roots but not theHartig net that is typical for ectomycorrhiza. Fungal gene expression was compared with the wood decay transcriptome. 1250 genes were more than twofoldupregulated compared to a glucose medium control. A large proportion of the upregulated genes (62 %) are unknown. Carbohydrate active genesrepresent only 3% of this gene set and genes with oxidoreductase activity, including monoxygenases represent 4% of the upregulated genes. This isconsiderably lower compared to saprotrophic growth on wood where carbohydrate active enzymes accounted for 26% and oxidative enzymes for 19%which dominated the gene expression on wood. Gene expression for genes involved in transportation was about the same, around 10% in this experimentand under wood decomposition. Several genes that indicate an interaction with a host were also upregulated. In conclusion, gene expression wasmarkedly different between a glucose medium, wood decomposition and growth on pine roots. This may be a signal of symbiosis, the effect on pineseedling growth was neutral. Thus we cannot conclude if the interaction is beneficial or negative to the host.561. You turn me on: Pyrenophora tritici-repentis genes differentially regulated early during infection of wheat. V. A. Manning 1 , I. Pandelova 1 , L. M.258

FULL POSTER SESSION ABSTRACTSCiuffetti 1,2 . 1) Botany and Plant Pathology, Oregon State University, Corvallis, OR 97331; 2) Center of Genome Research and Biocomputing, Oregon StateUniversity, Corvallis, OR 97331.Pyrenophora tritici-repentis (Ptr) is a necrotrophic fungal pathogen of wheat causal to the disease tan spot and host-selective toxins (HSTs) produced byPtr are the primary factors that contribute to virulence. One of these HSTs, Ptr ToxA, is a necrosis-inducing, proteinaceous HST that is also present in thewheat pathogen Stagonospora nodorum. Ptr ToxB is a chlorosis-inducing, proteinaceous HST produced by Ptr with active orthologues expressed in thebrome grass pathogen, Pyrenophora bromi. Despite the presence of active, orthologous HSTs produced by other fungi, Ptr appears to have an advantage insome wheat growing regions in the world, surpassing in disease relevance the other necrotrophic pathogens that contain orthologous HSTs. To begin tounderstand the molecular mechanisms that underlie this advantage, we used RNA-seq-based transcriptome analysis to identify Ptr genes that aredifferentially regulated in planta early in the disease cycle (at thirty hours post-infection) as compared with those genes that are expressed in culture.Functional annotations of the differentially expressed-in planta transcripts reflect the requirements of the pathogen for host penetration, cell walldegradation and the need to counteract the host response to infection; these include CAZymes, peptidases, transporters, and loci with predicted oxidoreductaseactivities including peroxidases. In addition, putative secondary metabolite clusters and Ptr-specific proteins are also differentially regulated.These findings provide the basis for understanding the roles of these proteins in virulence and the possibility of revealing common transcriptionalregulatory elements activated by interactions with the host.562. Characterisation of genes in Quantitative Trait Loci affecting virulence in the basidiomycete Heterobasidion annosum s.l. Ake Olson, Yang Hu, IngaBödeker, Malin Elfstrand, Mårten Lind, Jan Stenlid. Dept Forest Mycology/Pathology, SW Univ Agricultural Sci, Uppsala, Uppsala, Sweden.Heterobasidion annosum sensu lato (s.l.), is a devastating root rot pathogen on conifers present all over the northern hemisphere that causes losses of500 of million Euro per year for forest owners. The H. annosum s.l. consists of five phylogenetic distinct species with different but overlapping hostpreferences. The genome sequence of one isolates from H. irregulare and H. occidentale preferentially infecting pine and spruce species, respectively havebeen obtained. Analysis of the progeny of a genetic cross of the sequenced isolates resulted in a genetic linkage map of 15 groups representing almost thecomplete chromosome set-up. These groups have been aligned and anchored to the physical map of H. irregulare. Quantitative trait loci (QTL) forvirulence on one-year-old Pinus sylvestris and two-year-old Picea abies seedlings were identified and positioned on the map allowing a straight forwardidentification of virulence candidate genes. Gene content and sequence divergence of the QTL regions will be presented. Detailed expression analysis ofvirulence candidate genes with Q-PCR and protein localisation with immunohistochemistry will deduce their role during infection.563. Elevation of FPP synthase activity in Trichoderma atroviride results in higher biocontrol abilities. Sebastian Graczyk, Urszula Perlinska-Lenart,Wioletta Gorka-Niec, Patrycja Zembek, Sebastian Pilsyk, Grazyna Palamarczyk, Joanna S. Kruszewska. Laboratory of Fungal Glycobiology, Institute ofBiochemistry and Biophysics, Warsaw, Poland.In this study we present a new method to obtain the Trichoderma strains with enhanced antifungal and biocontrol activities. The method is based on theincrease synthesis of the mevalonate pathway products. In this pathway some metabolites are produced such as terpenoids, trichodermin, harzianum A,mycotoxin T2, lignoren, ergokonin A and B and viridin which are known from their antifungal and antibacterial activities. All these compounds aresynthesized from farnesyl pyrophosphate (FPP) which is itself synthesized by farnesyl pyrophosphate synthase encoded by ERG20 gene. FPP is also asubstrate for dolichol and ergosterol production which are indirectly engaged in antimicrobial action. In this study we increased production of FPP in T.atroviride by overexpression of the yeast ERG20 gene. We based on the assumption that the increased activity of FPP synthase would stimulate productionof all products of the mevalonate pathway. Five transformants showed higher activity of FFP synthase. Simple screening of the hydrolytic properties of theERG20 transformants revealed that they grew faster on plates with polycarbohydrates as carbon sources. Detailed studies showed higher cellulotytic andchitinolytic activity of enzymes secreted to the cultivation medium by the transformed strains. Antifungal activity was examined by cultivation of plantpathogen Rhizoctonia solani in the atmosphere of volatiles liberated by the transformants and the control. We also analyzed growth of Pythium ultimumon plates which were previously overgrown by Trichoderma strains and were filled with their metabolites. Both experiment showed significantly strongerinhibition of growth of the pathogens by the transformed strains compared to the control. Since the above experiments revealed enhanced antifungalproperties of the transformed strains we performed plant tests using the bean Phaseolus vulgaris L.. Transformed strains increased both, the germinationrate and the size of plants growing in soil infected by Pythium ultimum compared to the control strain. To conclude, an increased activity of themevalonate pathway caused higher activity of hydrolytic enzymes and increased production of volatiles and secondary metabolites and that way boostedantifungal and biocontrol activities of the Trichoderma ERG20 transformants.564. The life history of Ramularia collo-cygni. Maciej Kaczmarek 1,2 , James Fountaine 1 , Adrian Newton 3 , Nick Read 2 , Neil Havis 1 . 1) Crop and Soil Research,Scotland's Rural College, Edinburgh, United Kingdom; 2) Institute of Cell Biology, University of Edinburgh, Edinburgh, United Kingdom; 3) Cell andMolecular Sciences, The James Hutton Institute, Dundee, United Kingdom.The filamentous fungus Ramularia collo-cygni causes the late season disease of spring and winter barley called Ramularia Leaf Spot (RLS). It has becomean increasingly important problem for European farmers in the past decade and has recently been reclassified as a major disease of barley in the UK. Thelack of apparent varietal resistance to the disease has led to significant amounts of fungicide being applied to crops in north western and central Europe inorder to maintain green leaf area and prevent significant yield loss. These factors have contributed to an increasing focus on achieving a betterunderstanding of the fundamental biology of this elusive pathogen in order to develop more successful strategies of RLS management. Therefore, diseasedevelopment throughout the life cycle of the host barley plant has been analysed by the employment of transgenic R. collo-cygni isolate, expressing theGFP reporter molecule, and confocal microscopy. We have been able to examine the previously uncharacterised seed-borne stage and illustrate the modeof fungal transmission into barley seedlings. We have also analysed the potential sexual reproduction in the fungus by utilising a range of correlativetechniques, such as cryo-scanning electron microscopy, confocal microscopy and light microscopy. Here we describe for the first time the nature ofspeculated spermogonial stage called Asteromella and in addition, present preliminary evidence suggesting the existence of a perfect stage that, iffunctional, could resemble closely related Mycosphaerella species.565. Mechanical stress sensing in Epichloë fungal symbionts during colonization of grasses. Kahandawa G.S.U Ariyawansa 1 , Rosie E. Bradshaw 2 , Neil A.R.Gow 3 , Nick D. Read 4 , Richard D. Johnson 1 , Duane P. Harland 5 , Christine R. Voisey 1 . 1) AgResearch, Grasslands Research Centre, Palmerston North, NewZealand; 2) BioProtection Centre, Institute of Fundamental Sciences, Massey University, Palmerston North, New Zealand; 3) School of Medical Sciences,University of Aberdeen, United Kingdom; 4) Institute of Cell Biology, University of Edinburgh, United Kingdom; 5) AgResearch, Lincoln Research Centre,Christchurch, New Zealand.Epichloë festucae is an agronomically-important endophytic fungus that grows symbiotically within the intercellular spaces of temperate grass species27th Fungal Genetics Conference | 259

FULL POSTER SESSION ABSTRACTSsuch as Lolium perenne and L. arundinaceum. Colonization of host seedlings by E. festucae occurs when hyphae in the shoot apex invade developing hostleaves and extend via intercalary hyphal growth, a highly unusual mechanism of division and extension in non-apical compartments. We hypothesise thatintercalary hyphal growth is stimulated by mechanical stretch imposed by attachment of hyphae to elongating host cells, and that this stress is sensed bymechano-sensors located on the hyphal membranes. Genome analysis revealed that homologues of known mechano-sensors in Saccharomyces cerevisiaesuch as Mid1 (a stretch activated calcium ion channel), Wsc1 and Mid2 (cell wall integrity sensors) are present in the E. festucae genome. Genereplacement studies of mid1 and wsc1 in E. festucae reduced radial growth rate in axenic culture confirming the role of both genes in hyphal growth. Inaxenic culture both Dwsc1 and Dmid1 mutants were sensitive to fungal cell wall modifiers such as Calcofluor White, supporting their role in cell wallintegrity. Preliminary plant infection studies with Dwsc1 and Dmid1 mutants revealed a hyper-branched unsynchronized growth pattern within the host(Lolium perenne), and Dwsc1 also caused severe stunting in most plants suggesting a disruption in the symbiosis. A technique to stimulate intercalarygrowth under in-vitro conditions through mechanical stretch is being optimised to test the ability of Mid1, Wsc1 and Mid2 to sense mechanical stress andinitiate intercalary growth.566. Aspergillus flavus hypertrophy and hyphal entry by Ralstonia solanacearum is mediated by bacterial type three secretion system function. Joe ESpraker 1 , Nancy P Keller 2 . 1) Plant Pathology, University of Wisconsin Madison, Madison, WI; 2) Bacteriology, University of Wisconsin Madison, Madison,WI.Fungi and bacteria are two of the primary pathogens of plants, often infecting the same crops, however shockingly little is known of how theseorganisms interact independently of plant hosts. In examining the interaction between two economically important pathogens of peanut, Aspergillusflavus and Ralstonia solanacearm, a fungus and bacterium, respectively, we’ve shown that fungal hypertrophy is induced and that the bacterium is capableof entering these cells. The hypertrophic cells were imaged using calcofluor staining to show chitin cell wall structure. Bacterial invasion of these structureswas demonstrated using confocal microscopy of GFP labeled bacteria. Further, we demonstrate that bacterial mutants deficient in type three secretionsystems are incapable of eliciting the fungal hypertrophic response by culturing virtually isogenic bacterial type three secretion mutants. This is the firstreport of a well-known plant pathogenic bacterium eliciting fungal hypertrophy and invading hyphal cells. Current research is aimed at finding bacterialeffectors that may be facilitating this interaction and elucidating their mode of action.567. Vegetative hyphal fusion in epichloae endophytes. Jun-ya Shoji, Nikki D. Charlton, Sita R. Ghimire, Jin Nakashima, Kelly D. Craven. Plant BiologyDivision, The Samuel Roberts Noble Foundation, Ardmore, OK.Vegetative hyphal fusion establishes the interconnection of individual hyphal strands into an integrated network of a fungal mycelium. It is suspectedthat vegetative hyphal fusion plays many important roles such as in nutrient translocation, intramycelial signaling, and emergence of genetic diversity viahorizontal gene / chromosome transfer or interspecific hybridization. However, experimental support for these suspected roles is still largely lacking. Toinvestigate the role of hyphal fusion in fungal endophytes of epichloae, which form mutualistic symbiosis with grass hosts, we generated mutant strainslacking sftA, an ortholog of the hyphal fusion gene so in Epichloë festucae. The E. festucae DsftA mutant strains grew like the wild-type strain in culture butwith reduced aerial hyphae, and completely lacked hyphal fusion. The most striking phenotype of the E. festucae DsftA strain was that it failed to establisha mutualistic symbiosis with the tall fescue plant host (Lolium arundinaceum), and instead, killed the host plant within two months after initial infection.This suggests that hyphal fusion may have an important role in the establishment / maintenance of fungal endophyte-host plant mutualistic symbiosis. Tofurther investigate the importance of hyphal fusion in epichloae, frequency of hyphal fusion was quantified in different epichloae endophytes includingsexual isolates, asexual interspecific hybrids and asexual non-hybrids. A majority of sexual epichloae underwent frequent hyphal fusion, whereas hyphalfusion was less frequently found in asexual epichloae. Moreover, hyphal fusion was less common in asexual non-hybrid epichloae compared to asexualhybrids. Thus, it appears that the ability to undergo hyphal fusion correlates with the presence of the sexual cycle, and the hybrid status of epichloaeendophytes. Overall, our data provide evidence for the importance of hyphal fusion in establishment / maintenance of mutualistic symbiosis, andevolution of epichloae endophytes.568. Oxygen and the stomatal cue: Dissecting stomatal tropism in Cercospora zeae-maydis. R. Hirsch, B. Bluhm. Department of Plant Pathology,University of Arkansas Division of Agriculture, Fayetteville, AR.Cercospora zeae-maydis causes grey leaf spot of maize, one of the most widespread and destructive foliar diseases of maize in the world. Stomatalinfection is a critical, yet poorly defined, component of pathogenesis in C. zeae-maydis. At the onset of infection, the fungus senses and grows towardsmaize stomata, and then breaches the leaf surface by producing appressoria over stomatal pores. Directed growth toward distant stomata during infectionled us to hypothesize that C. zeae-maydis responded to an unknown chemical cue emanating from stomata. To elucidate mechanisms underlyinginfectious development in C. zeae-maydis, particularly stomatal tropism, a series of histological experiments were performed with epi-florescent andconfocal microscopy. Upon sensing maize stomata, C. zeae-maydis either reoriented hyphal tip growth towards stomata, or initiated new hyphaeoriginating from right-angle branches in close proximity to stomata. Hyphae exhibiting stomatal tropism were linear and lacked branches. Ontopographically accurate acrylic leaf replicas, C. zeae-maydis did not display stomatal tropism and failed to form appressoria upon encountering artificialstomata, which indicated that thigmotropic cues were not sufficient to elicit pre-penetration infectious development. However, in non-host interactions,C. zeae-maydis exhibited stomatal tropism and retained the ability to form appressoria over stomata, which suggested that a chemical cue emanating fromstomata elicited a chemotropic response in the fungus. Stomatal tropism and appressoria formation in C. zeae-maydis were impaired when atmosphericoxygen levels were disturbed, implicating the role of oxygen sensing in pathogenicity. This study characterized stomatal tropism during infection of maizeby C. zeae-maydis, directly implicated oxygen sensing as a component of pathogenicity, and provides a quantitative framework through which to studyfoliar pathogenesis and host/pathogen interactions in related systems.569. Host colonisation processes by symbiotic epichloid fungi are regulated through cAMP. Christine R. Voisey 1 , Damien J. Fleetwood 2 , Linda J. Johnson 1 ,Gregory T. Bryan 1 , Wayne R. Simpson 1 , Michael J. Christensen 1 , Suzanne J.H. Kuijt 1 , Kelly Dunstan 1 , Richard J. Johnson 1 . 1) Forage Biotechnology,AgResearch, Palmerston N, New Zealand; 2) School of Biological Sciences, The University of Auckland, Auckland 1142, New Zealand.The fungal symbiont, Epichloë festucae, colonises leaves of host grasses by ramifying through the shoot apical meristem (SAM) of the seedling, and theninfecting the leaf primordia. Hyphal infection of the SAM is dependent on apical growth, however after primordia have formed, leaf tissues undergo aphase of intercalary expansion, which the fungus, attached to host cells, must recapitulate to remain intact. E. festucae hyphae entering the leaf expansionzone switch from apical to intercalary growth, and extend in synchrony with the host until the leaf tissues mature. How colonising symbiotic fungiaccommodate the complexities of the plant developmental programme is currently unclear. Since cAMP signalling is often required for host colonisationby fungal pathogens, we disrupted the cAMP cascade by insertional mutagenesis of the E. festucae adenylate cyclase gene (acyA). Consistent with reports260

FULL POSTER SESSION ABSTRACTSon other fungi, disruption mutants had a slow radial growth rate in culture, and colonies were highly compact relative to controls. Furthermore, thehyphae were convoluted and hyper-branched suggesting that apical dominance had been disrupted. Nitro blue tetrazolium straining of hyphae showedthat cAMP disruption mutants were impaired in their ability to synthesise superoxide indicating that cAMP signalling is important for the production ofROS in culture in this species. This defect was reversed by re-insertion of a functional wild type acyA gene into mutant strains. Despite significant defects inhyphal growth and ROS production in culture, E. festucae DacyA mutants were infectious and capable of forming symbiotic associations with grasses,albeit at a lesser infection frequency than wild type. Plants infected with E. festucae DacyA mutants were indistinguishable from controls. However, as inculture, microscopic evidence showed that the mutant strains within the host were hyper-branched, and host tissues heavily colonised, indicating that thetight regulation over hyphal growth normally observed in developing and mature host tissues requires a functional cAMP signalling cascade. Furtherresearch is currently underway to understand how cAMP affects the hyphal growth transitions undertaken during host colonisation, particularly at thelevel of the cell cytoskeleton and hyphal cell wall synthesis.570. Role of VCP1 and SCP1 proteases in the mutitrophic behaviour of the nematophagous fungus Pochonia chlamydosporia. Nuria Escudero 1 ,Christopher R. Thornton 2 , Luis Vicente Lopez-Llorca 1 . 1) Laboratory of Plant Pathology, Multidisciplinary Institute for Environment Studies (MIES) RamónMargalef. University of Alicante, Alicante, SPAIN; 2) Food Security and Sustainable Agriculture, Biosciences, College of Life & Environmental Sciences,University of Exeter, Exeter. UK.Pochonia chlamydosporia (Goddard) Zare and Gams is a fungal parasite of female nematodes and eggs, which has been widely studied as a biologicalcontrol agent of cyst and root-knot nematode egg-shells. The nematode egg-shell is formed by several layers, including a chitinous layer composed of aprotein matrix embedding chitin microfibrils. Extracellular enzymes, such as serine porteases (e.g. VCP1), secreted by egg-parasitic nematophagous fungiare known to play an important role in egg infection. SCP1, a recently reported serine carboxypeptidase from P. chlamydosporia was found during plantroot endophytic colonisation by the fungus, its role in eggs parasitisim is unknown. We have investigated the role of VCP1 and SCP1 proteases in themutitrophic behaviour of the nematophagous fungus Pochonia chlamydosporia using immunological approaches using antiVCP1 and SCP1 polyclonalantibodies, these were raised against synthetic peptides of both proteases. ELISA and immunofluorescence have confirmed the production of bothproteases when Meloidogyne javanica eggs were used as inducer. P. chlamydosporia under starvation condition (water) also expressed both proteases. Itseems that the signal of SCP1 was more intense than of VCP1 under most conditions tested (eggs, protein substrate and starvation). Using proteomic,chitosan was previously found in our lab to induce VCP1 in P. chlamydosporia liquid cultures. Consequently, we have also evaluated the amount of VCP1and SCP1 in media with chitosan, to quantify the production of these proteases under multitrophic conditions. This study is casting light into the molecularaspects of the multitrophic behaviour of P. chlamydosporia. This will help to understand the biocontrol potential of the fungus and open newbiotechnological applications.571. Cellular development integrating primary and induced secondary metabolism in the filamentous fungus Fusarium graminearum. Jon Menke 1 ,Jakob Weber 2 , Karen Broz 3 , H. Corby Kistler 1,3* . 1) Department of Plant Pathology, University of Minnesota, St. Paul, USA; 2) Molekulare Phytopathologie,Universität Hamburg, Germany; 3) USDA ARS Cereal Disease Laboratory, St. Paul, MN, USA.Several species of the filamentous fungus Fusarium colonize plants and produce toxic small molecules that contaminate agricultural products, renderingthem unsuitable for consumption. Among the most destructive of these species is F. graminearum, which causes disease in wheat and barley and oftencontaminates the grain with harmful trichothecene mycotoxins. Induction of these secondary metabolites occurs during plant infection or in culture inresponse to chemical signals. Here we report that trichothecene biosynthesis involves a complex developmental process that includes dynamic changes incell morphology and the biogenesis of novel subcellular structures. Two cytochrome P-450 oxygenases (Tri4p and Tri1p) involved in early and late steps intrichothecene biosynthesis were tagged with fluorescent proteins and shown to co-localize to vesicles we call “toxisomes.” Toxisomes, the inferred site oftrichothecene biosynthesis, dynamically interact with motile vesicles containing a predicted major facilitator superfamily protein (Tri12p) previouslyimplicated in trichothecene export and tolerance. The immediate isoprenoid precursor of trichothecenes is the primary metabolite farnesylpyrophosphate. When cultures are shifted from non-inducing to trichothecene inducing conditions, changes occur in the localization of the isoprenoidbiosynthetic enzyme HMG CoA reductase. Initially localized in the cellular endomembrane system, HMG CoA reductase increasingly is targeted totoxisomes. Metabolic pathways of primary and secondary metabolism thus may be coordinated and co-localized under conditions when trichothecenesynthesis occurs.572. DNA double-strand breaks generated by yeast endonuclease I-Sce I induce ectopic homologous recombination and targeted gene replacement inMagnaporthe oryzae. T. Arazoe 1 , T. Younomaru 1 , S. Ohsato 1 , T. Arie 2 , S. Kuwata 1 . 1) Meiji University, Kanagawa, Japan; 2) Tokyo University of Agricultureand Technology, Tokyo, Japan.The filamentous fungus Magnaporthe oryzae causes the rice blast disease that is one of the most destructive fungal diseases of cultivated rice plants. Tocontrol this fungal disease, many resistant genes have been introduced into cultivated rice germplasm, however, breakdowns of the resistance often occurwithin several years by rapid evolution of the fungus. Therefore, studies on the evolutionary mechanisms of the fungus are important for elucidation of therapid evolution. We set out a novel detection/selection system of DNA double-strand breaks (DSBs)-mediated ectopic homologous recombination (HR)that is one of the evolutionary mechanisms. The system consists of two nonfunctional yellow fluorescent protein (YFP)/blasticidin S deaminase (BSD) fusiongenes as a donor and a recipient, and a yeast endonuclease I-Sce I gene as a DSB-inducer. In this system, ectopic HR can be detected and selected byrestorations of YFP fluorescence and blasticidin S (BS)-resistance at a single cell level. These donor and recipient genes were simultaneously integrated intothe M. oryzae genome and transformed lines were isolated. In the absence of the DSB-inducer, transformed lines showed relatively low frequencies of HRevents (>2.1%). On the other hand, by integration of the DSB-inducer gene into transformed lines, we could observe the frequencies of DSB-mediated HRraising up to ~40%. This result clearly showed that DSB into a certain gene induce ectopic HR events between the gene and its homologs. Accordingly, wefurther applied I-Sce I mediated DSB for TGR in M. oryzae. To detect TGR, we constructed simple system using donor and recipient genes. The recipientgene was integrated into the M. oryzae genome and transformed lines were isolated. To recipient gene integrated lines, the donor gene was introducedand restorations of YFP fluorescence and BS-resistance were evaluated. As we expected, the TGR frequencies were increased at least 37-folds by I-Sce I cotransformationas compared with those obtained without I-Sce I. This result provides a new method using DSB for improving the TGR frequency in M.oryzae. Taken together, it is strongly suggested that DSBs can drive genomic rearrangement and accelerate pathogenic variability in M. oryzae through theectopic HR between homologous sequences such as transposable elements and avirulence genes.573. Investigation of the Magnaporthe oryzae proteome and phosphoproteome during appressorium formation. William L. Franck 1 , Emine Gokce 2 ,Yeonyee Oh 1 , David C. Muddiman 2 , Ralph A. Dean 1 . 1) Plant pathology, NC State University, Raleigh, NC; 2) W.M. Keck FT-ICR Mass Spectrometry27th Fungal Genetics Conference | 261

FULL POSTER SESSION ABSTRACTSLaboratory, NC State University, Raleigh, NC.Magnaporthe oryzae, the causative agent of rice blast disease, infects plant leaves via formation of an appressorium which facilitates penetration of theleaf surface. In an effort to better understand the physiological changes accompanying the earliest stages of infection-related development, a nano-LCMS/MS-based global proteomics examination of conidial germination and cAMP-induced appressoria formation was undertaken at four distinctdevelopmental time points resulting in the identification of 3200 proteins. Label free quantification by spectral counting identified 591 proteins whoserelative abundance changed during germination. Furthermore, treatment of germinating conidia with cAMP to induce appressorium formation resulted inthe identification of 493 proteins whose relative abundance changed compared to untreated samples. In developing appressoria, changes in cell wallmodifying, transport, extracellular and plasma membrane localized proteins were observed. Proteomic analysis of a M. oryzae cAMP-dependent proteinkinase A (cpkA) mutant defective in appressorium formation following treatment with cAMP identified a subset of proteins whose regulation is dependenton cAMP signaling. A comparison of proteome and transcriptome data revealed little correlation between protein and transcript regulation. Finally, tobetter define the role of protein phosphorylation and the CPKA protein kinase during the development of appressoria, the phosphoproteome of M. oryzaeis being investigated. To date, a total of 980 phosphoproteins have been identified from conidia and experiments designed to identify changes in proteinphosphorylation during appressorium formation are in progress.574. Characterization of the binding site and downstream targets of the MST12 transcription factor in Magnaporthe oryzae. Guotian Li, Guanghui Wang,Jin-Rong Xu. Department of Botany and Plant Pathology, Purdue University, West Lafayette, IN.Rice blast caused by Magnaporthe oryzae is one of the most devastating diseases on rice. In M. oryzae, appressorium formation is regulated by PMK1mitogen-activated protein kinase (MAPK) pathway. Its orthologs are conserved a wide array of plant pathogenic fungi for regulating different plantinfection processes. In M. oryzae, one of the transcription factors functioning downstream from Pmk1 is Mst12 that is essential for appressoriumpenetration and invasive growth. The MST12-GFP transformant showed strongest GFP signals in appressoria and invasive hyphae. Pmk1 weakly interactedwith Mst12 in yeast two-hybrid assays. Overexpression of MST12 failed to complement the defects of the pmk1 mutant, suggesting that the activation ofMst12 by Pmk1 was required for its function. Site-directly mutagenesis analysis indicated that the MAPK-docking region and phosphorylation site wereessential for the function of MST12. However, mutant alleles of MST12 with point mutations in any one of the two tandem zinc finger domains were stillpartially functional, indicating their overlapping or redundant functions. Expression of MST12 functionally rescued the invasive growth defects of the yeastste12 mutant and EMSA analyses suggested that Mst12 had a binding site similar to the PRE sequence recognized by Ste12. During appressoriumformation, transcription levels of 222 genes were found to be significantly altered in the mst12 mutant (P

FULL POSTER SESSION ABSTRACTScopies of AVR-Pia. Screening and analysis of cosmid clones indicated that two copies of the DNA-type transposon Occan (Occan 9E12 and Occan 3A3 ) werelocated on the same chromosome and three copies of AVR-Pia were located in between the two Occan elements. Ina168m95-1 contains a conservedOccan element, named Occan m95-1 , between sequences homologous to the 5’-flanking region of Occan 3A3 and the 3’-flanking region of Occan 9E12 . Inaddition, sequence polymorphisms indicated a homologous recombination between Occan 3A3 and Occan 9E12 , which resulted in Occan m95-1 . Based on theseobservations, we propose the hypothesis that homologous recombination in the two Occan elements leads to the deletion of AVR-Pia in Ina168m95-1.578. The interactome of pathogenicity factors in the rice blast fungus Magnaporthe oryzae. Xiaoying Zhou 1 , Yang Li 1 , Keerthi Jayasundera 2 , Anton Iliuk 2 ,Andy Tao 2 , Jinrong Xu 1 . 1) Dept. of Botany and Plant Pathology, Purdue University, West Lafayette, IN; 2) Dept. of Biochemistry , Purdue University, WestLafayette, IN.Rice blast is a disease of significant economic impact worldwide and a model system for studying fungal-plant interactions. To date, over 100pathogenicity factors have been identified in Magnaporthe oryzae. However, there is only limited knowledge about their relationships. To betterunderstand molecular mechanisms regulating plant infection processes, it is critical to identify protein-protein interaction networks important forpathogenesis. In this study, we characterized the interactome of selected pathogenicity-related proteins. The affinity purification and proteomicsapproaches were used to identify proteins that interact with over 60 known pathogenicity factors, including components of important signaling pathways.Protein-protein interaction maps were established for these pathogenicity factors based on affinity purification data and information about their orthologsin yeast. Co-immunoprecipitation, BiFC, or yeast two hybridization assays were used to verify the interactions of selected genes. For a number ofpathogenicity factor-interacting genes, gene knock-out mutants were generated to determine their functions in pathogenesis. To our knowledge, resultsfrom this study represent the first study of protein-protein interaction networks of pathogenicity factors in plant pathogenic fungi in M. oryzae.579. Interaction between phenolic and oxidant signaling in Cochliobolus heterostrophus. Benjamin A Horwitz 1 , Samer Shalaby 1 , Olga Larkov 1 , MordechaiRonen 2 , Sophie Lev 3 . 1) Department of Biology, Technion - IIT, Haifa, Israel; 2) Department of Plant Science, Tel Aviv University, Ramat Aviv, Israel; 3)Centre for Infectious Diseases and Microbiology, University of Sydney at Westmead Hospital, Westmead, NSW 2145, Australia.The transcription factor ChAP1 is an ortholog of yeast YAP1 in the maize pathogen Cochliobolus heterostrophus. ChAP1 migrates to the nucleus uponexposure to oxidative stress, inducing antioxidant genes such as thioredoxin and glutathione reductase [1]. ChAP1 also localizes to nuclei on contact withthe leaf and during invasive growth. Though reactive oxygen species are encountered on the host, ChAP1 nuclear retention can occur without oxidativestress. One of the signals responsible is provided by phenolic compounds [1-3]. Using a genetically-encoded ratiometric reporter of the redox state, weshowed that leaf extract and phenolics, despite their antioxidant properties, promote nuclear accumulation of ChAP1. To study this dual role of ChAP1 weidentified genes expressed in response to phenolics. Intradiol dioxygenase CCHD1 is rapidly upregulated, independent of ChAP1 [2]. Coumaric acid causedrapid and simultaneous upregulation of most of the b-ketoadipate pathway genes. Deletion of CCHD1 provided genetic evidence that protocatechuic acidis an intermediate in catabolism of many aromatics [3]. The activity of a structure series showed complementary requirements for upregulation of CCHD1and ChAP1 nuclear retention. The ability to metabolize a compound and ChAP1 nuclear retention are inversely correlated. To find additional genesinduced by phenolics, microarrays designed from the predicted coding sequences of the C. heterostrophus genome [4] were hybridized to probes madefrom RNA of cultures exposed to coumaric acid, or controls. Expression of about 90 genes from different pathways primarily for metabolism, for example,the b-ketoadipate, quinic acid and shikimic acid pathways, as well as transporters from different families was altered in response to coumaric acid. Theability to respond to phenolics and detoxify or metabolize them via the b-ketoadipate pathway confers an advantage to plant pathogens, and explains thepresence of at least two response pathways detecting these compounds. [1] Lev et al. (2005) Eukaryot. Cell 4:443-454; [2] Shanmugam et al. (2010) Cell.Microbiol. 12:1421-1434; [3] Shalaby et al. (2012) MPMI 25: 931-940; [4] Ohm et al. (2012) PLoS Pathog 8: e1003037. Supported in part by the IsraelScience Foundation. We thank Michal Levin and Itai Yanai for help with microarray hybridization.580. Mode of Action of Chitosan: Antifungal and Gene Modulator from Natural Origin. Luis V. Lopez-Llorca. Laboratory of Plant Pathology, Department ofMarine Sciences and Applied Biology, Multidisciplinary Institute for Environmental Studies (MlES) Ramon Margalef. University of Alicante, E-03080Alicante, Spain. email: lv.lopez@ua.es.Chitin is an abundant, easily obtained and renewable natural polymer, second only to cellulose. Chitin is a main structural component of barriers (cuticlesand cell walls) of invertebrates (crustaceans, insects and nematodes) and fungi. Its deacetylated form, chitosan, has higher solubility and is known to haveinteresting biological properties. Chitosan, as a polycation, permeabilises the fungal membrane in an energy dependent manner. Chitosan kills orcompromises the growth of important plant and human fungal pathogens. Unlike these fungi, fungal parasites of invertebrates (FPI, mainlynematophagous and entomopathogenic fungi), widely used biological control agents in sustainable agriculture, are resistant to chitosan. Perhaps as aresult of coevolution with their hosts, FPI have evolved chitosan-resistant low-fluidity membranes (high content of saturated FFA) and produce efficientchitosan degrading enzymes. Besides, chitosan activates fungus development (e.g. conidiation) and expression of FPI pathogenicity factors such as serineproteases involved in the degradation of host barriers. Using chemogenomic platforms with yeast (sensitive to chitosan) we have identified chitosanputative gene targets. One of them, ARL1, a member of the Ras superfamily that regulates membrane trafficking, confers chitosan sensitivity as a deletionmutant and resistance when overexpressed in yeast. Yeast ARL1 overexpression in the presence of chitosan mainly caused down-regulation of genesinvolved in cell energy generation (mitochondrial biology, ATP metabolism, energy storage metabolites) and associated by-products (oxidative stress, ROS)and up-regulation of cell cycle progression (mitosis/meiosis, chromatin dynamics and sporulation) genes. Neurospora crassa conidia germination isparticularly sensitive to chitosan. Low nutrient content of media increases chitosan driven membrane permeabilisation and N. crassa sensitivity. UsingRNAseq we have found differential expression of genes involved in membrane permeability, cell energy/ROS generation and cell division as a response ofN. crassa conidia to chitosan. Concluding, we are using cell and molecular approaches to fully understand the multimodal action of chitosan to fully exploitit in biotechnological and health applications.581. Unraveling the metabolome: how zombie ant fungi heterogeneously control ant brains. Charissa de Bekker, David Hughes. Biology and Entomology,Center for Infectious Disease Dynamics,Pennsylvania State University, State College, PA.Fungal entomopathogens rely on cellular heterogeneity during the different stages of insect host infection. Their pathogenicity is exhibited through thesecretion of secondary metabolites. Infection strategies of this group of environmentally important fungi can thus be studied by analyzing theirmetabolome. Next to generalists such as Beauveria bassiana and Metarhizium anisopliae, specialist species exist that are able to control host behavior.One of the most dramatic examples is the death grip of ants infected by Ophiocordyceps unilateralis, where ants are being used as a vehicle and finally biteinto vegetation before dying, aiding fungal spore dispersal after death. To establish this the fungus must not only overcome the immune system of thehost, but also manipulate the brain and atrophy the muscles. To date, most work on manipulation of host behavior has described the ant’s behavior,27th Fungal Genetics Conference | 263

FULL POSTER SESSION ABSTRACTSleaving the molecular processes from the fungal point of view unresolved. To start unraveling the mechanisms underlying this phenomenon we arecombining metabolite profiling with an ex vivo insect tissue culturing system that allows us to study fungal metabolites secreted in different parts withinthe host. Using this technique we established that B. bassiana and M. anisopliae, and O. unilateralis heterogeneously react to brain and muscle tissue bysecreting a significantly different array of metabolites. The combination of these approaches with a concrete understanding of the host-parasiteinteraction in nature is allowing us to understand both the diversity of secondary metabolites as well as make discoveries regarding the temporal dynamicsthese fungi employ when releasing metabolites that affect the host. This project is financed by the Marie Curie International Outgoing Fellowships andPenn State University .582. Gene expression of fungal aldehyde dehydrogenases in ectomycorrhiza. Catarina Henke 1,2 , Kartrin Krause 1 , Erika Kothe 1 . 1) Friedrich SchillerUniversity of Jena, Institute of Microbiology, Microbial Communication, Neugasse 25, D-07743 Jena, Germany; 2) Max Planck Institute for ChemicalEcology, International Max Planck Research School, Hans-Knöll-Strabe 8, D-07745 Jena, Germany, chenke@ice.mpg.Ectomycorrhizal fungi form a mutual symbiosis with trees and perform important functions in the ecosystem, particularly improving plant growth,nutrient supply and plant protection against pathogens. The molecular level of the association between the basidiomycete fungus Tricholoma vaccinumand the specific host spruce (Picea abies) is studied to investigate the molecular mechanisms of interaction. Differential display analysis revealed a fungalaldehyde dehydrogenase encoding gene ald1 from the basidiomycete T. vaccinum specifically expressed in ectomycorrhiza during interaction with thecompatible host. Ald1 has a key function in the detoxification of alcohols and aldehydes occurring in mycorrhizal biotopes and is involved in production ofthe phytohormone indole-3-acetic acid. Gene regulation of ald1 is monitored via quantitative RT-PCR analyses. Transcription level was increased in themutualistic association and could be stimulated by different external supplements, namely alcohols and aldehydes. The ald1 overexpressing mutantsgenerated by Agrobacterium tumefaciens mediated transformation showed increased ethanol stress tolerance. Linkage between gene transcription leveland phenotypic characterization will elucidate deeper understanding of biological function, particularly of the possible pathways of auxin synthesis, andwill allow better understanding of aldehyde dehydrogenases in ectomycorrhiza.583. Interaction of ectomycorrhizal fungi with environment. Katrin Krause, Ines Schlunk, Erika Kothe. Institute of Microbiology, Friedrich SchillerUniversity, Jena, Thuringia, Germany.Ectomycorrhizal fungi play an important role in the biogeochemical cycle, act as decomposers and environmental indicators. Therefore they interact withdifferent compounds of the environment like secondary metabolites of plant and soil living microorganisms, heavy metals and xenobiotics. Differentenzymes and transporters are involved in these processes. One of these transporters, multidrug and toxic compound extrusion (MATE) mte1 of theectomycorrhizal fungus Tricholoma vaccinum, was upregulated during symbiosis. By heterologous expression of mte1 in Saccharomyces cerevisiae,different metals, xenobiotics and secondary metabolites were identified as substrates for the MATE transporter. Furthermore, a retrotransposon retroshowed an upregulation during the symbiosis with spruce. After long-term cultivation of the fungus an additional copy of the retrotransposon wasdetectable by Southern blot analyses in the fungal genome, showing transposition during co-cultivation with the plant. Since the fungus is prone toexperience some plant-derived defense, the induction of transposition might be caused by plant induced stress.584. Genetic exchange in an arbuscular mycorrhizal fungus; Rhisophagus irregularis. Pawel Rosikiewicz 1 , Ian Sanders 2 . 1) Department of Ecology andEvolution, University of Lausanne, Lausanne, Switzerland Pawel.Rosikiewicz@unil.ch; 2) Department of Ecology and Evolution, University of Lausanne,Lausanne, Switzerland, Ian.Sanders@unil.ch.Rhizophagus irregularis is a model species of an arbuscular mycorrhizal fungi (AMF). The AMF forms symbiotic relationship with roots of land plants,improving plant growth and protecting plants against parasites. R. irregularis is a particularly important species of AMF because it colonizes roots of mostof crop plants such as rice, potato and wheat. However, different isolates of this fungus can affect plant phenotype differently. Moreover, it recently hasbeen shown that two isolates of AMF can exchange genetic material, a process that can alter both, plant and fungal phenotypes. R. irregularis, is acoenocytic organism, which means that many nuclei coexist and can move in the common cytoplasm. The genetic exchange between two AMF isolatesoccurs via vegetative hyphal fusion. However, unlike in most fungi AMF produces multinucleate spores and it has been shown that each isolate of R.irregularis carries genetically different nuclei, which are maintained in successive AMF generations. What is unknown is the fate of parental nuclei after thegenetic exchange, how many parental nuclei are exchanged and whether the mix of nuclei is random. In addition the nuclei are exchange with thesurrounding cytoplasm. This lead to a question whether mitochondria from both parental isolates are transmitted to the offspring. In order to answerthose questions I performed an in vitro experiment, where 6 isolates of R. irregularis were grown in pairs and allowed to fuse and exchange theircytoplasm. Subsequently, spores from all in vitro cultures were collected and used to establish 215 potentially crossed AMF lines. Each AMF line wasestablished from a single spore. Fifty-seven of this newly produced AMF lines where genotyped, resulting in identification of 40 crossed AMF lines. Allgenotyped single spore lines carried only one mitochondrial haplotype. Moreover, all the progeny of a given pair of parental AMF isolates received themitochondria from the same parent.585. A surface hydrophobin in ectomycvorrhiza interaction. Dominik Senftleben, Katrin Krause, Erika Kothe. Friedrich-Schiller-Universitat, Jena, Germany.Hydrophobins are small-secreted proteins with low sequence homology. However, all proteins contain eight cysteines, which form disulfide bridges.There are two classes of hydrophobins, depending on their solubility, which have a broad range of functions such as cell wall integrity, coveringconidiospores, adhesion in pathogenic and ectomycorrhiza interactions. Therefore, hydrophobins as well as other genes in a mutual symbiosis aredifferential expressed. We showed this for hydrophobin tthyd1, which is up regulated in the Hartig’net during interaction of Tricholoma terreum with pine.We investigate hydrophobins in T. vaccinum, a widely spread basidiomycete (Agaricales - Tricholomataceae) which forms ectomycorrhiza with spruce. Theestablishment of a high compatible mycorrhiza needs in a co-culture system about one month, in comparison to a low compatible one, which needs aboutfour months. We know which hydrophobin T. terreum regulates specifically in both interactions. Is this also the case for T. vaccinum and can we improvethe low compatible interaction by heterologous or over expression? So far, we investigated five T. vaccinum hydrophobins and 17 in the Tricholoma genusin total. We also want to show via quantitative Real-Time PCR in which stage of the life cycle respectively symbiotic interaction T. vaccinum produceshydrophobins, what kind of role they play with respect to function in the symbiotic tissue and their regulation under heavy metal stress. Due to the factthat T. vaccinum is a k-strategist and the triggers for fruiting body development are unknown, over expression of hydrophobin protein are important tocharacterise their properties. Also in silico analyses are done such as protein-protein complexes with HADDOCK (High Ambiguity Driven protein-proteinDOCKing) for rodlet layer formation and phylogenetic trees (ITS, hydrophobin) to understand how the evolution of the host, fungus and its hydrophobinstook place. (Identification of a hydrophobin gene that is developmentally regulated in the ectomycorrhizal fungus Tricholoma terreum., Manke et al., App.and Environ. Microbiol., 2002 HADDOCK: a protein-protein docking approach based on biochemical and/or biophysical information. Bonvin et al., J. Am.264

FULL POSTER SESSION ABSTRACTSChem. Soc, 2003).586. Response of Alternaria brassicicola to the antifungal activity of isothiocyanates. Benoit Calmes, Jérôme Dumur, Thomas Guillemette, Nelly Bataillésimoneau,Phillippe Simoneau. Institut de Recherche en Horticulture et Semences UMR1345, Angers, France.Alternaria brassicicola is the causative agent of Brassicaceae black spot disease. This necrotrophic fungus causes glucosinolates degradation by plantmyrosinase during infection. Isothiocyanates (ITCs), the major breakdown compounds, have been shown to be exert some toxicity to various A.brassicicola isolates. We showed, by application of specific fluorescent probes, that ITCs cause ROS production, disrupt the mitochondrial membranepotential and trigger apoptosis in A. brassicicola cells. The generation of an oxidative stress following ITCs application was confirmed by the fact that KOmutants deficient for the transcription factors AP1 and SKN7 (both being involved in the oxidative-stress response) are hypersensitive to ITCs and havedecreased aggressiveness on glucosinolates accumulating host plants. Despite this host defense system, A. brassicicola is still able to complete itsinfectious cycle, indicating the existence of strategies to cope with this oxidative stress. Hypersensitivity to Al-ITC, Bz-ITC or Ph-ITC was observed in KOmutants deficient for distinct glutathione-S-transferases. Some of them were differentially expressed following exposure to ITCs and exhibited hightransferase activity with ITCs as substrate. The polyol mannitol has been proposed to act as an antioxidant agent and protect fungal cells by quenching ROSproduced by hosts in response to attack. We isolated the genes encoding the MPD and MDH enzymes, two essential enzymes of the mannitol metabolismin A. brassicicola, and used targeted gene disruption to create single and double mutants for each gene. Only mutants unable to accumulate mannitol inhyphae and conidia were sensitive to ITCs. Our results supported the involvement of fungal GST and mannitol metabolism in ITC-derived oxidative stress.GST participates in ITCs detoxification mechanisms and mannitol accumulation in protection against ROS. They highlight their importance with respect tothe ability of A. brassicicola to efficiently accomplish its pathogen life cycle despite exposure to plant-derived antifungal metabolites.587. Redox regulation of an AP-1-like transcription factor, YapA, in the fungal symbiont Epichloë festucae. Gemma M. Cartwright, Barry Scott, YvonneBecker. Molec Biosci, Massey Univ, Palmerston Nth, New Zealand.Reactive oxygen species (ROS) are emerging as important regulators required for the successful establishment and maintenance of the mutualisticassociation between the fungal endophyte Epichloë festucae and its grass host Lolium perenne. The generation of reactive oxygen species (ROS) by thefungal NADPH oxidase, NoxA has previously been shown to regulate hyphal growth of E. festucae in planta; a result that has led to the hypothesis thatfungal-produced ROS are key second messengers in the symbiosis. However, the highly reactive nature of these molecules dictates that cells possessefficient sensing mechanisms to maintain ROS homeostasis and prevent oxidative damage to cellular components. The Saccharomyces cerevisiae Gpx3-Yap1 and Schizosaccharomyces pombe Tpx1-Pap1, two-component H 2O 2 sensors, serve as model redox relays for coordinating the cellular response toROS. While proteins related to the Yap1 and Pap1 basic-leucine zipper (bZIP) transcription factors have been identified in a number of filamentous fungi,the components involved in the upstream regulation remain unclear. This study investigated the role of the E. festucae Yap1 homologue, YapA, andputative upstream activators GpxC and TpxA, homologues of Gpx3 and Tpx1, respectively, in responding to ROS. YapA is involved in responding to ROSgenerated at the wound site following inoculation into ryegrass seedlings. However, deletion of yapA did not impair host colonization indicatingredundancy in systems used by E. festucae to sense and respond to plant-produced ROS. In culture, deletion of E. festucae yapA, renders the mutantssensitive to only a subset of ROS and this sensitivity is influenced by the stage of fungal development. In contrast to the H 2O 2-sensitive phenotype widelyreported for fungi lacking the Yap1-like protein, the E. festucae yapA mutant maintains wild-type mycelial resistance to H 2O 2 but conidia of the yapAmutant are very sensitive to H 2O 2. Using a degron-tagged GFP-CL1 as a reporter, we found YapA is required for the expression of the spore specificcatalase, catA. Moreover, YapA is activated by H 2O 2 independently of both GpxC and TpxA, suggesting a novel mechanism of regulation exists in E.festucae. This work provides a comprehensive analysis of the role and regulation of the AP-1 transcription factor pathway in a filamentous fungal species.27th Fungal Genetics Conference | 265

FULL POSTER SESSION ABSTRACTS588. Genomic approaches to understand pathogenesis in the basidiomycete pathogen of food and energy crops, Rhizoctonia solani. Jonathan P.Anderson 1,2 , James K. Hane 1 , Rhonda Foley 1 , Cynthia Gleason 1 , Karam B. Singh 1,2 . 1) Plant Industry, CSIRO, Floreat, West Australia, Australia; 2) The UWAInstitute of Agriculture, The University Of Western Australia, Crawley, West Australia.Rhizoctonia solani is a broad-host-range necrotising fungal pathogen that is responsible for significant diseases to diverse crops. In Australia, R. solanimost notably causes bare patch of cereals and costs $77 million pa in direct losses, while internationally it is a significant problem for global riceproduction. In the absence of strong host resistance, an understanding of fungus pathogenesis underpins alternative approaches to enhance resistance incrop plants. While the majority of phytopathogens sequenced to date belong to the Ascomycotina, R. solani is a basidiomycete with the closest sequencedrelatives being biotrophic rust and smut fungi and saprophytic mushrooms, each possessing a lifestyle vastly different from R. solani. We have over-comecomplications associated with the multinucleate, heterokaryotic nature of R. solani to assemble a high quality consensus haploid genome of an AG8isolate. Transcriptomics assisted with genome annotation and identified putative pathogenesis genes. Several of these genes display host-specificexpression, while others show consistent infection-related expression across different anastomosis groups on different hosts. LC-MS basedproteogenomics identified proteins from three fractions; soluble hyphal proteins, membrane localised proteins and secreted proteins, from R. solanigrowing in-vitro or in wheat infection mimicking conditions. QPCR confirmed up-regulation of some of the corresponding genes in infected wheat rootscompared to R. solani grown in vitro, providing further support for a pathogenesis-related role. Functional testing of the role of candidate pathogenesisgenes is on-going. On the plant side of the interaction, large scale gene expression and mutant analyses revealed the high degree of resistance inArabidopsis (unlike its’ susceptible relative, canola) was dependent on reactive oxygen species and not jasmonic acid, ethylene or salicylic acid. Bycontrast, moderate resistance in Medicago truncatula was dependent on ethylene mediated defences, which when over-activated, lead to enhancedresistance. These findings suggest that different plant species employ different defences with differing effectiveness against the same pathogen and acollective understanding of the interplay of host and fungal responses may facilitate novel strategies for enhancing resistance.589. The Cpc1 (CpcA/Gcn4) regulator of the cross-pathway control of amino acid biosynthesis is required for plant infection of the vascular pathogenVerticillium longisporum. Susanna A. Braus-Stromeyer, Christian Timpner, Van Tuan Tran, Gerhard H Braus. Molecular Microbiology and Genetics, Georg-August-University, Goettingen, Germany.The plant pathogenic fungus Verticillium longisporum is the causal agent of early senescence and ripening in Brassica napus (oilseed rape, Canola) andother cruciferous crops. Verticillium wilts have become serious agricultural threats during the last decades. Verticillia infect host-plants through the rootsand colonize xylem vessels of the host-plant, which provide an environment with limited carbon sources. V. longisporum induces the cross-pathwaycontrol in the xylem fluid to cope with an imbalanced amino acid supply.The transcriptional activator gene VlCPC1 (similar to CpcA/GCN4) was knock-downed via RNA-mediated gene silencing and the expression of the twoCPC1 isogenes (VlCPC1-1, VlCPC1-2) in V. longisporum could be reduced up to 85%. The resulting mutants were more sensitive to amino acid starvationinduced by 5-methyltryptophane (5-MT). In plant infection assays, the silenced mutant showed significantly less symptoms such as stunting and earlysenescence. Knockouts of CPC1 in the haploid V. dahliae were sensitive to amino acid starvation and strongly reduced in symptom formation in their hostSolanum lycopersicum (tomato).The hybrid V. longisporum and the haploid V. dahliae are the first phytopathogenic fungi, which were shown to require CPC1 for infection andcolonization of their respective host plants oilseed rape and tomato.590. Fungal-Specific Transcription Factor AbPf2 Activates Pathogenicity in Alternaria brassicicola . Yangrae Cho 1 , Robin Ohm 2 , Igor Grigoriev 2 , AkhilSrivastava 1 . 1) Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Honolulu, HI; 2) United States Department of Energy JointGenome Institute, 2800 Mitchell Drive, Walnut Creek, CA 94598.Alternaria brassicicola is a successful saprophyte and necrotrophic plant pathogen. Molecular determinants of its life style shift between saprophyte andpathogen, however, are unknown. To identify these determinants we studied nonpathogenic mutants of a transcription factor-coding gene, AbPf2.Frequency and timing of germination and appressorium formation on host plants were similar between the nonpathogenic Dabpf2 mutants and wild-typeA. brassicicola. The mutants were also similar in vitro to wild-type A. brassicicola in vegetative growth, conidium production, and responses to chemicalstressors, such as a phytoalexin, reactive oxygen species, and osmolites. The mutants, however, did not penetrate host plant tissues, though their hyphaecontinued to grow on the plant surface. Transcripts of the AbPf2 gene increased exponentially soon after wild-type conidia encountered their host plants.A small amount of AbPf2 protein, monitored by fused green fluorescent protein, was located in both the cytoplasm and nuclei of young, mature conidia.The protein level decreased during saprophytic growth but increased several-fold during pathogenesis. Levels of both the proteins and transcripts sharplydeclined following colonization of host tissues beyond the initial infection site. When the transcription factor was expressed at an induced level in the wildtype during early pathogenesis, the expression of 106 fungal genes was down-regulated in the Dabpf2 mutants. Notably, 33 of the 106 genes encodedsecreted proteins, including eight putative effector proteins. Plants inoculated with Dabpf2 mutants expressed higher levels of genes associated withphotosynthesis, the pentose phosphate pathway, and primary metabolism, but lower levels of defense-related genes. Our results suggest that conidia ofA. brassicicola are programmed as saprophytes, but become parasites upon contact with their hosts. AbPf2 coordinates this transformation by expressingpathogenesis-associated genes, including those coding for effectors.591. WITHDRAWN592. Host-to-pathogen gene transfer facilitated infection of insects by a pathogenic fungus. Weiguo Fang, Xiaoxuan Chen. College of Life Sciences,Zhejiang University, Hangzhou, Zhejiang, China.Inspite being of great concern to human health and the management of plants and animals, the mechanisms facilitating host switching of eukaryoticpathogens remain largely unknown. The endophytic insect-pathogenic fungus Metarhizium robertsii evolved directly from endophytes and itsentomopathogenicity is an evolutionarily acquired characteristic. We found that M.robertsii acquired a sterol carrier (Mr-NPC2a) from an insect byhorizontal gene transfer (HGT). Mr-NPC2a increased the amount of ergosterol in hyphal bodies by capturing sterol from insect hemolymph, and thusmaintained cell membrane integrity and improved fungal survival rate. On the other hand, the reduction in sterol (substrate for molting hormonesynthesis) in insect hemolymph elongated larval stage, which allows the fungus to fully exploit host tissues and produce more conidia. This is first report ofHGT from host to a eukaryotic pathogen, and the host gene ultimately improved the infectivity of the pathogen.266

FULL POSTER SESSION ABSTRACTS593. Characterization of the CoPRF1 mutant of Colletotrichum orbiculare defective in evasion of host defense responses. Kaoru Tanaka, Yasuyuki Kubo.Graduate School of Life and Environmenrtal Sciences, Kyoto Prefectural University, Kyoto, Japan.Plant pathogens have co-evolved with their host plants which have evolved the defense system against their pathogens. It is generally accepted thatplants express basal immunity by the recognition of the pathogen-associated molecular patterns, but compatible pathogens suppress the plant basaldefense by secreting effector proteins. In our previous study, we have obtained several pathogenicity deficient insertional mutants in Colletotrichumorbiculare by Agrobacterium tumefaciens-mediated transformation (AtMT). Among them, in the mutant named YK4524 it was shown that a T-DNAinsertion disrupted a gene which presumably encodes an extracellular protein with signal peptide sequence. And BLAST search of the predicted sequencefound no significant homologous genes in published databases, suggesting that it is unique to C. orbiculare. So we named this gene CoPRF1 (Pathogenesisrelatedfactor). Target gene disruption mutants of coprf1 obtained by AtMT showed significant reduction in virulence on the host leaves. However,characteristics such as germination, appressorium formation and penetration hyphal formation of coprf1 mutants in vitro were normal, indicating thatCoPRF1 is not essential for infection related morphogenesis. On the other hand, penetration ability of mutants was attenuated on intact cucumbercotyledons, and the elongation of its invasive hyphae was slower compared with the wild type. To confirm the possibility that decreased virulence ofcoprf1 mutants was involved in plant defense responses, we inoculated coprf1 mutant on cucumber cotyledons of which defense responses was disturbedby transient heat-shock. As expected, the pathogenicity of coprf1 mutant was restored. Furthermore, expression analysis of CoPRF1 by RT-PCR showedthat CoPRF1 expressed in planta, but not in vitro culture. From these results, it was suggested that CoPRF1 would engage in evasion of host basalresistance at the host infection.594. CPS1 mutants in Coccidioides are avirulent and act as an attenuated vaccine in the valley fever mouse model. Hema P. Narra 1,4 , Lisa F. Shubitz 2,3 , M.Alejandra Mandel 1,3 , Leslie Gunatilaka 5 , Hien Trinh 2,3 , Marc J. Orbach 1,3 . 1) School of Plant Sciences, Univ. of Arizona, Tucson, AZ; 2) Veterinary Sciences andMicrobiology, Univ. of Arizona, Tucson, AZ; 3) Valley Fever Center for Excellence, Univ. of Arizona, Tucson, AZ; 4) Department of Pathology, Univ. of Texas,Medical Branch, Galveston, TX; 5) School of Natural Resources and the Environment, Univ. of Arizona, Tucson, AZ.Coccidioides species are mammalian pathogens endemic to the Southwestern US as well as parts of Mexico, Central and South America. The disease theycause, coccidioidomycosis, or valley fever, is considered an emerging infectious disease due to increases in reported cases over the past 10 years. Toidentify virulence factors of this pathogen that may be targets for therapeutics, we have identified and disrupted genes that are important forpathogenicity in both plant pathogens and other animal pathogens. Based on the work of Liu et al. (2003), we disrupted the Coccidioides ortholog of C.heterostrophus CPS1 in C. posadasii strain Silveira. CPS1 was originally identified as a potential non-ribosomal peptide synthase component, because itencodes a polypeptide with two AMP binding domains related to the adenylation domains in bacterial non-ribosomal peptide synthases. However it alsocontains a putative N-terminal DMAP1b domain. In mammals, this domain binds the DMAP1 transcriptional co-repressor that has been shown to bindregulatory proteins and is proposed to act as a co-repressor of transcription. The C. posadasii cps1 deletion strain is non-pathogenic in susceptible mice butdoes initiate the formation of spherules, the infectious form of Coccidioides. The mutant also forms spherules in vitro. Whether Cps1 plays a role as aregulator of virulence via the DMAP1b domain, or via production of a potential toxin is not known. This is being explored via RNA-seq analysis and isolationof secreted metabolites from both the wild type strain Silveira and the cps1 mutant. The cps1 mutant appears to have great potential as an attenuatedvaccine since it protects from infection; when susceptible mice are challenged with wild type C. posadasii after inoculation with the cps1 mutant, nearly allexperience extended survival of at least four weeks and have low fungal burdens. inoculation with the cps1 mutant, they are completely resistant toinfection. Lu, S. W., S. Kroken, B. N. Lee, B. Robbertse, A. C. L. Churchill, O. C. Yoder, and B. G. Turgeon. 2003. A novel class of gene controlling virulence inplant pathogenic ascomycete fungi. Proceedings of the National Academy of Sciences of the United States of America 100:5980-5985.595. Elucidating the response of wheat to the exposure of Stagonospora nodorum effectors. Lauren A. Du Fall, Peter S. Solomon. Research School ofBiology, Australian National University, Canberra, ACT, Australia.The dothideomycete Stagonospora nodorum is a necrotrophic fungal pathogen of wheat and is the causal agent of Stagonospora nodorum blotch (SNB).This disease is responsible for over $100 million of yield losses in Australia annually. Recent studies have shown that this fungus produces a number ofeffector proteins that are internalized into host cells of susceptible wheat cultivars. The mechanism by which these effectors induce tissue necrosis insusceptible hosts is yet to be fully elucidated. We have applied a multi-omics approach to elucidate the cellular processes leading to disease and provideinsight into the mode-of-action of these effectors. Gas chromatography-mass spectrometry analysis of primary polar metabolites has been undertaken ontissue extracts and apoplastic fluid from SnToxA infiltrated wheat. Results illustrate widespread perturbations in primary metabolism and reveal the firstdirect evidence of an increase in energy production in response to a pathogen effector. To further understand the host response to SnToxA at thesecondary metabolism level, samples were also analysed using liquid chromatography-mass spectrometry. Our data indicate SnToxA causes an increase indefence-related secondary metabolites. The effect of these metabolites on Stagonospora nodorum growth and sporulation in vitro and in planta hasidentified several compounds with novel anti-fungal properties. These complementary approaches have provided a novel insight into the contribution ofthe SnToxA effector protein to SNB in wheat.596. Nep1-like proteins of the downy mildew Hyaloperonospora arabidopsidis trigger immunity, but not necrosis, in the Arabidopsis host. Stan Oome 1,2 ,Adriana Cabral 1 , Simon Samwel 1 , Tom Raaymakers 1 , Guido Van den Ackerveken 1,2 . 1) Plant-Microbe Interactions, Utrecht University, Utrecht, Netherlands;2) Centre for BioSystems Genomics (CBSG), Wageningen, Netherlands.The genome of the downy mildew pathogen Hyaloperonospora arabidopsidis, an obligate biotrophic oomycete, encodes several necrosis and ethyleneinducingpeptide 1 (Nep1)-like proteins (NLP). The NLPs of H. arabidopsidis (HaNLPs) constitute a family of 12 genes and 15 pseudogenes, most of whichform a species-specific clade separate from NLPs of related Phytophthora species, suggesting that the family has recently expanded. The secreted HaNLPswere found to be nontoxic when tested on Arabidopsis or tobacco, in contrast to known necrosis-inducing NLPs, e.g. the P. sojae PsojNIP protein that iscytolytic and induces a strong cell death response in dicot plant tissues. Even HaNLP3, which is most similar to necrosis-inducing NLP proteins of otheroomycetes, and which contains all amino acids that are known to be important for necrosis-inducing activity, did not induce necrosis. Chimerasconstructed between HaNLP3 and the necrosis-inducing PsojNIP protein demonstrated that most of the HaNLP3 protein is functionally equivalent toPsojNIP, except for an exposed domain that prevents necrosis induction. The HaNLP genes are mostly expressed early during infection, suggesting analternative function of noncytolytic NLP proteins during biotrophic infection of plants. To investigate if HaNLP production in the host affects susceptibilityto infection, transgenic Arabidopsis plants were generated. Surprisingly, overexpression of HaNLP3, 5, 6, 9, and 10 resulted in plants with severely reducedgrowth. To be able to monitor NLP-effects on pathogen infection, in the absence of growth reduction, an Arabidopsis line with an estradiol-inducibleHaNLP3 construct was generated. DNA microarray analysis revealed that plant immune responses were strongly activated upon estradiol-induced HaNLP3expression. Furthermore, resistance to H. arabidopsidis infection was activated, suggesting that the plant is able to recognize the pathogen-associated27th Fungal Genetics Conference | 267

FULL POSTER SESSION ABSTRACTSHaNLP3 protein and mount an effective immune response. Our research is now focused on determining how Arabidopsis is able to respond to the HaNLPsand how the downy mildew pathogen can suppress the host immune response triggered by non-toxic NLPs.597. Genes important for in vivo survival of the human pathogen Penicillium marneffei. Harshini C. Weerasinghe, Michael J. Payne, Hayley E. Bugeja,Alex Andrianopoulos. Genetics, The University of Melbourne, Parkville, Victoria, Australia.Pathogenic fungi are having an increasing global impact in the areas of health, agriculture and the environment. As such it is essential to understand themechanisms that fungi employ to survive and grow within a host. The emergence of many new “opportunistic fungal pathogens” has to a great extentaltered the traditional view that pathogenicity was solely reliant on the inherent properties of the pathogen. In fact, the ability of a pathogen to causedisease in some hosts but not in others suggests that pathogenic determinants are complex and dynamic, and are largely dependent on specific pathogenhostrelationships. Despite this there are conserved aspects of the interactions between host and pathogen. For example., hosts employ innate immuneresponses as an almost immediate recognition and attack mechanism against invading pathogens. Penicillium marneffei is a temperature dependentdimorphic fungus, growing in a hyphal form producing conidia at 25°C and as a yeast form at 37°C. Despite its importance as an opportunistic pathogen,little is known about the biology and mechanism of infection of P. marneffei. The infectious agents (conidia) are believed to be inhaled, reaching thealveoli of the lungs, where they are phagocytosed by alveolar macrophages for elimination. At this point that P. marneffei switches growth to a pathogenicyeast cell form, and is able to withstand macrophage cytotoxic attacks to cause infection. In order to understand how P. marneffei responds to the host,RNA-seq analysis was used to create a transcriptomic profile of P. marneffei, when infected in murine macrophages. These results were compared to RNAseqdata from hyphal (25°C) and yeast (37°C) cells grown in vitro in order to identify genes that are specifically upregulated during infection. Based on thisanalysis a group of genes of varying functions were chosen for gene deletion studies and tested for defects in pathogenicity. Among these is a group ofPep1-like aspartic endopeptidases which are a uniquely expanded family in P. marneffei and that show reduced virulence in a macrophage model.598. Oxalate-minus mutants of Sclerotinia sclerotiorum via T-DNA insertion accumulate fumarate in culture and retain pathogenicity on plants.Liangsheng Xu 1 , Meichun Xiang 1 , David White 1 , Weidong Chen 1,2 . 1) Plant Pathology, Washington State University, Pullman, WA; 2) USDA-ARS, WashingtonState University, Pullman, WA 99164.Sclerotinia sclerotiorum is a ubiquitous necrotrophic pathogen capable of infecting over 400 plant species including many economically important crops.Oxalic acid production has been shown in numerous studies to be a pathogenicity factor for S. sclerotiorum through several mechanisms. During ourrandom mutagenesis study of S. sclerotiorum using Agrobacterium-mediated transformation, we identified three mutants that had lost oxalate production.Southern hybridization blots showed the mutation was due to a single T-DNA insertion, and plasmid rescue and DNA sequencing confirmed that the T-DNAinsertion site was located in the ORF of oxaloacetate acetylhydrolase (Ssoah, SS1G_08218) of S. sclerotiorum. The mutants did not change the color of apH-indicating medium (PDA amended with 50 mg/L bromophenol blue). The pH values of 6-day PDB culture filtrates were 1.8-2.0 for the wild type and 2.8-3.1 for the mutants. No oxalic acid was detected using HPLC in culture filtrates or in the mycelium of the mutants, but another acid compound wasaccumulated in culture filtrates of the mutants and detected by HPLC, and the compound was identified as fumaric acid using LC-MS. The mutants showedreduced vegetative growth on PDA and produced sclerotia that are beige in color and soft in texture. Artificial acidic conditions (pH 3.4 and 4.2) enhancedvegetative growth and promoted normal (black and hard) sclerotial formation of the mutants. Furthermore, the oxalate-minus mutants retainedpathogenicity on pea, green bean and faba bean in detached leaf assays and on intact plants of Arabidopsis thaliana, and their virulence levels were similarto that of the wild type strain on certain host plants, but varied depending on the plant species tested. The mutant had increased expression levels of cellwall-degrading enzymes such as polygalacturonases compared to the wild type strain during the process of infecting pea leaves. The results showed that alow pH condition is very important for growth and virulence of S. sclerotiorum on its wide range of host.599. Molecular characterization of fungi associated with superficial blemishes of potato tubers in Al-Qasim region, Saudi Arabia. Rukaia M Gashgari 1 ,Youssuf A. Gherbawy 2 . 1) Biology Dept, King Abdulaziz university, Jeddah, Saudi Arabia; 2) Biology Dept, Taif university, Taif , Saudi Arabia.Potato (Solanum tuberosum) becoming a more and more important foodstuff in the world. Also, the visual quality of fresh potatoes became a dominantcriterion and a significative economical issue in potato market. According the vegetative reproduction of this species, requirements for visual quality arealso needed for potato tubers. As an organ for reserve and propagation, the tuber grows underground and is in contact with soil-borne microorganisms,making it potentially exposed to blemishes. Some blemishes are due to known pathogens and others whose causes are unknown are called atypicalblemishes. Therefore, knowledge about the pathogens is needed to set up efficient control strategies and to help potato growers to better know thecauses of these blemishes and find technical solutions for improving the potato quality. Therefore, the objective of this proposed research study is thepossibility of using some modern methods of molecular diagnostics and rapid detection of the presence of fungal contaminants in potato blemishes in Al-Qasim (Saudi Arabia). Polygonal lesions was the most observed blemish type in the collected samples. One hundred and sixty isolates were collected fromdifferent types of blemishes recorded in this study. Fusarium , Penicillium, Ilyonectria, Alternaria and Rhizoctonia were the most common genera collectedfrom different blemish types. Using ITS region sequencing all collected fungi identified the species level. All Fusarium strains colled during this study wereuse to detect its pathogenicity against potato tubers. The inoculated fungi were re-isolated from the diseased potato tubers to prove the Koch’spostulates. This is the first comprehensive report on identity of major pathogenic fungi causing potato dry rot isolated from potato tuber blemishes inSaudi Arabia.600. Patterns of Distribution of Bacterial Endosymbionts in Lower Fungi. Olga Lastovetsky 1 , Xiaotian Qin 2 , Stephen Mondo 2 , Teresa Pawlowska 2 , AndriiGryganskyi 3 . 1) Microbiology Dept, Cornell University, Ithaca, NY; 2) Plant Pathology & Plant-Microbe Biology Dept, Cornell University, Ithaca, NY; 3)Biology Dept, Duke University, Durham, NC.Fungi are not typically known to have endosymbionts. However, some members of Glomeromycota and Mucoromycotina have recently been found toharbor bacteria in their hyphae and spores. The newly discovered association between Rhizopus microsporus (Mucoromycotina) and Burkholderia bacteria(betaproteobacteria) prompted us to search for endobacteria in other members of Mucoromycotina fungi. We screened a broad range of Mucoromycotinaisolates for the presence of bacterial endosymbionts using PCR with universal and Burkholderia-specific primers that targeted the 16S and 23S rRNAbacterial genes. Endobacteria were only found in certain strains of R. microsporus but in no other Rhizopus or Mucoromycotina isolates. A 28S rRNA genephylogeny of the screened fungal isolates revealed a clustering of bacteria(+) R. microsporus isolates away from bacteria(-) R. microsporus isolates. Toexplore this putative divergence within the R. microsporus lineage we are working on a multi-gene phylogeny of Rhizopus isolates, which is based onmultiple coding and non-coding regions.268

FULL POSTER SESSION ABSTRACTS601. Phylogenetic and genomic analysis of a novel, nematophagous species of Brachyphoris. S. Sharma Khatiwada, J. B. Ridenour, A. Thomas, J. Tipton, T.Kirkpatrick, B. H. Bluhm. University of Arkansas, Fayetteville, AR.Plant-parasitic nematodes are destructive pathogens of crops worldwide. The phase out of many chemical control methods has prompted a search forfeasible alternative control strategies. Nematophagous fungi are widely distributed in terrestrial and aquatic environments, and have evolved diversestrategies to parasitize nematodes. In this study, a previously characterized but unnamed nematophagous fungus (designated TN14) was taxonomicallyclassified and a draft genome sequence was obtained. Taxonomic identification of the fungus was conducted using the ITS1-5.8S-ITS2 rDNA sequences.Phylogenetic relationships were inferred with neighbor-joining and maximum likelihood methods. Based on the primary GenBank database search, the ITSregion of TN14 was compared with the ITS region of 41 taxa. From this analysis, the fungus is predicted to form a distinct monophylogenetic clade withBrachyphoris, a genus of nematophagous fungi related to Dactylella and Vermispora. Although some 200 species of nematophagous fungi are known,publicly available resources are very limited. Thus, we obtained a draft sequence of the TN14 genome via Roche-454 sequencing technology. Alignment ofover 90% of the sequenced reads revealed an estimated genome size of 100.1 MB, which is notably larger than the genomes of many other ascomycetes,including that of the only other sequenced nematophagous fungus, Arthrobotrys oligospora (40.07 Mb). Subsequent analyses of the genome of TN14 areproviding insight into molecular mechanisms underlying pathogenicity and the viability of TN14 as a potential bio-control agent in agricultural settings.602. The proteome of the traps of the nematode-trapping fungus Monacrosporium haptotylum . K-M. Andersson 1 , T. Meerupati 1 , F. Levander 2 , E.Friman 1 , D. Ahrén 1 , A. Tunlid 1 . 1) Microbial Ecology, Department of Biology, Lund University, Sweden; 2) Protein Technology, Department ofImmunotechnology, Lund University, Sweden.Nematode-trapping fungi have for a long time been seen as putative biological control agents against parasitic nematodes. A better knowledge on theinfection process will facilitate the development of these fungi as biological control agents and may also lead to the discovery of new nematicidal drugs.Monacrosporium haptotylum is a nematode-trapping fungus that captures nematodes using an adhesive trap called knob. In this study, proteins wereextracted from knobs and mycelium and analyzed using SDS-PAGE combined with LC/MS/MS. Peptides were matched against predicted gene models fromthe recently sequenced genome of M. haptotylum. Furthermore, the transcriptome in the knob during infection of nematodes were analyzed.The analysis showed that there was a large difference in the proteome of the knob compared to the mycelium. In total 336 proteins were identified. Aquantitative analysis showed that 54 proteins were expressed at significantly higher levels in the knobs versus the mycelium. Proteins containing apredicted secretion signals were overrepresented in knobs (knobs 41 %; mycelium 11 %). Five of the secreted proteins upregulated in knob were smallsecreted proteins (SSPs). Three of the SSPs were orphans since they showed no homology to the NCBI database and lack pfam domains. Interestingly, twoof them are upregulated in the transcriptome during infection of nematodes.Among the upregulated proteins were several putative cell-surface adhesins containing the carbohydrate binding domain WSC and repetitive regionsenriched in threonine/serine residues. Upregulated were also a diverse array of peptidases including serine endopeptidase (subtilisin), asparticendopeptidase, metalloendopeptidase, aminopeptidase and carboxypeptidase. Several proteins related to stress response and basic metabolism were alsoidentified in the trap proteome. During infection of nematodes, genes with the domains peptidase_S8 (subtilisin), DUF3129 and WSC are highlyupregulated in the knob.Taken together, our analysis shows that the trap cell has a unique proteome containing components that are involved in the early stages of infectionincluding adhesion and penetration of the nematode.603. Sequencing the in planta transcriptomes of Colletotrichum species provides new insights into hemibiotrophy. Richard J. O'Connell 1 , StéphaneHacquard 1 , Jochen Kleemann 1 , Emiel Ver Loren van Themaat 1 , Stefan Amyotte 2 , Michael Thon 3 , Li-Jun Ma 4 , Lisa Vaillancourt 2 . 1) Max Planck Institute forPlant Breeding Research, Cologne, Germany; 2) Department of Plant Pathology, University of Kentucky, Lexington, KY; 3) CIALE, Universidad de Salamanca,Villamayor, Spain; 4) Department of Biochemistry and Molecular Biology, UMASS Amherst, MA.Colletotrichum species cause devastating diseases on crop plants worldwide. Infection involves formation of a series of specialized cell-types associatedwith penetration (appressoria), growth inside living host cells (biotrophic hyphae) and tissue destruction (necrotrophic hyphae). To analyse thetranscriptional dynamics underlying these transitions, we used RNA sequencing to compare the transcriptomes of C. higginsianum infecting Arabidopsisand C. graminicola infecting maize. The early transcriptome is dominated by secondary metabolism and effector genes, suggesting both appressoria andbiotrophic hyphae are platforms for delivering protein and small molecule effectors to host cells. Genes encoding a vast array of wall-degrading enzymes,proteases and membrane transporters are up-regulated at the switch to necrotrophy, when the pathogen mobilizes nutrients from dead cells for growthand sporulation. However, the two species employ different strategies to deconstruct plant cell walls that are adapted to their host preferences. Thus, C.higginsianum activates more pectin-degrading enzymes during necrotrophy, whereas C. graminicola mostly activates hemicellulases and cellulases at thisstage. Remarkably, although appressoria formed in vitro are morphologically similar to those in planta, comparison of their transcriptomes showed >1,500genes are induced only upon host contact, suggesting that sensing of plant signals by appressoria dramatically reprograms fungal gene expression inpreparation for host invasion.604. Biological activities of natural products synthesized by the mammalian fungal pathogen, Histoplasma capsulatum. A. Henderson 1 , M. Donia 2 , M.Fischbach 2 , A. Sil 1 . 1) Microbiology and Immunology, UCSF, San Francisco, CA; 2) Department of Bioengineering and Therapeutic Sciences, University ofCalifornia, San Francisco, San Francisco, CA.Histoplasma capsulatum is a soil fungus that infects healthy mammalian hosts upon inhalation. Extrapolating from previous work, we hypothesized thatsmall-molecule natural products produced by Histoplasma are enriched for activity against host molecular targets. Using a bioinformatics approach, weidentified biosynthetic gene clusters in strain G217B containing genes required for natural product synthesis in other organisms: nonribosomal peptidesynthetases (NPS) and polyketide synthases (PKS). Experimentally, we found that partially purified compounds from Histoplasma culture supernatants areable to buffer supernatants against acidic challenge and promote macrophage lysis. Both activities are relevant to virulence in mammalian hosts. We arestructurally characterizing the relevant natural products using preparative HPLC, MS and NMR. In a complementary approach, we used RNA interferenceto target the complete set of NPS and PKS genes identified in the Histoplasma genome. We are using the resultant mutant strains to correlate biosyntheticgenes with small molecule production, and to assess the role of these genes in pathogenesis.605. From antagonism to synergism: roles of natural phenazines in bacterial-fungal interactions between Pseudomonas aeruginosa and Aspergillusfumigatus. He Zheng 1 , Fangyun Lim 2 , Jaekuk Kim 1 , Mathew Liew 1 , John Yan 1 , Neil Kelleher 1 , Nancy Keller 2 , Yun Wang 1 . 1) Northwestern University,Evanston, IL, USA; 2) University of Wisconsin-Madison, Madison, WI, USA.Secreted small molecules are increasingly recognized to mediate many types of bacterial-fungal interactions in nature and the clinical environment,27th Fungal Genetics Conference | 269

FULL POSTER SESSION ABSTRACTSwhich can have enormous impacts on human and ecosystem health. Despite their ubiquity and importance, very little is known about the molecularmechanisms underlying these interactions. To address this, we select to study the interactions between Pseudomonas aeruginosa and Aspergillusfumigatus, the ubiquitous opportunistic bacterial and fungal pathogens, respectively, via redox-active bacteria-secreted phenazines. We hypothesize thatthe functions of these molecules are multifactorial, dependent on genetic and environmental factors. By combining genetic, physiological, electrochemical,and metabolic profiling strategies, here we report that redox-active phenazines can mediate biofilm interactions between P. aeruginosa and A. fumigatusin multiple ways, ranging from antagonistic to synergistic. We find that phenazine production patterns are generally correlated with bacterial-fungalinteraction phenotypes, in a genetically- and temporarily-dependent manner. Further, fungi can convert the precursor phenazine-1-carboxylate (PCA)produced by bacteria into several other phenazines. These structurally related phenazines come in with characteristic physical-chemical propertiesincluding redox properties. Our most striking finding is to be able to draw connections between a phenazine’s structure and its mode of action. Under onegiven condition, some phenazines such as phenazine-1-carboxamide (PCN) can facilitate bacterial biofilm development by inhibiting fungal development;some others such as pyocyanin (PYO) show no apparent effect on fungal development; and 5-methyl-phenazine-1-carboxylic acid (5-Me-PCA) cansynergistically facilitate both bacterial and fungal biofilm developments. In addition, we find that changing the ambient redox and pH conditions can affecta phenazine’s mode of action, likely via influencing its redox activity. Taken together, our findings imply that phenazines-mediated bacterial-fungalinteractions have profound and diverse effects on multicellular behavior in competitive and mixed-species biofilm environments.606. Genomic analysis of Mortierella elongata and its endosymbiotic bacterium. Gregory Bonito 1 , Andrii Gryganskyi 1 , Christopher Schadt 2 , Dale Pelletier 2 ,Amy Schaefer 3 , Gerald Tuskan 2 , Jessy Labbé 2 , Sofia Robb 4 , Rebecca Ortega 1 , Francis Martin 5 , Mitchel Doktycz 2 , Kurt LaButti 6 , Matt Nolan 6 , Robin Ohm 6 , IgorGrigoriev 6 , Rytas Vilgalys 1 . 1) Duke University, Durham NC; 2) Oak Ridge National Laboratory, Oak Ridge TN; 3) University of Washington, Seattle WA; 4)University of California, Riverside CA; 5) Institut National de la Recherche Agronomique, Nancy France; 6) Joint Genome Institute, Walnut Creek CA.Mortierella belong to a group of basal fungi (Mortierellomycotina) common to soils and the rhizosphere and endosphere of many plant species.Mortierella species are known for rapid growth and abundant lipid production. Mortierella elongata is one species commonly isolated from forest soils andhealthy plant roots where it grows asymptomatically as an endosymbiont. Mortierella elongata is a heterothallic species but can also reproduce asexuallythrough chlamydospores and sporangiospores. Recent reports indicate that some isolates of M. elongata host endosymbiotic bacteria, which may betransmitted vertically via spores. However, it is still unclear whether all Mortierella species host endosymbionts or whether these are lineage-specificassociations. Given the geographically widespread distribution of Mortierella elongata and its ubiquitous presence in forest soils and plants we chose tosequence its genome through the JGI Forest Metatranscriptome CSP. We also sought to assemble the genome of the bacterial endosymbiont to addresswhether there are genomic signatures of co-adaptation or co-evolution in the genomes of Mortierella and its endosymbiotic bacterium, which may impactthe function and growth of Mortierella elongata. The 50 Mb genome of M. elongata was sequenced to 112x coverage. Of the 220,113 putative proteinsidentified in M. elongata, 109,093 appear to be unique (e.g. only ~50% have orthologs in other fungal species having sequenced genomes). The M.elongata genome appears to be enriched in genes related to tryptophan metabolism, siderophore group nonribosomal peptides, glucan 1,4-alphaglucosidases, and in lipid metabolism (e.g. sphingolipids, etherlipids, and glycerophopholids) compared to genome sequences of other basal fungi. Theendosymbiotic bacterium sequenced along with the M. elongata isolate is related to Glomeribacter (endosymbiont of Gigospora, Scutellospora, and otherGlomeromycota) within the Burkholdariales. The ~2.6 MB endosymbiont genome is larger than that of Glomeribacter but quite reduced compared to freelivingisolates of Burkholdaria. The reduced genome size of this bacterium, and the fact that it has thus far evaded pure culture isolation, supports the viewthat this is an ancient and obligate symbiosis.607. Diversity and Content of Maize Leaf Endophytes are Correlated With Maize Genotype. Alice C. L. Churchill 1* , Santiago X. Mideros 1 , Peter Balint-Kurti 2 , Surya Saha 1 , Rebecca J. Nelson 1 . 1) Department of Plant Pathology & Plant-Microbe Biology, Cornell Univ, Ithaca, NY; 2) USDA-ARS Plant ScienceResearch Institute, Department of Plant Pathology, North Carolina State University, Raleigh, NC 27695.All plants contain endophytes that have the potential to provide fitness benefits to their hosts by increasing tolerance to environmental stressors,boosting plant nutrition and growth, and providing increased resistance or tolerance to insect pests and plant pathogens. We are characterizingendophytic populations inhabiting aboveground maize tissues with the goal of associating maize genetic variation with the diversity, structure andconstitution of maize-associated microbial communities. Nine maize lines, representing a diverse subset of the founders of the NAM (Nested AssociationMapping) population, were grown at a single North Carolina field site in 2012 and assayed for culturable endophytic bacteria and fungi. Two distinct seedsources for each maize line were planted in a randomized experimental design, and three replicates per seed source were assayed, representing a total of54 samples. Leaf pieces were harvested just prior to pollination for each maize line, surface sterilized using standard endophyte isolation methodologies,and ground leaf extracts were cultured on four media that select for slow- and fast-growing fungi and copiotrophic, diazotrophic, and oligotrophicbacteria. Approximately 65% of the samples contained one or more phenotypically distinct, culturable bacteria, 28% contained one or more fungi, 22%contained both bacteria and fungi, and endophytes were undetectable in 28% of the samples. A greater number and diversity of fungi were cultured fromtropical maize lines than from temperate lines. Bacteria were isolated from all maize lines, with some lines exhibiting significantly greater microbialcommunity diversity than others. Several phenotypically similar bacteria and fungi were isolated from multiple maize lines. Microbial identity via 16S andITS sequencing, as well as identification of unculturable endophytes via whole genome metagenomic sequencing, are in progress. We are particularlyinterested in identifying members of the microbiome that modulate disease symptoms caused by maize leaf and ear pathogens. Hence, future studies willfocus on in vitro and in planta endophyte-pathogen interactions.608. Characterisation of epichloae endophytes from the Triticeae and their potential use in modern cereals. Wayne R Simpson, Marty J Faville, Roger AMoraga, Richard D Johnson. Agresearch Grasslands, Palmerston North, New Zealand.Epichloae endophytes infect grasses within the subfamily Pooideae including some within the tribe Triticeae. There have been no accounts of moderndomesticated Triticeae hosting epichloae endophytes but there have been reports in Elymus, Hordeum and other grasses within the tribe. Our goal is toisolate epichloae endophytes from the wild relatives of modern cereals and inoculate these into modern cereal crops. We surveyed populations of Elymusand Hordeum, primarily from Asia, and selected 29 Elymus and 13 Hordeum infected plants. We used simple sequence repeats (SSR) and b-tubulinsequencing to determine genetic similarity, hybrid status and closest non-hybrid ancestor. SSR data indicates 26 genetically distinct strains that fall into 5major clades. b-tubulin analysis shows that the majority of our isolates had Epichloë bromicola ancestry, with both hybrid and non-hybrid strainsidentified. Within the non-hybrid E. bromicola two major clades were identified. Of the hybrids we identified examples of E. bromicola x E. typhina and E.bromicola x E. amarillans. Although E. bromicola has been observed in Asian grasses we believe that this is the first report of an E. bromicola x E amarillanshybrid. Of the remaining isolates, we found examples of strains with E. yangzii ancestry (all non-hybrids) and E. elymi ancestry (both non-hybrid and270

FULL POSTER SESSION ABSTRACTSE.elymi x E. amarillans).609. The Interaction of Mycoplasma-related Endobacteria with their Arbuscular Mycorrhizal Fungal Host. Mizue Naito 1 , Teresa Pawlowska 2 . 1) Dept. ofMicrobiology, Cornell University, Ithaca, NY; 2) Dept. of Plant Pathology & Plant-Microbe Biology, Cornell University, Ithaca, NY.Arbuscular mycorrhizal fungi (AMF), comprising the monophyletic phylum Glomeromycota, are obligate biotrophs, and form symbiotic associations with80% of terrestrial plants. AMF associate symbiotically with the roots of plants, and are specialized in the transfer of nutrients from the soil to the planthost. In return for increased nutrient uptake, the plants supply AMF with up to 20% of their photosynthetically derived carbohydrates. Thus, AMFsymbiosis contributes significantly to global nutrient cycling and terrestrial ecosystems. AMF have been known to harbour two types of bacteria in theircytoplasm: (i) the Burkholderia-related Candidatus Glomeribacter gigasporarum and (ii) a Mycoplasma-related bacteria, which we refer to as Mycoplasmarelatedendobacteria (MRE). MRE live freely in the AMF cytoplasm, and have been found associated with all lineages of AMF worldwide. Virtually nothingis known about the MRE, such as their evolution, biological capabilities, and whether they are mutualists or parasites of their AMF hosts. In order tounderstand the nature of this symbiosis, and determine the role that the MRE play in arbuscular mycorrhizae, next generation sequencing (Roche 454 andIllumina) was performed on MRE isolated from 3 distinct AMF hosts, Claroideoglomus etunicatum, Funneliformis mosseae, and Racocetra verrucosa.Phylogenetic reconstruction and divergence dating using 22 conserved genes have revealed that MRE form a novel monophyletic subclade of theMycoplasmas and have diverged from their Mycoplasma relatives at least 400 million years ago, which may indicate the establishment of the MRE-AMFassociation to be quite ancient. Analysis of annotated genes have revealed novel proteins that are likely to play a role in interacting directly with the fungalhost. Preliminary data suggest that MRE are important in enabling the completion of the life cycle of their AMF hosts.610. The Velvet gene is required for mutualism between Epichloë festucae and perennial ryegrass. Mostafa Rahnama 1,2 , Richard Gardner 1 , DamienFleetwood 2 . 1) School of Biological Sciences, University of Auckland, Auckland, New Zealand; 2) Forage Improvement Group, AgResearch, Auckland, NewZealand.The velvet gene (veA or velA) is a key factor in the regulation of fungal development, biosynthesis of secondary metabolites and hyphal growth. Thisstudy aimed to determine the role of velA regulation in Epichloë festucae and its mutualistic interaction with the agriculturally important forage perennialryegrass (Lolium perenne). Infection of perennial ryegrass with an E. festucae mutant deleted in velA caused rapid seedling death in two thirds of infectedplants while remaining plants displayed a normal interaction phenotype, although after several weeks these plants also become stunted and died in anunusual delayed plant-interaction phenotype. No hypersensitive response was observed by microscopy, suggesting the response is not driven bypathogen-like effector proteins. Microscopic analysis showed different accumulation of polysaccharides between mutant and wild type strains. Themutant strain could grow in higher concentrations of calcolfluor and also there was different colony hydrophobicity between wild type and mutant strains.These different cell wall properties suggest a possible microbe associated molecular pattern (MAMP)-triggered defense response may be occurring inDvelA mutant associations. We are currently analysing the transcriptomes of wild type and mutant E. festucae/Lolium perenne symbiota to determine thevelA regulon and elucidate the mechanism of host death.611. An examination of phosphate solubilization and hormone production by two Penicillium species growing in the rhizoplane. Tim Repas 1,2 , DavidGreenshields 2 , Susan Kaminskyj 1 . 1) Department of Biology, University of Saskatchewan, Saskatoon, Saskatchewan, Canada; 2) Novozymes BioAg Ltd. 3935Thatcher Ave, Saskatoon, SK S7R1A3, Canada.Some soil microorganisms, including fungi, can enhance plant growth in the natural environment, however the mechanism(s) by which they promoteplant growth (PGP) are only partly known, and at any rate these will likely vary between organisms. Possible mechanisms for PGP include enhancednutrient uptake, hormone production, pathogen biocontrol, and increased water use efficiency. We are evaluating the potential of two rhizosphere fungi,Penicillium bilaiae (Pbil) and a novel isolate (Skj340) that is most closely related to Penicillium atramentosum. Both fungi can solubilize in liquid cultureeach of Fe-, Al-, and Ca-phosphates, which are commonly found in soil. Both strains were able to solubilize phosphate minerals equally well. However, Pbilproduces abundant organic acids, whereas Skj340 does not produce organic acids, nor even change the pH of the spent medium. Pbil has been tagged withred fluorescent protein (RFP); Skj340 has been stained with lactofuschin; both were imaged with confocal fluorescence. Both fungi are found on the rootsurface, and neither could be isolated from surface sterilized plants, thus it appears that these strains are not endophytic. We are currently evaluatingproduction of gibberellins and auxins from Pbil and Skj340 by assessing their ability to complement the phenotype of Arabidopsis mutants deficient inthese hormones. We will also assess whether either strain enhanced root hairs counts, which is expected to be correlated with both nutrient uptake andhormone activity.612. The role of Epichloe festucae RacA interacting proteins, PakA, PakB and RhoGDI, on cell polarity in culture and synchronized growth in Loliumperenne. Yvonne Becker, Carla Eaton, Isabelle Jourdain, Barry Scott. Institute of Molecular BioSciences, Massey University, Palmerston North, NewZealand.The fungal endophyte Epichloe festucae and its host Lolium perenne are an interesting model system to study signals and mechanisms involved inmutualistic symbiosis maintenance. Mutants defective in components of the ROS producing Nox complex show loss of synchronized growth of the fungusin the grass resulting in stunted, multi-tillered plants (Tanaka et al. 2006, Takemoto et al., 2006). The small GTPase RacA is crucial to activate the Noxcomplex in E. festucae and plays a crucial role in establishment and maintenance of polarized hyphal growth (Tanaka et al., 2008). The objectives of thisstudy were to determine whether key regulators of RacA in mammalian systems, the guanine nucleotide dissociation inhibitor (RhoGDI) and p21-activatedkinases (Paks), also regulate fungal RacA in order to control polarised growth in culture and Nox activity for maintenance of the symbiosis with perennialryegrass. We showed by yeast two-hybrid analysis that PakA (Cla4 homolog), PakB (Ste20 homolog) and RhoGDI interact with RacA, whereas the RhoGDIinteraction is compromised in a mutant of RacA (R73E) required for RhoGDI binding. Only partial complementation is achieved when RacA (R73E) isexpressed in the RacA deletion strain, indicating RhoGDI is important for controlling RacA function. Deletion of pakB had a mild effect on polarized hyphalgrowth in culture and wild-type growth in planta. Deletion of pakA had a severe effect on polarized hyphal growth in culture, with a reduction of radialgrowth and hyper-branching, a phenotype similar to the racA mutant but surprisingly plants infected with the pakA mutant had a wild-type interactionphenotype. The in planta results may reflect the fact that E. festucae grows by intercalary rather than tip growth in the intercellular spaces of perennialryegrass leaves.613. A Host-Induced Gene Silencing Approach to Control Mycotoxin Contamination in Corn. J. E. Smith, Y. B. Ramegowda, B. H. Bluhm. University ofArkansas, Fayetteville, AR.The fungal ear rot pathogens Aspergillus flavus and Fusarium verticillioides contaminate corn with aflatoxins and fumonisins, which pose severe health27th Fungal Genetics Conference | 271

FULL POSTER SESSION ABSTRACTSrisks and significantly limit grain marketability. Despite extensive breeding efforts, adequate resistance to mycotoxin accumulation has not beendeveloped. Additionally, tools currently available to control mycotoxin contamination are limited in number and efficacy. Recently, host-induced genesilencing (HIGS) has emerged as a way to manipulate gene expression in fungal pathogens via expression of pathogen-specific hairpin RNA (hpRNA) inplants. The goal of this research was to generate transgenic corn inhibiting mycotoxin accumulation via HIGS. To this end, a high-throughput workflow wasdeveloped to create and validate HIGS expression vectors. First, candidate fungal genes regulating mycotoxin biosynthesis were identified through variousapproaches, including expression profiling and functional genomics. Second, a one-step cloning process was developed to simultaneously create hpRNAencodingconstructs comprised of sense and antisense orientations of target fungal genes, separated by an intron from the Magnaporthe oryzae cutinasegene, flanked by the TrpC promoter and terminator. Constructs were validated by phenotypic assessment after transformation into either A. flavus or F.verticiliioides. Finally, constructs encoding hpRNAs that significantly reduced mycotoxin accumulation were cloned into a plant expression vector andtransformed into maize. Currently, silencing vectors have been created that target the a-amylase (AMY1) and hexokinase (HXK1) genes in F. verticillioidesand the polyketide synthase (aflC) and hexokinase (kxk) genes in A. flavus, and transgenic corn plants have been created. Thus, this research will advancethe current understanding of HIGS in maize and will ultimately provide new tools to control mycotoxin contamination of corn.614. CDiT1, a novel type of proteinaceous toxin secreted by the necrotrophic pathogen of the roots of tomato Pyrenochaeta lycopersici. Pierre-HenriClergeot 1 , Herwig Schuler 2 , Ejvind Mørtz 3 , Maja Brus 1 , Simina Vintila 1 , Sophia Ekengren 1 . 1) Vaxtfysiologi, Stockholms Universitet, Stockholm, Sweden; 2)Karolinska Institutet, Stockholm, Sweden; 3) Alphalyse A/S, Odense, Denmark.During the 24th Fungal Genetics Conference in Asilomar in 2009, we reported the isolation by Fast Protein Liquid Chromatography of a putativeproteinaceous toxin of 18 kDa, secreted in liquid medium by the corky root rot pathogen of tomato, the filamentous ascomycete Pyrenochaeta lycopersici.This molecule, CDiT1, was thought to be the cause of cell death observed in tomato leaves after infiltration of culture filtrates into their apoplast. Furthercharacterization of CDiT1 revealed that it is secreted as a dimer and encoded by a single gene, whose expression peaks during tomato root infection.Infiltration into leaves of various hosts of P. lycopersici of recombinant CDiT1 purified by affinity confirmed its phytotoxic, but differential activity.Especially, currant tomato (Solanum pimpinellifolium) proved to be tolerant to a higher concentration of recombinant CDiT1 than cultivated tomato (S.lycopersicum). This correlates with the observation that roots of currant tomato are less prone to intracellular infection by a transformant of the fungusexpressing a reporter gene than those of cultivated tomato. Affinity-purified recombinant CDiT1 was also used in cut root assays to confirm lethal activityof the toxin on tomato root cells. Finally, searches made by sequence similarity in the genomes of other pathogenic Pleosporales showed that CDiT1 has aputative orthologue in the cereals pathogens Stagonospora nodorum, Pyrenophora teres f.sp. teres and Pyrenophora tritici-repentis, species known forsecreting proteinaceous toxins contributing to virulence as well. In conclusion, our data validate the experimental approach of characterizing moleculessecreted by Pyrenochaeta lycopersici and inducing disease-related symptoms after infiltration into host leaves, this in order to highlight potential geneticresistance against corky root rot in related species or varieties (see also Clergeot et al. 2012, Phytopathology, 102:878-891).615. Nonhost-specific phytotoxicity of the polyketide-derived toxin solanapyrone A produced by Ascochyta rabiei and Alternaria solani. W. Kim 1 , L.Tymon 1 , D. Johnson 1 , W. Chen 2 . 1) Plant Pathology, Washington State University, Pullman, WA; 2) USDA-ARS, Grain Legume Genetic and PhysiologyResearch Unit, Pullman, WA.Solanapyrone A is a polyketide-derived metabolite produced by Ascochyta rabiei and Alternaria solani, which are the most destructive necrotrophicpathogens of chickpea and potato/tomato, respectively. They belong to the Order Pleosporales within the Class Dothideomycetes, but are phylogeneticallydistantly-related. All isolates of the two fungi tested so far are capable of producing solanapyrone A in synthetic media, which may imply that it isindispensable for their life cycle. However, very little is known about the genetics of solanapyrone A production and its role in pathogenesis and their lifecycle. Recently, solanapyrone biosynthesis gene cluster was identified in Al. solani. Six genes (Sol1 - Sol6) form the gene cluster, spaning about 20 kb of thegenome. Among them, Sol5 gene encodes a Diel-Alderase which catalyzes the final step of solanapyrone biosynthesis pathway. Knockout of the Sol5 genein both A. rabiei and Al.solani resulted in the production of three compounds (presumably solanapyrone precursors), instead of solanapyrone A. Colony ofsol5 mutants showed expansive growth in agar medium until covering the entire plates, in contrast to restricted growth of colony of their correspondingwild-type progenitors. The restricted growth of the wild type strains is likely due to solanapyrone toxicity. Phytotoxicity of solanapyrone A was examinedwith various plant species including their natural host plants. Solanapyrone A produced similar size of necrotic lesions on all plant species tested. On theother hand, one of the putative solanapyrone precursors with the same molecular weight of solanapyrone A caused much smaller lesion only around thewounds of application sites. These results indicate that solanapyrone A is a nonhost-specific phytotoxin because it caused similar degree of lesions on hostand nonhost species.616. Crosstalk of the unfolded protein response and regulatory pathways controlling pathogenic development in Ustilago maydis. Kai Heimel 1 , JohannesFreitag 2 , Martin Hampel 1 , Julia Ast 2 , Michael Bölker 2 , Jörg Kämper 3 . 1) Department of Molecular Microbiology and Genetics, Georg-August-University,Göttingen, Germany; 2) Genetics Department, Philipps-University, Marburg, Germany; 3) Genetics Department, Karlsruhe Institute of Technology (KIT),Karlsruhe, Germany.Development of eukaryotic pathogens is accompanied by dramatic changes in morphology, lifestyle, nutrient acquisition and growth behavior.Consequently, pathogens require robust control systems to adapt to changing host environments and to maintain cellular physiology and homeostasis.The unfolded protein response (UPR) is a conserved eukaryotic signaling pathway counteracting endoplasmic reticulum (ER) stress during situations ofincreased demands on the secretory pathway. We identified and characterized the homologs of the central UPR regulators, Hac1 and Ire1 in the biotrophicfungus U. maydis. The UPR is tightly interlinked with the b mating-type-dependent signaling pathway that regulates pathogenic development. Exact timingof UPR is required for virulence and premature activation interferes with the b-dependent switch from budding to filamentous growth. A smut-specific C-terminal extension of the U. maydis Hac1 homolog, Cib1, mediates direct interaction with Clp1, an essential component of the b-mediated signalingcascade. This interaction leads to stabilization of Clp1, increased ER stress resistance and thus prevents deleterious hyperactivation of the UPR duringbiotrophic growth of U. maydis. Since Clp1 expression is decisive for cell cycle release and fungal proliferation in planta we suggest that UPR activationserves as a checkpoint to time developmental progression and secretion of effector molecules, which promote the establishment of the biotrophicinteraction.617. Transcriptional profiling of the APSES genes in Trichophyton rubrum during growth on human nail. Elza A. S. Lang 1 , Nalu T. A. Peres 1 , Maíra P.Martins 1 , Tiago R. Jacob 1 , Pablo R. Sanches 1 , Larissa G. Silva 1 , Antonio Rossi 2 , Nilce M. Martinez-Rossi 1 . 1) Department of Genetics, Ribeirao Preto School ofMedicine, University of Sao Paulo, Brazil; 2) Department of Biochemistry and Immunology, School of Medicine of Ribeirao Preto, University of Sao Paulo,Brazil.272

FULL POSTER SESSION ABSTRACTSThe dermatophyte Trichophyton rubrum is worldwide spread and is the most prevalent causative agent of clinical cases of skin and nail mycoses inhumans. Adhesion, invasion and colonization of keratinized host tissues are crucial for the success of the infection process and depend on the modulationof genetic responses during host-pathogen interactions. The APSES transcription factors are exclusive of the fungi kingdom and have been reported to playimportant roles in cell growth, differentiation, pathogenicity and virulence in several fungi species. In this work, in silico analyses of T. rubrum genomerevealed the presence of genes encoding distinct proteins containing the APSES domain, suggesting that these proteins may play different roles in the cell.A high number of genes potentially regulated by the APSES regulators were identified by in silico analyses of 1000nt upstream of the annotated ORFs of T.rubrum. Transcriptional profiles of the APSES genes were analyzed during growth of T. rubrum on human nail or keratin, as the sole source of nutrients. Invitro infection of human nail was also evaluated by light microscopy. The results revealed that the transcription levels of the APSES genes are modulatedduring the human nail infection process and keratin degradation. Taken together, our findings suggest that the APSES genes of T. rubrum may beimplicated in host-pathogen interactions. Financial support: FAPESP, CAPES, CNPq, FAEPA.618. Carbohydrate binding proteins of two Leptosphaeria pathogens of Brassica napus. Rohan G T Lowe 1 , Bethany Clark 1 , Angela Van de Wouw 1 , AndrewCassin 1 , Jonathan Grandaubert 2 , Thierry Rouxel 2 , Barbara Howlett 2 . 1) School of Botany, University of Melbourne, Melbourne, Victoria, Australia; 2) INRA-Bioger, Campus AgroParisTech, Thiverval-Grignon, France.Effectors include small secreted proteins (SSPs) produced by pathogens to modify or subvert defence responses of the host organism. Leptosphaeriamaculans “brassicae”, the foremost pathogen of Brassica napus (canola), has 651 genes predicted to encode SSPs. The related species L. biglobosa"canadensis" more aggressively infects Brassica cotyledons, but causes fewer stem cankers. This difference in symptomology may be due to a differentresponse from the host innate immune system. Compared to the L. maculans “brassicae” v23.1.3 reference genome, L. biglobosa has a relatively compactgenome (30 Mbp) lacking the characteristic AT-rich, gene-poor repeats of L. maculans. We are using comparative transcriptomics to identify genesinvolved in early stages of infection. RNAseq analysis revealed that >300 L. maculans “brassicae” genes are highly upregulated (>100-fold) 7 days afterinfection compared to in vitro growth. These genes are enriched for SSPs, which comprise 25 of the top 100, but only 5% of the total gene complement. Amajor class of SSPs are carbohydrate active enzymes (CAZY), some of which play a role in evasion of chitin-triggered immunity in plants. L. maculans“brassicae”and L. biglobosa "canadensis" have different complements of the chitin-associated CAZY domains, CBM18 and CBM50 (aka LysM). Both ofthese domains bind chitin or peptidoglycan and may be found with chitinase domains in the same protein. The genomes of both Leptosphaeria speciesencode a similar number of CBM18 domains (27 and 29, respectively). L. maculans “brassicae” has predominantly multi-domain CBM18 proteins that arenot highly upregulated 7 days after infection, whilst L. biglobosa "canadensis" has more abundant single domain proteins, two of which are highlyupregulated (>100-fold) at 7 days post inoculation. Homologs of the well-characterised CBM50/LysM protein, ECP6, from Cladosporium fulvum are presentin both species, and a triplication of the N terminal LysM domain has occurred in L. maculans “brassicae”. The composition and regulation of CBM18 andCBM50-containing genes in Leptosphaeria may be involved in determining the degree of chitin-triggered immunity in the host canola, and concomitantly,the success of the pathogen. Silencing of key CAZY genes in L. maculans “brassicae” is underway, with a focus on members of the CBM50/LysM family.619. Domains for plant uptake of Ustilago maydis secreted effectors. Anupama Ghosh, Armin Djamei, Shigeyuki Tanaka, Regine Kahmann. Max PIanckInstitute for Terrestrial Microbiology, Department of Organismic Interactions, Karl-Von-Frisch-Strasse 10, D-35043 Marburg, Germany.The genome of the corn smut fungus Ustilago maydis codes for a large repertoire of secreted effectors. Some of them play crucial roles for virulence andestablishment of the biotrophic phase. The chorismate mutase Cmu1 is one such secreted translocated effector of U. maydis. cmu1 deletion strains areattenuated in virulence that is attributed to higher salicylate levels in plants infected with the mutant strain, most likely through alterations in thechanneling of chorismate from the plastids to the cytosol. Here we identify the motif in Cmu1 that is necessary for the translocation of the protein acrossthe plant plasma membrane and present a mutational analysis of this region. To test for uptake we assayed the ability of mutant proteins to complement acmu1 mutant strain as well as the retained ability to complement the growth defect of a Daro7 strain of S. cerevisiae in minimal medium. By deletionanalysis a region of 20 amino acids adjacent to the signal peptide was shown to be essential for the translocation. Microscopic analysis of maize tissueinfected with U. maydis strains expressing Cmu1-mcherry fusion proteins with or without the probable uptake motif revealed that the 20 amino acid motifallows binding of the protein to an as yet unknown plant plasma membrane component. We hypothesize that the translocation of Cmu1 across the plantplasma membrane is a two step process; initiated by binding followed by translocation across the membrane. In addition, we present results where the 20amino acid motif is substituted by motifs from other effectors.620. Lipid metabolism influences virulence in Ustilago maydis. Scott Lambie, Matthias Kretschmer, Jim Kronstad. Michael Smith Laboratories, Universityof British Columbia, Vancouver, BC, Canada.Plant tissues and surfaces are a source of lipids which act as a potential carbon source and signals for the pathogenic development of the biotrophic smutfungus Ustilago maydis. This pathogen is an excellent model for the molecular genetic analysis of lipid use during disease and responds to lipids with amorphologic transition from budding to filamentous growth. In addition, the fungus possesses both peroxisomal and mitochondrial b-oxidation pathways,and numerous putative phospholipases (PLs), to exploit lipid carbon sources for nutritional and signalling purposes. We have shown that bothmitochondrial and peroxisomal b-oxidation is important for the utilization of fatty acids and the pathogenic development of U. maydis and may thereforerepresent a potential target to combat crop disease caused by fungal pathogens. In U. maydis, deletion of components of these pathways influencedmating, lead to a decrease in virulence, caused a defect in fatty acid metabolism, a loss of acetate metabolism and the accumulation of toxicintermediates. To further explore the role of the b-oxidation pathway during morphogenesis and pathogenic development, we have investigated theeffects of several non-steroidal anti-inflammatory drugs (NSAIDs), which are known to interfere with b-oxidation functions at various stages. Diclofenacinhibited the usage of fatty acids of different chain length and saturation state as sole carbon sources and had an influence on the filamentation efficiencyof those fatty acids. Further it showed fungicidal activity by inducing apoptosis, and influenced mating and pathogenic development. In an attempt tofurther elucidate the role of lipid utilization during infection we are investigating the role of PLs as a potential mechanism by which host-derived lipidsignals or fatty acids for subsequent b-oxidation are generated. We present an analysis of 17 candidate PL genes identified by genome mining, as well as apreliminary functional characterization of these genes during mating and infection. Overall, our work demonstrates the utilization of host-lipids by U.maydis as an important nutritional and signalling source that is required for pathogenic development. Furthermore, we have begun to elucidate theunderlying mechanisms involved which represent a potential target to combat crop diseases caused by fungi.621. Functional characterization of the putative cell surface receptor for hydrophobicity, Msb2, in Ustilago maydis. Marino Moretti, Daniel Lanver, IrinaL. Schmidt, Regine Kahmann. MPI for Terrestrial Microbiology, Marburg, Germany.Msb2 is a transmembrane mucin protein involved in plant surface sensing in U. maydis. Msb2 deletion mutants are defective in sensing the hydrophobic27th Fungal Genetics Conference | 273

FULL POSTER SESSION ABSTRACTSleaf surface which is a prerequisite for the differentiation of infection structures. Consequently, msb2 mutants are attenuated in virulence (Lanver et al.,2010). The molecular mechanism leading to an activation of Msb2 and the downstream MAP kinase cascade is so far unknown. In yeast Msb2p isprocessed by the aspartyl protease Yps1p leading to an active cell-associated form and a secreted glycosylated part which has an inhibitory function in thefull length protein. In U. maydis Msb2 is also processed, but so far there is no evidence that this leads to an activation of surface sensing. Using a yeastmutant lacking five aspartyl proteases we could demonstrate that yeast Yps1p is able to cleave U. maydis Msb2. In addition, by using this heterologoussystem, two U. maydis aspartyl proteases were identified that were weakly able to cleave Msb2. The respective genes were deleted in the solopathogenicstrain SG200 and its Dmsb2 derivative expressing Msb2-HA-GFP. Possible phenotypic alterations in virulence as well as in Msb2 processing will bemonitored. In addition, a synthetic codon-adapted YSP1 gene has been introduced in the above-mentioned U. maydis strains to analyze the effects of anincrement in Msb2 cleavage on biological activity of the protein. Finally, the extracellular domain of Msb2 was subjected to a mutational analysis toidentify regions with a presumed positive regulatory function. Lanver, D., Mendoza-Mendoza, A., Brachmann, A. and Kahmann, R. (2010). Sho1 and Msb2-Related Proteins Regulate Appressorium Development in the Smut Fungus Ustilago maydis. The Plant Cell 22, 2085-2101.622. The U. maydis effector Pit2 inhibits maize cysteine proteases to suppress host defense. Andre Mueller 1 , Sebastian Ziemann 1 , Steffi Treitschke 2 ,Daniela Abmann 1 , Gunther Doehlemann 1 . 1) MPI for Terrestrial Microbiology, Karl-von-Frisch-Strabe 10, 35043 Marburg, Germany; 2) Fraunhofer ITEM-R,Biopark I, Josef-Engert-Strabe 9, 93053 Regensburg, Germany.The basidiomycete Ustilago maydis is the causal agent of smut disease in maize. Infected plants show tumor formation in all infected aerial parts asprominent symptoms. As a biotroph pathogen, U. maydis depends on living plant tissue and hence efficient suppression of plant immunity is required.Therefore, infectious hyphae secrete effector proteins that interfere with specific components of the plant immune system. One such secreted effectorproteinis Pit2 (Protein important for tumor-formation 2), which, in a previous study, was found to be essential for tumor formation in infected plants [1].Instead of tumors, necroses can be observed at infection sites indicating that plant defense and cell death reactions are triggered in Dpit2 infections [1].Using a combination of yeast-two-hybrid- and protease activity assays, we could show that Pit2 acts as an inhibitor of apoplastic plant cysteine proteaseswhose activity is directly linked with salicylic acid (SA)-associated plant defenses. Sequence comparisons with Pit2 orthologs from related smut fungiidentified a conserved 14 amino acid motif. Mutation of this motif leads to a loss-of-function of Pit2 and consequently to avirulence of U. maydis,suggesting that the protease inhibition by Pit2 is essential for plant infection. Moreover, synthetic peptides of the conserved motif show full activity asprotease inhibitor. Interestingly, expression of only this motif in U. maydis partially restores virulence of the Dpit2-mutant, substantiating the importantrole of this novel protease inhibitor in suppression of host immunity. [1] Doehlemann et al. 2011. Mol Micobiol 81: 751-766.623. The Ustilago maydis MAP Kinase signaling pathway: Identification of direct MAP kinase targets by phospho-peptide enrichment. Vikram Naik 1 ,Gerold J.M. Beckers 2 , Wolfgang Hoehenwarter 3 , Regine Kahmann 1 . 1) Max Planck Institute for Terrestrial Microbiology, Marburg, Germany; 2) PlantBiochemistry and Molecular Biology Group, RWTH-Aachen University, Aachen; Germany; 3) Department for Molecular Systems Biology, Faculty of LifeSciences, University of Vienna, Vienna, Austria.In the plant pathogenic fungus Ustilago maydis three MAP kinase modules have been identified mostly via their homology to genes in Saccharomycescerevisiae. The module consisting of the MAP kinase kpp2, the MAP kinase kinase fuz7 and the MAP kinase kinase kinase kpp4 controls pheromonesignalling and plays an essential role in mating and pathogenicity. Kpp2 is involved in filamentation and appressorium development while the MAP kinase,Kpp6, which also acts downstream of Fuz7, is required for appressorial penetration of plant epidermal cells. Our goal is to identify crucial virulence factorswhich act directly downstream of the MAP kinases Kpp2 and Kpp6. For this we generated a strain in which MAP kinase signaling can be induced byexpressing a constitutively active version of the MAPKK Fuz7 (Fuz7DD) under an inducible promoter in the presence or absence of kpp2 and kpp6. We thenused a two-step chromatographic procedure combining phosphoprotein enrichment using Al(OH)3-based metal oxide affinity chromatography (MOAC),followed by tryptic digest of enriched phosphoproteins, and TiO2-based MOAC for phosphopeptide enrichment. This enabled detection of low abundantphosphorylated peptides using LC-MS/MS and allowed direct identification and site-specific quantification of phosphorylated peptides that differentiallyaccumulated after MAP kinase activation in wild type and mutant cells. LC-MS/MS analysis of the phosphopeptide fraction obtained after the two-stepMOAC yielded 111 putative substrates of Kpp2 and Kpp6 MAP kinases in three replicate experiments. Of these 20 differentially phosphorylated proteinswere chosen for subsequent functional analyses. We are presently generating deletion mutants of these genes in compatible U. maydis strains that carrydifferent a and b alleles and in a solopathogenic strain. In addition, we are analysing the expression pattern of the chosen genes during the differentdevelopmental stages of U. maydis. Results on the role of these U.maydis genes on signaling and pathogenicity will be presented.624. See1 : A novel organ specific effector in the Ustilago maydis - maize interaction. Amey Redkar 1 , Christoph Hemetsberger 1 , Ziba Ajami-Rashidi 1 ,Virginia Walbot 2 , Gunther Doehlemann 1 . 1) Max Planck Institute for Terrestrial Microbiology, Department of Organismic Interactions, Karl von FrischStrasse 10, Marburg, 35043 Germany; 2) Department of Biology, Stanford University, Stanford, California. 94305-5020 USA.Ustilago maydis is a biotrophic smut fungus which infects all aerial organs of its host plant maize. The disease progression and development of infectionis by reprogramming of the plant tissue which ultimately results in formation of tumors. This tumor induction is likely being triggered by small secretedproteins by the fungus, so called effectors. Given the fundamental differences between the different maize organs that are colonized by U. maydis, wehypothesized that the fungus deploys organ specific effectors to manipulate physiology and development of specific host tissues (1). To further investigatethe role of individual organ specific effectors in modulating biotrophy, we in the present study identified a novel secreted protein, termed See1 (Seedlingefficient effector 1) that is strongly induced in seedling leaves but only weakly expressed in tassels and ears. U. maydis deletion mutants for see1 show astrong reduction of tumor formation in maize seedlings but not in floral tissues. Laser scanning confocal microscopy shows that the mutant hyphaesuccessfully enter the leaf tissue but might be blocked during pre proliferation stages in the mesophyll tissue of the leaf. Moreover, by labeling replicatingDNA by 5-ethynyl-2’-deoxyuridine (EdU) we observed that maize seedling colonized by Dsee1 do not show mitotic activity during infection, while celldivision in leaves is specifically induced in wildtype infected host cells. In contrast, the Dsee1 mutant induces normal tumor formation in tassels and alsoshows the stable cell division rate in colonized anthers. Overexpression of see1 causes a hypervirulent phenotype only in the vegetative parts of the tassel,which are not transformed to tumors in wild type infections. To localize See1 during the disease progression we are applying confocal microscopy with livecell imaging using mCherry-tagged See1 protein. Most importantly, we are aiming for the identification of see1 interaction partners to link the observedphenotypes with its molecular function to understand its organ-specific function for U. maydis virulence. (1) Skibbe D*, Doehlemann G*, Fernandes J,Walbot V. (2010) Maize tumors caused by Ustilago maydis require organ-specific genes in host and pathogen. Science 328:89-92.625. Investigation of unconventionally secreted proteins in Ustilago maydis. Stefanie Reissmann 1 , Sina Krombach 1 , Florian Bochen 1 , Till Ringel 1 , SaskiaKreibich 1 , Thomas Brefort 1 , Kerstin Schipper 1,2 , Matthias Mann 3 , Regine Kahmann 1 . 1) Organismic Interactions, Max Planck Institute for Terrestrial274

FULL POSTER SESSION ABSTRACTSMicrobiology, Marburg, Hessen, Germany; 2) Heinrich Heine University Düsseldorf, Institute for Microbiology, Universitätsstrabe 1, 40225 Düsseldorf; 3)Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.Secreted fungal proteins play a crucial role during the biotrophic interaction between the smut fungus Ustilago maydis and its host plant Zea mays. Inthe last decade it has been well established that proteins without a signal peptide can also be targeted to the outside of the cell in an ER/ Golgiindependent manner. We want to identify such unconventionally secreted proteins in U. maydis and investigate their potential function as pathogenicityfactors. Our approach is based on affinity purification of tagged candidate proteins, detected in the apoplastic fluid of infected maize leaves. Four oftwelve candidate proteins tested so far could be detected in culture supernatants. One candidate protein, Um11938, displays 55 % amino acid similarity tothe human sterol carrier protein 2 (SCP2) and we were able to demonstrate that it co-localizes intracellularly with peroxisomes. Mammalian SCP2 isdetected in peroxisomes but also found in the cytoplasm. It interacts with a variety of phospholipids as well as cholesterol and has been implicated in nonvesicular cholesterol transport and in regulating lipid rafts (Schroeder et al., 2007). um11938 deletion strains of U. maydis are severely compromised invirulence and peroxisomal localization of Um11938 is required to fulfill its function as a pathogenicity factor. um11938 deletion strains display no growthdefect on minimal media supplemented with different fatty acids as sole carbon source, suggesting that the Um11938 protein is not involved in bona fideb-oxidation. The results of ongoing experiments aimed to differentiate whether the pathogenicity relevant function of Um11938 is performedextracellularly or within the fungal peroxisomes will also be presented.Schroeder F., Atshaves B.P., McIntosh A.L., Gallegos M., Storey S.M., Parr R.D., Jefferson J.R., Ball J.M. and Kier A.B. Sterol carrier protein-2: New roles inregulating lipid rafts and signaling, Biochim. Biophys. Acta 1771 (2007) 700-718.626. Identification of a key regulator for the developmental switch leading to sporogenesis in Ustilago maydis. Marie Tollot, Regine Kahmann.Organismic Interactions, MPI Marburg, Marburg, Hessen, Germany.Ustilago maydis is a biotrophic pathogen of maize. Its life cycle begins with the mating of two compatible haploid sporidia to form a dikaryotic infectiousfilament. After penetrating the plant cuticle, the dikaryon spreads inside the plant tissues and induces the formation of tumors. At a defined time ofdevelopment, the sporogenesis program begins in tumor tissue: the hyphae start to fragment into individual cells that eventually differentiate into matureteliospores. The isolation of several mutants affected in spore formation has shown that a tight regulation of the cAMP signaling pathway and the activityof two transcriptional regulators Hda1 and Rum1, potentially functioning in the same chromatin modifying complex, are required. We have identified anew regulator for the sporogenesis program of U. maydis. It belongs to a family of transcriptional regulators that are characterized by the presence of aWOPR DNA binding domain. The best studied member of this family is Wor1 from Candida albicans which plays a key role in the switch from the nonpathogenicto the pathogenic form of the fungus. The U. maydis wor1 homologue um05853, was deleted in the compatible haploid strains FB1 and FB2.The deletion mutants were able to mate and to infect maize as efficiently as the wild-type. However, although the tumor rate was similar, no spores couldbe detected in plants infected by the deletion mutants. Confocal microscopy of the mutant dikaryon revealed that hyphal fragmentation and consequentlyspore maturation were not occurring. The hyphae were still spreading at a time when the wild type dikaryon had already formed mature spores. To furtherinvestigate the role of um05853, we expressed the gene in a haploid strain where the filamentous program can be induced in axenic culture via expressionof a functional b-heterodimer. The cells failed to switch to filaments, started to enlarge and showed septation, with each section containing one nucleus.This result suggests that Um05853 is able to counteract the b function and trigger fragmentation. Preliminary results show that Um05853 mightdownregulate several b-dependent genes including the gene encoding the master regulator Rbf1. We are currently identifying targets of Um05853 using amicroarray approach and expect that these results will highlight how Um05853 controls spore formation.627. Genetic characterization of virulence in the Pyrenophora teres f. teres - barley pathosystem. Timothy L. Friesen 1,2 , Rachel A. Shjerve 2 , Justin D. Faris 1 ,Robert S. Brueggeman 2 . 1) Cereal Crops Research Unit, USDA-ARS, Fargo, ND; 2) Department of Plant Pathology, North Dakota State University, Fargo, ND.Pyrenophora teres f. teres is a necrotrophic fungal pathogen that causes net form net blotch (NFNB) on barley throughout the world. Several resistancesources have been identified but few are effective against all P. teres f. teres pathotypes, indicating that the pathogen has an arsenal of effectors that areinvolved in disease induction. Genetic analysis identified two barley genotypes, Rika and Kombar, that each harbor unique dominant susceptibility geneslocated at the centromeric region of barley chromosome 6H. P. teres f. teres isolate 15A is virulent on Kombar but avirulent on Rika whereas P. teres f.teres isolate 6A is virulent on Rika but avirulent on Kombar. Based on the necrotrophic effector model, 15A and 6A each secrete unique effectors that aredirectly or indirectly interacting with genes on chromosome 6H in Kombar and Rika, respectively. A linkage map was generated using a mappingpopulation developed from a sexual cross between 15A and 6A, and 118 progeny were phenotyped on barley genotypes Rika and Kombar. Two majorvirulence QTL derived from 15A contributed to virulence on Kombar, and two additional unique virulence QTL derived from 6A contributed to virulence onRika. All susceptibility loci in the host mapped to the same region on barley chromosome 6H. Therefore, it is likely that at least four necrotrophic effectorspresent throughout the P. teres f. teres genome are interacting with host susceptibility genes located in one region of barley chromosome 6H. Theseresults strongly indicate that the NFNB-barley compatibility is at least partially due to necrotrophic effector-host susceptibility gene interactions that resultin disease induction. Currently, we are using a genotype by sequencing (GBS) approach to generate a saturated map to identify candidate genes in the QTLregions in order to clone and characterize the effector genes involved in the NFNB interaction.27th Fungal Genetics Conference | 275

FULL POSTER SESSION ABSTRACTS628. Illumina-based genetic linkage map for wheat leaf rust. David L. Joly 1,2 , Barbara Mulock 3 , Christina A. Cuomo 4 , Barry J. Saville 2 , Brent D. McCallum 3 ,Guus Bakkeren 2 . 1) Pacific Agri-Food Research Centre, Agriculutre and Agri-Food Canada, Summerland, British Columbia, Canada; 2) Forensic ScienceProgram and Environmental & Life Sciences Graduate Program, Trent University, Peterborough, ON, Canada; 3) Cereal Research Centre, Agriculture andAgri-Food Canada, Winnipeg, MB, Canada; 4) Broad Institute of Massachusetts Institute of Technology and Harvard University, Cambridge, MA 02142.Few genetic maps have been made for rust fungi; yet they are useful in identifying candidate loci for phenotypic traits or in unravelling chromosomalarrangements. This lack of maps is, in part, due to the obligate biotrophic nature of rusts and the difficulties in manipulating their life cycle in a way thatenables controlled crosses. Recently, the genome sequence of a wheat leaf rust (Puccinia triticina) isolate was determined and this prompted thesequencing of additional isolates using next-generation sequencing technologies. This has dramatically increased the amount of sequence informationavailable at a substantially decreased per base cost. Fifty-seven F2 progeny of a wheat leaf rust sexual cross between race 9 (SBDG) and race 161 (FBDJ)were sequenced using Illumina. In order to generate a high-resolution genetic linkage map, genome-wide single-nucleotide polymorphisms (SNPs) wereidentified. Employing the genome sequence information from the two parents and the F1 isolate, more than 25,000 SNPs were selected and used togenerate a genetic linkage map. Although they were obtained from different isolates, the genetic map and the reference genome were integrated,allowing the creation of pseudomolecules. Those represent a strong improvement over the currently fragmented status of the reference genome.Moreover, at least 9 seedling and 2 adult-plant avirulence genes were shown to segregate in this F2 population and candidate genes identified using thegenetic map are currently being investigated.629. The deletion of the Histoplasma capsulatum RYP1 homolog in Coccidioides posadasii is avirulent. M Alejandra Mandel 1,3,4 , Hien Trien 2,3,4 , AmrithaWickramage 1 , Lisa Shubitz 2,3,4 , Marc Orbach 1,3,4 . 1) School of Plant Sciences, University of Arizona, Tucson, AZ; 2) Department of Veterinary Sciences andMicrobiology, University of Arizona, Tucson, AZ; 3) The Bio5 Institute, University of Arizona, Tucson, AZ; 4) Valley Fever Center for Excellence, Tucson, AZ.Coccidioides spp. are mammalian fungal pathogens endemic to the desert southwestern US, parts of México and Central and South America that causethe respiratory disease coccidioidomycosis, or valley fever. These dimorphic fungi grow as filamentous saprotrophs in soil, but when a spore is inhaled bythe host and localizes to the lung, it switches from polar to isotropic growth resulting in the development of a spherule. In Histoplasma capsulatum, Ryp1is a master switch required for the transition from the filamentous to the infectious yeast phase, and thus is essential for virulence. We have performed awhole-gene deletion of the RYP1 homolog in Coccididoides posadasii strain Silveira to determine whether it plays a similar role in virulence in thispathogen. Phenotypic effects were observed in both the filamentous and the parasitic phases of C. posadasii. During filamentous growth, there is areduction in colony size, and defects in sporulation. The mutant is avirulent in our susceptible mouse model. Our results indicate that Ryp1 is a masterswitch in different fungal models. Although avirulent, the ryp1 mutant is not able to induce a protective response when used to vaccinate mice prior towild type infection.630. Cpkk2, a MEK from Cryphonectria parasitica is necessary for maintenance of CHV1 virus infection. M. Moretti, M. Rossi, M. Ciuffo, S. Abba', M.Turina. IVV, CNR, Torino, Italy.We have recently obtained and characterized the knock out strains of the three MEKs present in the Cryphonectria parasitica genome, Cpkk1, Cpkk2 andCpkk3, homologues of yeast Mkk1p/Mkk2p, Ste7p and Pbs2p, respectively. We tried to infect each of the knock-out strain with Cryphonectria hypovirus 1(CHV1), a mycovirus causing hypovirulence: Dcpkk1 and Dcpkk3 were easily infected by CHV1 through anastomosis, but we failed to infect Dcpkk2. Wethen showed that hyphal fusion was prevented in such knock-out strain: for this reason we attempted at infecting the Dcpkk2 strain with two alternativeprotocols that overcome the hyphal phusion impairment: stable transformation of protoplasts with a cDNA infectious clone and transfection of protoplastswith viral RNA transcripts obtained in vitro from a cDNA infectious clone. We originated infected strains with both protocols using wild-type C. parasiticaprotoplasts, whereas no stable infected strain was obtained starting from Dcpkk2 protoplasts, which, on the contrary, could be transformed with theempty vector carrying only the resistance gene for selection. Given the uniqueness of such result, we are now trying to show what is the specific molecularimpairment that prevents CHV1 maintenance in Dcpkk2 strain. A proteomic approach was undertaken using 2-DE MALDI-TOF MS/MS and shotgun coupledto LC-MS/MS to compare the WT and Dcpkk2 strains. A number of metabolic pathways are heavily impacted in the mutant. Of interest, proteins involvedin folding, transport and trafficking, are up-regulated suggesting an altered protein turnover. Defence machinery is also up-regulated, indicating that thefungus perceives a stress situation. Moreover, a strong down-regulation of proteins involved in energy production and conversion was detected, indicatinga possible reduction of the energetic metabolism. Among them are some GAPDH isoforms. Given the recent discovery of the role of GAPDH in viralreplication complexes of RNA viruses, we obtained anti-GAPDH antibodies in order to study its possible role in CHV1 viral replication.631. Deep RNAseq of wheat leaf infection by M. graminicola identifies phase-specific in planta expressed genes and varying transcriptionalcontributions of fungal chromosomes. Jason J Rudd 1 , Juliet Motteram 1 , Mark Derbyshire 1 , Keywan Hassani-Pak 2 , Bob Dietrich 3 , Arvind K Bharti 4 , Andrew DFarmer 4 , Ambrose Andongabo 2 , Mansoor Saqi 2 , Mikaël S Courbot 5 . 1) Rothamsted Research, Department of Plant Biology and Crop Science, Harpenden,Hertfordshire, AL5 2JQ, UK; 2) Rothamsted Research, Department of Computational and Systems Biology, Harpenden, Hertfordshire, AL5 2JQ, UK; 3)Syngenta Biotechnology, Inc., 3054 East Cornwallis Road, Durham, NC 27709, USA; 4) National Center for Genome Resources (NCGR), Santa Fe, NM 87505,USA; 5) Syngenta Crop Protection Münchwilen, Schaffhauserstrasse, 4332 Stein, CH.Mycosphaerella graminicola is the causal agent of Septoria tritici blotch disease of wheat. Infection of leaves by M. graminicola involves a characteristiclong period of symptomless intercellular growth of at least 8-10 days prior to the formation of necrotic leaf lesions. The genome sequence of the modelisolate of M. graminicola, IPO323, was recently published by the research community in conjunction with the JGI and has been shown to contain 21chromosomes. We have performed a deep RNAseq analysis to investigate fungal gene expression in vitro (in Czapek-Dox (CDB) and Potato Dextrose broth)and throughout phases of plant infection: day 1 (d1) germination on the leaf surface, day 4 (d4) slow growth in the absence of symptoms within the leaf,day 9 (d9) symptoms of disease become visible, day 14 fungal growth rate increases and finally day 21 when the fungus is sporulating asexually in fullynecrotic plant tissue. Sequencing was performed on the Illumina Hiseq platform. The RNA-seq data was analysed using the Tuxedo tools (Trapnell et al.,2012). Tophat2 was used to map the reads against the M. graminicola genome. Transcript abundance (in FPKM) was determined using Cufflinks. Significantchanges in transcript expression across all 21 pairwise comparisons were determined using cuffdiff (FDR

FULL POSTER SESSION ABSTRACTSchromosomes, which may be consistent with their being dispensable for asexual plant infection.632. A genomic analysis of the infection strategies employed by Phoma medicaginis a necrotrophic fungal pathogen of alfalfa and the model legumeMedicago truncatula. Angela H. Williams 1,4 , James K. Hane 2 , Robert D. Trengove 3 , Karam B. Singh 2 , Richard P. Oliver 4 , Judith Lichtenzveig 4 . 1) MurdochUniversity, Perth, Australia; 2) CSIRO Plant Industry, Perth, Australia; 3) Separation Science and Metabolomics Laboratory, Murdoch University, Perth,Australia; 4) Department of Environment and Agriculture and the Australian Centre for Necrotrophic Fungal Pathogens, Curtin University, Perth, Australia.Phoma medicaginis is a necrotrophic plant pathogen that causes black spot of alfalfa (Medicago sativa) and the closely related model legume Medicagotruncatula. It is a member of the Didymellaceae family, a distinct clade within the order Pleosporales which includes some of the most importantpathogens of legume crops. We present here the first genome assembly of P. medicaginis and the results of investigations into the host-pathogeninteraction, focusing on identification of necrotrophic effectors (NEs) using a combination of proteogenomic and transcriptomic analyses. A draft genomeassembly was constructed using Illumina paired-end reads, de novo assembled into 952 nuclear scaffolds totaling 31.4 Mbp, with ~27 x coverage andencoding ~10,500 predicted proteins (>50 amino acids) . Of these, ~1,000 are predicted to be secreted. Peptide sequencing via mass spectrometry wasconducted in order to validate the gene set and characterise the protein content of intracellular and necrosis-inducing secreted fractions. This enabled theconfirmation of 554 predicted genes and identified 162 proteins in the necrosis-inducing secreted fraction. To further validate the predicted gene set andexamine differences in gene expression, the transcriptome was sequenced via RNA-seq at four important lifestyle phases. These included: 1) 1-5 days postinfection of M. truncatula; 2) vegetative growth in vitro; 3) sporulation in vitro and 4) during growth in media where the culture filtrate produces necrosisand chlorosis when infiltrated into the plant. Close to 10,000 genes were expressed under one or more of these conditions with ~ 3,000 showingdifferential expression between the in planta and in vitro samples. The combination of proteogenomic and transcriptomic analyses has enabled thevalidation and fine-tuning of the majority of de novo predicted gene models. Several novel genes were identified via manual annotation of RNA-seq data.We have previously demonstrated that the genome is manipulable via Agrobacterium-mediated transformation which means that the functions ofpotential effector genes can be readily investigated. Collectively these data form a valuable resource from which a short list of effector candidates wasderived and genes involved in the pathogenicity mechanisms of Didymellaceae fungi against their legume hosts were predicted.633. Two G protein-coupled receptors, GprC and GprD, regulate density-dependent development in Aspergillus flavus. Katharyn J. Affeldt, Nancy P.Keller. University of Wisconsin-Madison, Madison, WI.Aspergillus flavus is an opportunistic pathogen of several plant hosts, including maize. This interaction is mediated in part by oxygenatedpolyunsaturated acids, or oxylipins, that are produced by both the fungus and the plant host. Although much has been learned about the synthesis ofthese oxylipins, how the fungus perceives them remains unknown. We hypothesize that G protein-coupled receptors (GPCR) are responsible for receivingand transducing oxylipin signals in A. flavus. We have deleted and overexpressed two GPCRs, gprC and gprD, and found that they are important inregulating density-dependent development, which is thought to involve oxylipin signaling. Specifically, depletion of both gprC and gprD locks the fungusinto a low-density state, even when grown at high density. Furthermore, this mutant is unable to respond to spent medium of a wild type high-densityculture. Inoculation of these mutants on corn kernels will ask whether GprC and GprD are important for pathogenicity, and heterologous expression ofGprC and GprD in Saccharomyces cerevisiae is being used to address questions concerning direct ligand-receptor activation.634. Characterization of genes encoding putative secreted proteins during pathogenesis in Magnaporthe oryzae. Seongbeom Kim, Kaeun Kim, Sook-Young Park, Jaeyoung Choi, Junhyun Jeon, Yong-Hwan Lee. Department of Agricultural Biotechnology, Seoul National University, Seoul, South Korea.The repertoire of secreted proteins defines the nature of interactions between microbe and host at the molecular level. Thus, cataloging andcharacterizing the list of secreted proteins from a given pathogen is a pivotal step in understanding molecular mechanisms of pathogenesis. Unlikebacterial and Oomycete pathogens, however, only a limited number of secreted proteins has been identified and analyzed in plant pathogenic fungi. Herewe set out to identify and characterize new secreted proteins in the rice blast fungus. SingalP program predicted a total 1,885 genes encoding secretedproteins in M. oryzae. We prioritized 15 genes, MoSPE1 to MoSPE15, with T-DNA mutants available for in-depth analysis. To reveal their roles inpathogenicity, gene deletion mutants were generated and characterized their functionality. Deletion of MoSPE1 rendered the fungus non-pathogenic,while deletion of MoSPE3, MoSPE6, and MoSPE15 resulted in reduced virulence. Rice sheath inoculation of DMospe1 and DMospe15 showed that defectsin pathogenicity could be attributed to the inability to grow inside plant tissues, suggesting their implication in interaction with rice. In addition, the twogenes were indeed up-regulated during invasive growth in rice. Proteins encoded by MoSPE1, MoSPE6 and MoSPE15 were capable of being secreted inyeast secretion trap system. We believe that our work would reveal novel function of secreted proteins, providing new insight into fungal pathogenesis.635. The biosynthesis of oxalate is entirely dependent on oxaloacetate acetylhydrolase in Sclerotinia sclerotiorum . X. Liang 1 , D. Liberti 2 , M. Li 3 , Y.-T.Kim 4 , R. Wilson 1 , J. Rollins 1 . 1) Plant Pathology Department, University of Florida, 1453 Fifield Hall, Gainesville, FL, 32611-0608; 2) Nunhems NetherlandsBV, PO Box 4005, Haelen 6080 AA, Netherlands; 3) Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, FL; 4)Environmental Biotechnology Research Centre, 125 Gwahak-ro, Yuseong-gu, Daejeon 305-806, Republic of Korea.Sclerotinia sclerotiorum (Lib.) de Bary is one of the most devastating necrotrophic fungal plant pathogens worldwide and its successful infection involvesthe accumulation of oxalate (up to 10 mM) in plant tissues. Oxaloacetate acetylhydrolase (EC 3.7.1.1), catalyzing the hydrolytic cleavage of oxaloacetate toform acetate and oxalate, has been shown to be the key enzyme catalyzing oxalate biogenesis in Aspergillus niger, Botrytis cinerea and Cryphonectriaparasitica. To dissect the genetic regulation of oxalate biogenesis and pathogenesis of S. sclerotiorum, the S. sclerotiorum oxaloacetate acetylhydrolasegene Ss-oah1 was functionally characterized. Previously we demonstrated that oxalate accumulation in S. sclerotiorum is under strong alkaline induction.Strikingly Ss-oah1 gene expression is regulated in the same manner; neutral pH strongly induces the accumulation of Ss-oah1 transcripts and this pHinduction is completely suppressed in the Ss-pac1 knock out mutant. Ss-oah1 knock out mutants fail to accumulate oxalate in culture and during plantinfection and these phenotypes are restored by complementation with the wild type gene. These data demonstrate that Ss-Oah1-catalyzed oxaloacetatehydrolysis is solely responsible for oxalate production in S. sclerotiorum. On all tested host plants, Ss-oah1 knock out mutants are dramatically reduced invirulence and induce a strong host defense response. On leaves, Ss-oah1 knock out mutants produce limited dark brown-green lesions compared with thespreading, necrotic, light brown lesions produced by the wild type. Host tissue bordering the lesion is clearly defined with a thin, dark zone and while theuninfected leaf tissue becomes yellow and senescent the colonized area often retains chlorophyll reminiscent of “green islands”. In sum, our experimentaldata establish the key function of oxaloacetate acetylhydrolase in oxalate biogenesis and pathogenesis in S. sclerotiorum and indicate that the oah1oxalate minus mutant retains some aspects of virulence but cannot suppress host defense.27th Fungal Genetics Conference | 277

FULL POSTER SESSION ABSTRACTS636. In vivo efficacy of antifungal treatment of Aspergillus terreus infections and the influence on host immune response in Galleria mellonella.Elisabeth Maurer 1 , Neill Browne 2 , Kevin Kavanagh 2 , Cornelia Lass-Flörl 1 , Ulrike Binder 1 . 1) Division of Hygiene and Medical Microbiology, Medical UniversityInnsbruck, Innsbruck, Tirol, Austria; 2) Medical Mycology Unit, Department of Biology, National Institute for Cellular Biotechnology, NUI Maynooth,Ireland.Background Infections with Aspergillus (A.) terreus are of major concern, due to its high likelihood of dissemination and its intrinsic resistance toamphotericin B (amB). The reason for this resistance is not known yet and the exact mode of amB action is still not fully understood. Recently, threeclinical isolates have been found to be amB susceptible in vitro. In order to investigate for differences in virulence of the respective isolates, and to test theamB efficacy in vivo, we used Galleria (G.) mellonella as an alternative model. Methods Virulence of amB resistant and amB susceptible A. terreus isolateswas compared in the invertebrate model G. mellonella. Further, we performed in vivo infection studies with combined antifungal therapy, and additionallywe investigated the potential effect of A. terreus infection and antifungal treatment on the G. mellonella immune system. Proteomic analysis of larvalhaemolymph, haemocyte counts and post-treatment infection studies were performed according to Kelly & Kavanagh 2011. Results Larval survival ratesdiffered for the various isolates tested, resulting in highest mortality rate for one amB susceptible isolate. Increase in survival was seen for all testedstrains, when larvae where treated with voriconazole. Treatment with amB only showed success in the groups infected with amB susceptible strains.Antifungal administration in larvae resulted in an increased number of circulating haemocytes. Proteomic studies showed different protein expression of anumber of proteins which have immune function. Pre-treatment of larvae with different antifungals also increased their resistance to Staphylococcus (S.)aureus infection, indicating a general ability of antifungals to prime the insect's immune system.637. Recognition and response to non self in Podospora anserina: a model of the fungal immune system. Marina Lamacchia, Annick Breton, AsenDaskalov, Frédérique Ness, Muhammad Khalid Salamat, Martine Sicault-Sabourin, Sven Saupe, Mathieu Paoletti. Institut de Biologie et GénétiqueCellulaire, UMR 5095 CNRS et Université Victor Segalen Bordeaux, Bordeaux, France.Recognition and response to non self, whether conspecific (between individuals from the same species) or heterospecifc (individuals from anotherspecies) is essential to many aspects of life including development, symbiosis and protection against pathogens. However distinction between thesemodes of recognition and responses is somehow blurred and can overlap. For instance in plants and animals Pathogen Recognition Receptors (PRRs) canoccasionally lead to auto-immune diseases in absence of pathogens. The NLR and NBS-LRR STAND proteins (a class of signal transduction proteins) aremajor PRRs in plants and animals, but these receptors remain largely unidentified in fungi. In Podospora anserina vegetative incompatibility (VI), aconspecific non self recognition process, leads to cell death and autophagy. VI is determined by interaction of het-c, encoding a glycolipid transfer protein,with members of the hnwd gene family encoding for STAND proteins. hnwd gene family members display the hallmarks of PRR encoding genes, includingfast evolution promoting production of a repertoire of receptors and ability to initiate a cell death reaction. het-c is also showing signs of fast evolution.We hypothesized that these genes are involved in pathogen recognition and that recognition of heterospecific non self would initiate a response similar tothe VI reaction. In this context, VI can be considered as an autoimmune disease. We undertook the task of deciphering the response of P. anserina toheterospecific non self, focusing our efforts on the description of the cellular response and the identification of fungal PRRs. We show that P. anserina’sresponses to another fungal species (Epicoccum nigrum), or to bacteria such as Serratia entomophila or Pseudomonas putida largely overlap the VIresponse at all levels investigated so far, including cellular morphology and cytology, requirement of autophagy and induction of the expression of a set ofgenes. We also provide evidence that het-c encoding the GLTP contributes to the response to non self and argue that this protein may be targeted bypathogen’s effectors. We develop efforts to identify PRRs involved in the initiation of these responses.638. Increased late blight resistance in HIGS potato lines targeting a P. infestans gene. N. Temme, C. Blumenhagen, A. Schwarzer, L. Weimer, K. Prenzler,T. Sauter, M. Pflugmacher, D. Stahl. KWS SAAT AG, Einbeck, Germany.Worldwide potato harvests are strongly diminished due the late blight disease caused by the oomycete Phytophthora infestans. The fungus-likeeukaryote and its interaction with its host plants has been extensively investigated during the last decades whereas diverse research projects focus on itsinfection processes. P. infestans colonizes potato as well as tomato plants and thereby differentiates haustoria. Those barriers between host cells andinvading pathogens are capable for exchange of nutrients, minerals but also of macromolecule like RNA molecules as shown for haustoria of parasiticplants. In oomycetes the exchange of effector protein from the pathogen to its host has been demonstrated. The movement of RNAi signals was shown inthe interaction of parasites with their host plants and can be used to target not only plant genes but also genes of plant invading organism in a mechanismcalled host-induced gene silencing (HIGS). This technique has been applied for gene silencing in plant parasites as well as in nematodes and fungi. Inoomycetes the RNA silencing is used as a standard method to characterize genes either by transient or by stable gene silencing and enzymes of the RNAimachinery have been identified. However, no efficient HIGS of oomycetes could be observed so far. We defined a P. infestans gene expressed duringdiverse developmental and infection stages of the oomycete as a HIGS target and could show that in planta expression of a HIGS hairpin constructtargeting this particular gene in transgenic potato lines can be employed for late blight control. Our results present the appropriate processing oftransformed HIGS hairpin constructs to siRNAs, their efficient function to silence the specific target gene sequence as shown in the reporter gene assaysand subsequently reduced infection levels and diminished disease spreading on those transgenic HIGS potato lines in the field.639. WITHDRAWNPopulation and Evolutionary Genetics640. Fertility in Aspergillus fumigatus and the identification of an additional ‘supermater’ pair. Céline M. O'Gorman 1 , Sameira S. Swilaiman 1 , Janyce A.Sugui 2 , Kyung J. Kwon-Chung 2 , Paul S. Dyer 1 . 1) School of Biology, University of Nottingham, University Park, Nottingham NG7 2RD, United Kingdom; 2)Molecular Microbiology Section, Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes ofHealth, Bethesda, Maryland, USA.Aspergillus fumigatus is an opportunistic human pathogen that causes a range of allergic and invasive diseases in severely immunocompromisedindividuals, with a very high mortality rate typically in excess of 50%. A functional sexual cycle was discovered in 2009 and a highly fertile ‘supermater’ pair,AFB62 and AfIR928, was later identified from a collection of 50 isolates. Here we describe the results of a larger, worldwide fertility screen and present anadditional ‘supermater’ pair. A set of 126 clinical and environmental A. fumigatus isolates were crossed against two Irish reference strains of each matingtype. A subset of the eight most-fertile strains was then tested in all pairwise combinations. The pairing of isolates 47-169 x 47-154 had consistently high278

FULL POSTER SESSION ABSTRACTSmating efficiency and outcrossing ability after four weeks, therefore it was chosen as an additional ‘supermater’ pair for community use in mating projects.It is important to have alternative tester strains to allow for unexpected mating differences when crossing isolates of diverse genetic origins. This isbecause factors such as heterokaryon incompatibility (het) loci and single nucleotide polymorphisms, can considerably influence sexual compatibility. Theworldwide fertility screen found that approximately 85% of isolates are sexually fertile, indicating that sexual reproduction should be possible in naturewhen suitable environments are present. Next, the plasticity of sexual crossing conditions was tested, to determine whether they could be manipulated toincrease fertility in crosses involving low-fertility strains of interest. A range of environmental and growth conditions were examined, including incubationtemperature, CO 2 level, and oatmeal agar type. Fertility levels were significantly affected by certain parameters. Work is ongoing to integrate these factorsto further optimize fertility in the ‘supermater’ pairs.641. Understanding the dynamic plant pathogen Ramularia collo-cygni at both the sequence and field level. James Fountaine, Peter Hoebe, MaciejKaczmarek, Marta Piotrowska, Neil Havis. Crop and Soils Systems Research, Scotland's Rural College, Edinburgh, Scotland, United Kingdom.The fungus Ramularia collo-cygni is the major biotic agent involved in Ramularia Leaf Spot (RLS). The fungus produces necrotic lesions on leaves,primarily after flowering takes place in the host plant. Despite being initially reported on crops in Italy in the late 19th Century RLS only became aneconomic pathogen of barley in the late 20th Century. The geographical spread of the disease now covers much of Europe, North and South America andNew Zealand. Research in the last decade, using molecular tools, has helped elucidate the life cycle of the fungus and has indicated a seed borne stage.These tools have also allowed detailed testing of spring barley archive samples, which has revealed a significant increase in pathogen levels since the1990’s. The pathogen appears to develop rapid resistance in the field to fungicides and mutations conferring resistance have also been detected in thearchive samples. Ramularia collo-cygni is currently classified as a member of the Mycosphaerella genera and sequence data derived within our groupsuggests a genetic similarity between R. collo-cygni, Mycosphaerella graminicola and M. fijiensis. These sequences focus primarily on the genes associatedwith the target sites for fungicides, such as Beta tubulin, Cytochrome b, Succinate dehydrogenase and eburicol 14a-demethylase (CYP51) genes. Thispresentation will demonstrate our current knowledge of this fungal pathogen and highlight the newly obtained genome and transcriptomic datagenerated by the combined approach of illumiina/solexa and Roche/454 sequencing. This combined approach has enabled the assembly of a completegenome sequence. The finished assembled genome of R. collo-cygni is 30.2 Mb and is currently to be found in 355 contigs. The complete annotation of thisgenome is currently underway using the FGENESH 2.6 software to generate first consensus gene calls. This approach will allow for comparative genomeanalysis in related genomes which will help to address the biology of R. collo-cygni in areas such as pathogenicity, population genetics and fungicideresistance. These advances should enable a greater understanding of the complex relationship between the fungus and host plant and furthermore, assistin the development of environmentally sound strategies to control this increasingly important disease of barley production systems.642. Comparing germination dynamics of conidia and ascospores from natural isolates of Neurospora crassa. Kolea C.K. Zimmerman 1 , Dan Levitis 2 , AnnePringle 1 . 1) Organismic and Evolutionary Biology, Harvard University, Cambridge, MA; 2) Max Planck Institute for Demographic Research, Rostock,Germany.Many organisms experience high mortality during the first stages of growth. This is especially relevant in fungi because of the large ratio betweenpropagules produced and the number of those propagules that germinate and grow into a mature mycelium. Furthermore, in many organisms, asexuallyreproduced progeny have higher survival rates compared to sexually reproduced progeny. We have developed a high throughput pipeline using flowcytometry to analyze spore germination dynamics and have applied this pipeline to study the germination differences between asexual and sexual spores.Specifically, we applied this pipeline to study variation in germination of Neurospora crassa conidia and ascospores from 32 natural isolates and crossesamong these isolates, respectively. Using publicly available sequence data, we computed all pairwise genetic distances between the 32 strains and crossedstrains of varying genetic distance from each other to determine the effects of parent relatedness on ascospore viability. We found that viability of asexualspores is higher than viability of sexual spores in most cases and there is no clear linear relationship between genetic relatedness and ascospore viability.Future work will include experiments to evaluate the ability of sexual recombination to purge deleterious mutations. The data from these experiments willhelp us quantify the relative costs and benefits of asexual vs. sexual reproduction in Neurospora crassa and inform evolutionary theories on the evolutionof sex.643. Cryptococcus gattii and the origins of outbreaks. Tien Bui 1 , Anna Foley 1 , Leona Campbell 1 , Patrick Brunner 2 , Bruce McDonald 2 , Dee Carter 1 . 1) Schoolof Molecular Bioscience, University of Sydney, NSW 2006, Australia; 2) Institute of Integrative Biology, ETH, 8092 Zurich, Switzerland.Cryptococcus gattii and its sibling species C. neoformans cause cryptococosis in humans and a range of animal. Strains within these species fall into anumber of distinct molecular genotypes, and these vary in their ecology, geographic distribution, and various virulence-associated phenotypes. In C. gattii,molecular type VGI is found worldwide, usually in warmer regions, and causes sporadic infection in apparently healthy people and animals. VGII is morerestricted in distribution, and while it also causes sporadic infection it is responsible for significant outbreaks that have expanded its geographic range intotemperate areas. VGIII infections occur predominantly in immunocompromised hosts in the southern California region, and cases of VGIV infection are sofar restricted to southern Africa, with a single case from India. Our interests lie in understanding the ecology and evolution of C. gattii in the environment,and how these relate to its ability to cause infection and outbreaks of disease. We have found the level of sexual recombination varies by molecular type,and that while in general the C. gattii population structure is sexual, this varies by genotype and in VGII is punctuated by periodic, clonal lineages. Here werefine our analysis using extended MLST data, haplotype networks and coalesence theory. We find the level of diversity among global VGI and VGIIgenotypes is highly constrained and comparable to some recently evolved plant pathogens, while the more geographically restricted VGIV genotype issubstantially more diverse. Outbreak VGII clones are highly derived with an apparent history of expansion and extinction events. Evidence for purifyingselection occurs at the master regulator of mating type for VGII, suggesting recombination is important in the generation of outbreak lineages.644. Evolution of the mating type locus in the species within and closely related to the pathogenic Cryptococcus species complex. Sheng Sun, JoshGranek, Joseph Heitman. Molecular Genetics & Microbiology, Duke University, Durham, NC.Cryptococcus amylolentus is the most closely related sister species to the pathogenic Cryptococcus species complex that includes the common humanpathogenic fungi Cryptococcus neoformans and Cryptococcus gattii. We recently reported that C. amylolentus has a tetrapolar mating system, in whichmating type is determined by two unlinked mating type (MAT) loci (A and B) that are located on different chromosomes. This is in stark contrast to themating systems in the species within the pathogenic Cryptococcus species complex, where all the species are bipolar and have a contiguous large (>100 kb)MAT locus. Thus, analyzing the tetrapolar MAT loci of C. amylolentus could provide insights into how the derived bipolar MAT locus in the pathogenicCryptococcus species complex evolved. In this study, we first provide a fully detailed characterization of both alleles for each of the two C. amylolentusMAT loci, illustrating expansion of the A MAT locus in C. amylolentus, as well as the chromosomal rearrangements between the alleles from the opposite27th Fungal Genetics Conference | 279

FULL POSTER SESSION ABSTRACTSmating types. Additionally, by analyzing meiotic progeny of C. amylolentus, we found evidences that both MAT loci are linked to their respectivecentromeres. Using both genetics and genomics techniques, we further narrowed down the candidate centromeric regions to be located within 150 and100 kb from the A and B MAT loci, respectively. Furthermore, genome comparison between C. neoformans and C. amylolentus showed that the majority ofcentromeres in C. neoformans are flanked by sequences from different chromosomes in C. amylolentus, indicating that ectopic recombination withincentromeric regions may frequently lead to chromosomal translocations. We propose a model in which the large bipolar MAT locus that is present in thepathogenic Cryptococcus species complex originated through ectopic recombination in the centromeres (possibly mediated by common repetitivesequences present in the centromeric regions). This process brought together the two MAT loci of the ancestral tetrapolar mating system onto the samechromosome, and subsequent chromosomal rearrangements (inversions and transpositions) resulted in the current state of the MAT locus seen in thederived bipolar pathogenic Cryptococcus species.645. Evolutionary history and genetic diversity of Exobasidium sp., the cause of an emerging disease of blueberry. Marin Brewer, Ashley Turner.Department of Plant Pathology, University of Georgia, Athens, GA.Emerging fungal diseases, usually the result of pathogen introductions, the evolution of virulent races, or adaptation to new niches, are an increasingthreat. Exobasidium fruit and leaf spot of blueberry has rapidly increased in incidence in the southeastern USA over the past two years. We took aphylogenetic approach to understand the evolutionary history of this fungus. We sequenced the LSU-rDNA region from nine isolates collected from fruit orleaf spots of Vaccinium spp. from Georgia and North Carolina. Additionally, sequences from GenBank with high similarity to the emerging parasite andfrom Exobasidium spp. parasitizing other Vaccinium spp. in North America were obtained. The sequences were assembled, aligned and subjected tophylogenetic analyses. Results indicated that Exobasidium sp. from blueberry in the southeastern USA is unique and distinct from Exobasidium sp. thatcauses a leaf spot on lowbush blueberry in the northeastern USA and Canada. Both species, however, are genetically different from other Exobasidiumspp. that cause diseases on cranberry and blueberry, and from E. vaccinii from V. vitis-idaea. Results also suggested that within the Southeast the parasiteis not genetically differentiated based on blueberry host species or cultivar, host tissue (fruit or leaf), or geographic region. To further investigate diversityand population structure of the parasite in the Southeast we sequenced ITS for 80 isolates from diverse host species, cultivars, and locations. We obtained75 unique sequences, which is an extremely high level of diversity and is unexpected for any fungus let alone one causing an emerging disease. The highdiversity indicates that the fungus causing Exobasidium fruit and leaf spot in the Southeast is not an evolutionarily young species and that the recentincrease in incidence is not a result of increased aggressiveness or a recent host switch. Analyses of genetic differentiation of the ITS sequences confirmour findings with LSU-rDNA that isolates within the Southeast are not differentiated by host species or cultivar, host tissue, or geographic region,suggesting that a single population is causing this disease of blueberry across the Southeast. We hypothesize that an environmental change is responsiblefor the recent emergence of this disease.646. Microsatellite markers reveal population structure and genetic diversity in the blueberry pathogen Monilinia vaccinii-corymbosi. Kathleen MBurchhardt, Marc A Cubeta. Department of Plant Pathology, North Carolina State University, Raleigh, NC.The ascomycete Monilinia vaccinii-corymbosi (Mvc) is a widespread fungal pathogen of blueberry (Vaccinium spp.) in North America. Both asexual andsexual spore production are required within a season for the fungus to complete its life cycle. Overwintered infected fruit (mummies) produce apotheciathat release aerially dispersed ascospores which infect newly emerging blueberry shoots, resulting in blighting of infected tissues followed by productionof conidia. Insect pollinators deposit conidia on flowers that infect the ovary through the gynoecial pathway, leading to fruit mummification. The primaryobjective of our research was to use population genetics-based approaches to examine genetic diversity, structure, and gene flow among populations ofMvc throughout the United States. A total of 437 samples from 18 blueberry fields in 10 states (one field in GA, MA, ME, MI, MS, NJ, NY, OR, and WA and 9fields in NC) were analyzed with 10 microsatellite markers. Population genetic analyses supported population structure and high intraspecific geneticdiversity, with 203 unique multilocus haplotypes (MLHs) identified from the samples. However, there were differences in genetic diversity and populationstructure based on locality and host species. Low genetic diversity and selfing were suggested based on analysis of samples of infected shoots or fruitcollected from rabbiteye (V. virgatum) varieties in MS, GA, and five fields in NC. Only three unique MLHs were identified from analyzing the 141 samplescollected from the seven fields, with two of the unique MLHs detected within four and five of the fields, respectively. At least 12 unique MLHs weredetected within all other fields except OR, with all MLHs being exclusive to their field of origin. Samples from the 10 fields were collected from eitherinfected shoots of rabbiteye, northern highbush (V. corymbosum), or southern highbush (V. corymbosum x V. darrowii), or from infected fruit of northernhighbush or lowbush (V. angustifolium). Analysis of molecular variance and the software STRUCTURE supported significant genetic differentiation amongthese fields, indicating restricted gene flow. The majority of microsatellite markers were in linkage equilibrium within the fields, suggesting randommating. Future research will examine the potential for host specialization of isolates of Mvc.647. Genetic diversity of Australian Pyrenophora tritici-repentis isolates using microsatellites. Caroline Moffat, Pao Theen See, Rick Dolling, RichardOliver. Department of Environment & Agriculture, Curtin University, Perth, WA, Australia.Pyrenophora tritici-repentis, the causal agent of tan spot of wheat, is an economically significant necrotrophic fungal pathogen. In Australia, tan spot isthe most damaging wheat disease, resulting in yield losses of $212 million per annum. The disease was first recorded in Australia in the 1950s, some tenyears after it was initially reported on wheat in the USA. Here, we examine the genetic diversity of a collection of Australian P. tritici-repentis isolates usingmicrosatellites. We discuss relatedness and structure, and consider the findings in the broader context of biogeography.648. WITHDRAWN649. Fungal community composition analysis by Internal Transcribed Spacer (ITS) sequencing using Illumina MiSeq. Robin A. Ohm 1 , Julien Tremblay 1 ,Kanwar Singh 1 , Feng Chen 1 , Claude Murat 4 , Matthias Hess 1,2,3 , Francis Martin 4 , Susannah G. Tringe 1 , Igor V. Grigoriev 1 . 1) US DOE Joint Genome Institute,Walnut Creek, CA., USA; 2) Systems Biology & Applied Microbial Genomics Laboratory, Washington State University, USA; 3) Chemical and BiologicalProcess Development Group, Pacific Northwest National Laboratory; 4) Lab of Excellence ARBRE, Tree-Microbes Interactions Department, INRA, Nancy,France.Fungal species identification and community surveys relied for a long time on Internal Transcribed Spacer (ITS) sequencing using the Sanger platform.Later, 454 (Roche) pyrosequencing was used for the same purpose, capturing shorter ITS1 or ITS2 fragments, or more recently the entire ITS region usinglonger 454 XLR reads. The Illumina sequencing platform has now largely surpassed 454 in terms of read quantity and quality (e.g., HiSeq2000 yields of upto 600 Gb in a single run) but the length of produced reads (up to 150 bp in HiSeq2000) is insufficient for ITS analysis. Illumina’s newly-introduced MiSeqsequencing platform can produce paired-end 250 base reads in a single day run, which, when combined, would cover most of either ITS1 or ITS2 regions.280

FULL POSTER SESSION ABSTRACTSAt the US DOE Joint Genome Institute we tested the Illumina MiSeq platform for the analysis of fungal community composition in forest soil and cowrumen, and we developed a workflow for the subsequent data analysis. We surveyed fungal populations in these environments by targeting the ITS2region. These amplicons were sequenced with an Illumina MiSeq instrument from both 5’ and 3’ ends with a 2x250 bases sequencing configuration. Thiswas followed by in silico assembly using their shared overlapping part, where possible. The UNITE database of fungal ITS sequences was used as areference database to classify the sequenced amplicons. As a classification method, both a naive Bayesian classifier (from the Ribosomal Database Project)and BLAST are explored. Our results suggest that the fungal population surveys on MiSeq successfully recapture known biological results and shouldprovide a useful tool for fungal community characterization.650. Estimation of genetic diversity of Ramularia collo-cygni populations using nuclear SSR markers to infer its potential to adapt to environmentalchanges. Marta Piotrowska 1 , Fiona Burnett 1 , Peter Hoebe 1 , Richard Ennos 2 , James Fountaine 1 . 1) Crop and Soil Research Group, Scotland’s Rural College,Edinburgh, EH9 3JG, United Kingdom; 2) Institute of Evolutionary Biology, University of Edinburgh, Edinburgh, EH9 3JT, United Kingdom.Ramularia collo-cygni (Rcc) is a fungal pathogen of barley (Hordeum vulgare) but it can also infect other cereal crops such as wheat (Triticum aestivum),rye (Secale cereale) and oats (Avena sativa). Its economic impact has increased in the last two decades, when Rcc started to have an economic impact ongrower’s yields. Rcc has been present as a major barley pathogen in Scotland, since 1998. Quinone outside Inhibitor (QoI) fungicides were widely used tocontrol the disease, but between 2001/2002 the first resistant strains appeared. Presently Succinate Dehydrogenase Inhibitors (SDHIs) are widely used andrecommended as one of the most effective fungicide treatments against Rcc and currently all of the available data suggests that Rcc is still sensitive to allSDHI fungicides. However, Rcc has presently been exposed to SDHI fungicides for a number of growing seasons and the risk of fungicide resistancedevelopment is probably high. In this study we use newly designed SSR markers to describe the diversity of Rcc populations at field scale and understandits ability to adapt to environmental changes (i.e. fungicide applications). Using SSR markers we aim to obtain information about the distribution of geneticvariation within and between Rcc populations and predict if clonal and/or sexual reproduction is taking place. Populations that are characterised by sexualor mixed reproduction systems over the growing season, may have higher adaptive potential than clonal populations, and thus could develop fungicideresistance more quickly. To study genetic variability in Rcc populations we developed 12 SSR markers and initially tested 10 isolates from 7 locations acrossthe world: Austria, Switzerland, Czech Republic, Denmark, France, Great Britain and New Zealand. Eleven variable pentanucleotide repeat loci have beenchosen for further testing. Further analysis was performed on a Scottish site, where 60 isolates were hierarchically sampled, and a Czech site where 30isolates were sampled. Preliminary data collected from 10 isolates sampled worldwide indicates variability among Rcc populations that can not beexplained by its geographical location alone.651. Alkaloid genotype profiling of tall fescue endophytes to determine influence of ancestral progenitors. J.E. Takach, C.A. Young. Forage ImprovementDivision, The Samuel Roberts Noble Foundation, Ardmore, OK.Epichloid endophytes, comprised of Epichloë and asexual Neotyphodium species, associate with cool-season grasses such as the agronomically importantforage tall fescue (Lolium arundinaceum syn Festuca arundinacea). This mutualistic symbiosis provides the plant host with protection from animal andinsect herbivory through the production of multiple classes of bioactive alkaloids (ergot alkaloids, indole-diterpenes, lolines, and peramine) by theendophyte partner. Many Neotyphodium species, including the endophytes present in tall fescue (N. coenophialum, Festuca arundinacea taxonomicgroups FaTG-2 and FaTG-3) arise from interspecific hybridization events and contain genomic information from multiple ancestral progenitor species. Assuch, hybrid Neotyphodium species are capable of producing multiple classes of alkaloids and can contain multiple copies of the loci from required foralkaloid production. Significant genetic and chemotypic diversity has been reported for tall fescue endophytes but few studies have assessed this diversityat a population level. The incidence and diversity of tall fescue endophytes present in extant tall fescue seed collections was evaluated using PCR-basedgenotype profiling of seed from a set of 97 tall fescue accessions obtained from the Germplasm Resource Information Network (GRIN). A total of 71endophyte-infected accessions were identified from both Continental (summer-active) and Mediterranean (summer-dormant) tall fescue germplasm.Genotype profiles from the GRIN tall fescue collection were compared to previously characterized tall fescue endophytes in order to predict the speciesand probable chemotype. Variation based on presence and absence of genes within the loci required for each alkaloid indicated likely chemotypic diversityamong and between species. The copy number of selected alkaloid genes was determined by sequence analysis of PCR amplicons. The ancestralprogenitor origins of mating type and alkaloid genes were inferred from phylogenetic analyses of partial gene sequences. These results support priorevidence that multiple alkaloid gene copies are the result of inheritance, not post-hybridization gene duplication, and suggest that multiple independenthybridization events have occurred during the evolutionary life history of tall fescue endophytes.652. Evolution of the pan-secretome among lineages of Magnaporthe oryzae attacking different host-plants. E. Fournier 1 , E. Ortega-Abboud 1,2 , L.Mallet 3,4 , H. Chiapello 3,5 , C. Guérin 3 , F. Rodolphe 3 , A. Gendrault 3 , J. Kreplak 4 , J. Amselem 4 , M-H. Lebrun 6 , T. Kroj 1 , D. Tharreau 2 . 1) INRA, BGPI lab, INRA,Montpellier, cedex 5, France; 2) CIRAD, BGP lab, TA 54K, 34398 Montpellier; 3) INRA, MIG lab, 78352 Jouy-en-Josas, France; 4) INRA, URGI lab, 78026Versailles, France; 5) INRA, BIA lab, 31326 Castanet-Tolosan, France; 6) INRA, BIOGER lab, 78850 Thiverval-Grignon, France.Over the past decade, considerable advances have been made in the understanding of the role of fungal effectors, and especially small secreted proteins(SSPs), in the infectious process. NGS technologies offer powerful tools to study, at the genomic scale, how deep are SSPs involved in the adaptation offungal populations to different host plants. We addressed this question in the plant pathogenic fungus Magnaporthe oryzae, the agent of blast on rice andother Poaceae. This species encompasses isolated genetic lineages specifically attacking different hosts. In the GEMO project, we sequenced eight strainsof M. oryzae representing different genetic groups pathogenic of different species of Poacees (5 strains attacking rice Oryza sativa, 1 attacking wheatTriticum sp., 1 attacking foxtail millet Setaria sp., 1 attacking finger millet Eleusine sp.), and one strain of the sister species M. grisea (attacking fonio milletDigitaria sp). The nine genomes have been sequenced using NGS technologies (454 and Solexa/Illumina) and assembled by the Genoscope (Evry, France).We included the public reference strain of M. oryzae 70-15 in our analyses. Gene annotation and orthology predictions have been carried out. We alsoannotated transposable elements and assessed the amount of horizontal transfers. Here we present the characterization of the repertoires of SSPs in thenine genomes, established using classical predictors of peptide signals (SignalP), transmembrane domains (TMHMM), GPI anchors (PrediGPI) andsubcellular location assignment (TargetP). These lists were then curated using two complementary approaches: systematic tBlastn searches of the SSPpredicted in each genome against the nine genomic databases of the project (including its own), and gene mining through the RNAseq analysis of the inplanta transcriptome of one of the strain. We will compare these lists with orthology predictions to analyze the core-secretome and the dynamics ofgains/losses/duplications of SSPs in the different lineages. We will also address the question of co-localization of SSPs with transposable elements. Finallywe will search for signatures of adaptive evolution in SSPs.653. Exploiting the high evolutionary potential of Leptosphaeria maculans minimises severity of blackleg disease of canola. Steve J. Marcroft 1 , Angela P.27th Fungal Genetics Conference | 281

FULL POSTER SESSION ABSTRACTSVan de Wouw 2 , Barbara J. Howlett 2 . 1) Marcroft Grains Pathology, Grains Innovation Park, Horsham, 3400, Vic., Australia; 2) School of Botany, theUniversity of Melbourne, 3010, Vic., Australia.Blackleg caused by Leptosphaeria maculans, is the most important disease of Brassica napus (canola) worldwide. Field populations of this sexuallyoutcrossing fungus rapidly adapt to selection pressure from extensive sowing of varieties with major gene resistance and can ‘overcome’ resistance. Thishigh evolutionary potential of the fungus is reflected in its genome structure. Effector genes are embedded in AT-rich, gene- poor regions withtransposable elements that have been degenerated by Repeat Induced Point (RIP) mutations. Thus effectors are easily gained, lost or inactivated. For thelast decade we have monitored virulence of blackleg populations and disease severity of varieties in field trials across Australia. In 2003 after two seasonsof extensive sowing, blackleg resistance of a set of varieties ‘broke down’ in the Eyre Peninsula, South Australia, causing 90% yield losses and withdrawal ofthese varieties from sale. By 2005, virulence of populations towards these varieties declined appreciably. Thus the blackleg-canola interaction behaves in a‘Boom and Bust’ manner. Analysis of isolates collected before and after the resistance breakdown showed that deletions, RIP mutations and amino acidsubstitutions accounted for rapid evolution of four linked effectors, including the avirulence gene complementary to the resistance gene that had beenovercome. After this resistance breakdown, Eyre Peninsula farmers sowed varieties with a different source of resistance. However in November 2011significant levels of disease in trial sites and commercial paddocks were observed, which led to a warning in February this year that these varieties shouldnot be sown. Growers heeded this advice and sowed varieties with different resistance sources. Our prediction of a resistance ‘breakdown’ was vindicated,as this variety had high disease levels in field trials on Eyre Peninsula, but not in other canola-growing regions. Commercial crops of other varieties on EyrePeninsula had only low levels of disease. By sowing other varieties, not only have farmers have been saved $20 million (based on conservative estimates ofarea sown, predicted yield loss and current canola prices), but seed companies have been able to sell the ‘at risk’ varieties in other canola-growing regions,where resistance breakdown was not predicted.654. Experimental demonstration of Crozier's paradox in fungi. Eric Bastiaans, Alfons J.M. Debets, Duur K. Aanen. Plant Science Group, WageningenUniversity, Wageningen, Netherlands.Kin selection can favour cooperation between individuals. This requires assortment between genetically related individuals and genetic kin recognition isthe predominant means to achieve this. However, Crozier realised that the diversity of kin-recognition alleles necessary for kin recognition, observed inmany social organisms, poses a paradox: common alleles will receive more cooperation than rare alleles, and therefore increase in frequency, thus erodinggenetic kin recognition diversity. We provide experimental evidence for Crozier’s theoretical prediction using somatic fusion between fungal individuals(mycelia) as a model for cooperation. Using fusion mutants and incompatible strains, we first show that fitness is strongly correlated with the degree offusion, which demonstrates that fusion between mycelia is mutually beneficial. We then experimentally demonstrate Crozier’s prediction that positivefrequency-dependent selection erodes kin-recognition diversity.655. A completely unknown lifecycle in mushrooms: cyclical inbreeding and haplo-diploidy. Duur K. Aanen 1 , Tim Möhlman 1 , Eric Bastiaans 1 , BartNieuwenhuis 1 , Bertha Koopmanschap 1 , Thomas W. Kuyper 2 . 1) Plant Science Group, Wageningen University, Wageningen, Netherlands; 2) Department ofSoil Quality, Wageningen University, Wageningen, The Netherlands.Mycena galericulata (Basidiomycota, Agaricales) occurs in two forms, a clampless with two-spored basidia and a clamped with four-spored basidia. It isgenerally accepted that the two-spored form is haploid asexual (apomictic), and the four-spored form sexual (dikaryotic and heterothallic). In order tostudy the interrelationship between both forms, we performed mating tests and phylogenetic and genetic analyses of a sample of both forms. Surprisingly,our results are inconsistent with any currently known life-cycle. While the four-spored form is heterothallic indeed, we show that the two-spored form isdiploid, and produces diploid spores via intra-tetrad selfing. However, the absence of genetic differentiation between both forms, and the high degree ofheterozygosity in the two-spored form, indicate that the two-spored form frequently arises from the four-spored. We hypothesise that the two-sporedform can again give rise to four-spored forms. Consistent with this, we discovered that a small percentage of fruiting bodies has both two-spored and foursporedbasidia.656. Diversity and evolution of ABC proteins in basidiomycetes. Andriy Kovalchuk 1 , Yong-Hwan Lee 1,2 , David Hibbet 3 , Fred O. Asiegbu 1 . 1) Department ofForest Sciences, University of Helsinki, Finland; 2) Department of Agricultural Biotechnology, Seoul National University, Seoul, Korea; 3) Department ofBiology, Clark University, Worcester MA 01610, USA.ABC proteins constitute one of the largest families of proteins. They are implicated in a wide variety of cellular processes ranging from ribosomebiogenesis to multidrug resistance. With the advance of fungal genomics, the number of known fungal ABC proteins increases rapidly, but the informationon their biological functions remains scarce. In this work, we extended our previous analysis of fungal ABC proteins to include recently genome sequencedspecies of basidiomycetes. We performed an identification and initial cataloguing of ABC proteins from 23 new species representing 10 orders from withinthe class of Ascomycotina. To identify gene loci encoding ABC proteins in the fungal genomes, multiple tblastn and blastp searches against selectedgenomes were performed at the website of the Fungal Genomics Program of the Department of Energy Joint Genome Institute (JGI). Sequences ofCoprinopsis cinerea ABC proteins representing all known subfamilies were used as queries. Phylogenetic analysis was performed with the programpackage MEGA5 using neighbor-joining, minimum evolution and maximum likelihood algorithms and bootstrapping with 500 replicates. ABC proteins ofeach species were separated into subfamilies by their comparison with S. cerevisiae, C. cinerea, C. neoformans and U. maydis proteins. Set of ABC proteinsidentified in basidiomycetes and ascomycetes were compared, and their common features and principal differences are discussed. Two groups of ABCproteins specific for basidiomycetes were identified. Results of the survey should contribute to a better understanding of evolution of ABC proteins in fungiand support further experimental work on their characterization.657. Co-evolution and life cycle specialization of plant cell wall degrading enzymes in a hemibiotrophic pathogen. Patrick C. Brunner 1 , Stefano F. F.Torriani 1 , Daniel Croll 1 , Eva H. Stukenbrock 2 , Bruce A. McDonald 1 . 1) Integrative Biology, ETH Zurich, Zurich, Switzerland; 2) Max Planck Institute forTerrestrial Microbiology, Marburg, Germany.Co-evolution of species has long been recognized as a driving force in generating and maintaining biodiversity. Co-evolution is an ubiquitousphenomenon investigated in prey and predator, plant and herbivore, or mutualistic interrelationships. However, signatures of co-evolution are likely to bestrongest in host-pathogen systems because of the strong selective pressures that each can exert directly on the other. While traditional studies mainlysought phenomenological evidence for co-evolution, more recent approaches look directly at the molecular/gene level. We hypothesized four mainscenarios for host-pathogen co-evolution and predicted the corresponding genetic signatures. We combined comparative genomics, transcriptomics andselection analyses to investigate genes that are likely affected by co-evolution to assign them to one of these scenarios. Zymoseptoria tritici is animportant fungal pathogen on wheat and has two closely related sister species Z. pseudotritici and Z. ardabiliae that infect wild grasses. This recently282

FULL POSTER SESSION ABSTRACTSemerged host-pathogen system provides a rare opportunity to investigate the dynamics of gene evolution by natural selection within and between speciesand on different hosts. Here, we focused on evolution of plant cell wall degrading enzymes (PCWDEs) secreted by the fungus. We found widespreaddifferential transcription among different members of the same gene family, challenging the idea of functional redundancy and suggesting instead thatspecialized enzymatic activity occurs during different stages of the pathogen life-cycle. We also found that natural selection has significantly affected atleast 19 of the 48 identified PCWDEs. The majority of genes showed signatures of purifying selection, typical for the scenario of conserved substrateoptimization. However, six genes showed diversifying selection that could be attributed to either host adaptation or host evasion. This information can beused to determine which genes are the most appropriate targets for subsequent wet lab experimentation to elucidate enzymatic function during relevantphases of the pathogen life cycle.658. Recombination landscape of the plant pathogenic fungus Zymoseptoria tritici (syn. Mycosphaerella graminicola). D. Croll, M. Lendenmann, E.Stewart, M. Zala, B.A. McDonald. ETH Zurich, Zurich, Switzerland.Recombination is a fundamental process driving the evolution of genomes. The rate of recombination influences the level of genetic variation inpopulations and the efficacy of selection. Furthermore, heterogeneity in recombination rates along chromosomes shapes the genetic architecture ofphenotypic traits. Hence, the evolution of virulence and other traits in pathogenic fungi critically depends on the rate of recombination. Despite theimportance of recombination, the rate and homogeneity of recombination in fungal chromosomes is poorly understood. We analyzed 60 progeny from acontrolled sexual cross between two isolates of the wheat pathogen Zymoseptoria tritici (syn. Mycosphaerella graminicola). We genotyped parental strainsusing whole-genome resequencing and we generated progeny genotypes by restriction site-associated DNA sequencing (RADseq). We obtained a total of46’037 single nucleotide polymorphisms (SNP) segregating among the progeny. Based on the physical and genetic map locations of the markers, weshowed that recombination rates were strongly heterogeneous along chromosomes and were influenced by gene density and GC content. We locatedmultiple chromosomal hotspots of recombination that were interspersed by large segments of low recombination rates. Furthermore, we found thatchromosomal regions that were enriched in SNP and indels showed lower recombination rates compared to less diverged regions. The local variation inrecombination rates in Z. tritici may have significant effects on the evolutionary potential of different genomic compartments. Hence, heterogeneity inrecombination rates may play an important role in the evolution of virulence.659. The evolution of Sfp1 mediated, cell size control in Ascomycete fungi. Toni M. Delorey 1 , Jenna M. Pfiffner 1 , Sushmita Roy 2 , Jay Konieczka 1 , Dawn A.Thompson 1 , Aviv Regev 1 . 1) Broad Institute, 7 Cambridge Center, Cambridge, MA 02139; 2) Wisconsin Institute for Discovery (WID), 330 N. Orchard St,Madison, WI 53715.Divergence in gene regulation can play a major role in evolution. We used a phylogenetic framework to measure mRNA profiles in 15 yeast species andreconstruct the evolution of their modular regulatory programs. We found that modules diverge with phylogenetic distance, with prominent regulatorychanges accompanying changes in lifestyle and ploidy. Gene paralogs have significantly contributed to this regulatory divergence. To explore the role oftrans regulator duplication, we examined Sfp1, as gain or loss of the Sfp1 binding site underlied regulatory rewiring of carbon metabolism. In S. cerevisiae,Sfp1, a TOR target, activates transcription of “growth” genes. S. cerevisiae, sfp1D mutants have smaller cells and slower growth, suggesting that thesephenotypes are intertwined. However, we show that duplication of SFP1 in other yeast species has resulted in sub- and neo-functionalization of regulatoryprograms controlling growth rate and cell size. In particular, in S. castellii , the two Sfp1 paralogs have subfunctionalized; one controls cell size while theother controls growth. Therefore, we hypothesize that Sfp1 regulation of ribosome biogenesis underlies growth rate while cell size is mediated by adifferent, unidentified function. To better understand Sfp1-mediated cell size control, we used a two-tiered analysis system of comparing gene expressionand ChIP Seq data to distinguish indirect or direct Sfp1 targets. Expression programs and phenotypes of sfp1D mutants were analyzed in S. cerevisiae, C.glabrata, S. castellii, K. lactis and S. pombe. To identify putative cell size regulators, we examined differentially expressed orthologs in species where sfp1Dmutants had a small size phenotype (S. cerevisiae, C. glabrata and one paralog of S. castellii), and excluded genes involved in ribosomal biogenesis andthose differentially expressed genes in species where sfp1 mutants grew slower but had normal cell size (K. lactis and S. pombe).We found 17 overlappingorthologs including a promising candidate for cell size regulation; the S. cerevisiae ortholog, Ard1, involved in telomeric silencing, and cell cycle control.Finally, we found that Sfp1 binds to the SCH9 promoter in S. cerevisiae and S. paradoxus. Sch9 is a kinase and mutants have reduced cell size. From thesefindings, we present a novel model for cell size regulation.660. Cryptic population subdivision, sympatric coexistence and the genetic basis of local adaptation in Neurospora discreta. Pierre Gladieux, DavidKowbel, Christopher Hann-Soden, John Taylor. Department of Plant and Microbial Biology, University of California, Berkeley, CA.Identifying the genes for ecologically relevant traits is a central challenge in empirical population genetics. Species distributed across strongenvironmental gradients are excellent models to discover and identify the genetic targets of local selection as they are more likely to experience spatiallyheterogeneous selection pressures leading to local adaptation of ecologically important traits. We studied the origin of ecological differentiation in N.discreta phylogenetic species 4 (PS4), a species with a broad latitudinal distribution. We Illumina-sequenced the complete genomes of 52 individualsrepresenting 8 collections sites in Alaska, New Mexico, Washington, California, and Western Europe (average sequencing depth: 52X). Reads were mappedto the N. discreta PS4 reference genomes, and analyses were based on a final set of ca. 1.2 million high-quality SNPs. Phylogenetic analyses identified fourwell-supported clades. Papua New-Guinea individuals formed the most basal clade. Individuals from Alaska and Europe on the one hand, and from NewMexico on the other hand grouped into sister clades, and individuals from California were basal to these two clades. Individuals from Washington, sampledwithin the same site, grouped with either the New Mexico individuals, or the California individuals, indicating the coexistence in sympatry of two divergentpopulations. The observed pattern of population subdivision is being used as a reference to identify genes departing from the genome-wide background,and showing increased divergence consistent with divergent selective pressures, or decreased divergence consistent with gene-flow. Our findingsemphasize the need to continue exploration to uncover divergent populations of Neurospora, and place N. discreta, along with N. crassa, among thehandful of species that have the attributes to serve as outstanding evolutionary and ecological model organisms.661. WITHDRAWN662. Evolutionary genomics of NRPS gene clusters in Beauveria and its allies. J.-G. Han 1 , J. Oh 3 , M.-W. Hyun 2 , B. Shrestha 1 , G.-H. Sung 1 . 1) MushroomResearch Division, Rural Development Administration, Suwon 441-707, Republic of Korea; 2) College of Pharmacy, Chung-Ang University, Seoul 156-756,Republic of Korea; 3) Department of Microbiology and Institute of Basic Sciences, Dankook University, Cheonan 330-714, Republic of Korea.Beauveria is an ascomycetous asexual genus that comprises of 12 species of insect pathogens and linked to its teleomorphic stage of Cordyceps. Amongspecies of Beauveria, Beauveria bassiana is economically important for its use as biological control agent and produces several secondary metabolites suchas beauvericin and bassianolide, which are the causal metabolites of entomopathogenicity. Genes involved in these secondary metabolites are including27th Fungal Genetics Conference | 283

FULL POSTER SESSION ABSTRACTSnonribosomal peptide synthetase (NRPS) that are often physically clustered. To more understand the origins and evolution of these genes, we performedgenome-wide comparative analyses among B. bassiana and its allied species including B. pseudobassiana, B. sungii, Cordyceps militaris, C. pruinosa, Isariatenuipes, and I. farinosa after assembling these genomes. Initially, all the potential gene clusters for secondary metabolites were predicted using byantiSMASH. In B. bassiana, 16 NRPSs and 3 NRPS-PKS hybrid modules were estimated, while in C. militaris, 5 NRPSs and 3 hybrids were found.Interestingly, the presence/absence of NRPSs and PKSs provides a clue in generating their evolutionary hypotheses (e.g., horizontal gene transfer and genefusion). For example, bassianolide synthetase were specifically found in the species referred to Beauveria with the syntenic conservation of genesencoding calreticulin, cyclophilin, DNA replication complex, IdgA domain protein, and phenol 2-monooxygenase. The inferred phylogeny based on theacyltransferase (AT) domain showed that ATs of beauvericin synthetase gene belong to two independent lineages, suggesting that beauvericin synthetaseis a fusion gene which is formed from two previously separated genes. It is also suggested that bassianolide synthetase in Beauveria spp. are possiblyacquired by horizontal gene transfer (HGT) from distantly related fungi.663. Neurospora presents a model for the evolution of mating systems. Christopher Hann-Soden, Pierre Gladieux, John Taylor. Plant and MicrobialBiology, UC Berkeley, Berkeley, CA.The study of plants and animals has yielded a host of theories for the evolution of outbreeding and selfing mating systems, yet these theories have yet tobe extended to microbial organisms, where they might be more easily challenged. Within the genus Neurospora there is evidence of multiple shifts toselfing (homothallic) from outcrossing (heterothallic) ancestral states. However, the most well-studied clade, the members of which produce brightlycolored macroconidia, contains no homothallic members, and there is only one known clade of aconidial heterothallic Neurospora from which homothallicNeurospora could have evolved (Nygren et al. 2011, Glass et al. 1990). We hypothesized that sampling of heterothallic Neurospora has been biased towardmembers that produce brightly colored macroconidia or perithecia in isolation, thereby ignoring aconidial heterothallic species. Collections of crypticheterothallic Neurospora would allow comparison of the evolutionary forces acting on closely related pairs of selfing and outcrossing species. To this end,we have isolated 1 new strain of heterothallic Neurospora and 9 new strains of homothallic Neurospora from soil from two locations in California. Basedon partial sequences of nik-1 (Nygren et al. 2011) we constructed a phylogeny of Neurospora including the new strains. Surprisingly, the new heterothallicNeurospora was found to be almost identical to the only other aconidial, heterothallic Neurospora individuals (Glass et al. 1990), despite over 2,300 km ofseparation. Seven of the eight sequenced homothallic strains were found to be most closely related to Neurospora novoguineensis, but the remainingstrain was more closely related to the aconidial, heterothallic strains. These preliminary results suggest that additional sampling to obtain sufficiently largepopulations of related, aconidial heterothallic and homothallic Neurospora would facilitate studies of the transition from outbreeding to selfing as well asstudies of the genetic basis of the development of macroscopic, brightly colored conidia. References, Glass, N. L., Metzenberg, R. L. & Raju, N. B.Experimental Mycology 14, 274-289 (1990). Nygren, K. et al. Molecular phylogenetics and evolution 59, 649-63 (2011).664. Profiling conditionally dispensable chromosomes of the plant-pathogenic fungus Zymoseptoria tritici (syn. Mycosphaerella graminicola). RonnyKellner 1 , Veronika Schott 1 , Stephan Poppe 1 , Rachel Brem 2 , Eva H. Stukenbrock 1 . 1) Max Planck Institute for Terrestrial Microbiology, Fungal Biodiversity,Karl-von-Frisch-Strasse 10, 35043, Marburg, Germany; 2) University of California, Department for Molecular Cell Biology, 176 Stanley Hall, Berkeley, CA94720-3220, USA.Conditionally dispensable chromosomes (cDCs) are common genomic features in many parasitic ascomycetes. The presence of cDCs entails a highamount of intraspecific genomic variation that is inherited in a non-Mendelian manner. Because genes located on cDCs have in some species been shownto play a role in pathogenicity cDCs may promote rapid adaptive evolution in response to host defenses. With up to eight cDCs the genome of the wheatpathogen Zymoseptoria tritici (syn. Mycosphaerella graminicola) contains by far the largest known proportion of dispensable elements among allascomycetes. In comparison to the core chromosomes, cDCs of Z. tritici are smaller, have on average less and shorter genes with a lower GC content and ahigher amount of paralogous sequences and repetitive elements. Hitherto, the functional relevance of cDCs for Z. tritici remains unclear.In this study we elucidate the relevance of cDCs in Z. tritici by assessing expression profiles during in-planta and axenic growth. Because our RNAseqdataset covers both host and non-host interactions we broaden the perspective of our approach by insights into host-specific expression profiles. In orderto verify the current genome annotation we mapped all RNAseq reads to the genome of Z. tritici and predicted gene transcripts. By combining our resultswith the latest genome annotation we set up a new transcript list, which was used in further analyses. We demonstrate an overall significantly lowertranscription of genes located on cDCs relative to genes located on core chromosomes. In addition, cDCs encode several unique genes that are expressedunder certain conditions. We identify duplicated genes using a blast approach and show differential gene expression between paralogs on cDCs and corechromosomes. To link the transcription of cDC genes to specific stages in the interaction of Z. tritici and wheat we focus on a gene family of three paralogswhere two genes encode secreted proteins. We quantified their expression via qPCR at seven different time points of the interaction. In summary, ourstudy suggests the functional relevance of single cDC genes and the relevance of cDCs for gene innovation and adaptive evolution in Z. tritici.665. Take a walk on the wild side: evolutionary consequences of resistance to apple scab introgressed from a wild host. C. Lemaire 1 , T. Leroy 1 , M. deGracia 2 , C-E. Durel 2 , M. Templeton 3 , J. Bowen 3 , V. Caffier 2 , B. Le Cam 2 . 1) IRHS, University of Angers, Angers, France; 2) IRHS, INRA, Beaucouzé, France; 3)Plant & Food Research, Auckland, New Zealand.Theories on emergence of pathogens are dominated by ecological hypotheses. For instance overcoming of a host resistance gene is often described as aconsequence of the spreading of a mutant that invades resistant hosts from infected susceptible cultivars. However invasion of virulent populations fromwild habitats are often neglected. Though taking into account the occurrence of a preexisting allopatric virulent population greatly impacts conclusionsmade about emergence of new virulence and the nature of barriers to reproduction with the avirulent population. Indeed under allopatric scenario, thesebarriers are not expected to be only adaptive. Here, we intended to decipher the evolutionary history of the European populations of the apple scabpathogen Venturia inaequalis enable to overcome the Rvi6 resistance gene introgressed in Malus x domestica from the crabapple Malus floribunda. Usingmicrosatellite and sequence data in an Approximate Bayesian Calculation (ABC) framework, we demonstrate that virulence corresponding to a newresistance gene introgressed in agrosystems was previously present in the wild within a population that has diverged since several thousands years. Weshow that deployment of the corresponding resistance gene in agrosystems can then act as gateways for virulent populations to invade orchards. At lastwe show that secondary contact followed by mating between invading and resident population can reveal genetic incompatibilities (Dobzhansky-MüllerIncompatibilities) accumulated during divergence between the two populations. These incompatibilities induce hybrid depression by negative epistasis.This study based on state-of-the-art tools of population genomics, phenotyping and genetic mapping is the first to demonstrate the occurrence of intrinsicpost-zygotic barriers in pathogens revealed by a secondary contact. Overall, it points out the risk of generating new “hybrid” populations harbouring newpathogenic traits by introgression of resistance genes in agrosystems by breeders.284

FULL POSTER SESSION ABSTRACTS666. Genomic footprint of adaptive divergence in Ophiostoma montium, a fungal symbiont associated with the mountain pine beetle. J. F. Mao, B.Dhillon, C. Tsui, K. Ritland, R. Hamelin. Faculty of Forest Sciences, University of British Columbia, Vancouver, BC, Canada.Ophiostoma montium is the most common fungus associated with the mountain pine beetle, the insect responsible for the destruction of 18 million haof pine forests in Canada. In order to determine the evolutionary histories of O. montium populations, high-throughput genome sequencing was used touncover the genetic changes that accompany divergence as lineages colonize different conifer hosts in different climatic regions. 36 fungal isolates fromdifferent hosts and different environments were sequenced. De novo genome assembly from one isolate was used as reference to call variants (922,000SNPs and 126,000 Indels). Multiple analyses including whole genome variants, genetic distance, population structure and Identity by descent (IBD)identified three population lineages, corresponding to the various hosts and three geographic groups: US, Rocky, and North Canada (NC). Faster linkagedisequilibria (LD) decays were observed in both Rocky and NC groups, indicating an increase in genomic recombination and/or high effective populationsize. Presence of genomic regions with high LD and negative Tajima's D was evidence that population size expansion (after bottleneck or a selectionsweep) and/or purifying selection occurred in the NC group. Loci in the genome contributing to both host shift and climatic transition were also identifiedby multiple lineage-specific genomic scans for selection. Additionally, genome recombination events were recovered for lineages experiencing differentdemographic histories. Our study highlights the value of whole genome sequences both in evolutionary dynamics and genetics of plant pathogens.667. Ecological context in symbioses: when is your enemy also your friend? Georgiana May 1 , Paul Nelson 2 . 1) Dept Ecol, Evol, Behavior,#100, UnivMinnesota, St Paul, MN; 2) EEB graduate program University of Minnesota St. Paul MN.Most plants are rife with fungal symbiotic partners with many of these having little apparent effect on the host's health and fitness. In this work, weexplore the degree to which the outcome of interactions between an endophytic fungus, pathogen and plant host depend on ecological context. Inparticular, we ask whether interactions between the endophyte of maize, Fusarium verticilliodes, with the pathogen Ustilago maydis, depend on hostresistance to the pathogen. In the case of a host susceptible to the pathogen, the two fungal species should meet frequently, and compete over hostresources, potentially driving greater virulence to the host in one or the other fungal species. In the case of a host resistant to the pathogen, theendophyte might be a "bystander" to the pathogen, because the two meet too infrequently to drive their co-evolutionary interaction. We show evidencethat the two fungal species have evolved stronger antagonistic interactions in maize susceptible to the pathogen, and further, that this might beassociated with greater virulence by the pathogen. Results of modeling will also be presented from which we predict longer term evolutionary trajectoriesfor this 3-way interaction.27th Fungal Genetics Conference | 285

FULL POSTER SESSION ABSTRACTS668. Population shifts and mating-type heterokaryosis in Aspergillus flavus. Rodrigo A. Olarte 1 , Bruce W. Horn 2 , Carolyn J. Worthington 1 , Rakhi Singh 1 ,Ignazio Carbone 1 . 1) Department of Plant Pathology, North Carolina State University, Raleigh, NC; 2) National Peanut Research Laboratory, AgriculturalResearch Service, U.S. Department of Agriculture, Dawson, GA.Aspergillus flavus is a heterothallic fungal pathogen of many economically important crops worldwide. We sampled A. flavus strains from a cornfield inRocky Mount, North Carolina, USA. Plots were inoculated at tasselling with either A. flavus AF36 or NRRL 21882 (=Afla-Guard) nonaflatoxigenic biocontrolstrains, both of which are mating type MAT1-2. Subsequently, aflatoxigenic strain NRRL 3357 (MAT1-1) was applied to all plots, including control plots notinoculated with biocontrol strains. Sclerotia were harvested from infected corn ears and ninety single-ascospore isolates were obtained from ascocarpsoriginating from plots treated with AF36 and NRRL 21882. In addition, eighty A. flavus isolates were collected from soil one month after planting (beforebiocontrol application) and one year after biocontrol application, for a grand-total of 250 isolates. PCR amplification revealed grouping of isolates intothree distinct mating-type classes: MAT1-1, MAT1-2 and MAT1-1/MAT1-2. An overwhelming majority (54%) of isolates sampled prior to biocontroltreatments were heterokaryotic for mating type (MAT1-1/MAT1-2), but was shifted to only 9% of isolates from soil after biocontrol treatments; 39% ofisolates obtained from ascospores were heterokaryotic, with the remaining comprising either MAT1-1 or MAT1-2. Multilocus genotyping indicated thatascospores might have originated from Afla-Guard as a putative parent; there was no evidence of AF36 or NRRL 3357 in ascospores or in pre- or posttreatmentsoil samples, which may explain the genetic structure of the indigenous population. The vertical transmission of MAT1-1/MAT1-2 to progenyascospore isolates suggests that heterokaryosis can be maintained in subsequent generations. Furthermore, matings were performed to determinefunctionality of these MAT1-1/MAT1-2 strains and all isolates tested were strictly functional as MAT1-2. Further characterization of heterokaryons andtheir frequency in A. flavus populations may be important in understanding the adaptation of these fungi to changing environmental conditions and couldlead to better and more effective biocontrol strategies specific to a geographic region. Understanding population structure is the key to unlocking thesecrets of a successful biocontrol strain.669. Structural variation of trichothecene mycotoxins has resulted from multiple evolutionary processes in the fungal order Hypocreales. R. H. Proctor 1 ,A. M. Stanley 1 , M. G. Malmierca 2 , N. J. Alexander 1 , S. Gutiérrez 2 , S.P. McCormick 1 . 1) Bacterial Foodborne Pathogens and Mycology, USDA ARS NCAUR,Peoria, IL; 2) Universitary School of Agricultural Engineers, University of León, Ponferrada, Spain.Trichothecenes are secondary metabolites produced by fungi in at least six genera of the order Hypocreales. These metabolites are of concern becausethey are toxic to humans and other animals and can accumulate in grain used for food and feed. They also contribute to plant pathogenesis of Fusariumand to biological control activity of Trichoderma. Although all trichothecenes share the same molecular skeleton, a tricyclic structure with a12,13-epoxide,different genera produce trichothecenes that differ in patterns of oxygenation and acylation. To investigate how such structural variation has evolved, weexamined 1) variation in gene function and content in homologs of the trichothecene biosynthetic gene (TRI) cluster and 2) phylogenetic relationships ofTRI genes among trichothecene-producing genera. The results suggest that the ancestral hypocrealean TRI cluster consisted of at least seven genes,including the enzyme-encoding genes TRI4 and TRI5 responsible for synthesis of the trichothecene skeleton, the regulatory genes TRI6 and TRI10, and thetransporter gene TRI12. Phylogenetic analyses indicate that oxygenation of carbon atom 4 (C-4), which occurs in all trichothecene-producing genera, likelyarose when different genera acquired different C-4 hydroxylase genes: e.g. TRI11b in Trichoderma and Myrothecium and TRI13 in Fusarium. In contrast, C-3 oxygenation, which occurs in only one genus, likely arose by a change in function of TRI4, a gene that exists in all genera. These results, and those fromstudies of Fusarium and Trichoderma, indicate that structural variation of trichothecenes has arisen by recruitment, changes in function, and deletion ofTRI genes during evolution of the Hypocreales.670. Evidence for birth-and-death evolution and horizontal transfer of the fumonisin mycotoxin biosynthetic gene cluster in Fusarium. R.H Proctor 1 , F.Van Hove 2 , A. Susca 3 , G. Stea 3 , M. Busman 1 , T. van der Lee 4 , C. Waalwijk 4 , A. Moretti 3 , T.J.. Ward 1 . 1) Bacterial Foodborne Pathogens and Mycology, USDAARS NCAUR, Peoria, IL; 2) Earth and Life Science Institute, Université catholique de Louvain, Louvain, Belgium; 3) Institute of Sciences of Food Production,National Research Council, Bari, Italy; 4) Plant Research International B.V., Wageningen, The Netherlands.In fungi, genes required for synthesis of secondary metabolites are often clustered. The FUM gene cluster is required for synthesis of fumonisins, a familyof toxic secondary metabolites produced predominantly by species in the Fusarium (Gibberella) fujikuroi species complex (FFSC). Fumonisins are a healthand agricultural concern because their consumption is epidemiologically associated with multiple diseases in humans and other animals. Among FFSCspecies, the FUM cluster is discontinuously distributed but uniform in gene order and orientation. In this study, we demonstrate that the FUM clusterexists in at least four different genomic contexts within the FFSC and that phylogenetic relationships derived from analyses of FUM cluster genes arecorrelated with genomic context, but are inconsistent with species relationships inferred from analyses of primary-metabolism genes. In addition, analysesof synonymous site divergence suggested that FUM cluster divergence predated divergence of the FFSC. These results are not consistent with transspeciesevolution of ancestral cluster alleles or with interspecies hybridization, but suggest duplication of the cluster within an FFSC ancestor andsubsequent loss and sorting of paralogous clusters in a manner consistent with the birth-and-death model of evolution previously described for multigenefamilies. A model based on horizontal gene transfer (HGT) could also explain these observations, but seems unlikely because it requires independenttransfer events from multiple unknown donors to multiple FFSC recipients. However, analyses of phylogenetic relationships and synonymous sitedivergence provided strong evidence that F. oxysporum strain FRC O-1890 acquired the FUM cluster via a relatively recent HGT event from F. bulbicola or aclosely related species within the FFSC. These results indicate that, as with other secondary metabolite clusters, species phylogenies do not provide anadequate picture of the complex evolutionary history of the FUM cluster within Fusarium.671. Chemotype predominance in Fusarium graminearum is not directly affected by the use of the fungicides trifloxystrobin and isopyrazam. MatiasPasquali, Tiphaine Dubos, Friederike Pogoda, Lucien Hoffmann, Marco Beyer. Environment and Agro-biotechnologies Department, CRP GABRIELLIPPMANN, Belvaux, Luxembourg.Mitochondrial respiration inhibitors are effective fungicides used to control wheat diseases in Europe. Since the beginning of the monitoring ofchemotype diversity in wheat fields in Luxembourg, we are trying to identify factors involved in the shift of chemotype prevalence (nivalenol vs. 15ADONvs. 3ADON). In this study we investigated whether the use of fungicides belonging to the respiratory inhibitors complex II and III (that are used in wheatfields for treating other diseases) may play a role in selecting a specific F. graminearum chemotype. Strains from Luxembourg and from a world collectionwith isolation dates ranging from 1969 to 2011 were chemotyped by genetic means and then analysed for their sensitivity to trifloxystrobin (inhibitor ofrespiratory complex III) and isopyrazam (inhibitor of respiratory complex II) using a microplate in vitro test on conidia. The maximum level of inhibitionwhich could be obtained by trifloxystrobin ranged from 14 to 65% for the 55 strains analyzed with no complete inhibition up to a concentration of 3mM.Fortyone isolates tested for their sensitivity towards isopyrazam were insensitive with the average rate of inhibition converging towards 28%. For bothfungicides, EC50 values did not significantly depend on the chemotype, suggesting that these two fungicides do not exert a direct pressure on the selection286

FULL POSTER SESSION ABSTRACTSof chemotypes in F. graminearum. Our study also suggests that F. graminearum seems to be significantly insensitive to respiratory complex inhibitors (IIand III). Molecular mechanisms involved in the insensitivity are under investigation.672. Evolution of races within f.sp lycopersici of Fusarium oxysporum. BV. Chellappan, PM. Houterman, M. Rep, BJC. Cornelissen. Molecular PlantPathology, University of Amsterdam, SILS, Science Park 904, 1090 GE Amsterdam, The Netherlands.Three physiological races (1, 2 and 3) of Fusarium oxysporum f.sp lycopersici (Fol) have been identified based on their inability to infect tomato cultivarscarrying Fol resistance genes (I, I-2 or I-3, respectively). We wished to unravel the molecular mechanisms underlying the evolution of Fol races. It isgenerally assumed that race 2 evolved from race 1 by loss of AVR1 and that race 3 evolved from race 2 by a point mutation in AVR2, thus overcoming I andI-2 mediated resistance, respectively. We have sequenced a genomic region of approximately 100 kb containing AVR1 in race 1 isolate Fol004 andcompared it to the sequenced genome of race 2 isolate Fol4287. A genomic fragment of 30.5 kb containing AVR1 was found to be missing in Fol4287.Further analysis suggests that race 2 evolved from race 1 by deletion of this 30.5 kb fragment, most likely due to recombination between helitronsbordering the fragment. A worldwide collection of Fol isolates was subjected to PCR analysis of the AVR1 genomic region, including the two borderinghelitrons. The results suggest that, based on the deletion event that led to loss of AVR1, Fol isolates can be divided into distinct lineages that coincide withtheir geographical origin. Our results also suggest that transposable elements played a major role in the evolution of races within f.sp lycopersici ofFusarium oxysporum.673. Detection of Mitochondrial DNA Heteroplasmy in the progeny of crossed genetically divergent isolates of Arbuscular Mycorrhizal Fungi. MaryamNadimi, Ivan de la Providencia, Gabriela Rodriguez, Denis Beaudet, Moahmed Hijri. IRBV, Biological Sciences Dep., University of Montreal, Montreal, QC,Canada.Nonself fusion and nuclear genetic exchange has been documented in arbuscular mycorrhizal fungi (AMF) particularly in Glomus irregulare, which is acommon and widespread species. However, mitochondrial transmission accompanying nonself fusion of genetically divergent isolates remains unknown.We developed a series of crossing experiments between different isolates of G. irregulare, harboring genetically divergent mitochondrial DNA (mtDNA)haplotypes. We tested the hypothesis that heteroplasmy (i.e. mixture of genetically different mtDNA in a common cytoplasm) occurs in the progenies ofthe crossed isolates. Three isolates of geographically distant locations were used to investigate nonself fusions and mtDNA transmission in the progeny. Tobe able to trace the mtDNA haplotypes, we sequenced two mtDNAs of two G. irregulare isolates (DAOM-240415 and DAOM-234328) additional to thecurrent available isolate DAOM-197198. We developed isolate-specific markers in variable regions of intergenic mtDNAs (cox3-rnl) of these isolates. Threecrossing combinations in pre-symbiotic and symbiotic phases were performed. Interestingly, nonself fusion frequency was low and was usually associatedwith irregular shape and aborted spores, although normal spores were also observed. Ten progeny spores per crossing combination were genotyped usingisolate-specific markers. We showed the evidence that nonself fusion occurs between isolates originated from different continents both in pre-symbioticand symbiotic phases. Genotyping patterns of individual spores from the progenies clearly showed the presence of markers of the two parental mtDNAhaplotypes. Our results demonstrated the occurrence of mtDNA heteroplasmy in the progeny of crossed isolates. This raises the questions whethermtDNA heteroplasmy is transient or persistent in AMF? What are their consequences in evolution of AMF? Are there any conflicts of the presence ofmtDNA heteroplasmy within an individual? Further studies on vegetative compatibility and incompatibility and putative sex machinery in AMF will providenew information to explore and solve these questions and thereby advance our understanding of the evolution of AMF.674. Evolution of mode of infection in the rice blast fungus and allied species. Ning Zhang 1 , Shuang Zhao 1 , Jing Luo 1 , Guohong Cai 1 , DebashishBhattacharya 2 , Bradley Hillman 1 . 1) Plant Biology and Pathology, Rutgers Univ, New Brunswick, NJ; 2) Ecology, Evolution and Natural Resources, RutgersUniv, New Brunswick, NJ.The family Magnaporthaceae contains devastating fungal cereal and grass pathogens, such as Pyricularia oryzae (Magnaporthe oryzae, rice blast fungus),Magnaporthiopsis poae (Magnaporthe poae, summer patch pathogen of turf grasses), and Gaeumannomyces graminis (take-all fungus of various cerealsand grasses), which are popular model organisms in fungal biology and host-pathogen interaction studies. Despite their ecological and economicimportance, the phylogenetic relationships among the constituent species remain ambiguous due to the lack of convincing morphological characters andpaucity of molecular data for the majority of the non-model species in the family. In this study, our multilocus phylogeny suggests that both Magnaportheand Gaeumannomyces are polyphyletic genera. Therefore, a new genus, Magnaporthiopsis is proposed based on phylogeny and morphology. Thephylogeny also provides insights into fungal biology and pathogenesis. Pyricularia oryzae formed a basal clade, while Magnaporthiopsis poae andMagnaporthiopsis rhizophila formed another well-supported clade with Magnaporthiopsis incrustans (G. incrustans), G. graminis and Nakataea sigmoidea(Magnaporthe salvinii). The basal species infects both root and aerial parts of plant host, while the aerial infection capacity seems to be lost in the taxa ofthe latter clade. The study indicates that anamorphic and ecological features are more informative than the teleomorphic characters in definingmonophyletic groups among these taxa. In addition, we performed genome sequencing for 6 species in Magnaporthaceae: Magnaporthiopsis rhizophila,Magnaporthiopsis incrustans, Harpophora maydis, Nakataea sigmoidea, Ophioceras dolichostomum, and Pseudohalonectria lignicola, in order to conductphylogenomic and comparative genome analyses for both pathogenic and non-pathogenic members of this family.675. WITHDRAWN676. Population genomic analysis reveals a complex evolutionary history of Neurospora tetrasperma. Padraic Corcoran 1 , Fen Chen 2 , Martin Lascoux 1 ,Peixiang Ni 2 , Hanna Johanesson 1 . 1) Uppsala University, Uppsala, Sweden; 2) BGI, Hong Kong, Hong Kong.Fungal population genomics as a field of inquiry has seen a rapid growth in the recent years, with ability to sequence the genomes of multiple strains offungi sampled from many populations. These studies have aimed at utilizing a population genomic approach to understanding the evolutionary forces thathave had the greatest effect in shaping the genomes of fungal species. In this study, we extend the use of population genomics to help understand theevolutionary history of Neurospora tetrasperma. Neurospora tetrasperma has been the focus of much research in recent years, with most effort devotedto the study of its predominantly non-recombining mating type chromosomes. Here we present results of analysis on the whole genome resequencing of86 homokaryotic strains of N. tetrasperma, sampled from locations in England, New Zealand and Louisiana, USA, together with one strain each of the closeheterothallic species N. hispaniola and N. sitophila. These genomes were sequenced to a mean depth of between 25 to 30X coverage. Phylogenetic andpopulation structure analysis of the genomewide SNP data produced identified that the sequenced strains of N. tetrasperma belong to 5 previouslyrecognised lineages of N. tetrasperma. Comparisons of the multiple N. tetrasperma genomes with the N. sitophila, N. hispaniola and N. crassa genomesconfirmed previous observations on introgression, but also revealed signatures of introgression between the English population of N. tetrasperma and N.hispaniola. Furthermore, comparisons between the genomes of the two homokaryons isolated from each heterokaryon revealed a large number of27th Fungal Genetics Conference | 287

FULL POSTER SESSION ABSTRACTSdifferences across all chromosomes in the UK population of N. tetrasperma, while the other lineages show much fewer heteroallelic sites. Analysis of themating type chromosome reveals considerable variation in the sizes of putative regions of suppressed recombination in the 5 lineages of N. tetraspermainvestigated. We also present findings on the genome wide patterns of polymorphism within populations and divergence between populations, with theaim of dissecting which regions of the genome have been recently acted on by natural selection. The initial results gained in this study are showing thatmultiple N. tetrasperma populations have complex histories that have left strong signatures in their genomes.677. Rapid evolution of female-biased genes: a novel example from the eukaryotic model organism Neurospora crassa. Hanna Johannesson, CarrieWhittle. Evolutionary Biology, Uppsala University, Uppsala, Sweden.In animals and plants, sex-biased gene expression plays a major role in gene evolution. In particular, reproductive genes with male-biased expressiontend to exhibit rapid protein evolution and reduced codon bias as compared to female-biased or unbiased genes. Minimal data are available for fungi.Here, we demonstrate that sex-biased expression is associated with gene evolution in the filamentous fungus Neurospora crassa, but in contrast toanimals and plants, the rapid evolution occurs for female-biased genes. Based on analyses of >25,000 expressed sequence tags (ESTs) from male (conidial),female (protoperithecial) and vegetative (mycelial) tissues, we show that reproductive genes with female-biased expression exhibit faster proteinevolution and reduced optimal codon usage than male-biased genes and vegetative genes. Furthermore, our data suggest that female-biased genes arealso more apt to experience selective sweeps. The sex-biased expression effects are observable at the species and population level. We argue that therapid molecular evolution of female-biased genes is best explained by sexual selection via female-female competition, but could also result from matechoiceand/or directional natural selection.678. Fungal Community Dynamics During Biomass Degradation in the Cow Rumen Determined by ITS Sequencing. Hailan Piao 1,2 , Julien Tremblay 3 , RobinOhm 3 , Kanwar Singh 3 , Fernanda Haffner 4 , Stefan Bauer 4 , David Culley 2 , Kenneth Bruno 2 , Kerrie Barry 3 , Feng Chen 3 , Scott Baker 2,5 , Roderick Mackie 6 ,Susannah Tringe 3 , Igor Grigoriev 3 , Matthias Hess 1,2,3,5* . 1) Washington State University, Richland, WA; 2) DOE Pacific Northwest National Laboratory,Richland, WA; 3) DOE Joint Genome Institute, Walnut Creek, CA; 4) Energy Biosciences Institute, UC Berkeley, Berkeley, CA; 5) Environmental MolecularScience Laboratory, Richland, WA; 6) University of Illinois, Urbana-Champaign, IL.The microbial community that inhabits the cow rumen is composed of Archaea, Bacteria and Eukarya and is well known for its biomass-degrading ability.In order to understand this ecosystem at the whole-systems level it is important to monitor the dynamics of the individual community members.Sequencing of the 16S rRNA gene has been used intensively to obtain insights into the ecology of the prokaryotic fraction of the microbial rumencommunity. To obtain insights into the ecology of the fungal fraction of the rumen community and its dynamics during biomass-degradation, we amplifiedthe ITS2 region from fungi that colonized corn stover and switchgrass during rumen incubation. Amplicons were generated from rumen-incubatedswitchgrass and corn stover and rumen fluid at six different time points and from two different host animals. Sequencing on Illumina’s MiSeq platformresulted in a total of 10,675,384 sequences with an average read length of ~240bp amounting to a total of >2.6 Gbp of sequence information. Succeedingsequence analysis revealed a fungal community of low complexity, with two phyla as the dominant players. Members of the phylumNeocallimastigomycota were absent on the pre-incubated biomass and appeared to colonize both corn stover and switchgrass throughout the incubationprocess. Members of the phylum Ascomycota were less dominant (

FULL POSTER SESSION ABSTRACTSthe loci on chromosome five in two of the populations. The reduced pair-wise linkage disequilibrium with increasing distance cannot be attributed tomutation alone, and thus the high intrachromosomal recombination is most likely due to meiotic recombination following outcross in these populations.Recombination hot spots and cold spots were detected. Mating type loci in 59 isolates of two populations were genotyped using PCR with allele-specificprimers. About 40% of the isolates showed both MAT1-1 and MAT 1-2 idiomorphs, as expected for a homothallic species. However, the remaining 60% ofthe isolates had only the MAT1-2 idiomorph as detected by the allele-specific PCR. Although the nature of the absence of the MAT1-1 idiomorph remainsto be determined, the results showed variations in mating type alleles in natural populations, suggesting that some of the isolates may not be trulyhomothallic.681. Population genomics of Suillus brevipes. Sara Branco, John Taylor, Tom Bruns. Plant and Microbial Biology, University of California, Berkeley, CA.Environmental heterogeneity may result in divergent selection, which in turn may lead to local adaptation resulting in populations showing habitat-baseddiscontinuous variation. Very little is known on the patterns of adaptive divergence and local adaptation or underlying selective forces in ectomycorrhizalfungal populations. We are studying the population genomics of Suillus brevipes (Peck) Kuntze, an ectomycorrhizal fungus associated with pine trees. Ourgoals are 1) to test the existence of barriers to gene flow across populations, and 2) detect genomic regions exhibiting selection sweeps. We sequencedthe whole genomes of 30 S. brevipes individuals from two populations, one from a coastal Bishop pine forest in Mendocino Co (CA) and another from ahigh altitude Lodgepole pine forest in Yosemite National Park (CA). One isolate was selected as the reference strain and its genome was assembled denovo through a collaboration with the Joint Genome Institute. All other individuals were aligned to this reference and single nucleotide polymorphismsdetected across populations. These markers are being used to find regions with evidence of positive natural selection, including possible islands ofintrogression, increased rates of non-synonymous substitutions, accelerated rates of divergence, and gene duplications. Results from our study will make asignificant contribution for understanding patterns of neutral and functional variation as well as gene flow and selection in ectomycorrhizal fungi.682. Ugly, understudied and undertreated: population genomics of the most common human fungal pathogen - the dermatophyte Trichophytonrubrum. M. Gajdeczka, W. Li, J. Heitman. Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC.Trichophyton rubrum causes athlete’s foot, nail infections and ringworm in about twenty percent of the general population. Despite an annotatedgenome and hundreds of clinical isolates, the global genomic diversity and population biology of T. rubrum are not well understood. Additionally, T.rubrum cannot be crossed, genetic manipulation is difficult, and animal infections with T. rubrum are acute and highly inflammatory (rather than chronicand relatively mild as in humans). These factors have made it difficult to study virulence in T. rubrum and have limited the development of effectivechemotherapies. This study will test three hypotheses: (1) clonal reproduction and limited dispersal have resulted in undiscovered local sequence variationand linkage disequilibrium (LD); (2) observed variation supports an out-of-Africa isolation-by-distance migration model; and (3) patterns of sequencediversity in sex- and virulence-associated genomic regions support the significant evolutionary roles of reproductive clonality and close interaction withhumans. Worldwide isolates appear reproductively clonal and remarkably genetically monomorphic. Five common MLST markers reveal no variationamong 50 European, Asian and North American strains. We Sanger sequenced 7.5 kb of non-coding intergenic regions in ten T. rubrum isolates collectedover a span of ten years. These sequences revealed no polymorphisms useful for differentiating strains. Lastly, VNTR genotyping by our lab and othergroups revealed two broad geographic types, though these are based on a few highly variable loci subject to homoplasy. We are assembling wholegenome sequences of 32 geographically and clinically diverse strains to characterize sequence diversity more completely. The genome obtained by theBroad Institute will be used as a reference. We will obtain measures of diversity, polymorphism, LD, population differentiation and divergence. Thesemeasures will be used to compare isolates genome-wide, determine population structure and infer evolutionary and demographic forces contributing toobserved patterns of variation. Locus-specific markers will be developed based on standing variation to genotype additional strains. Furthermore, we willtest for variants associated with hypothesized virulence mechanisms involved in chronic T. rubrum infection.683. Poppr: an R package for genetic analysis of populations with mixed reproduction. Zhian N. Kamvar 1 , Niklaus J. Grünwald 1,2 . 1) Botany and PlantPathology, Oregon State University, Corvallis, OR; 2) USDA-ARS Horticultural Research Laboratory, Corvallis, OR.Analysis of populations with mixed reproductive systems, including a blend of sexual and clonal reproduction, remains a challenge. We developed an Rpackage implementing existing approaches for analysis of mixed populations. R is a multi-platform, open source, statistical environment that has gainedpopularity over the past few years. While there are a plethora of packages in R that perform population genetics analyses, many standard analysismethods for populations with mixed modes of reproduction remain hard to accomplish. Poppr aims at providing functions to facilitate rapid analyses ofthis data, particularly including methods for analysis of recombination (index of association), clone-censored analysis of full datasets in a hierarchicalmanner over all levels of sampling, genotypic diversity analyses, and distance analyses. As implemented, poppr requires minimal commands withconvenient summary functions while providing compelling graphics. Unlike many platform dependent, standalone programs, poppr can be used for batchprocessing of data including all kinds of population genetic data (dominant/codominant; microsatellites, AFLP, SNPs). Poppr is available as a beta releasefor testing upon request and continues to be improved.684. The heterothallic fungus Cercospora beticola contains fragments of both mating type genes. Melvin D. Bolton 1 , Zhaohui Liu 2 . 1) USDA - ARS, Fargo,ND; 2) North Dakota State University, Department of Plant Pathology, Fargo, ND.In most heterothallic Ascomycota, the ability to reproduce sexually is determined by a single mating type locus (MAT1) represented by two idiomorphsknown as MAT1-1 and MAT1-2 that are required to control nonself-recognition and mating of compatible partners. To investigate the MAT1 locus in theheterothallic fungus Cercospora beticola, we performed Southern analyses of both mating types using each mating type gene (MAT1-1-1 and MAT1-2-1) asa probe. Surprisingly, several bands of similar size were observed in both mating types using either probe, suggesting that multiple loci contain sequencesof both MAT genes in the C. beticola genome. We screened a BAC library derived from a MAT1-2 isolate using both MAT gene probes to identify clonescontaining MAT gene sequences. Sequence analysis of four BAC clones confirmed that one BAC contained the true MAT1-2 idiomorph, while the otherthree contained fragments of both MAT genes in close proximity (8 to 1,075 bp) to each other. To investigate whether this holds true in the oppositemating type, we sequenced the identical regions in a MAT1-1 isolate. Sequence analysis confirmed that the MAT1-1 isolate contained one full MAT1-1idiomorph as well as additional regions that contained fragments of both MAT genes. In all cases, introns normally present in MAT genes were not presentin the MAT1-1-1 or MAT1-2-1 gene fragments in either mating type. Although heterothallism has been suggested to be the ancestral state in theDothideomycetes, the identification of fragments of both mating types in a single isolate suggests that homothallism is the ancestral state in C. beticola.685. Population structure from mountain to coast of two Lophodermium endophytes; a case study comparing rare and common fungal species in pineneedles. Ryoko Oono 1,2 , Laurel Koch 1 , A. Betsy Arnold 3 , Georgiana May 4 , François Lutzoni 1 , Ignazio Carbone 2 . 1) Department of Biology, Duke University,27th Fungal Genetics Conference | 289

FULL POSTER SESSION ABSTRACTSDurham, NC; 2) Department of Plant Pathology, North Carolina State University, Raleigh NC; 3) School of Plant Sciences, University of Arizona, Tucson AZ;4) Department of Ecology, Evolution, and Behavior, University of Minnesota, St. Paul, MN.Many foliar fungal endophytes are transferred horizontally among plant hosts and exhibit various degrees of host specificity. For example, endophyticLophodermium species (Rhytismatales) are commonly found in needles of certain pine species but infrequently in broad leaves of angiosperm hosts. Suchendophytes may have populations or subspecies that are genetically structured according to their host preference, climate regimes or geographic ranges,which might be associated with increased fitness for plants hosting specific mutualistic endophytic fungi. Hence, we set out to explore the populationstructure of Lophodermium spp. within mature foliage of loblolly (Pinus taeda) and Virginia pine (Pinus virginiana) in southeastern U.S., covering hostranges from the mountains of Appalachia to the coasts of the Atlantic. We analyzed the nuclear ribosomal internal transcribed spacer (ITS1/5.8S/ITS2) andthe intragenic spacer (IGS) regions as well as three protein-coding genes (actin, calmodulin, chitin synthase I) of the isolated Lophodermium strains. Thesein-depth population genetic analyses of endophyte species provide insight into the capacity of multilocus genetic markers that resolve on differentevolutionary time scales to capture sub-species level structure and to identify criteria for species delimitation. This multilocus analysis identified a rarecryptic species, which possibly explains the high genetic diversity of the Lophodermium spp. Genetic and geographic distance were correlated in the rarecryptic species, but not in the common species. We found no evidence for host species preference for the cryptic species vs. the common species. Ourfindings suggest that genetic exchange and recombination can be limited by dispersal if the species is rare.686. Fungal pathogen and endophyte genetics within the context of forest community dynamics. M.-S. Benitez 1 , M. H. Hersh 2 , L. Becker 1 , R. Vilgalys 3 , J. S.Clark 1,3 . 1) Nicholas School of the Environment, Duke University, Durham, NC; 2) Department of Biology, Eastern Michigan University, Ypsilanti, MI; 3)Department of Biology, Duke University, Durham, NC.Fungal pathogens play important roles in forest community dynamics, particularly through negative-density dependent regulation. Negative-densitydependence regulation is hypothesized to be regulated by the presence of host-specific pathogens. Studies on forest pathogens, however, indicate thepredominance of generalist seedling pathogens, capable of infecting more than one host species. To understand the mechanisms through which“generalist” pathogens contribute to forest-community dynamics we conducted extensive surveys of seedling pathogens in temperate hardwood forestsof the eastern U.S.A. Species in the genera Colletotrichum and Ilyonectria were among the most commonly isolated and recovered amplicon sequencefrom seedlings of multiple host species showing disease symptoms. Further, co-infection by both Colletotrichum and Ilyonectria species decreases hostsurvival, as quantified by posterior model probabilities. To investigate molecular mechanisms associated with multi-host generalism and co-infection, andto determine whether these “generalist” pathogens are distinct species or species-complexes, the genomes of three common species in our dataset (e.g.C. fioriniae, C. gloesporoides and Ilyonectria europea) were sequenced. The largest genome of the three belonged to Ilyonectria at 63.66 Mb, which alsocontained the highest number (22,250) of genes. The smallest genome belonged to C. fioriniae with 50.04 Mb and 15,777 genes. Genome size and numberof predicted genes appears expanded, confirming their role as seedling pathogens. For instance, three out of four polysaccharide lyase (PL) enzymedomains found in fungal genomes, are enriched in these three species. PL enzymes are relevant in plant pathogenicity since they may contribute to initialstages of host penetration. The genome sequence of these fungal groups will serve as a reference set for population level studies to address hostspecificityand local adaptation within our isolate database.687. Discovery of Sexual Reproduction in the Black Aspergilli. Heather L. Darbyshir 1 , Peter JI. van de Vondervoort 2 , Paul S. Dyer 1 . 1) School of Biology,University of Nottingham, Nottingham, NG7 2RD United Kingdom; 2) DSM Biotechnology Center, PO Box 1, 2600 MA Delft, The Netherlands.The black aspergilli are members of the genus Aspergillus that are typically characterized by the production of dark or black asexual conidia (classified assection Nigri). The group includes Aspergillus niger, which is of particular industrial importance because of its safe use status and ability to produce a widerange of enzymes and organic acids. All members of the black aspergilli have previously only been known to reproduce by asexual means. However, as aresult of combined molecular and cultural experimental studies it can now be revealed that at least one member of the black aspergilli, Aspergillussclerotiicarbonarius, is able to complete a sexual cycle. Wild type isolates of A. sclerotiicarbonarius were found to retain the ability to form sclerotia,structures associated with both dormancy and sexual reproduction, and strains of complementary MAT1-1 and MAT1-2 could be identified based on thepresence of mating-type genes. Crossing strains of opposite mating type, and an extended period of incubation, resulted in the production of sclerotiacontaining multiple ascocarps, with asci and viable ascospores, within the matrix of a sclerotium. This is consistent with past studies of phylogeneticallyrelated species in the Aspergillus section Flavi (teleomorph genus Petromyces). Progeny analysis is being undertaken based on data arising fromcomparative genome sequencing of parental isolates, mating-type distribution and phylogenetic analysis. The discovery of a heterothallic sexual cycle in A.sclerotiicarbonarius provides insights into the evolution of asexuality in the black aspergilli. It is hoped that ongoing molecular genetic studies into theearly sexual morphogenesis may provide an insight into the regulation of sexual reproduction in the black aspergilli.688. Culture-based survey of soil fungi from bat hibernacula. Jeffrey M. Lorch 1 , Daniel L. Lindner 2 , Andrea Gargas 3 , Laura K. Muller 4 , Andrew M. Minnis 2 ,David S. Blehert 4 . 1) University of Wisconsin - Madison, Madison, WI, USA; 2) US Forest Service, Northern Research Station, Center for Forest MycologyResearch, One Gifford Pinchot Drive, Madison, WI, USA; 3) Symbiology LLC, Middleton, WI, 53562, USA; 4) US Geological Survey - National Wildlife HealthCenter, Madison, WI, USA.Bat white-nose syndrome (WNS), a fungal disease now spreading in eastern North America, is causing unprecedented mortality among hibernating bats.To investigate fungal communities present in bat hibernacula, we identified culturable fungi present in soil samples from 24 bat hibernation sites in theeastern United States. Isolates were characterized by sequencing regions of ribosomal DNA (internal transcribed spacer and partial intergenic spacer). Weisolated Geomyces destructans from soil samples collected in hibernacula within the known range of WNS, and we found a wide diversity of Geomycesspecies, comprising around one third of all isolates. Many of these Geomyces species, along with numerous potentially novel lineages, appear to beundescribed.Other Topics689. Understanding the cellular basis of Azole resistance in Aspergillus fumigatus. Michael J. Bromley, Marcin Fraczek, Rebecca Collins, Emma Davies,Paul Bowyer. Translational Med, Univ Manchester, Manchester, United Kingdom.Resistance of Aspergillus fumigatus to the azole class of antifungals is becoming a major problem in Europe and is being driven by two factors, theprolonged exposure (several months to several years) of patients to azoles and the extensive use of agricultural azoles driving environmental resistance.Our understanding of the mechanisms that govern azole resistance in filamentous fungi is limited. While some clinical resistant isolates harbor mutations290

FULL POSTER SESSION ABSTRACTSin the azole target, lanosterol 14 a-demethylase (cyp51A), more than 50% do not. We have used a combination of whole genome sequencing,transcriptomics, transposon based mutagenesis and high throughput directed mutagenesis to identify novel mechanisms that may explain the resistanceobserved in these strains. I will summarize and discuss our progress to date and present a worrying mechanism that results in both pan-azole andamphotericinB resistance.690. Chemically Induced Haploinsufficiency Screens to Identify Drug Mechanism of Action in Aspergillus Fumigatus. D. A. Macdonald 1 , A. E. Johns 1 , M.Eberle 2 , P. Bowyer 1 , D. Denning 1 , M. J. Bromley 1 . 1) Institute of Inflammation and Repair, Respiratory & Allergy Centre, University of Manchester,Manchester, United Kingdom; 2) Applied Microbiology, Institute for Applied Life Sciences, University of Karlsruhe, Hertzstrae 16, 76187 Karlsruhe,Germany.Current drugs used to treat Aspergillus infections are limited and suffer from a variety of shortcomings including low efficacy, toxicity and increasingresistance. Despite the discovery of numerous promising drug targets, few lead compounds have been discovered by target based approaches. This can beexplained, in part, by the ‘druggability’ of a target as some compounds which demonstrate promising activity against an enzyme are not active against thewhole cell or are toxic to humans. Consequently most of the antimicrobials presently on the market were originally discovered by random screening ofcompounds against whole cell screens. A solution to this problem is to identify gene targets utilizing compounds that already show antifungal activity andhave clean toxicity profiles.Chemical genetic profiling aids identification of drug mechanism of action as a diploid strain lacking a single copy of a drug’s target is hypersensitive tothat drug. Heterozygote S. cerevisiae and C. albicans libraries have been used to identify the mechanism of action of several promising compounds;however, this has been hindered in A. fumigatus by the complexity in generating an adequate set of heterozygous strains. A high-throughput targetedgene KO method for A. fumigatus has been established by employing fusion-PCR to generate targeted gene disruption cassettes, optimizing the commontransformation protocol for A. fumigatus high-throughput gene disruption, and utilising a diploid Ku80 - /Ku80 - mutant to facilitate more reliablehomologous recombination. Preliminary efforts have produced 46 heterozygous KO strains and subsequently, the feasibility of chemical genetichaploinsufficiency studies in filamentous fungi has been demonstrated with several compounds. High-throughput methods of chemical genetic profiling bypooling multiple heterozygous KO strains into a single culture is currently being validated and preliminary data is promising. This will enable highthroughputmethods for surveying the genome of A. fumigatus for new drug targets and supports unveiling the mechanisms of action of antifungal drugs.691. Antifungal Pisum sativum defensin 1 Induces a non-Apoptotic Death in Aspergillus nidulans. Caroline M Fernandes 1 , Luciano N Medeiros 1 , Landi VGCostilla 1 , Hilda Petrs-Silva 1 , Patrícia A de Castro 2 , Gustavo H Goldman 2 , Eleonora Kurtenbach 1 . 1) Federal University of Rio de Janeiro, Rio de Janeiro, Rio deJaneiro, Brazil; 2) Sao Paulo University, Ribeirao Preto, Sao Paulo, Brazil.Psd1 is a basic, cysteine-rich plant defensin isolated from Pisum sativum seeds which inhibits the growth of a broad range fungi species. Defensins arealso non-toxic to mammalian cells, highlighting their potential as antifungal drugs. We have shown that FITC-labelled Psd1 was internalized in F. solanihyphae, interacting with cyclin F and leading to fungal cell cycle arrest. This internalization seemed to be dependent of glucosylceramide (CMH, ofcerebroside monohexoside), once C. albicans cells lacking the ceramide synthase are 25 % less susceptible to Psd1 than the parental strain. Fungal andmammalian CMH are structurally divergent, as the former presents a C8-unsaturation and C9-methylation on the sphingoid base, which could possiblydrive Psd1 selectivity. In this work, we investigated the cell death mechanisms triggered by Psd1 in Aspergillus nidulans and the contribution of CMHstructure to Psd1-induced fungal death. We characterized, through fluorescence microscopy, several apoptotic events, such as intense formation ofreactive oxygen species (ROS), metacaspase activation and DNA strand breaks. Although A. nidulans hyphae treated with 20 mM Psd1 for 24 hoursexhibited severe cell injury, no apoptosis-phenotype was observed. We also investigated whether Psd1 incubation would lead to membranepermeabilization typical of a necrotic death. To this, A. nidulans cells were maintained in the presence or absence of the peptide and the membranedamage was evaluated through Propidium Iodide (PI) staining. We observed 15 % PI positive cells in the suspension treated with Psd1, in contrast to 2 % incontrol culture. To investigate the role of fungal CMH and its structural modifications to Psd1-induced cell death, we constructed strains lacking theglucosylceramide synthase (ANID_08806), sphingolipid DD8-desaturase (ANID_04592) and sphingolipid C9-methylase (ANID_05688 and ANID_07375)genes. Phenotype analysis showed impaired growth of strains deficient in ANID_08806 and ANID_04592 in comparison to the parental strain. Furtherinvestigation will be conducted to characterize Psd1 antifungal activity and apoptosis or necrosis induction in the mutant strains. Unraveling themechanisms of cell death induced by antifungal peptides may lead to the identification of new targets that drive antimycotic selectivity.692. Is fungal secondary metabolism regulated by competing insects? Annika Regulin 1 , Nancy Keller 2 , Frank Kempken 1 . 1) Department of Botany,Christian-Albrechts University, Kiel, Germany; 2) Department Medical Microbiology and Immunology, Dept of Bacteriology, UW-Madison, USA.Fungi synthesize an astonishing variety of secondary metabolites, some of which belong to the most toxic compounds in the living world. Even thoughlittle is known about the benefit of these metabolites, the ability to regulate the secondary metabolism might be seen as an evolutionary adaptation.Presumably fungi regulate secondary metabolites (e.g. mycotoxin) in response to confrontation with natural competitors like insects to guarantee efficientexploitation of environmental resources (1-3). Admittedly it should be mentioned that secondary metabolites are not the only defence mechanisms offungi (4). In order to enlighten the biological function of these secondary metabolites with reference to chemical defence reactions of insect-fungalinteractions, we utilized complementary approaches of experimental ecology and functional genomic techniques. The vinegar fly Drosophila melanogasterand its natural antagonist Aspergillus nidulans are used as an ecology model system. To analyse fungal up- or down regulated target genes in theinteraction of A. nidulans with Drosophila larvae microarray analysis was performed. This led to the identification of secondary metabolite genes up- ordown-regulated under these conditions. Quantitative RT-PCR was employed to analyze secondary metabolite gene expression at different time points.Fungal single, double and triple mutations of identified up-regulated genes are currently analyzed in confrontation assays to identify potentialmodifications in gene expression and the survival rate of larvae concerning to chemical defense reaction of fungus-insect interaction compared to wildtype. This could reveal insights about the biological function of secondary metabolite genes and clusters such as stc and mdp.(1.) Rohlfs, M., Albert, M., Keller, N. P., and Kempken, F. (2007) Biol Lett 3, 523-25. (2.) Kempken, F., and Rohlfs, M. (2010) Fungal Ecol 3, 107-14. (3.)Rohlfs, M., Trienens, M., Fohgrub, U., and Kempken, F. (2009) in "The Mycota XV. (Anke, T., Ed.), Springer Heidelberg, New York, Tokyo, pp. 131-51 (4.)Kempken, F. (2011) Mol Ecol 20, 2876-77.693. Eisosome distribution and localization in the meiotic progeny of Aspergillus nidulans. A. Athanasopoulos 1 , H. Boleti 2 , C. Scazzocchio 3 , V.Sophianopoulou 1 . 1) Institute of Biosciences and Applications, Microbial Molecular Genetics Laboratory, National Center for Scientific Research,Demokritos (NCSRD), Athens, Greece; 2) Intracellular Parasitism Group, Molecular Parasitology Laboratory, Department of Microbiology and Light27th Fungal Genetics Conference | 291

FULL POSTER SESSION ABSTRACTSMicroscopy unit, Institut Pasteur Hellenique, Athens, Greece; 3) Department of Microbiology, Imperial College, London, United Kingdom.In the model filamentous fungus Aspergillus nidulans, PilA and PilB, two homologues of the Saccharomyces cerevisiae eisosome proteins Pil1/Lsp1, andSurG, a strict orthologue of Sur7, are assembled and form tightly packed structures in conidiospores. A. nidulans differs from the Saccharomycotina in thatit has the ability to reproduce through two different types of spores, conidiospores and ascospores, cells which have a radically different morphology andare formed through completely different developmental pathways. Ascospores are formed only after the completion of meiosis inside asci, conidiosporesarise from mitotic budding of specialized cells (phialides). We thus investigated eisosome composition and distribution in ascospores. Our results showthat core eisosome proteins PilA, PilB and SurG are not expressed in hülle cells or early ascospores, but are expressed in mature ascospores. PilA formsstatic punctate structures at the plasma membrane as does PilB (with higher concentration at the areas where the two halves of ascospores are joinedtogether), while SurG was localized both at the membrane and perinuclearly. In germlings originating from ascospores the punctate structures wereshown to be composed only of PilA. In germinated ascospores PilA foci did not colocalise with the punctate structures of AbpA, a marker for sites ofclathrin-mediated endocytosis. In the presence of myriocin -a specific inhibitor of sphingolipid biosynthesis- PilA-GFP foci of ascospore germlings were lessnumerous and their distribution was significantly altered. In this study we also investigated one of the two A. nidulans orthologues of Nce102, a proteinthat determines the structure and function of membrane microdomains in S. cerevisiae. In quiescent conidia localization of the closest orthologue,AnNce102 is detected in PilA plasma membrane associated foci as well as in 3-5 round-shaped intracellular structures. In early hyphae, a cytoplasmicfraction of Nce102 is additionally detected in highly dynamic structures that resemble Golgi equivalents. Deletion of core eisosomal genes causesmislocalization of Nce102 from the plasma membrane to these cytoplasmic structures. Ongoing experiments are investigating AnNce102 localization inresponse to sphingolipid biosynthesis and the nature of the intracellular compartments where it is located.694. Fungal-bacterial interactions: Bacillus subtilis forms biofilm on Aspergillus niger hyphae. Isabelle Benoit 1,2 , Marielle H. van den Esker 3 , MiaomiaoZhou 1 , Oscars P. Kuipers 3 , Ronald P. de Vries 1,2 , Ákos T. Kovács 4 . 1) Fungal Physiology, CBS-KNAW, Utrecht, Utrecht, Netherlands; 2) Microbiology & KluyverCentre for Genomics of Industrial Fermentation, Utrecht University, Utrecht, The Netherlands; 3) Molecular Genetics Group, Groningen BiomolecularSciences and Biotechnology Institute, University of Groningen, Groningen, The Netherlands; 4) Terrestrial Biofilms, Institute of Microbiology, FriedrichSchiller University, Jena, Germany.Pure cultures of the filamentous fungi Aspergillus niger and Aspergillus oryzae and the Gram-positive bacterium Bacillus subtilis are widely used inindustry for the production of pharmaceuticals, food ingredients and enzymes. However both by design and by accident, industrial fermentation can alsoinvolve mixed populations of micro-organisms. Moreover, in natural biotopes, these organisms live in mixed communities and have complex interactionsranging from competition to symbiosis. B. subtilis, in specific conditions, is capable of forming beneficial biofilms on surfaces and interfaces from plantroots to metal surfaces. In this study, co-cultivations of A. niger and A. oryzae together with B. subtilis were performed. A. oryzae inhibits B. subtilis growthwhile a bacterial coating was observed on A. niger hyphae. Microscopic and transcriptomic approaches were combined to study this fungal-bacterialinteraction example.695. Co-cultivations of fungi: microscopic analysis and influence on protein production. Isabelle Benoit 1,2 , Arman Vinck 2 , Jerre van Veluw 2 , ThijsGruntjes 1,2 , Han A.B. Wösten 2 , Ronald P. de Vries 1,2 . 1) Fungal Physiology, CBS-KNAW, Utrecht, Utrecht, Netherlands; 2) Microbiology & Kluyver Centre forGenomics of Industrial Fermentation, Utrecht University, Utrecht, The Netherlands.During their natural life cycle most fungi encounter other microorganisms and live in mixed communities with complex interactions, such as symbiosis orcompetition. Industrial fermentations, on purpose or by accident, can also result in mixed cultures. Fungal co-cultivations have been previously describedfor the production of specific enzymes, however, little is known about the interactions between two species that are grown together. Aspergillus niger andAspergillus oryzae are two of the most important industrial fungi worldwide and both have a long history of strain improvement to optimize enzyme andmetabolite production. We have co-cultivated the wild type strains of these two Aspergilli with each other as well as the XlnR knock out strains. XlnR is atranscription factor inducible by the presence of xylose and responsible for the regulation of a variety of genes encoding plant polysaccharide degradingenzymes. The morphology and mechanism of the interaction of these cultures on wheat bran is addressed using microscopy and proteomics.696. Improving heterologous protein production in Aspergillus vadensis . Ourdia Bouzid 1,2 , Ronald P. de Vries 1,2 . 1) Microbiology & Kluyver Centre forGenomics of Industrial Fermentation, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands; 2) CBS-KNAW Fungal Biodiversity Centre,Uppsalalaan 8, 3584 CT, Utrecht, The Netherlands.Aspergillus vadensis is a good candidate for heterologous protein production, because it produces very low levels of extracellular proteases and does notacidify the medium. To improve protein production in A. vadensis two strategies were tested: (i) identification of new promoters for high gene expression,and (ii) overexpression of the xylanolytic regulator, XlnR. Six new A. niger constitutive promoters were selected and compared to the gpdA promoter usingan arabinofuranosidase (abf) encoding gene from Fusarium oxysporum as a reporter. Several of the new promoters resulted in higher Abf activity thangpdA. For the second strategy, A. vadensis was transformed with xlnR, xlnD (encoding beta-xylosidase) and faeA (encoding feruloyl esterase) alone, andwith combinations of xlnD and xlnR, and faeA and xlnR. Southern blot profiles confirmed the presence of multiple copies of the genes in the transformants.XlnD and FaeA activities were measured and were compared to the control strain. This demonstrated that increased copy numbers of faeA and xlnD had amuch larger effect on the corresponding activities than increased copy numbers of xlnR. These data demonstrate that the new promoters in combinationwith high copy integration of the target genes can result in higher protein production by A. vadensis. Highlights from this study will be presented.697. Production and characterization of esterases from Chaetomium thermophilum and their applicability in biomass conversion. Xiaoxue Tong, PeterBusk, Morten Grell, Lene Lange. Section for Sustainable Biotechnology, Department of Biotechnology, Chemistry and Environmental Engineering. AalborgUniversity Copenhagen, Denmark.Xylan is the dominating hemicellulose constituent of plants and the most abundant renewable polysaccharide in nature after cellulose. Xylan and itshydrolysis products are potential resources for nutraceuticals, cosmetics, foods, bioalcohol, and industrial fine chemical production. Feruloyl esterase andacetyl xylan esterase are required for complete enzymatic hydrolysis of xylan due to its highly heterogeneous nature. The aim of this study was to produceand characterize esterases from the thermophilic fungus Chaetomium thermophilum. The esterase genes were identified by a novel bioinformatics toolPPR (Peptide pattern recognition, Busk & Lange, 2011). A Feruloyl esterase gene (CtFaeA) and a xylan esterase gene (CtAxeA) were successfully expressedin the yeast Pichia pastoris. They were purified to homogeneity from the culture supernatants. The effect of temperature and pH on the activity andstability of the esterases, as well as their substrate specificities, were studied. Both CtFaeA and CtAxeA displayed broad thermal stability and pH stability.Moreover, both esterases were active on hydrolysis of wheat arabinoxylan. These results show that Chaetomium thermophilum has a high capacity fordegradation of xylan in addition to its already described cellulolytic potential. Furthermore, the robust esterases from Chaetomium thermophilum have292

FULL POSTER SESSION ABSTRACTSpotential application in biomass bioconversion to e.g. higher value chemicals or biofuels.698. Antioxidant adaptation by Eugenol and its derivatives and their affect on the expression of virulence in candida species. Aijaz Ahmad, NikhatManzoor. Bioscience, Jamia Millia Islamia, New Delhi, Delhi, India.Present work investigates the antifungal activity and mode of action of eugenol (EUG), and its three derivatives- methyl eugenol, thymol and carvacrol.EUG and its derivatives were tested for antifungal activity by standard methods of CLSI. These varied in their mechanism of action depending upon theperiod of exposure. Short exposures of 5-15 minutes resulted in reduced H+ efflux by the H+-pump. From our studies we conclude that EUG and itsderivatives induce production of free radicals which stimulates the enzyme SOD. An increased SOD activity resulted in an increase in the concentration ofH2O2 which further stimulates the peroxide eliminating enzyme, primarily GPx. It is noteworthy that the levels of GSH an essential substrate of GPx weredrastically reduced by the test compounds and this reduction gets even greater as increased levels of H2O2 decrease the activity of G6PDH which providesreducing equivalents to GR, an enzyme that recycles GSH from GSSG. Decreased G6PDH activity aids further in the reduction of GSH. Again, reducedavailability of GSH explains decreased GPx activity. Another enzyme to eliminate H2O2 is catalase, which triggers a cellular response leading to an increasein its activity. Hence increase in the activity of two important antioxidant enzymes SOD and catalase, clearly demonstrates an increase in the concentrationof ROS when the Candida were exposed to the EUG and derivatives. These enzymatic responses were not enough to defend the cell completely against ahigh rise in ROS and therefore did not meet the required cellular antioxidant demand. Ultimately, the outburst of free radical production led to severe lipidperoxidation. Cell death on exposure to EUG and its derivatives hence may be due to (i) decrease in the rate of H+efflux (ii) reduced ergosterol content (iii)Induction of oxidative stress in the cell (iv) These processes impair membrane structure and function which form lesions. Infection process of Candida ischaracterized by crucial pathogenicity markers. The process of germ tube induction followed by the secretion of hydrolytic enzymes help in the invasion ofthe host cells. The expression profile of selected genes associated with Candida virulence by RT-PCR showed a reduced expression of HWP1, SAP1 andPLB2 genes in Candida treated with EUG and its derivatives.699. Elevation of chitin is linked with multiparallel mechanisms in response to C. albicans cell wall stress. F. Nogueira, L. Walker, C. Munro, N. Gow.Institute of Medical Sciences, University of Aberdeen, Aberdeen, United Kingdom.The role of the MAPK, Ca 2+ /calcineurin and cAMP/PKA signal transduction pathways in regulating the Candida albicans cell wall stress response wasinvestigated. A library of mutants lacking receptors, signalling elements and transcription factors were screened for alterations in the ability to respond toa range of cell wall stressing agents, including CaCl 2, Calcofluor White and caspofungin. Pre-treatment of wild-type cells with CaCl 2 and CFW, activates theCa 2+ /calcineurin and PKC pathways, leading to an increase in chitin content, and reduced susceptibility to caspofungin. Although elevation of cell wall chitincontent often resulted in decreased sensitivity to caspofungin, we show here that some strains with increased chitin levels remained sensitive tocaspofungin. The results show that elevation of chitin is a common property of a range of mutants that are affected in coordinating cell wall stresspathways, but that multiple mechanisms are likely to operate in maintaining the robustness of the C. albicans cell wall.700. Prezygotic and postzygotic control of uniparental mitochondrial DNA inheritance in Cryptococcus neoformans. Rachana Gyawali, Xiaorong Lin.Biology, Texas A&M University, College Station, TX.Uniparental inheritance of mitochondrial DNA is pervasive in non-isogamic higher eukaryotes during sexual reproduction and postzygotic and/orprezygotic factors are shown to be important in ensuring such inheritance pattern. Although the fungus Cryptococcus neoformans undergoes sexualproduction with isogamic partners of opposite mating types a and a, most progeny derived from such mating events inherit the mitochondrial DNA fromthe a parent. The homeodomain protein complex Sxi1a/Sxi2a, formed in the zygote after a-a cell fusion, was previously shown to play a role in thisuniparental mtDNA inheritance. Here, we defined the timing of the establishment of the mtDNA inheritance pattern during the mating process anddemonstrated a critical role in determining the mtDNA inheritance pattern by a prezygotic factor Mat2. Mat2 is the key transcription factor that governsthe pheromone sensing and response pathway, and it is critical for the early mating events that lead to cell fusion and zygote formation. We show thatMat2 governs mtDNA inheritance independent of the postzygotic factors Sxi1a/Sxi2a, and the cooperation between these prezygotic and postzygoticfactors help achieve stricter uniparental mitochondrial inheritance in this eukaryotic microbe.701. SIS, a sex genome defense mechanism operating in Cryptococcus neoformans. Xuying Wang, Sabrina Darwiche, Joseph Heitman. Department ofMolecular Genetics and Microbiology, Duke University Medical Center, Durham, NC.Cryptococcus neoformans is a human fungal pathogen that undergoes a dimorphic transition from yeast to hyphae during a-a opposite-sex mating and a-a unisexual reproduction (same-sex mating). Infectious spores are generated during both processes. We previously identified a sex induced silencing (SIS)pathway in the C. neoformans serotype A var. grubii lineage, in which tandem transgene arrays trigger RNAi-dependent gene silencing at a high frequencyduring a-a opposite-sex mating, but at an ~250-fold lower frequency during asexual mitotic vegetative growth. Here we report that SIS also operatesduring a-a unisexual reproduction. A self-fertile strain containing either SXI2a-URA5 or NEO-URA5 transgene arrays exhibited an elevated silencingfrequency during solo and unisexual mating compared with mitotic vegetative growth. We also found that SIS operates at a similar efficiency on transgenearrays of the same copy number during either a-a unisexual reproduction or a-a opposite-sex mating. URA5-derived small RNAs were detected in thesilenced progeny of a-a unisexual reproduction and RNAi core components were required, providing evidence that SIS induced by same-sex mating is alsomediated by RNAi via sequence-specific small RNAs. This study, together with our previous finding of SIS in a-a opposite-sex mating of the C. neoformansserotype A var. grubii lineage, demonstrates that SIS is a conserved process between the divergent C. neoformans serotype A and serotype D siblingspecies. In each case, our data show that the SIS RNAi pathway operates to defend the genome via squelching transposon activity during the sexual cycles.Thus, our discovery of SIS brings a fresh perspective to meiotic silencing involving the upregulation of RNAi pathways as a strategy to guard genomicintegrity during sex. More importantly, the presence of SIS in both a-a unisexual reproduction and a-a opposite-sex mating indicate that SIS may betriggered by the shared pheromone sensing Cpk1 MAPK signal transduction cascade. Ongoing studies focus on defining at a mechanistic level how the SISRNAi pathway is initiated, including identifying new components involved in SIS.702. Effects of the use of biocontrol agent (Phlebiopsis gigantea) on fungal communities of Picea abies stumps. E. Terhonen 1 , H. Sun 1 , M. Buée 2 , R.Kasanen 1 , L. Paulin 3 , F. Asiegbu 1 . 1) University of Helsinki, Department of Forest Sciences, P.O.Box 27, FIN-00014, University of Helsinki, Finland; 2) INRA,UMR 1136 INRA/Nancy Université Interactions Arbres/Microorganismes, INRA-Nancy, 54280 Champenoux, France; 3) DNA Sequencing and Genomics Lab,Institute of Biotechnology, University of Helsinki, P.O. Box 56, FIN-00014 Helsinki, Finland.The saprotrophic fungus Phlebiopsis gigantea has for several years been used as a biocontrol agent against pathogen Heterobasidion annosum. Thispathogen is the major cause of root rot disease in conifers that results in economic losses estimated at 50 million euros to Finnish forestry. A major27th Fungal Genetics Conference | 293

FULL POSTER SESSION ABSTRACTSproblem is that, although the effectiveness of P. gigantea as a bio-control agent has empirically been shown, the long term biological effect of this funguson conifer trees as well as on other soil micro-flora has not been empirically proven. We investigated the impact of P. gigantea treatment on stumpmycobiota using metagenomic pyrosequencing approach as this has not been done before. Samples from forest sites pre-treated with P. gigantea for 1, 6and 13 years ago were collected, DNA was isolated and pyrosequenced. Similarly samples were also collected from untreated stumps within the sameforest sites. Sequences were quality trimmed using Mothur software. After trimming we had 26 398 sequences from 53 117. For the extraction of the fullITS1 of the nuclear ITS region FungalITSextractor was used and these sequences were clustered at 97% similarity using cdhit-454 with the most abundantsequence types serving as cluster seeds. The most frequent sequence type in each cluster was used for the BLAST searches against NCBI BLASTN and ³ 97%similarity across the entire length of the pairwise alignment was taken to indicate conspecificity. Differences between control and treated stumps weretested statistically with Paired-Sample T-test (SPSS 19). Also diversity indexes and similarity indexes between controls and treated were calculated usingEstimates 8.2.0. After one year of the clear-cut we found from Phlebiopsis gigantea-treated stumps 107 different fungal OTUs and from non-treatedstumps 119 fungal OTUs, from which they shared 102 OTUs. After 6 years we observed from treated stumps 118 fungal OTUs and from non-treated 134fungal OTUs and they shared 99 OTUs. After 13 years we found from treated stumps 131 OTUs and from non-treated 139 OTUs and shared OTU numberwere 109. However there were no statistical differences between control and treatment. Based on our results our primary conclusion is that stumptreatment should continue as there is no obvious adverse effect on the other stump mycobiota.703. Diversity of yeast and mold species in different cheeses. Nabaraj Banjara 1 , Kenneth Nickerson 2 , Heather Hallen-Adams 1 . 1) Food Science andTechnology, University of Nebraska-Lincoln, Lincoln, NE; 2) Biological Sciences, University of Nebraska-Lincoln, Lincoln, NE.The yeast and mold diversity from different commercial cheeses collected from local markets (Lincoln, NE, USA) was studied using microbial counting andmolecular biology approaches. Twenty one distinct types of cheese samples were investigated. Briefly, 10 grams of each cheese sample was homogenizedin distilled water, serially diluted from 10 to 10 -6 , grown in Yeast Extract Glucose Chloramphenicol Agar (YGC) and the population was counted fromdilution plates. Yeasts and molds were identified by amplification and sequence analysis of the nuclear ribosomal RNA genes, using ITSIF and TW13primers. Debaryomyces hansenii was the predominant fungal species in most of the cheeses (59% of samples at up to 5.7 x 10 6 CFU/ gram). Other fungiisolated included D. fabryi, D. prosopidis, D. subglobosus, Penicillium roqueforti and Candida sake. In our samples, farmstead cheese had the highest (2.1 x10 8 CFU/gram) and Swiss cheese the lowest (8x10 3 CFU/gram) fungal population.704. Heterologous expression and characterization of soil organic matter-specific proteases secreted by the ectomycorrhizal fungus Paxillus involutus.Morten N. Grell 1 , Linas Pupelis 1 , Tomas Johansson 2 , Firoz Shah 2 , Lene Lange 1 , Anders Tunlid 2 . 1) Section for Sustainable Biotechnology, Department ofBiotechnology, Chemistry and Environmental Engineering, Aalborg University Copenhagen, Denmark; 2) Microbial Ecology Group, Department of Biology,Lund University, Sweden.Paxillus involutus (Batsch) Fr. (Basidiomycetes; Boletales) is widely distributed in the Northern hemisphere, and is one of the best-studiedectomycorrhizae fungi, especially with respect to its ecology and physiology. In a study on the mechanisms by which Paxillus involutus degrade complexorganic matter extracted from plant litter material, transcriptomes were sequenced using the 454 technology and NimbleGen microarrays produced. Anumber of genes encoding extracellular enzymes showed an increased transcript level during degradation of soil organic matter (SOM), as compared withgrowth on a defined medium (MNM). They were suggested to constitute a Fenton-like, radical-based biodegradation system that disrupts the organicmatter-protein complexes thereby mobilizing embedded nitrogen (Rineau et al. 2012, Environm. Microbiol. 14, 1477-1487). Supporting this, a number ofprotease genes were found to have a significantly increased SOM/MNM expression value and to be upregulated during growth on protein-rich substrates.Most highly expressed were aspartic, metallo, and serine proteases. This is in agreement with biochemical analysis of SOM degradation. To study substratespecificity and regulation of selected proteases in detail these are expressed in the yeast Pichia pastoris.705. Effector proteins in fungal defense against fungivorous nematodes: Targets and functional significance. Therese Wohlschlager 1 , StefanieSchmieder 1 , Alex Butschi 2 , Paola Grassi 3 , Alexander Titz 4 , Stuart Haslam 3 , Michael Hengartner 2 , Markus Aebi 1 , Markus Künzler 1 . 1) Institute of Microbiology,ETH Zürich, Switzerland; 2) Institute of Molecular Life Sciences, University of Zürich, Switzerland; 3) Division of Molecular Biosciences, Imperial College,London, United Kingdom; 4) Department of Chemistry, University of Konstanz, Germany.The defense of fungi against fungivores is largely based on the production of intracellular toxins. A significant proportion of these toxins are peptides andproteins that are synthesized by the ribosome and stored in the cytoplasm. Protein toxins include lectins that target specific glycoepitopes in the intestineof the fungivore upon ingestion and kill the fungivore by a yet unknown mechanism. In our laboratory, we focus on the functional characterization offungal protein toxins that are directed against nematodes. We use the model nematode Caenorhabditis elegans to identify the targets and to study thetoxicity mechanism of these fungal defense effector proteins in the nematode. In addition, we employ the fungivorous nematodes Aphelenchus avenaeand Bursaphelenchus willibaldi to study the diversity, the functional significance and the transcriptional regulation of these proteins in the fungus.Recently, we identified a nematotoxic lectin from the mushroom Laccaria bicolor that is homologous to animal lectins involved in innate immunity againstbacteria. We found that the nematotoxicity of the lectin is based on its specific binding to methylated fucose residues on nematode N-glycans. Amonganimals, this epitope is only present in worms and molluscs but not in insects or vertebrates. We performed affinity chromatography of C. elegans wholeworm protein extracts using the L. bicolor lectin and other nematotoxic fungal lectins recognizing protein-bound glycans. The results of this analysissuggest that these lectins target the same set of glycoproteins in the nematode intestine and may confer toxicity by a common mechanism. In order toaddress the functional significance of these proteins for fungal defense against fungivorous nematodes, we expressed some of the fungal proteinsdisplaying toxicity towards C. elegans, in the filamentous ascomycete Ashbya gossypii. These transformants were fed to A. avenae and the propagation ofthe fungivorous nematode on the various transformants was determined. Expression of some effector proteins significantly inhibited propagation of thenematode suggesting that these proteins have a role in fungal defense against these organisms. Experiments addressing the relative fitness of the variousA. gossypii transformants upon selective pressure of feeding by A. avenae are under way.706. Microfluidic platforms for monitoring interactions between fungi and bacteria. Martina A. Stöckli 1 , Claire E. Stanley 2 , Pauli T. Kallio 1 , MarkusKünzler 1 , Andrew J. deMello 2 , Markus Aebi 1 . 1) Microbiology and Immunology, ETH Zurich, Zurich, Switzerland; 2) Chemistry and Applied Biosciences, ETHZurich, Zurich, Switzerland.Bacteria and fungi share many microhabitats where they interact with each other. These interactions are diverse and play important roles in certainhuman infections, as well as in the biological control of plant diseases. The basis of these interactions, however, is not well understood, as theirinvestigation is technically challenging. Conventional microscopic approaches are limited, in that they can only monitor interactions at a particular time294

FULL POSTER SESSION ABSTRACTSpoint and are not able to follow dynamic interactions. Herein, the development of a microfluidic platform, to study the interplay of individual hyphae withbacteria in a confined compartment, is detailed. A microfluidic device, comprised of single, interconnected microchannels, is filled with medium. Thedevice is manufactured from the polymer, polydimethylsiloxane, which is bonded to a glass layer. Importantly, this polymer possesses some desirablecharacteristics, making it compatible with experiments of biological nature, for example it is oxygen permeable, and allows optical detection from 240 nmto 1100 nm. The fungus can easily grow into the microchannels from an agar plug that is placed next to a lateral opening. Subsequently, the device can beco-inoculated with bacteria through a separate inlet, allwoing changes in morphology, growth rate, and interaction patterns of the same hyphae upon theaddition of bacteria to be investigated over time. As an example, the interaction of the basidiomycetous model organism, Coprinopsis cinerea, and thegram-positive bacterium, Bacillus subtilis, which are both found in dung of herbivores, has been studied. Using these microfluidic platforms, it has beenobserved that B. subtilis cells attach to hyphae in an end-on manner. The frequency of attachment within hyphae and between different hyphae varies.Furthermore, the tips of the hyphae are not colonised by the bacterial cells. In summary, this approach can be used to study phenotypic changes ofbacteria and fungi over specific time periods. In future, it is envisaged that this data will be combined with fungal and bacterial genetic approaches.707. WITHDRAWN708. The French Fusarium Collection: a living resource for mycotoxin research. L. Pinson-Gadais, M. Foulongne-Oriol, N. Ponts, C. Barreau, F. Richard-Forget. INRA, UR1264-MycSA, 71 avenue Edouard Bourlaux, F-33883 Villenave d’Ornon, France.Fusaria are responsible for prejudicial diseases on cereal crops worldwide, such as crown rot and Fusarium head blight. Beyond economic losses due toinfection symptoms, these pathogens can produce several types of mycotoxins that are harmful to livestock and humans. They are extremely diverse atthe intra-specific levels in terms of types as well as quantities of toxins that a strain can produce. Developing appropriate strategies to limit contaminationwith Fusarium mycotoxins requires a greater knowledge about this variability. We have collected a large number of toxinogenic Fusarium strains. Ourassortment now includes about 800 strains, mostly from the species graminearum, culmorum, verticilloides, proliferatum, and temperatum. Species wereidentified based on morphology and real-time PCR. More than half of our strains were further characterized for toxin production using biochemical and/orreal-time PCR-based tools. We isolated about 70 F. graminearum strains from either wheat or maize grains originating from different French cerealproduction areas. Our results show a high representation of 15-acetyldeoxynivalenol-producing strains in our French samples. Within the samechemotype, we observe a large variability in toxin production levels. The F. graminearum strains were characterized with microsatellite markers and showa large genetic diversity. Two groups were delineated according to their genetic background, roughly corresponding to strains isolated from Europe in onehand and America in the other hand. Our results are also in agreement with the fact that only F. graminearum sensu stricto strains seem to be detected inFrance so far. The demonstrated genetic and phenotypic diversity provides a sound ground for countless downstream studies such as genetic associationand quantitative genetics to understand the determinism of toxin production. Such information should be doubtlessly considered in plant breeding effortsand other disease management strategies aimed at reducing the mycotoxin risk in food and feeds. Our collection is a valuable tool to improve ourunderstanding of toxigenic diversity in Fusarium species. It is managed through a database gathering all information collected on each strain, alreadyavailable upon request and soon publically available as a web-based interface.709. Chemical genetics: Discovery of novel fungicides and their targets in the phytopathogen Fusarium graminearum. G. Subramaniam, C. Mogg.Agriculture Canada, Ottawa, ON, Canada.Chemical genetics screen is based on the ability of small chemical molecules to bind to biological molecules and alter their function. Screening ofpharmaceutical libraries has revealed novel molecules effective against cancer and other diseases. We have adopted similar approach and identify bioactivecompounds that will block the growth and development of F. graminearum. We have developed a 96-well format to monitor the growth of F.graminearum in liquid media. The fungus is tagged with a green fluorescent protein (GFP) and the growth is monitored by the measurement offluorescence of the GFP. This format facilitates high throughput screening for small molecules that could potentially disrupt the growth of the fungus. Asproof of concept, we screened ~560 compounds from the TimTec NDL-3000 natural product collection (TimTec LLC, Newark, DE, USA) and identifiedseveral compounds with anti-Fusarium properties. One compound identified form our screen, “Antofine” was purified from Vincetoxicum rossicum andwas used in subsequent studies, to identify targets in the fungus. We used the gene deletion library of the budding yeast Sacchromyces cerevisiae toidentify targets for Antofine. GeneMANIA (http://www.genemania.org), an online multiple association network integration algorithm was used to uncoverinformation pertaining to genetic and physical interactions of these targets. Our efforts to identify targets in Fusarium against Antofine will be discussed.710. Functional characterization of an Aspergillus flavus polyketide synthase gene necessary for the synthesis of a sclerotium-specific pigment. J.W.Cary 1 , P. Harris-Coward 1 , K.C. Ehrlich 1 , P. Dowd 2 , S. Shantappa 3 , A.M. Calvo 3 . 1) US Department of Agriculture, ARS-SRRC, New Orleans, LA; 2) USDepartment of Agriculture, ARS-NCAUR, Peoria, IL; 3) Northern Illinois University, DeKalb, IL.The filamentous fungus, Aspergillus flavus, produces the toxic and carcinogenic, polyketide-derived family of secondary metabolites termed aflatoxins(AFs). In addition to the AF biosynthetic gene cluster, analysis of the A. flavus genome has identified 55 gene clusters predicted to be associated withsecondary metabolism. To date, very few of the metabolites produced by these clusters have been identified. Secondary metabolism is controlled byglobal regulators such as LaeA and VeA. In a veA knockout mutant we identified a significantly down-regulated polyketide synthase (PKS) gene belongingto cluster 27. Although the metabolite produced by this cluster was unknown, in silico cluster analysis predicted that cluster 27 would consist of the PKSgene and four other genes. qRT-PCR analysis confirmed that expression of the cluster 27 PKS (pks27) gene was down-regulated in the veA mutant.Inactivation of the pks27 gene resulted in loss of the dark pigment associated with A. flavus sclerotia. Sclerotia are survival structures produced bycondensation of mycelia and function as propagules in the field. Conidial pigmentation did not appear to be affected in the pks27 knockout strain. TLC andHPLC analysis of sclerotial extracts identified the cluster 27 metabolite as asparasone A. Insect feeding studies using wild-type and mutant sclerotiaindicated that the pigment may be acting as a feeding deterrent. To our knowledge this is the first report on the identification of a gene that encodes asclerotium-specific pigment. The pigment likely plays a role in sclerotial resistance to insect feeding and possibly other environmental stresses.711. Functional Analysis of the Pleurotus ostreatus Manganese-Peroxidase Gene Family. Tomer Salame, Doriv Knop, Dana Levinson, Oded Yarden,Yitzhak Hadar. Microbiology and Plat Pathology, Hebrew Unversity, Rehovot, Israel.Mn amendment to P. ostreatus cultures enhances degradation of recalcitrant aromatic compounds. Manganese peroxidase (MnP) isoenzymes are keyplayers in these processes. The MnP gene family is comprised of five Mn -dependent peroxidases (mnp3, 6, 7, 8 and 9) and four versatile-peroxidases(mnp1, 2, 4 and 5; VPs). In liquid medium, Mn amendment resulted in a drastic up-regulation of the predominantly expressed mnp3 and mnp9, and downregulationof mnp4. To obtain direct evidence for the role of these enzymes, we produced genetically-modified (knockout, knockdown and/or over-27th Fungal Genetics Conference | 295

FULL POSTER SESSION ABSTRACTSexpression) strains in mnps and studied their degradation capacity. The compounds studied were: azo-dyes such as orange II and reactive black,recalcitrant pharmaceutical compounds found in treated waste water such as Carbamazepine and lignocellulosic agricultural waste. We engineered atransformant, constitutively expressing mnp4 a VP naturally repressed by Mn (designated OEmnp4) under the control of the b-tubulin promoter. Now,despite the presence of Mn in the medium, OEmnp4 produced mnp4 transcript as well as VP activity as soon as four days after inoculation. OEmnp4decolorized the azo-dyes two days earlier relative to the wild type in Mn amended medium. RNAi silencing targeting mnp3 resulted in a delay in thedecolorization capacity which occurred concomitantly along with a marked reduction of the expression level of all mnps, particularly mnp3 and mnp9. Thisobservation supported the conclusion that MnPs are involved in the process but could not determine the specific contribution of the different genes to theoutcome. Therefore we produced a Dku80 strain, exhibiting a 100% homologous DNA recombination rate, to enable specific gene replacement.Subsequently, homokaryon mnp2, 3, 4 and 9 knockout strains were produced. In Mn amended GP, orange II decolorization was not significantly inhibitedby any of these strains, indicating on functional redundancy. In Mn deficient GP, inactivation of mnp4 proved that it encodes the key VP responsible forMn dependent and Mn independent peroxidase activity, as well as resulted in reduction of the azo dye reactive black 5 decolorization capacity. The toolsand protocols developed increase the amenability of P. ostreatus to genetic manipulations and expand options for gene function analyses.712. Temperature- and pH characteristics of endo-cellulases in Rhizophlyctis rosea. Bo Pilgaard 1 , Frank Gleason 2 , Peter Busk 1 , Lene Lange 1 . 1) Section forsustainable biotechnology, Aalborg University- Copenhagen, Denmark; 2) School of Biological Sciences, University of Sydney, New South Wales, Australia.The zoosporic true fungi of the order Chytridiales are of special interest since they constitute a central root position of the entire fungal kingdom.Enzymes of the anaerobic zoosporic rumen fungi have been studied quite extensively, but only scarce information is available about enzymes from aerobiczoosporic true fungi from soil. We have previously confirmed the presence of cellulases in Rhizophlyctis rosea (AUS13), which was isolated from soils of theSydney Basin. Other studies have shown that this fungus can survive and grow within a wide pH-range (Gleason et. al 2010) and high temperatures(Gleason et. al 2005). We investigate the cellulolytic potential of Rhizophlyctis rosea in several cellulase assays and characterize the enzymatic propertieswith respect to temperature and pH stability of the enzymes. Moreover, we are sequencing the Rhizophlyctis rosea genome in order to find the genes forcellulose-degrading enzymes (GH6, GH7, GH45, GH6) that are present in Chytrids as compared to the cellulases of these families found in other fungalgroups.713. Applying unconventional secretion of the endochitinase Cts1 to export heterologous proteins in Ustilago maydis . J. Stock, P. Sarkari, M. Knopp, S.Jankowski, S. Bergmann, M. Feldbrügge, K. Schipper. Institute for Microbiology, Heinrich-Heine University Düsseldorf, 40204 Düsseldorf, Germany.The demand on the biotechnological production of proteins for pharmaceutical, medical and industrial applications is steadily growing. Not every proteincan easily be produced by the existing platforms. For the production of such challenging proteins, we aim to establish a novel expression system in the wellcharacterized eukaryotic microorganism Ustilago maydis. In this fungus, secretion of the endochitinase Cts1 depends on mRNA transport alongmicrotubules, which is mediated by the key RNA-binding protein Rrm4. We recently demonstrated that Cts1 secretion occurs via a novel unconventionalroute. We used b-glucuronidase (Gus) as a reporter for unconventional secretion. This bacterial enzyme is inactivated by N-glycosylation during its passagethrough the conventional eukaryotic secretory pathway. By contrast, in our system Gus was exported in its active form by fusion to Cts1 confirming itssecretion by an unconventional route. Furthermore, we showed that this secretory mechanism can be exploited for the export of active heterologousproteins. As a proof-of-principle for economically important biopharmaceuticals we expressed an active single-chain antibody. Importantly, the novelprotein export pathway circumvents N-glycosylation which is advantageous in many applications, for example to avoid undesired immune reactions inhumans. Currently, the system is optimized with respect to product yield by e.g., reducing the proteolytic activity in the culture supernatants. Thus, theunconventional Cts1 secretion machinery has a high potential for the production of biotechnologically relevant proteins.714. A new method for fungal genetics: flow cytometry of microencapsulated filamentous microcolonies. L. Delgado-Ramos 1 , A. T. Marcos 1 , X. Peñate 1 ,M. S. Ramos-Guelfo 1 , L. Sánchez-Barrionuevo 1 , F. Smet 2 , D. Canovas 1 , S. Chávez 1 . 1) Dept of Genetics, Univ of Sevilla, Spain; 2) Union Biometrica, Geel,Belgium.Genetic analysis of non-filamentous microorganisms is facilitated by the isolation of consistent, well-defined colonies on solid media and the handling ofindividual cells by flow cytometry. In contrast, some filamentous fungi are hard to be analyzed using these procedures; in particular by flow cytometry. Thecombination of single spores microencapsulation and large particle flow cytometry is a possible alternative for the analysis of filamentous fungi.Microencapsulation allows the early detection of fungal growth by monitoring the development of hyphae from encapsulated individual spores. Myceliumproliferation inside the microcapsules can be detected using COPAS large particle flow cytometry. Here we show the successful application of the FlowFocusing® technology to the microencapsulation of filamentous fungi in monodisperse alginate microspheres, using Aspergillus and Trychoderma as modelsystems. Using a Cellena® Flow Focusing microencapsulator, we managed to produce monodisperse microparticles containing individual spores and todevelop microcolonies of these fungi upon germination in the appropriate conditions. Proliferation inside the particles was monitored by microscopy andlarge particle flow cytometry without requiring fluorescent labeling. Sterility was preserved during the microencapsulation procedure, preventingundesired contaminations. Conditional mutants were utilized to demonstrate the feasibility of the method. This procedure allows for the handling,screening and analysis of clonal colonies in liquid culture. Examples of applications will be provided.715. DNA methylation dynamics during development in the rice blast fungus. Junhyun Jeon 1 , Jaeyoung Choi 2 , Gir-Won Lee 2 , Sook-Young Park 3 , AramHuh 1 , Ralph Dean 4 , Yong-Hwan Lee 1,2,3,5 . 1) Department of Agricultural Biotechnology, Seoul National University, Seoul 151-921, South Korea; 2) FungalBioinformatics Laboratory, Seoul National University, Seoul 151-921, Korea; 3) Center for Fungal Pathogenesis, Seoul National University, Seoul 151-921,Korea; 4) Center for Integrated Fungal Research, Department of Plant Pathology, North Carolina State University, Raleigh, NC 27607, USA; 5) Center forFungal Genetic Resources, Seoul National University, Seoul 151-921, Korea.Cytosine methylation is an important epigenetic modification of DNA that is involved in genome defense and transcriptional regulation in eukaryotes. Inmammals and plants, many roles of DNA methylation depend on dynamic changes of DNA methylation pattern. In fungi, DNA methylation is consideredprimarily as a stable mark for silencing transposable elements. Here we used genetic manipulations and high-throughput bisulphite sequencing on themodel plant pathogenic fungus, Magnaporthe oryzae to elucidate the dynamics and mechanics of DNA methylation during pathogenic development. Wefound that genome-wide reprogramming of DNA methylation in and around genes occurs during progression of fungal development and that suchreprogramming is important for normal development. RNA-seq analysis showed that DNA methylation is associated with transcript abundance of genes incontext-dependent manner. Our study reveals that DNA methylation in fungi could be a dynamic epigenetic entity that has assumed new roles indevelopmental processes other than genome defense.296

FULL POSTER SESSION ABSTRACTS716. pH-enotype array: a novel phenomics platform for filamentous fungi. Jaejin Park 1 , Yong-Hwan Lee 1,2 . 1) Department of Agricultural Biotechnology,Seoul National University, Seoul 151-921, Korea; 2) Center for Fungal Pathogenesis, Center for Fungal Genetic Resources, Plant Genomics and BreedingInstitute, and Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Korea.Rapid increase of genome information and application of high-throughput mutagenesis technology allowed large-scale gene characterization infilamentous fungi. However, phenotype screening is a bottleneck as it entails time-consuming and labor-intensive processes. Thus there is a demand for anovel platform for high-throughput phenotype screening. Although several microplate-based assays are currently available, their application on organismsgrowing in filamentous form has been met with considerable difficulty due to uneven distribution of cells. As a solution, we developed pH-enotype array -a new phenomics platform in filamentous fungi. This platform is based on the pH change in the culturing media, reflecting viability and physiological statusof cells. The pH in culturing media is continuously measured by a microplate spectrophotometer using two pH indicators, bromocresol purple and phenolred. The validity of the system was comprehensively evaluated with Magnaporthe oryzae strains and we confirmed that the growth responses to variousstresses or nutrient conditions can be characterized reliably within 24 hours using the optimized medium. This phenomics platform would provide a novel,high-throughput phenotype screening method in filamentous fungi and promote phenotype standardization for comparative functional genomic studies.717. GFP analysis of meiotic recombination in Neurospora Dmsh-2 homozygotes. P Jane Yeadon, Frederick J Bowring, David E A Catcheside. Sch BiologicalSci, Flinders Univ, Adelaide, South Australia, Australia.We have confirmed that, as expected for a mismatch repair gene with both vegetative and meiotic roles, the phenotype of a msh-2 deletion inNeurospora crassa is recessive. Chromatid data indicates that deletion of msh-2 increases allelic recombination at his-3 by a factor of 1.6 with no effect oncrossing over in the lys-4 to ad-3 interval in which lies both his-3 and the recombination hotspot cog. Although analysis of a small number of octads froman msh-2 deletion cross suggested that the only non-Mendelian segregation is post-meiotic, the ease by which octads can be scanned for recombinationevents under a fluorescent microscope when GFP is inserted close to cog allowed us to reveal the wide range of recombination outcomes normally hiddenby the activity of Msh-2. We report that a degree of mismatch repair is retained in the absence of Msh-2 and that recombination initiated by cog exhibits astrong bias for repair in the direction of restoration rather than conversion. In contrast to recombination events in budding yeast, symmetric heteroduplexappears to be frequent in the his-3 region, suggesting variation between recombination pathways utilised in the two species.718. Residual recombination in Neurospora crassa spo11 mutant homozygotes occurs during meiosis. Frederick J Bowring, P Jane Yeadon, David E ACatcheside. Sch Biological Sci, Flinders Univ, Adelaide, South Australia, Australia.We have previously shown that although most genomic regions of Neurospora spo11 RIP mutants lack meiotic recombination, crossing over in the his-3region persists at close to wild-type levels. However, this residual recombination could conceivably occur after meiosis, during a transient period of partialdiploidy. Using crosses heteroallelic for his-3 mutations, we have shown that in spo11 RIP homozygotes, as in wild type, stable His + progeny are generated athigh frequency. We have utilised mutations in either end of a histone H1-GFP fusion gene, inserted between the recombination hotspot cog and his-3, toshow that the frequency at which GFP + spores arise in a cross homozygous for spo11 + is comparable to the frequency of His + spores, and that glowingnuclei first appear during pachytene, as expected for a product of meiotic recombination. In similar spo11 deletion homozygotes, GFP + spores also arise athigh frequency and glowing nuclei are also first seen at pachytene. Thus, spo11 mutant homozygotes experience both crossing over and allelicrecombination during meiosis, suggesting there is a spo11-independent mechanism for initiation of recombination at his-3.719. Controlled synthesis of gold nanoparticles by Neurospora crassa extract and their SERS properties. Katrin Quester 1 , Ernestina Castro-Longoria 1 ,Miguel Avalos-Borja 2,3 , Alfredo Rafael Vilchis-Nestor 4 , Marco Antonio Camacho-López 5 . 1) Departamento de Microbiología, Centro de InvestigaciónCientífica y de Educación Superior de Ensenada. Ensenada, B.C. México; 2) Centro de Nanociencias y Nanotecnologia, UNAM. Ensenada, B.C. México; 3)División de Materiales Avanzados, IPICyT. San Luis Potosí, S.L.P. México; 4) Centro Conjunto de Investigación en Química Sustentable, UAEMex. Toluca,Estado de México, México; 5) Laboratorio de Investigación y Desarrollo de Materiales Avanzados, Sección de Espectroscopía Raman, Facultad de Química-UAEMex, Unidad Rosedal. Toluca, Estado de México, México.Nanotechnology, the study of the controlling matter of an atomic and molecular scale, has emerged as an interesting and important scientific field andthe controlled synthesis of nanostructures from different chemical composition as well as their shape, size, and dispersity are important areas of research.The so-called ‘green chemistry’ or ‘nanobiotechnology’ employs microorganisms to fabricate nanostructures and has the benefit of improving thebiocompatibility of nanomaterial, however the control over average particle size and uniform particle morphology is required but still a challenge. Thiswork bases on the use of Neurospora crassa, a non-pathogenic filamentous fungus with rapid growth rate, for the biosynthesis of gold nanoparticles ofcontrolled size and shape. Briefly, the fungal extract was incubated with the gold precursor solution at different conditions of temperature, pH and time ofreaction. The best results obtained were from incubations at 60°C; at pH 3, particles of different shapes (e.g. spheres, triangles, hexagons, pentagons,rhombs and bars) were formed while at pH 5.5 and pH 10 small quasi-spherical particles were formed with size ranges of 6 to 21 nm and 3 to 12 nm,respectively. High resolution transmission electron microscopy (HRTEM) using a FEI Tecnai F30 transmission electron microscope confirmed the crystallineand elemental character of the gold nanoparticles. The synthesized gold nanoparticles of different shapes were shown to possess excellent surfaceenhancedRaman scattering (SERS) enhancement ability relative to quasi-spherical gold nanoparticles. Small quasi-spherical nanoparticles of 3 to 12 nmenhances the Raman signals of methylene blue about 2 times, those of 6 to 23 nm enhance the Raman signals about 25 times whereas nanoparticles ofdifferent shapes with a broad size range enhances the Raman signals of methylene blue about 40 times. Results are promising and show that these goldnanoparticles might have potential applications for biological sensing and labeling systems.720. Cellulase production in Neurospora crassa. M. Reilly, L. Glass. Energy Biosciences Institute, University of California, Berkeley, CA.The filamentous fungus Neurospora crassa is a well-studied model organism that is frequently isolated from the environment in association with burnedvegetation. Enzyme activities related to the metabolism of plant cell wall material have been observed in N. crassa, but the overall cellular response of theorganism to such a recalcitrant carbon source remains poorly defined. In order to investigate the elements involved in fungal growth on cellulose, weutilized the near full genome collection of single gene knockout strains that has been generated under the auspices of the Neurospora Genome Project.Strains with deletions in loci thought likely to play a role in cellulase activity or production - including the known secretome of N. crassa when cultured onplant biomass, homologs of the Saccharomyces cerevisiae secretion apparatus, and proteins predicted to traverse the secretory pathway - were singledout for analysis. This subset of the N. crassa deletion collection was cultured on a cellulosic substrate and the levels of secreted protein and cellulaseactivity were compared to wild-type. A number of hyper- and hypo-secretion mutants have been identified. Sequence analysis suggests that while many ofthe loci encode fungal-specific proteins of undetermined functions, some of the deleted genes likely act in transcription, protein synthesis, andintracellular trafficking. Early work with the latter found that their hyper- or hypo-secretion phenotypes were a specific response to cellulose, but not27th Fungal Genetics Conference | 297

FULL POSTER SESSION ABSTRACTSlimited to the production of cellulolytic enzymes. The continuing characterization of these mutants will enhance our understanding of the ability of N.crassa to utilize the complex carbon sources present in its natural environment.721. Identification and Functional analysis of New Neurospora crassa Nonself Recognition Loci. Jiuhai Zhao, Charles Hall, Elizabeth Hutchison, DavidKowbel, Juliet Welch, N. Louise Glass. Department of Plant and Microbial Biology, University of California, Berkeley USA 94720.Self/nonself recognition is a ubiquitous and essential function for many organisms. In filamentous fungi, self/nonself recognition is conferred by geneticdifferences at het (heterokaryon incompatibility) loci. The genes that mediate HI (heterokaryon incompatibility) exhibit characteristic evolutionarysignatures, including balancing selection and trans-species polymorphisms. Recent analyses show that genes containing a HET domain are involved in HI,making HET domain genes good candidates for identifying new het loci. In this study, we utilized RNA-seq data from a population of 110 Neurospora crassastrains to look for HET domain genes that were highly polymorphic, have multiple alleles, and show balancing selection, and trans-species polymorphisms.Using this approach, we identified 19 of the 62 HET domain genes in N. crassa that fit the criteria for a het locus. Further, we showed that one of these HETdomain genes, NCU09037, functions as a het locus.722. Combinatorial cationic and oxidative stresses promote the killing of Candida albicans cells by human neutrophils. Alistair J P Brown 1 , DespoinaKaloriti 1 , Mette Jacobsen 1 , Zhikang Yin 1 , Anna Tillmann 1 , Miranda Patterson 2 , Deborah A Smith 2 , Emily Cook 3 , Tao You 4 , Iryna Bohovych 1 , Celso Grebogi 4 ,Neil A R Gow 1 , Janet Quinn 2 , Ken Haynes 3 . 1) School of Medical Sciences, University of Aberdeen, Aberdeen, United Kingdom; 2) Institute for Cell andMolecular Biosciences, Faculty of Medical Sciences, Newcastle University, United Kingdom; 3) School of Biosciences, College of Life & EnvironmentalSciences, University of Exeter, United Kingdom; 4) Institute for Complex Systems and Mathematical Biology, School of Natural and Computing Sciences,University of Aberdeen, United Kingdom.Candida albicans is an opportunistic pathogen of humans. It is thought to have evolved as a relatively harmless commensal. C. albicans is a frequentcause of mucosal and skin infections (thrush). However, our immune system normally blocks potentially lethal systemic infections of the bloodstream andinternal organs. Neutropenic patients are prone to systemic candidiasis because they lack the effective defenses provided by circulating neutrophils. Wehave shown that the efficient killing of C. albicans by human neutrophils is mediated by the potent combinations of stresses they impose on the invadingfungus, rather than specific individual stresses. In particular, exposure to the combination of reactive oxygen species and cation fluxes kills C. albicanssynergistically. We have explored the mechanistic basis for this synergistic killing using genomic exploration and molecular dissection in C. albicanscombined with dynamic mathematical modeling. Signalling via the Hog1 stress activated protein kinase and the Cap1 AP-1-like transcription factor isinhibited by combinatorial oxidative and cationic stresses, and as a result, their downstream gene targets are not induced. This prevents the activation ofnormal oxidative and cationic stress adaptation and repair mechanisms. In particular, hydrogen peroxide detoxification mechanisms are inhibited byelevated salt concentrations. This leads to the accumulation of intracellular reactive oxygen species, and ultimately to accelerated necrotic death. Ectopicexpression of key detoxification mechanisms in C. albicans cells decreases the efficacy of killing by human neutrophils.723. Phosphoproteomic analysis of the aquatic fungus Blastocladiella emersonii during germination. J. Crestani, S. Lopes Gomes. Biochemistry, IQ, USP,Sao Paulo, Brazil.The aquatic fungus Blastocladiella emersonii presents an interesting life cycle with two cell differentiation stages, the germination and the sporulation,during which drastic morphological and biochemical changes are observed. During germination, protein synthesis is inhibited, a transient increase of cAMPlevels is observed, activation of PKA is detected, as well as mobilization of cellular glycogen and an efflux of calcium. These results suggest a possiblephosphorylation/dephosphorylation control, probably through cell signaling networks, which were not characterized up to now. Therefore, the presentwork aims to elucidate these signaling mechanisms by using a phosphoproteomic analysis of B. emersonii during the germination. Firstly, we compared thephosphoproteomic profile from two distinct cell types of B. emersonii, the zoospores and the germling cell (at 45min of germination) by using twodimensionalgels stained with Pro-Q Diamond phosphoprotein dye. The comparison revealed about 82 phosphoproteins from germling cells and 44phosphoproteins from zoospores. These preliminary results suggest that phosphorylation events may be involved during early germination. To detect thesignaling processes and the proteins involved in these events we utilized IMAC-IMAC phosphoproteomic methodology to obtain an enrichedphosphopeptide sample from each stage of germination and early vegetative growth (0, 25, 45, 60 and 90 min). Enriched phosphopeptide samples will bedetected by using a LTQ Velos Orbitrap mass spectrometer; filtered using DTASelect and analyzed using Ascore and Debunker. The results of this work willcontribute to improve the knowledge of the cellular regulatory processes in this early diverging fungus. Financial support: FAPESP and CNPq.724. Towards an accurate genome: high-throughput proteogenomic validation of Stagonospora nodorum genes via sub-cellular proteomics. KejalDodhia 1 , Robert Syme 1 , Thomas Stoll 2 , Marcus Hastie 2 , James Hane 3 , Angela Williams 3 , Eiko Furuki 1 , Jeffrey Gorman 2 , Richard Oliver 1 , Kar-Chun Tan 1 . 1) TheAustralian Centre for Necrotrophic Fungal Pathogens, Environment & Agriculture, Curtin University, Perth, Bentley 6102, Australia; 2) Protein DiscoveryCentre, Queensland Institute of Medical Research, Herston, Qld 4029, Australia; 3) Plant Industry, Commonwealth Scientific and Industrial ResearchOrganisation, Private Bag No, 5, Wembley WA 6913, Australia.Stagonospora nodorum is the causal agent of stagonospora nodorum blotch on wheat. S. nodorum was the first of the Pleosporales fungi to have itsgenome sequence published and genes annotated. However, in silico gene annotation can be erroneous. Therefore, experimental evidence is oftenneeded to refine gene annotations. Proteogenomics is an emerging high-throughput technique, which is a “direct-to-genome mapping” techniquewhereby the mass spectra from protein analyses are mapped onto the predicted gene set and/or the 6-frame whole genome translation. In this study, weperformed a comprehensive proteogenomic analysis of the secreted, intracellular and cell-wall/membrane sub-proteomes of S. nodorum using a twodimensionalliquid chromatography (2D-LC) LTQ Orbitrap MS approach. This study has verified a total of 3580 genes from all sub-proteomes. Of these, 113had not been experimentally verified previously. When combined with previous proteomic data, 4377 (35% of the total predicted gene set) genes wereverified. In addition, all mass spectra were matched to a 6-frame genome translation database to identify evidence of gene model conflicts. The study hasfound that 2629 genes showed evidence of frame conflicts and extensions of coding exons into annotated introns or untranslated regions. At least 43potential new genes were identified.725. Evolutionary Imprint of Fungal PKS-NRPS Catalytic Domains. Daniela Boettger 1 , Holger Bergmann 2 , Barbara Kühn 1 , Ekaterina Shelest 2 , ChristianHertweck 1 . 1) Department Biomolecular Chemistry, Leibniz Institute for Natural Product Research and Infection Biology - Hans Knöll Institute,Beutenbergstrasse 11a, 07743 Jena, Germany; 2) Department of Systems Biology/Bioinformatics, Leibniz Institute for Natural Product Research andInfection Biology - Hans Knöll Institute, Beutenbergstrasse 11a, 07743 Jena, Germany.Fungal polyketide synthase-nonribosomal peptide synthetase (PKS-NRPS) hybrid enzymes produce a broad array of ecologically and medicinally relevant298

FULL POSTER SESSION ABSTRACTSnatural products. To date, only a dozen gene clusters could be matched to the requisite PKS-NRPS pathways and the programming of the multifunctionalenzymes is still enigmatic. The (heterologous) expression of chimeras of PKS (lovastatin synthase, LovB) and NRPS (cytochalasin synthetase, CheA) inAspergillus terreus did not result in the production of polyketide-amino acid hybrid molecules and suggests a potential incompatibility of a fungal highlyreducing PKS (hrPKS) with the NRPS component of fungal PKS-NRPS hybrids. Furthermore, the heterologous expression of a shortened CheA (truncatedafter the C domain) in A. oryzae did not lead to cyclized products. To rationalize the unexpected outcome of the gene fusion (and shortening) experiments,we accomplished extensive bioinformatic analyses of fungal PKS-NRPS hybrids and LovB-type PKS. Hence, a noncanonical function of C-terminalcondensation (C) domains in truncated PKS-NRPS homologues and an evolutionary imprint of the PKS-NRPS domains, which reflect the evolutionaryhistory of the entire megasynthase, was inferred. Moreover, the participation of not only the adenylation (A) domain but also the C domain to amino acidselection was shown to be likely. These findings shed new light on the complex code of this emerging class of multifunctional enzymes and will greatlyfacilitate future combinatorial biosynthesis and pathway engineering approaches towards natural product analogues.726. Secondary metabolism and development is mediated by LlmF control of VeA subcellular localization in Aspergillus nidulans. Jonathan M. Palmer 1 ,Jeffrey Theisen 1 , Rocio Duran 2 , Scott Grayburn 2 , Ana Calvo 2 , Nancy Keller 1 . 1) Medical Microbiology and Immunology, Univ Wisconsin, Madison, WI; 2)Biological Sciences, Northern Illinois University, DeKalb, IL.Secondary metabolism and development are linked in Aspergillus through the conserved regulatory velvet complex composed of VeA, VelB and LaeA.The founding member of the velvet complex, VeA, shuttles between the cytoplasm and nucleus in response to alterations in light. Here we describe a newinteraction partner of VeA identified through a reverse genetics screen looking for LaeA-like methyltransferases in Aspergillus nidulans. One of the putativeLaeA-like methyltransferases identified, LlmF, is a negative regulator of sterigmatocystin production and sexual development. LlmF interacts directly withVeA and the repressive function of LlmF is mediated by influencing the localization of VeA, as over-expression of llmF decreases the nuclear to cytoplasmicratio of VeA while deletion of llmF results in an increased nuclear accumulation of VeA. We show that the methyltransferase domain of LlmF is required forfunction, however LlmF does not directly methylate VeA in vitro. This study identifies a new interaction partner for VeA and highlights the importance ofcellular compartmentalization of VeA for regulation of development and secondary metabolism.727. Overproduction of phleichrome by synthetic inducers and cloning of polyketide synthase genes in phytopathogenic fungus Cladosporium phlei. K.-K. So 1 , N.-L. Nguyen 1 , J.-M. Kim 2 , Y.-S. Jang 1 , Y.-S Jeong 1 , D.-H. Kim 1 . 1) Institute for Molecular Biology and Genetics, Center for Fungal pathogenesis,Chonbuk National University, Jeonju, Jeonbuk, South Korea; 2) Department of Bio-Environmental Chemistry, Wonkwang University, Iksan, Jeonbuk, SouthKorea.Phleichrome pigment produced by a Cladosporium phlei is a pathogenic toxin of timothy plant (Phleum pretense). Phleichrome reacts with oxygenmolecules following light activation to produce highly toxic reactive oxygen species. Phleichrome is structurally similar to elsinochrome and several other4,9-dihydroxyperylene-3,10-quinone fungal toxins. Phleichrome has a huge potential to be used as photodynamic agent for treatment of cancer and viralinfection. Using the UV mutagenesis method we were able to obtain two mutant strains that overproduced phleichrome in different culture conditionscompared with the wild type strain. In addition, we synthesized two different diketopiperazines as inducers and confirmed that diketopiperazinessignificantly enhanced phleichrome biosynthesis in a dose dependent manner. To gain insight into the metabolic pathway of phleichrome production, weperformed to clone and sequence several polyketide synthase (PKS) genes. Among the three representative types of PKS, two, one, and one gene forreducing-, partially reducing-, and non-reducing type PKS, respectively, were cloned, sequenced, and characterized. Biological characterization of thesegenes is underway to determine its role in the production of phleichrome and open the possibility of metabolically engineering this pathway foroverproduction of the desired substance.728. Symbiotic fungal endophytes that confer tolerance for plant growth in saline soil. Zakia Boubakir, Elizabeth Cronin, Susan GW Kaminskyj. Biology,Univ Saskatchewan, Saskatoon, Saskatchewan, Canada.Fungal endophytes are plant symbionts, and appear to be ubiquitous in plants growing in natural soils. Pioneering work by RS Redman and RJ Rodriguezshowed that class II fungal endophytes confer tolerance to harsh growth environments. These endophyte strains are expected to enhance plant growthand improve nutrient uptake under normal and saline conditions, although currently the mechanism(s) is unknown. Soil salinity is one of the most seriousagricultural problems that restrict plant growth and crop yield in many areas of the world. Saskatchewan has large areas of salinized soils as well as manysaline lakes. In this study, we are characterizing endophyte fungi isolated from salinized soils in southern Saskatchewan. These include potash minetailings, which are ~ 95 % NaCl. In the spring of 2012 we collected 90 plant samples from 9 sites, from which we isolated ~450 endophyte fungi. Here, wewill present a preliminary characterization of isolate Skj422.08. This strain has been shown to confer NaCl tolerance for tomato and wheat that weregrown in soil mix watered with fresh water, then stressed with 200 mM or12 g/L NaCl (tomato), or 300 mM or 18g/L NaCl (wheat). Skj422.08 alsoimproved growth when tomato and wheat were grown from seed in 150 mM or 9 g/L NaCl. This project is funded in part by Mosaic Co, a major producerof potash in Saskatchewan, as well as phosphate and micronutrients in other parts of North America, and worldwide.729. Stable cesium and radiocesium response of Schizophyllum commune. Matthias Gube 1 , Alix Günther 2 , Flemming Katrin 2 , Linde Jörg 3 , Raff Johannes 2 ,Kothe Erika 1 . 1) Microbial Communication, Institute for Microbiology, Friedrich Schiller University of Jena, Thuringia, Germany; 2) Helmholtz-CentreDresden-Rossendorf, Germany; 3) Hans-Knöll-Institute for Natural Product Research, Jena, Germany.Radioisotope contamination poses a threat to both ecosystem functioning and public health. Compared with plants, fungi can accumulate much higheramounts of heavy metals and radionuclides in their fruiting bodies. This was seen after the reactor accident of Chernobyl in 1986, when it became clearthat fungal uptake of radionuclides such as 60Co, 90Sr, and most importantly 137Cs may reach harmful levels if consumed. It is thus of crucial importanceto study radionuclide uptake into fungi to evaluate subsequent migration in the environment, thus allowing for meaningful ecotoxicological riskassessment. Usually, this is being performed by analysing stable isotopes of the same elements, whenever these are available. Due to their lowercriticality, this reduces costs and risk of associated experiments. It is generally assumed that the effects of both are identical. However, comparativeanalyses have seldom been performed, and never with fungi. Ionizing radiation is known to cause effects ranging from DNA and Protein damage toincreased oxidative stress and possibly apoptosis in a number of organisms. Especially in gene expression studies, certain reactions such as regulation ofprotein and nucleic acid repair mechanisms might thus deviate between stable and radioisotope exposure. Thus, differential gene expression of the fullysequenced model organism S. commune was analysed using MACE (Massive Analysis of CDNA Ends) under treatment with stable 133Cs and the b-decaying 137Cs and compared with untreated samples. Differential gene expression analysis points out factors responding to either radiation or Cs ions,thus differentiating between metal ion stress and radiation effects. While several carbohydrate metabolism genes and especially hydrophobin genes arespecifically regulated following radiation, most expected responding factors, such as genes involved in stress response or ion and water transport, are27th Fungal Genetics Conference | 299

FULL POSTER SESSION ABSTRACTSregulated after treatment with both 133Cs and 137Cs.730. The completion of meiosis in Ustilago maydis requires an Ndt80 ortholog. C. E. Doyle 1 , H.Y. K. Cheung 1 , B. J. Saville 1,2 . 1) Environmental & LifeSciences Graduate Program, DNA Building, Trent University, 2140 East Bank Dr., Peterborough, ON, Canada; 2) Forensic Science Program, DNA building,Trent University, 2140 East Bank Dr. Peterborough, ON, Canada.Meiosis in the model fungal plant pathogen Ustilago maydis requires growth in the plant host; as such, control of meiosis responds to cues receivedduring pathogenic development. To begin investigating this process, an ortholog of the Saccharomyces cerevisiae meiotic control protein, Ndt80 (nondityrosine) was identified in Ustilago maydis. It was hypothesized to control progression through meiosis and has been designated mcg1 (meiosis controlgene 1). To assess its role in meiosis, mcg1 deletion mutants were constructed in compatible U. maydis haploid strains by replacing the gene with differentselectable markers. This allowed the impact of mcg1 deletion on pathogenesis, teliospore development and the completion of meiosis to be determined.Infections with compatible Dmcg1 strains were fully pathogenic, but teliospores produced from these crosses displayed a distinctive, abnormalmorphology and meiotic segregation assays indicated that they germinated without undergoing meiosis. This suggests that Mcg1 is involved in theregulation of meiosis and teliospore formation. To investigate this possibility that Mcg1 accomplishes this function by acting as a transcription factor, theupstream region of U. maydis genes was searched for variants of sites known as middle sporulation elements (MSE, the binding site of S. cerevisiae Ndt80).89 genes with upstream MSEs were screened by RT-PCR, using RNA from dormant teliospores of wild-type and Dmcg1 strains. The results suggested thattranscript levels for 43 of these genes differed in wild-type, relative to Dmcg1 teliospores, which indicated that the expression of these genes was affected,either directly or indirectly, by Mcg1. Further screens using RT-qPCR allowed the confirmation of genes with increased transcript levels, as well as thosewith decreased transcript levels, in the Dmcg1 teliospores relative to the wild-type teliospores. The upstream regions of these genes were screened for thepresence of conserved sequence elements. In parallel, Mcg1 was aligned with putative orthologs to identify conserved regions. Based on these alignments,mcg1 genes containing targeted mutations were synthesized. Together, these analyses begin the determination of how in planta transitions in U. maydisdevelopment are controlled.731. Impact of changes in the target P450 CYP51 enzyme associated with altered triazole-sensitivity in the Wheat pathogen Mycosphaerellagraminicola. Eileen Scott 1 , Regula Frey 2 , Helge Sierotzki 2 , Michael Csukai 1 . 1) Syngenta, Biological Sciences, Jealotts' Hill International Research Centre,Bracknell, United Kingdom; 2) Syngenta Crop Protection Munchwilen AG, Research Biology Centre, Schaffhauserstrasse, Stein, Switzerland.The triazoles are a widely used class of fungicides, targeting the cytochrome P450 sterol 14a-demethylase Cyp51. They are hence also known as the 14a -demethylase inhibitors, the 14-DMIs. Despite heavy use of this chemical class in the field over a considerable period of time, catastrophic resistance hasnot been observed in the economically important plant pathogen M. graminicola. Rather, there has been a slow shift toward reduced sensitivity. A largenumber of mutations in the Cyp51 gene have been previously associated with this shift in sensitivity to DMIs, although other resistance mechanisms suchas alteration in sterol biosynthesis and fungicide uptake and efflux may also play a role. There have been attempts to correlate changes in resistance levelswith specific Cyp51 mutations in field isolates. However, due to the genetic diversity of Mycosphaerella , the possible effect of non-target site mutationsand issues with expression in exogenous fungi making solid conclusions has been problematic.In order to accurately assess the contribution of each of the target site substitution mutations found in the field associated with 14-DMI resistance wehave introduced mutations individually and in combination into the endogenous Cyp51 gene in a uniform genetic background (M. graminicola genomesequenced strain, IPO323). Here, we present the findings of the comparative efficacy of varying triazole structures against this comprehensive collection ofmutants.732. Molecular Evolutionary Analysis and Synteny of Fungal GAL Genes. Julien S Gradnigo 1 , C. L Anderson 2 , R. A Wilson 3 , E. N Moriyama 1,4 . 1) School ofBiological Sciences, University of Nebraska - Lincoln, Lincoln, NE; 2) Department of Computer Science and Engineering, University of Nebraska - Lincoln,Lincoln, NE; 3) Department of Plant Pathology, University of Nebraska - Lincoln, Lincoln, NE; 4) Center for Plant Science Innovation, University of Nebraska- Lincoln, Lincoln, NE.In many fungal species (including Saccharomyces, Candida, Schizosaccharomyces and related genii), genes involved in successive steps of a metabolicpathway are often physically clustered in the genomes. Within genes involved in the Leloir pathway for galactose catabolism, such clustering is consideredto facilitate niche adaptation via rapid gene inactivation. This pathway involves three structural genes - GAL1, a galactokinase, GAL7 a uridylyl transferaseand GAL10, a bifunctional protein with two epimerase domains. The products of the GAL80, GAL4 and GAL3 genes - a co-repressor, activator and coactivator- cooperatively regulate expression of the structural genes. GAL1 and GAL3 are highly similar (>90% identity) and likely arose from an ancientduplication event. GAL1, 7 and 10 are known to cluster in many divergent fungal lineages, including Saccharomyces, Candida, Schizosaccharomyces, andCryptococcus. To further investigate potential syntenic patterns in a wider range of fungal lineages including filamentous species, we identifiedorthologous GAL proteins from over 60 fungi. An initial set of orthologue candidates was generated using a combination of BLAST, reciprocal BLAST andprofile hidden Markov model searches. Sequences meeting the percent identity and coverage thresholds established for each protein were subsequentlyaligned using MAFFT. We then reconstructed maximum likelihood phylogenies and, where necessary, compared predicted secondary structures toproduce the orthologue dataset. Location information was obtained from the respective source database (NCBI, JGI or the BROAD Institute). Weconfirmed that GAL1, GAL7 and GAL10 are not clustered in all 53 species of filamentous fungi we studied. While in 7 species closely related to S. cerevisiaeas well as C. albicans, as previously reported, the two GAL10 domains exist as a single fused protein, they exist as separate proteins in Yarrowia lipolyticaand not at all in Ashbya gossypii. In all filamentous fungi we examined, these domains exist independently. We found widespread duplication of bothdomains, and are examining the evolutionary origins of GAL10 proteins and the timing of domain duplication and acquisition events. As GAL10 proteinsparticipate in both the first and final steps of the Leloir pathway, such duplication may promote catabolic efficiency.733. Meiotic Drive: A Single Gene Conferring Killing and Resistance in Fungal Spore Killer. Pierre Grognet 1,2* , Fabienne Malagnac 1,2 , Hervé Lalucque 1,2 ,Philippe Silar 1,2 . 1) Univ Paris Diderot, Sorbonne Paris Cité, Laboratoire Interdisciplinaire des Energies de Demain, 75205 Paris CEDEX 13 France; 2) UnivParis Sud, Institut de Génétique et Microbiologie, Bât. 400, 91405 Orsay cedex, France.Meiotic drives (MD) are nuclear genetic loci ubiquitous in eukaryotic genomes that cheat the Mendel laws by distorting segregation in their favor. Allknown MD are composed of at least two linked genes, the distorter that acts as a toxin by disrupting the formation of gametes, and the responder thatacts as an antitoxin and protects from the deleterious distorter effects. In fungi, MDs are known as Spore Killers (SK). In the model ascomycete Podosporaanserina, MD has been associated with deleterious effect during ascospore formation of the Het-s prion and in Neurospora crassa a resistance gene(responder) to the Sk-2 and Sk-3 distorters has been identified. MDs are easily studied in P. anserina thanks to the ascus structure as SKs are identified bythe presence of 2-spored asci in crosses between strains. Here, we identify and characterize by targeted deletion in P. anserina Spok1 and Spok2, two MD300

FULL POSTER SESSION ABSTRACTSelements. We show that they are related genes with both spore-killing distorter and spore-protecting responder activities carried out by the same allele,unlike other known MD. These alleles act as autonomous elements and exert their effects in any region of the genome. Moreover, Spok1 acts as aresistance factor to Spok2 killing. As Spok1 and Spok2 belong to a multigene family, these Spore Killer genes represent a novel kind of selfish genes thatproliferate in population through meiotic distortion.734. Alkaliphilic fungi from soda lakes and soda soils. Alexey A. Grum-Grzhimaylo 1 , Alfons J.M. Debets 1 , Marina L. Georgieva 2 , Elena N. Bilanenko 2 . 1)Wageningen University, Wageningen, The Netherlands; 2) Lomonosov Moscow State University, Moscow, Russia.Filamentous fungi growing optimally at pH exceeding neutral values have received little scientific attention and generally it is believed that onlyprokaryotic organisms are able to survive harshly elevated ambient pH values. To date, only a handful of alkaliphilic fungi (i.e. fungi growing optimally atpH > 9) have been reported. The few studies devoted to fungi growing at high pH lack a systematic molecular phylogenetic analysis. Our study aims toreveal the taxonomic distribution of alkaliphilic filamentous fungi we isolated from soils at different sites near soda lakes. We intend to test if thealkaliphilic trait has occurred independently throughout the fungal kingdom or rather once in a single lineage. Soda lakes and soils with pH ranging from 8to as high as 11 are believed to be the natural habitats for alkaliphiles. The high pH is maintained mainly due to strong buffering capacity of carbonate saltspresented there. We used alkaline agar medium (the pH is buffered at around 10) with antibiotic as a selective medium for screening for potentialalkaliphilic fungi in the collected soil samples. By these means we isolated 99 ascomycetous strains which were capable of growing, to different extents, atpH 10. Two thirds of the total number turned out to be anamorphic fungi displaying only asexual sporulation (mostly Acremonium-like) while only 19strains were holomorphic homothallic being able to develop the full life cycle. Seventeen isolates produced only sterile mycelium without reproductivestructures under laboratory conditions. We sequenced five genes (SSU rDNA, LSU rDNA, RPB2, TEF1alpha, ITS region of rDNA) to pinpoint the taxonomicpositions of all isolated. After phylogenetic reconstructions all our strains had tendency to group in two different lineages within the Ascomycota. The firstlineage is the Plectosphaerellaceae family (insertae sedis within subphylum Hypocreomycetidae) which harbors 39 alkaliphilic isolates while the secondlineage is Emericellopsis-clade (order Hypocreales, subphylum Hypocreomycetidae) within the Acremonium cluster containing 30 strains. The remaining 30isolates are presumably alkalitolerant members of the Pleosporinae and Sordariales lineages known to be ubiquitous soil fungi.735. A new method for gene mining and enzyme discovery. Y. Huang 1,2,3 , P. Busk 1 , M. Grell 1 , H. Zhao 2,3 , L. Lange 1 . 1) Section for Sustainable Biotechnology,Department of Biotechnology, Chemistry and Environmental Engineering, Aalborg University Copenhagen, Denmark; 2) Environmental Microbiology KeyLaboratory of Sichuan Province, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, Sichuan 610041, PR China; 3) University of theChinese Academy of Sciences, Beijing 100049, PR China.Peptide pattern recognition (PPR) is a non-alignment based sequence analysis principle and methodological approach, which can simultaneouslycompare multiple sequences and find characteristic features. This method has improved the understanding of structure/function relationship for enzymeswithin the CAZY families, which would make it easier to predict the potential function of novel enzymes, creating bigger promises for industrial purposes.Mucor circinelloides, member of the former subdivision Zygomycota, can utilize complex polysaccharides such as wheat bran, corncob, xylan, CMC andavicel as substrate to produce plant cell wall degrading enzymes. Although the genome of M. circinelloides has been sequenced, only few plant cell walldegrading enzymes are annotated in this species. In the present project, PPR was applied to analyze glycoside hydrolase families (GH family) and miningfor new GH genes in M. circinellolides genome. We found 19 different genes encoding GH3, GH5, GH6, GH7, GH9, GH16, GH38, GH43, GH47 and GH125 inthe genome. Of the three GH3 encoding genes found, one was predicted by PPR to encode a b-glucosidase. We expressed this gene in Pichia pastoris andfound that the recombinant protein has high b-glucosidase activity (4884 U/mL). In this work, PPR provided targeted short cut to discovery of enzymeswith a specific activity. Not only could PPR pinpoint genes belonging to different GH families but it did also predict the enzymatic function of the genes.736. Occurrence of dsRNA mycovirus (LeV-FMRI 2427) in edible mushroom Lentinula edodes and its meiotic stability. J.-M. Kim 1 , S.-H. Yun 2 , M.-S Yang 2 ,D.-H. Kim 2 . 1) Department of Bio-Environmental Chemistry, Wonkwang University, Iksan, Jeonbuk, South Korea; 2) Institute for Molecular Biology andGenetics, Center for Fungal Pathogenesis, Chonbuk National University, Jeonju, Jeonbuk, South Korea.The dsRNA was first found in the malformed cultures of Lentinula edodes strain FMRI 2427, one of three most popular sawdust cultivating commercialstrains of shiitake. This dsRNA was also found in the healthy-looking fruiting bodies and actively growing mycelia. Cloning of partial genome of dsRNArevealed the presence of RdRp sequence of a novel L. edodes mycovirus (LeV) and sequence comparison of the clone amplicon showed the identicalsequence to the known RdRp genes of LeV found in strain HKA. Meiotic stability of dsRNA was examined by the measuring the ratio of the presence ofdsRNA among the sexual monokaryotic progenies. More than 40% of monokaryotic progenies still contained the dsRNA indicating the persistence ofdsRNA during sexual reproduction. Comparing mycelial growth of monokaryotic progenies suggested that, although variations in growth rate existedamong progenies, there appears no direct relationship of mycovirus infection to the growth rate.737. Analysis of fungal communities associated with grapevine wood diseases, based on fungal ITS pyrosequencing. Nicolas Lapalu 1,2 , AngéliqueGautier 2 , Laetitia Brigitte 1,2 , Jessica Vallance 3 , Emilie Bruez 3 , Joelle Amselem 1,2 , Hadi Quesneville 1 , Valérie Laval 2 , Marc-Henri Lebrun 2 , Patrice Rey 3 . 1) INRA-URGI, Versailles, France; 2) INRA, BIOGER, Thiverval Grignon, France; 3) INRA, Santé Végétal, Bordeaux, France.The Grapevine Trunk Diseases (GTDs) are the most common diseases of grapevine wood inducing a slow decay leading to plant death. Due to theenvironmental impact, chemical treatments are no longer authorized, and prevention or trunk removal are the last available control methods. Fightingagainst these slow evolving diseases requires a better knowledge of fungal and bacterial communities associated with GTDs. Our approach is based onfungal species identification using ITS (Internal Transcribed Spacer) sequences obtained by pyrosequencing (Roche 454) of grapevine wood samples. DNAswere extracted from different parts of grapevine trunks and amplified using fungal specific ITS primers. A workflow was set up to analyze pyrosequencingdata, allowing taxonomic assignment with a database extracted from Genbank and curated with the FungalITSextractor (Nilsson H et al. 2010). Thepipeline links tools, including cleaning and extracting ITS sequences to limit the impacts of sequencing errors on clustering and assignation steps. Then,Operational Taxonomic Units (OTUs) detection and taxonomic assignments were performed with the QIIME package (Caporaso JG et al. 2010). Samplesfrom different vineyards (infected or not), with several dates of sampling, were analyzed. Different ITS PCR primers and technical replicates were testedusing controls corresponding to the mixture of fungal DNAs from diverse known species. These controls highlighted interests and limits of PCR ampliconspyrosequencing and the relevance of the bioinformatics methods to extract accurate data to fit to the context of taxonomy.738. The Antidepressant Sertraline Provides a Promising Therapeutic Option for Neurotropic Cryptococcal Infections. Bing Zhai, Cheng Wu, Linqi Wang,Matthew Sachs, Xiaorong Lin. Biology, Texas A&M University, TAMU-3258, TX.Therapeutic treatment for systemic mycoses is severely hampered by the extremely limited number of antifungals. Treatment for fungal infections in the27th Fungal Genetics Conference | 301

FULL POSTER SESSION ABSTRACTScentral nervous system is further compounded by the poor CNS penetration of most antifungals due to the blood-brain barrier. Only a few fungistatic azoledrugs, such as fluconazole, show reasonable CNS penetration. Here we demonstrate that sertraline (brand name Zoloft), the most prescribedantidepressant, displays potent antifungal activity against Cryptococcus neoformans, the major causative agent of fungal meningitis. In the in vitro assays,this neurotrophic drug is fungicidal to all natural Cryptococcus isolates tested at clinically relevant concentrations. Furthermore, sertraline interactssynergistically or additively with fluconazole against Cryptococcus. Importantly, consistent with our in vitro observations, sertraline alone reduces thebrain fungal burden at a comparable efficacy as fluconazole in a murine model of systemic cryptococcosis. It works synergistically with fluconazole inreducing the fungal burden in brain, kidney, and spleen. In contrast to its potency against Cryptococcus, sertraline is less effective against strains ofCandida species and its interactions with fluconazole against Candida strains are often antagonistic. Therefore, our data suggest the unique application ofsertraline against cryptococcosis. To understand the antifungal mechanisms of sertraline, we screened the whole genome deletion collection ofSaccharomyces cerevisiae for altered sertraline susceptibility. Gene ontology analyses of selected mutations suggest that sertraline perturbs translation. Invitro translation assays using fungal cell extracts show that sertraline inhibits protein synthesis. Taken together, our findings indicate the potential ofadopting this antidepressant in treating cryptococcal meningitis.739. Analysis of gene expression of proteases of Trichoderma spp during confrontation with plant pathogens in vivo. Valdirene Monteiro 1 , Eva Couto 1 ,Fenix Oliveira 1 , Marcela Suriani 2 , Raphaela Georg 2 , Cirano Ulhoa 2 . 1) Universidade Estadual de Goiás, Anápois, Brazil; 2) Universidade Federal de Goiás,Goiânia, Brazil.The genus Trichoderma with teleomorfismo in Hypocrea comprises a group of saprophytic fungi and micoparasitas widely used as biological controlagents of soil borne plant pathogens. Among the mechanisms proposed for the biocontrol of Trichoderma species are mycoparasitism by production of cellwall degrading enzymes of plant pathogens, antibiotic production volatile and non-volatile competition for nutrients, rhizosphere competent andinduction of defense responses in plants. In recent years, several efforts have been made to better understanding of molecules involved inmycoparasitism. In this study we assessed the expression of protease genes of Trichoderma species by qRT-PCR and the gene for alpha-tubulin asnormalizer. Differential expression was assessed from antagonism of Trichoderma virens and Trichoderma harzianum during contact and after contact withthe host hyphae of fungal pathogens Rhizoctonia solani and Sclerotinia sclerotiorum. The "primers" were designed based on the sequences of the genesencoding these enzymes deposited in the database DOE Joint Genome Institute (http://www.jgi.doe.gov/). The protease genes of both species wereaspartate protease, Carboxypeptidase, cysteine protease, Subtilisin peptidase, trypsin, protease Serino metallopeptidase. Initial results showed that duringthe contact of Trichoderma virens and Rhizoctania solani for metalloprotease gene is 80% higher than that expressed in the same condition forTrichoderma harzianum. When observing the condition after contact between T.virens and Sclerotinia sclerotiorum the metalloprotease gene does nothave its expression as required presenting least 60% expressed in relation to genes aspartate, carboxy, and subtilisin serine proteases. The expression wasvery different between the clashes of Trichoderma and plant pathogens as well as for contact times. Leading us to believe that specific classes of proteasesare required at different stages of contact of Trichoderma and pathogen. Person's test was performed to analyze the correlation between the expressionof these genes and other genes of proteases involved in mycoparasitism as glucanases and chitinases.740. Analysis of differential protein profile during antagonism of Trichoderma harzianum and Sclerotinia sclerotiorum. Valdirene Monteiro 1 , RobertoSilva 3 , Cirano Ulhoa 2 . 1) Universidade Estadual de Goiás, Anápois, Anápolis, Brazil; 2) Universidade Federal de Goiás, Goiânia, Brazil; 3) University of SãoPaulo.Trichoderma sp. are effective biocontrol agents for several soil-borne fungal plant pathogens including Fusarium sp., Rhizoctonia solani, and Sclerotiniasclerotiorum some species are also known for their abilities to enhance systemic resistance to plant diseases as well as overall plant growth. The biocontrolexercised by Trichoderma can occur by means of several antagonistic mechanisms such as nutrient competition, antibiotic production and mycoparasitism.Mycoparasitism has been proposed as the major antagonistic mechanism displayed by Trichoderma sp. In the present study were examined thedifferential production of proteins in three different conditions were antagonism. These conditions were T. harzianum vs mycelium of fungal pathogensSclerotinia sclerotiorum; Trichoderma harzianum vs esclerotia of fungal pathogens Sclerotinia sclerotiorum and Trichoderma harzianum vs leaves beansinfected with Sclerotinia sclerotiorum. The differential expression of proteins in different conditions of antagonism was analyzed by two-dimensionaldifference gel electrophoresis (2D-DIGE). Initial analyzes showed profiles varied between interactions. The proteins which were expressed were sodifferent and trypsin-like SM1 between conditions T.harzianum and bean leaves and T. harzianum and mycelium. There is a higher expression ofglucanases and chitinases in the condition of the interaction between T. harzianum and sclerotia. It was observed the presence of three proteins predictedmolecular weight of 15kDa that are present in all tested conditions but with different expression levels. These and other proteins are in phase sequencing.741. Effector proteins in fungal defense against fungivorous nematodes: Diversity and regulation of expression. D.F. Plaza 1 , S.S. Schmieder 1 , C.E.Stanley 2 , S. Bleuler-Martinez 1 , A.J. deMello 2 , M. Aebi 1 , M. Künzler 1 . 1) Institute of Microbiology, ETH Zürich, Wolfgang-Pauli-Str.10, 8093 Zürich; 2) Institutefor Chemical and Bioengineering, ETH Zürich, Wolfgang-Pauli-Str.10, 8093 Zürich.The regulation of defense mechanisms of fungi is poorly understood. So far, research has focused mainly on the regulation and production of toxicsecondary metabolites and peptides. Several lines of evidence also implicate cytoplasmic lectins, many of which are up-regulated during fruiting bodydevelopment, in fungal defense. We could demonstrate that many of these lectins are toxic, not only to model organisms such as Caenorhabditis elegansand Drosophila melanogaster, but also to fungivorous nematodes. In order to gain more insight into the diversity and regulation of the defense responseof fungi against predatory nematodes, we studied the genome-wide gene expression in the model organism Coprinopsis cinerea upon challenge with thefungivorous nematodes Aphelenchus avenae and Bursaphelenchus willibaldi, using next generation RNA-seq. Preliminary transcriptome data shows thatthe sets of upregulated genes differ between the two nematodes tested, suggesting a nematode-specific defense response. Whereas B. willibaldi inducedthe expression of genes hypothetically involved in secondary metabolite production, such as a non-ribosomal peptide synthase and a terpenoid synthase;A. avenae induced previously characterized nematotoxic fungal lectins, such as CGL2, CCL1 and a chimeric RicinB-fold protein. To characterize thisinducible defense response in more detail, we developed a reporter system in C. cinerea consisting of the cgl2 promoter coupled to the gene encoding thered fluorescent protein dTomato. The reporter C. cinerea strain was grown into a microfluidics device with compartmentalized channels and confrontedwith A. avenae. This system allowed us to microscopically monitor the induction of the effector molecule within the mycelium over time. Preliminaryresults indicate that the induction can spread within hyphae but is locally confined. The expression of the reporter protein dTomato, driven by the cgl2promoter, was first detected after 10 h of nematode predation. In conclusion, our findings show, first, that the defense of C. cinerea against fungivorousnematodes is regulated at the level of transcription. Second, we could demonstrate that the transcriptional induction of defense effector proteins inresponse to predation is not only locally confined but also predator-specific.302

KEYWORD LISTABC proteins ................................................ 656Acremonium chrysogenum ......................... 471Aegerolysin .................................................. 226Agaricus bisporus .......................................... 27Agrobacterium ......................... 69 221 271 354............................................... 476 495 537 598Annotation .................... 34 69 222 223 224 225.................................. 226 227 228 229 230 244.................................. 279 283 284 285 286 304.................................. 308 313 335 338 454 604Appressorium ........................ 69 70 71 140 477.................................. 478 479 480 573 593 603Arabidopsis thaliana .................................... 551Arnium arizonense ...................................... 321Asbya gossypii ............................................. 135Ashbya gossypii ............................. 122 134 158Aspergillosis ............................... 3 481 482 483Aspergillus flavus ........................... 347 566 633Aspergillus fumigatus ....... 1 2 3 4 5 72 73 74 75........................ 76 77 226 231 427 355 356 357.................... 358 359 456 689 690 482 484 485............................................... 486 487 488 640Aspergillus nidulans ............. 6 7 8 9 54 78 79 80.......................... 81 82 83 84 85 86 87 88 89 90...................... 91 92 93 94 95 96 97 98 214 223.................... 228 232 233 340 360 361 362 363............................. 364 365 366 367 368 369 44.......................... 452 463 714 691 692 693 726Aspergillus niger ......... 10 11 68 99 100 101 102........................... 103 228 370 371 694 695 696Aspergillus oryzae ........... 57 12 13 104 223 234............................................... 235 450 372 695Aspergillus sp. ............................................. 687Aspergillus terreus, Galleria mellonella ....... 636Aspergillus vadensis, heterologousexpression ................................................... 696Autophagy ................................................... 167BUD-2, BUD-5, polarity establishment ........ 181Barley ............................................ 564 489 641Batrachochytrium ........................................ 164Biofuels ....................................................... 720Biological control ......................................... 702Biomass ................. 14 37 45 47 60 205 236 237.................... 306 310 373 424 463 466 697 678Biotrophic ........... 490 491 492 493 494 596 603Blastomyces dermatitidis ............................ 441Botrytis cinerea ........... 15 105 106 107 108 238........................................ 374 375 495 496 497Calcium imaging ............................................ 75Candida albicans............ 16 17 109 110 111 239.................... 224 340 376 377 378 379 434 698........................... 699 498 499 500 501 502 503Carbohydrate catabolism ............................ 375Cell Cycle ................... 105 112 113 114 115 116............... 137 158 78 206 215 267 380 386 693Cell Wall ................. 18 19 20 21 22 3 4 110 117............. 118 119 120 121 122 146 164 189 193............ 86 87 88 101 103 212 240 241 381 382........................................ 360 699 503 504 485Cellulases..................................................... 712Centromeres ............................................... 242ChIP-chip ..................................................... 378ChIP-seq ............. 175 242 383 384 385 423 659Chitin ................... 18 20 21 22 101 505 522 618Chitosan ...................................................... 580Chlamydospores .......................................... 499Chromatin ............................................ 175 333Chromosomes ............................................. 664Chrysosporium ............................................ 679Circadian Clock .......... 386 387 388 389 390 444Coccidioides ................................................ 342Cochliobolus sativus, NPS,........................... 319Cohliobolus lunatus ....................................... 67Colletotrichum gloeosporioides .................. 346Conidiation ............. 43 123 124 147 90 100 298.................................. 309 341 433 435 369 642Coprinopsis cinerea ....................... 218 219 337Coprinopsis cinerea; Transformation;Tryptophan ................................................. 470Corn ................................. 506 534 568 613 633Cre-loxP ....................................................... 273Cryptococcus neoformans ..........23 24 114 125............. 126 127 243 244 245 246 312 391 392............. 393 394 395 396 397 398 399 458 700.................... 701 507 508 509 510 639 643 644DNA Repair ......... 115 137 188 337 398 399 401............................................................. 443 572Development ....... 25 127 128 129 130 131 132............. 133 134 135 136 166 169 171 185 194.................. 79 85 89 93 94 97 212 215 276 400...... 446 364 450 462 469 715 733 575 616 626Dicer ............................................................ 469Diversity .......... 26 36 247 260 702 511 512 513............. 645 646 647 648 649 650 651 678 682Dothideomycetes ........................................ 579Double Strand Break ................................... 472Drug Target Discovery, ................................ 690EFFECTOR RXLR PI3P ENTRY ........................ 530ER stress ...................................................... 104Ecology .......... 36 136 351 703 704 705 706 692...... 729 501 514 515 540 551 607 649 655 660Education ................................351 352 353 539Effector ..................... 138 165 248 249 250 270............. 336 402 403 476 492 505 506 516 517...... 518 519 520 521 522 523 524 525 526 527...... 528 529 530 531 532 533 534 477 557 483...... 576 577 486 588 593 596 622 624 625 627...................................................... 489 652 653Enzyme analysis .......................................... 704Epichloae ..................................................... 608Epichloë festucae ........................................ 569Epigenetic ................................................... 403Epigenetics .................................................. 385Esterase, Peptide Pattern Recognition ........ 697Evolution ........... 16 48 50 222 249 251 252 253...... 254 255 256 257 258 259 270 280 289 292...... 294 301 302 305 308 317 324 347 349 469...... 732 502 535 536 572 606 642 643 644 646...... 652 653 654 655 656 657 658 659 660 661...... 662 663 664 665 666 667 668 669 670 672...................................................... 675 677 681Frequency ................................................... 702Fruiting Body .......... 25 27 107 108 132 133 139202 207 219 258 287 404 437 454 707 537 687Functional prediction .................................. 343Fungal PKS-NRPS ......................................... 725Fungal defense against nematodes ............. 741Fungicide resistance .................................... 641Fusarium ....... 28 140 141 229 260 261 262 263...... 264 232 318 326 405 406 407 408 409 447708 538 539 540 541 542 543 544 599 669 670Fusarium graminearum ...... 38 142 143 144 145...... 146 147 195 230 265 266 267 268 269 410....... 411 709 529 545 546 547 548 549 550 671Fusarium oxysporum 148 149 248 270 551 552........................... 553 554 555 556 557 558 672G protein Signaling ...................................... 184G protein signaling ...................................... 145GFP ............. 130 163 178 196 103 217 302 346.................................. 366 471 717 718 537 564Gene Deletion ... 24 1 120 132 143 144 150 151............. 152 81 82 198 99 207 239 262 271 272....... 273 235 330 354 451 371 459 460 710 690....... 691 733 539 542 559 577 598 615 630 672Gene Expression ...... 15 27 29 30 31 32 37 6 46.... 51 57 124 203 104 221 248 274 275 276 279300 307 312 315 325 339 346 373 374 376 378380 382 383 384 388 389 392 394 396 397 400401 403 407 408 412 413 414 415 416 417 418419 420 421 422 423 424 425 426 427 428 429430 431 432 433 434 435 357 442 360 363 366367 455 462 465 372 471 473 697 701 704 711712 713 739 741 508 521 544 557 560 561 562..............563 580 583 590 602 613 489 657 659Genome Database ...................................... 340Genome Sequencing . 236 225 245 247 249 250252 259 260 261 266 269 277 278 279 280 281282 283 284 285 286 287 288 289 290 291 292297 301 233 306 320 323 234 343 347 350 724..............490 494 601 609 656 674 676 679 641Genome Structure ...................................... 290Geomyces ................................................... 688Glycan ......................................................... 705Glycogen ..................................................... 553Haustoria .................................................... 299Heterobasidion irregulare; Laccase;Coprinopsis ................................................... 64Histoplasma capsulatum ............................. 468Hyphae ....... 33 134 72 86 87 88 99 416 434 706......................................... 714 510 565 566 648Hyphal Fusion ...... 148 153 154 174 179 185 92.................................. 273 293 567 584 654 673Hyphal Growth .. 20 22 33 105 111 117 118 122136 142 150 152 155 156 157 158 159 160 161... 74 168 171 186 187 193 80 82 83 95 209 213..............217 422 541 559 565 568 569 610 612Hypoxia ....................................................... 420Innate Immunity ......................................... 548Inquiry......................................................... 353Interaction .................................................. 560Lentinula edodes ........................................ 736Light ... 25 106 73 211 258 294 399 406 409 431......................................... 435 436 437 446 473Lignocellulosic feedstocks ........................... 303Lipases ........................................................ 552Magnaporthe oryzae .. 116 138 70 165 166 196221 240 222 296 297 298 438 715 716 476 492478 480 572 573 574 575 576 577 578 634 652.................................................................... 674Maize ................. 540 579 607 619 621 622 623Malassezia .................................................. 350Mannosylerythritol lipid ............................... 62Marine ........................................................ 252Mass Spectrometry .. 7 46 247 299 310 355 724Mat genes ................................................... 206Mating Type Locus (MAT) ........................... 644Meiosis................................................. 205 730Meiotic recombination ........................ 717 718Membrane .... 130 167 192 80 214 438 693 580Metabolism ............ 15 16 19 23 30 32 33 34 3527th Fungal Genetics Conference | 303

KEYWORD LIST......... 36 37 40 11 13 61 63 67 282 296 315 383................................................ 423 444 558 620Metabolomics ......... 2 38 54 55 10 385 581 595Metarhizium robertsii ................................. 592Methylation 149 168 266 397 439 440 443 445.................................................................... 715Microarray ........... 61 102 238 326 432 362 694Microtubule ................................................ 374Mitochondria ....................................... 170 673Monascus pilosus, laeA ................................. 58Morphogenesis .... 35 109 111 113 116 121 131.... 71 150 155 72 169 170 171 172 173 176 181......... 189 191 197 91 200 300 307 391 441 468............................................................. 509 536Mucor, Argonaute, RNA silencing ............... 426Mycorrhiza ......... 227 301 327 442 513 515 527.................... 560 582 583 584 585 609 673 675Mycosphaerella........................................... 332Mycosphaerella graminicola, Fungicide,Resistance ................................................... 731Nematophagous fungi, Nematodes, Coevolution..................................................... 535Neurospora .......... 39 163 173 174 175 176 177....... 178 179 302 418 467 654 660 663 676 677Neurospora crassa ....... 40 41 42 43 44 117 118....... 121 128 131 153 154 159 162 168 172 180181 182 183 184 185 186 187 188 189 190 191192 193 198 213 241 303 304 338 384 386 387388 389 390 401 419 421 430 440 443 444 445.................... 446 475 717 718 719 720 721 642Nitrate assimilation ....................................... 41Nitric Oxide, Denitrification ........................ 538Noncoding ................................................... 263Oomycete ........................................63 119 532Organ specificity ......................................... 624Organic acids ................................................. 11Oxidative Stress .. 40 43 45 52 106 123 172 180194 195 196 311 404 410 698 722 729 546 558...................................................... 579 586 587PPR; Mucor circinelloides;β-glucosidase ..... 735Pathogenicity ...... 24 4 67 113 119 166 197 238244 253 261 272 275 281 288 320 330 332 336....... 339 402 420 439 465 698 699 709 722 491495 496 497 498 500 505 507 508 510 512 514516 518 520 522 523 525 526 477 479 480 536541 542 543 544 545 546 547 549 550 554 555556 561 562 482 568 575 578 485 581 588 589590 591 592 593 594 595 596 597 598 602 604617 618 621 627 629 632 634 637 638 653 686Perithecium ................. 10 128 180 204 532 663Peroxisome ..................................................... 8Peroxisome, ApsB ......................................... 98Phenomics .................................................. 716Phosphate ................................................... 611Phosphoproteomics .................................... 723Photoactivated Localization Microscopy(PALM) .......................................................... 95Phycomyces ................................................ 473Phylogenetic ... 34 39 209 253 254 256 257 305306 318 328 331 349 725 732 513 514 600 601.................... 608 645 648 662 674 678 679 685Phytophthora .................................533 638 661Phytophthora infestans............................... 112Platform ........................................................ 31Podospora anserina .................................... 453Polyketide Synthase .................................... 727Population.......... 245 342 703 708 694 731 501304511 600 643 645 646 649 650 655 661 665 666.............. 668 640 676 680 681 682 683 685 686Potato ............................................ 543 599 638Proteasome .................................................. 44Protein Degradation ................................... 487Protein Kinase ........ 42 110 115 129 177 183 96198 240 267 307 308 322 414 439 361 370 479.............................................................. 547 623Protein Localization ..... 30 5 49 12 112 141 153...... 162 163 164 165 74 169 186 191 81 84 295....... 365 457 464 726 496 524 525 570 571 486Protein Stability .......................................... 190Protein production ............................... 428 432Protein-Protein Interactions ..... 49 160 178 184.... 83 94 199 204 208 210 309 351 440 445 457........................... 458 504 556 578 484 585 616Proteomics ...... 21 7 46 47 135 100 299 310 325....... 328 723 724 695 740 507 570 573 602 630Pyrenochaeta lycopersici ............................ 614RNA-seq ........ 63 154 200 236 237 241 225 230259 274 275 282 231 300 311 312 313 314 315316 402 426 427 447 448 456 466 689 491 517......................................... 561 597 599 603 632RNAi ............................................... 125 243 416RNAi, genome defense, transposon, sex .... 701Ramularia.................................................... 564Ramularia collo-cygni .................................. 650Recombination ........................................... 658Recombination, outcrossing ....................... 680Response to non self................................... 637Saccharomyces cerevisiae ... 28 53 194 317 709Sclerotinia, oxalate, virulence ..................... 635Secondary metabolism .26 28 31 5 6 9 48 49 50.. 51 52 53 54 59 12 68 93 202 276 284 285 288309 311 318 234 341 409 411 417 359 364 449450 451 452 464 372 710 725 726 571 488 604................................................ 651 662 669 670Secondary metabolite .2 38 8 55 56 57 58 59 62.. 79 283 286 319 320 405 410 447 448 370 461.................... 708 692 727 586 594 595 615 671Secreted protein ......................................... 634Septoria tritici ............................................. 631Septum ....................................................... 173Sexual Development ... 59 107 108 125 133 139143 144 146 151 170 199 200 201 202 203 204205 206 207 208 210 211 218 219 243 251 268233 321 350 393 394 395 400 404 411 369 453.................... 454 455 700 668 640 677 684 687Signal Transduction .... 47 109 120 123 124 126.... 127 129 71 142 157 162 75 174 176 177 179.... 182 183 190 85 97 199 201 203 208 209 210246 294 322 345 393 414 430 431 436 438 359361 367 368 456 457 458 722 723 503 545 548......................................... 555 565 574 633 637Single Nucleotide Polymorphism ... 211 323 334.............................................................. 335 658Sordaria macrospora .................................. 139Spore Killer ................................................. 733Ste12 ........................................................... 574Susceptibility............................................... 265Symbiosis ........ 26 48 255 278 281 314 324 327417 459 728 490 493 511 515 521 527 528 559566 567 569 584 587 600 606 607 608 609 610.................................. 611 612 651 667 675 685Synteny ....................................................... 732Telomerase ................................................... 39Telomere-Linked Helicase ........................... 256Tomato ................................... 728 554 589 614Toxin ......... 55 56 145 293 345 460 461 705 741................................. 529 571 583 613 614 615Transcription ........ 44 333 376 387 408 412 422............................................................. 429 441Transcription Profiling ..... 155 287 375 379 407...... 437 362 449 371 711 729 478 617 631 664Transcription factor ... 9 56 126 76 187 89 91 96212 218 237 239 262 268 274 277 296 298 373382 391 395 405 412 413 418 419 421 424 355365 451 453 455 459 462 463 464 465 466 468...... 689 730 504 550 587 590 616 617 626 629Transcriptome60 140 147 102 246 263 277 278293 231 303 313 314 316 322 325 326 327 328...... 329 334 381 436 442 467 493 519 610 618Transformation ............................................ 354Translation.............................. 216 390 363 467Transport .... 41 61 156 161 192 92 213 214 216............................................... 329 428 713 619Transposable Elements ................................ 280Trichoderma .................................................. 60Trichoderma , protein production ................. 29Trichoderma harzianum ................ 316 739 740Trichoderma reesei...................................... 413Trichoderma, Biocontrol .............................. 563Trichoderma, PKS, ....................................... 341USER Friendly cloning .................................. 271Ustilago maydis ...... 35 62 137 152 160 215 216...... 330 331 381 713 730 619 620 621 622 623...................................................... 624 625 626Verticillium .................................................. 589Virulence ....... 167 76 195 197 229 251 264 291319 379 396 358 448 498 499 509 516 518 519528 530 533 534 552 553 562 483 570 484 487488 586 591 592 594 597 620 625 627 628 629............................................... 630 636 639 665Wheat .......... 265 460 728 512 524 549 628 631Whole Genome Sequencing ..... 50 224 227 254255 290 297 304 305 317 321 323 324 329 331332 333 334 335 336 337 338 339 342 343 517...... 526 588 601 606 628 632 666 681 682 686YFP ............................................................... 141alkaliphiles, biodiversity, moleculartaxonomy .................................................... 734amanitin ........................................................ 51biodegradation ....................................... 52 711bioenergy ...................................................... 13calcium ........................................................ 182carbohydrate structure; polysaccharidesynthases ....................................................... 19carbon metabolism ........................................ 32carotenoids.................................................. 406catalase-peroxidase encoding genes ........... 272cell-to-cell movement.................................. 138chromatin .................................................... 380chromosome transfer .................................. 149community genetics .................................... 667encapsulation, flow citometry ..................... 714enzymatic hydrolysis of PET .......................... 65functional unknown genes .......................... 235fungal meningitis, antidepressant,Cryptococcus ............................................... 738fungi-ant interactions .................................. 581fungicide ...................................................... 671genomics ..................................................... 257heme ............................................................... 1heterokaryon incompatibility ...................... 721

KEYWORD LISThigh temperature growth (37°C) ................. 639hrizontal chromosome transfer ................... 148hydrophobin ................................................ 585hypoxia, antifungal susceptibility ................ 481innate immunity .......................................... 523interaction ................................................... 706intron .......................................................... 392introner, spliceosomal intron, intron gain ... 344iron .............................................................. 358iron regulation ............................................. 357lichen-forming fungi, PKS genes, Cladoniametacora ....................................................... 66lignolysis ........................................................ 45mRNA degradation ........................................ 23marine fungi .................................................. 68miRNA ......................................................... 415mitochondria ........................................ 188 289mitochondrial inheritance ........................... 700mushroom breeding, sporulation ................ 707natural selection.......................................... 657nuclear division ........................................... 114nutrient immunity ....................................... 500oomycete .................................................... 520oomycete, Phytophthora, actin ................... 220peptide ........................................................ 691porous substrates ........................................ 161prion; STAND ............................................... 345quantification .............................................. 576repetitive elements ..................................... 348roxithromycin ................................................ 70sclerotium ................................................... 710sidB, RNAi, Aspergillus nidulans .................. 474small RNA .................................................... 349smallRNA, Transposon, ................................ 475smut fungi ................................................... 291statin resistance ............................................ 53stress ........................................................... 151taxonomy, wood_diseases, ITS.................... 737thigmotropism............................................. 159twintron ...................................................... 232vacuole ........................................................ 217veA, Gliotoxin .............................................. 356velvet ............................................................. 90water ........................................................... 156white-opaque switch, biofilms .................... 37727th Fungal Genetics Conference | 305

AUTHOR LISTAAanen, Duur K. .......... 654, 655Abba', S. .................... 167, 630Abba, S. ............................. 327Abbey, Darren ................... 502Abdeljalil, Salma ................ 412Abduljabar, Hamzah .......... 172Abe, A. .............................. 577Abe, Keietsu ...... 367, 368, 456Abe, M. ............................. 437Abe, Tomoko ..................... 472Aβmann, Daniela............... 622Abreu-Goodger, C. ............ 435Abt, Beate ......................... 471Adan, O. C. G. ............ 156, 161Adan, Olaf ......................... 136Addicott, Ethan ................... 40Aebi, M. ............................ 741Aebi, Markus ..... 219, 705, 706Aerts, A. ............................ 332Affeldt, Katharyn ............... 486Affeldt, Katharyn J. ............ 633Agafonov, Oleg .................... 41Aguirre, Jesus .................... 180Ah-Fong, Audrey M. V. ...... 112Ahmad, Aijaz ..................... 698Ahmad hussin, N. ................ 17Ahmed, Yasar Luqman ........ 79Ahrén, D. ........................... 602Ahrendt, Steven ................ 294Ahuja, Manmeet ................. 54Aït-Benkhali, J. .......... 321, 453Ajami-Rashidi, Ziba............ 624Alam, Md. Kausar .............. 360Albermann, Sabine E. ........ 405Alcazar-Fuoli, L. ................. 484Aleman-Duarte, M. I. ........ 521Alexander, Johnson ........... 377Alexander, N. J. ................. 669Ali, Hazrat............................ 55Alkan, Noam...................... 277Almeida, Fausto Bruno DosReis ..................................... 18AlSheikh, Khlood S. ........... 351Altelaar, A. F. Maarten ...... 100Amanianda, Vishu K. ......... 485Amano, K. .......................... 480Ames, Brian ........................... 5Amir, Sherman .................. 479Ampomah, Osei ................ 119Amselem, J. ....................... 652Amselem, Joelle ........ 222, 737Amyotte, Stefan ................ 603Amyotte, Stephan ............. 248Anasontzis, George E. ....... 325Anchieta, Amy ................... 300Anderluh, Gregor .............. 226Andersen, Mikael R. .............. 6Anderson, C. L. .................. 732Anderson, Jonathan P. ...... 588Anderson, Matthew Z. ...... 376Anderson, Ryan G. ............ 533Andersson, K.-M................ 602Andes, David ..................... 356Andongabo, Ambrose ....... 631Andrews, Jared ................. 339Andrianopoulos, Alex 212, 597Angelard, C. ...................... 675Anglin, Sarah Lea ................ 78Anishchenko, I. M. ............ 305Anita, Sil ............................ 591Antignani, Vincenzo .......... 483Antonieto, Amanda............. 47Antoniêto, Amanda C. C. ... 413Antoniw, John ................... 229Antony, C. ......................... 134Anyaogu, Diana ..................... 7Anyaogu, Diana C. ............... 31Ao, Jie ............................... 118Aouini, Lamia .................... 329Aoyama, Miki .................... 463Araujo-Palomares, C. ........ 181Arazoe, T. .......................... 572Are, Tina Marie Monge ....... 41Arentshorst, Mark ............. 102Arie, T. .............................. 572Ariyawansa, Kahandawa G. S.U. ...................................... 565Arnaud, Martha ................ 223Arnaud, Martha B. ............ 340Arnold, A. Betsy ................ 685Arnold, A. Elizabeth .......... 679Aro, N. ............................... 432Aro, Nina K. ......................... 29Arredondo, F. .................... 530Arredondo, Felipe D. ......... 415Arredondo, Felipe R. ......... 295Arruda, L. K. ...................... 504Asano, K. ........................... 577Asiegbu, F. ........................ 702Asiegbu, Fred O. 349, 648, 656Åsman, Anna K. M............. 402Ast, Julia ...................... 62, 616Atanasova, Lea .................. 341Atanasova-Penichon, V. ...... 38Athanasopoulos, A. ........... 693Atkin, Audrey L.................. 498Atkinson, S. ....................... 147Attanayake, Renuka Nilmini680Attard, Agnes .................... 477Au, Chun Hang .................. 287Avalos, J. ........................... 406Avalos, Javier .................... 141Avalos-Borja, Miguel ......... 719Averettec, Anna ................ 350Avrova, Anna O. ................ 402Axtell, Michael .................. 551Azeddine, Saad ................... 15Azurmendi, Hugo .............. 528BBååth, Jenny ....................... 61Bach, Jocelyne .................... 15Baek, J.-H. ......................... 414Baek, M. ............................ 386Bahn, Yong-Sun ......... 246, 393Bailey, Andy M. ........... 57, 537Baker, Scott ... 13, 54, 306, 678Baker, Scott E. ................... 341Baker, S. E. ................ 282, 310Bakkeren, G. ..................... 331Bakkeren, Guus ......... 299, 628Baldin, Clara ...................... 231Baldwin, Thomas .............. 538Balesdent, Marie-Hélène .. 403Balint-Kurti, Peter ............. 607Baller, Joshua A................. 376Balmas, Virgilio ................. 262Ban, A. ................................ 12Banerjee, D. ........................ 23Bangera, Gita .................... 352Banjara, Nabaraj ............... 703Banno, S. ........................... 183Banno, Shinpei .................. 131Banta, Travis ....................... 78Banuett, Flora ................... 169Barker, Bridget .................. 355Barker, Bridget M. .... 342, 485Barra, James ....................... 78Barral, Jose ....................... 390Barreau, C. ........ 410, 447, 708Barry, Kerrie ...................... 678Bartnicki-García, S............. 173Bartnicki-García, Salomón 121Barton, Hazel ...................... 36Basenko, Evelina Y. ........... 443Bass, B. ............................. 147Bastiaans, Eric ........... 654, 655Bastidas, Robert J. .... 109, 416Bataillé-simoneau, Nelly ... 586Bauer, S. ........................... 303Bauer, Stefan .................... 678Baumgartner, Kendra ....... 537Bayram, Ö. ........................ 117Bayram, Oezguer .......... 79, 93Bayram, Özgür ............ 94, 487Bazafkan, Hoda ................. 201Beaudet, Denis ................. 673Beaurepaire, C. ................. 378Beausiegle, S. .................... 332Beauvais, Anne ..................... 3Bec, Sladana ..................... 490Bech, Lasse ......................... 60Becker, L. .......................... 686Becker, Yvonne ......... 587, 612Beckers, Gerold J. M. ........ 623Beckmann, Nicola ................. 1Beffa, R. ............................ 240Beimcik, Eva ...................... 299Bekker, M. ........................ 156Bélanger, R. R. .................. 331Belfort, Marlene ............... 257Bell, A. E. ........................... 321Bellgard, Matthew ............ 261Bello, Marco ....................... 43Bell-Pedersen, Deborah .... 383Belzile, F. ........................... 331Benitez, M.-S. ................... 686Benoit, Isabelle ......... 694, 695Benz, J. P. .......................... 303Bergmann, Holger ............. 725Bergmann, S. .................... 713Bergquist, Peter ................ 425Berkes, Charlotte .............. 591Berman, Judith ......... 376, 502Berne, Sabina ...................... 67Bernhards, Yasmine .......... 204Bernillon, S. ........................ 38Berry, Daniel ....................... 48Berthier, Erwin ................. 486Bertolini, Maria Célia ........ 419Bertolini, M. C. ........... 42, 384,................................. 388, 421Bertuzzi, M. ...................... 484Bertuzzi, Margherita ........... 75Betenbaugh, Michael ........... 7Bettgenhaeuser, Jan ........... 75Betz, Ruben ...................... 527Beyer, Marco ............ 262, 671Beyhan, Sinem .................. 468Bharti, Arvind K. ............... 631Bhatnagar, Deepak ............. 69Bhattacharya, Debashish .. 674Bi, Qing ............................. 534Bidart, Frédérique ............ 293Bienkowski, Damian ......... 493Bignell, Elaine ..................... 75Bignell, E. M. ..................... 484Bilanenko, Elena N. ........... 734Billard, Alexis ...................... 15Billmyre, R. Blake .............. 243Billon-Grand, Geneviève ... 495Bilmyre, Blake ................... 125Binder, Ulrike ............ 481, 636Binkley, Gail ...................... 340Binkley, Jonathan ............. 340Bin Terhem, Razak .... 107, 108Birch, Paul R. J. ................. 532Bitas, Vasileios .................. 551Bittner, Noëlle .................. 679Blanc, Guillaume ............... 278Blanchette, Robert ........... 306Blanchette, Robert A. ......... 52Blatzer, Michael ................ 471Blehert, David S. ............... 688Bleuler-Martinez, S. .......... 741Blicher, Lene H. ..................... 6Bloch Jr., Carlos .................. 46Bloemendal, Sandra ... 49, 199Bloom, Amanda L. M. ....... 397Blosser, Sara J. .................. 355Bluhm, B. .......................... 568Bluhm, B. H. .............. 601, 613Blumenhagen, C. .............. 638Blumhoff, Marzena L. ......... 11Blümke, Antje ................... 529Bochen, Florian ................. 625Boddey, Justin A. .............. 532Bödeker, Inga ................... 562Boenisch, Marike .............. 140Boenisch, Marike Johanne......................................... 545Boeren, Sjef ...................... 248Boettger, Daniela .............. 725Bohlmann, Joerg ............... 288Böhm, Julia ....................... 202Bohovych, Iryna .......... 16, 722Bok, Jin Woo ............... 54, 452Boleti, H. ........................... 693Bölker, Michael ... 62, 152, 160,................................. 457, 616Bollmann, Stephanie R. .... 415Bolton, Melvin .................. 251306

AUTHOR LISTBolton, Melvin D. ....... 516, 684Bonito, G. ......................... 305Bonito, Gregory ................ 606Borhan, Hossein ............... 280Borkovich, Katherine . 162, 178Borkovich, Katherine A. .... 190Bormann, Jörg ... 545, 546, 547Bose, I. .............................. 114Boshoven, Jordi ................ 526Boshoven, Jordi C. ............ 516Boubakir, Zakia ................. 728Bourque, S. ....................... 371Boussau, Bastien .............. 306Bouzid, Ourdia .................. 696Bovenberg, Roel ................. 55Bowen, J. .......................... 665Bowen, Joanna .. 249, 517, 526Bowman, Barry J. .............. 217Bowman, Emma Jean ....... 217Bowring, Frederick J. . 717, 718Bowyer, P. ................. 488, 690Bowyer, Paul .................... 689Boyd, Kendra .................... 348Bracher, Franz .................. 481Bradshaw, Rosie E. .... 336, 565Braesel, Jana ....................... 50Braithwaite, Mark ............ 493Brakhage, A. ..................... 484Brakhage, Axel .................. 231Brakhage, Axel A... 2, 357, 358,.................................. 359, 487Branco, S. ......................... 314Branco, Sara ..................... 681Brand, Alex ....................... 111Brandström Durling, Mikael ............................................ 259Brandt, Raphael ................ 179Brandt, Ulrike ................... 370Braunberger, Katharyn ..... 449Braus, Gerhard ....... 79, 93, 94,......................................... 536Braus, Gerhard H. ...... 487, 589Braus, G. H. ....................... 117Braus-Stromeyer, Susanna .............................................. 536Braus-Stromeyer, SusannaA. ...................................... 589Bravo Ruiz, G. A. ............... 552Bredeweg, E. .................... 384Bredeweg, Erin L............... 383Brefort, Thomas ............... 625Brem, Rachel .................... 664Brestelli, John ................... 338Breton, Annick .................. 637Breuil, Colette .................. 288Brewer, Marin .................. 645Brigitte, Laetitia ................ 737Brock, M. .......................... 482Bromley, Michael J. .......... 689Bromley, M. J. ............ 488, 690Brown, Alistair .................. 110Brown, Alistair J. P. ......16, 722Brown, Christopher .......... 493Brown, Daren ................... 306Brown, Daren W. .............. 318Brown, Jessica C. S. ........... 244Browne, Neill .................... 636Brownlee, Christopher ...... 679Broz, Karen ........................ 571Brückner, Elena ................. 545Brueggeman, Robert S. ..... 627Bruel, C. ............................. 374Bruez, Emilie ..................... 737Brun, S. .............................. 453Brunk, Brian ...................... 338Brunner, Michael ........ 44, 446Brunner, Patrick ................ 643Brunner, Patrick C. .... 448, 657Bruno, Kenneth ..... 13, 54, 678Bruns, T. D. ........................ 314Bruns, Tom ........................ 681Brus, Maja ......................... 614Bryan, Gregory T. .............. 569Brzywczy, J. ....................... 362Budge, Susan ............... 16, 110Budkewitsch, G. ................ 371Bueche, Joanna A. ............. 198Buée, M. ............................ 702Buée, Marc ..........................45Bugeja, Hayley E. ....... 212, 597Bühler, Nicole ......................80Bui, Tien ............................ 643Bulone, Vincent ...... 19, 20, 21,............................ 22, 119, 275Burai, J. .............................. 371Burchhardt, Kathleen M. ... 646Burggraaf, Maria A. .............99Burnett, Fiona ................... 650Buscot, F. ........................... 327Bushley, Kathryn E. ... 311, 320Busk, P. .............................. 735Busk, Peter ........ 548, 697, 712Busk, Peter K. .................... 343Busk, Peter Kamp ................60Busman, M. ....................... 670Butler, Geraldine ............... 239Butschi, Alex ...................... 705Butts, Arielle ...................... 244CCabral, Adriana .................. 596Cabrera, Ilva Esther ........... 162Caddick, Mark X. ....... 223, 363Caffier, V. .......................... 665Cai, Guohong ............. 279, 674Calcagno-Pizarelli, A. M. .... 484Callahan, Damien ................56Callegari, Eduardo ............. 236Calmes, Benoit .................. 586Calo Varela, Silvia .............. 416Calvo, A. M. ....................... 710Calvo, Ana ......................... 726Calvo, Ana M. .................... 356Calvo-Byrd, Ana M............. 364Calzada, Arturo.................. 137Camacho-López, MarcoAntonio ............................. 719Camborde, L. ..................... 520Campbell, Leona ........ 507, 643Candido, T. .................. 42, 388Candido, Thiago S. ............. 419Cano-Domínguez, N. ......... 181Cano-Dominguez, Nallely .. 180Canovas, D. ....................... 714Cao, Han ............................ 290Capaul, R. .......................... 122Capelluto, Daniel ............... 528Carbone, Ignazio . 69, 668, 685Cardenas, Maria E. ............ 416Cardinale, Francesca ......... 408Carlough, J......................... 530Carreras-Villaseñor, N. ..... 433,.......................................... 435Carrington, James C. ......... 415Carter, Dee ........ 448, 507, 643Cartwright, Gemma M. ..... 587Cary, J. W. ......................... 710Cassin, Andrew.................. 618Castanheira, Sónia M. ....... 215Casto-Longoria, Ernestina . 180Castrillo, Marta ................. 141Castrillo Jimenez, Marta .... 406Castro, Lilian ....................... 47Castro, Lílian S. .................. 413Castro-Longoria, E. ............ 181Castro-Longoria, Ernestina 719Catcheside, David E. A. ..... 717,.......................................... 718Cerqueira, Gustavo C. 223, 340Cerutti, A. .......................... 520Cervantes, M. .................... 426Cha, Joonseok ................... 390Chacko, Nadia ................... 391Chae, Keon-Sang ........... 85, 93Chae, Michael ........... 188, 390Chang, Chia-Chen .............. 182Chang, Jinhui ............. 287, 307Chang, P.-K. ....................... 309Chang, Shu-lin ..................... 54Chankhamjon,Pranatchareeya ..................... 2Charlton, Nikki D. 26, 511, 567Chatterjee, G. .................... 114Chatterjee, Sayantani ........ 375Chattoo, Bharat B. ............. 575Chaudhary, Suman ............ 211Chávez, S. .......................... 714Chayakulkeeree, Methee .. 126Chellappan, Biju ................ 248Chellappan, BV. ................. 672Chemnitz, Jan .................... 549Chen, C. L. ......................... 205Chen, Dan.............................. 3Chen, Donquan ................. 300Chen, Fen .......................... 676Chen, Feng ................ 649, 678Chen, Julian J.-L. .................. 39Chen, Ling ......................... 274Chen, She .......................... 390Chen, Su-Der ....................... 25Chen, W. ........................... 615Chen, Weidong ......... 598, 680Chen, Xiaoxuan ................. 592Chen, Y. ............. 305, 312, 314Cheng, Xuanjin .......... 287, 469Cherif, Chetouhi ................ 265Cheung, H. Y. K. ................. 730Cheung, Man Kit................ 287Cheverton, A. .................... 484Chiang, Yiming .................... 54Chiapello, H. ...................... 652Chiapello, Helene .............. 222Chibucos, Marcus.............. 223Ching-yu, Lu ...................... 169Chitnavis, Madhura ........... 348Chitrampalam, Periasamy . 455Cho, A.-R. .......................... 145Cho, Yangrae ..................... 590Choi, G. J. .......... 144, 145, 195Choi, H.-K. ......................... 276Choi, Jaehyuk .... 221, 296, 297Choi, Jaeyoung . 221, 297, 349,.................................. 634, 715Choi, Jeahyuk .................... 298Chonofsky, M. ................... 214Chonofsky, Mark ................. 92Chowdhury, Tahmeena ..... 109Christensen, Michael J. ..... 569Christmann, Martin ............. 94Chu, Ashley ....................... 460Chujo, Tetsuya .................. 417Chung, Chia-Lin ................. 506Chung, Dawoon ................ 355Chung, Hyunjung .............. 298Churchill, Alice C. L. ........... 607Cirino, Robert .................... 142Ciuffetti, L. M. ................... 561Ciuffetti, Lynda ......... 336, 460Ciuffo, M. .......................... 630Clarissa, Nobile ................. 377Clark, Bethany ................... 618Clark, Helen....................... 518Clark, Helen R.................... 483Clark, J. S. .......................... 686Clarke, David ..................... 365Clarke, David F. ................. 366Clavé, Corinne ................... 293Clergeot, Pierre-Henri219, 614Coelho, P. S. R. .................. 504Cohrs, Kim ......................... 106Coile, Clifford ...................... 78Collemare, Jérôme ............ 344Collet, James ....................... 13Collins, Rebecca ................ 689Connolly, Lanelle R. ... 242, 385Connolly, Leona ................ 239Cook, Emily ....................... 722Cooke, D. E. L. ................... 661Cooke, Ira .......................... 526Cooper, Michael.................. 35Cooper, Sarah ................... 449Coppin, E. .................. 321, 453Coradetti, Samuel T. ......... 418Corcoran, Padraic.............. 676Cornelissen, BJC. ............... 672Cornelissen, Maxim .......... 375Corral Ramos, C. ............... 553Corrochano, LM ................ 473Corrochano, Luis ............... 211Corrochano, Luis M. .......... 446Costilla, Landi VG .............. 691Cotty, Peter ....................... 347Courbot, Mikaël S. ............ 631Couto, Eva ......................... 739Cowen, Leah ..................... 110Cox, Russell J. ...................... 5727th Fungal Genetics Conference | 307

AUTHOR LISTCramer, Robert A. ..... 355, 485Craven, Kelly D. ........... 26, 567Crenshaw, Nicole J. ........... 539Crespo, Rhaisa ................... 266Crestani, J. ......................... 723Croll, D. ............................. 658Croll, Daniel ............... 448, 657Cronin, Elizabeth ............... 728Csikasz-Nagy, A. ................ 386Csukai, Michael ................. 731Cubeta, Marc A. ................ 646Cullen, Dan ........................ 247Culley, David ............... 13, 678Cuomo, Christina .............. 339Cuomo, Christina A. .......... 628Cupertino, F. ............... 42, 421Cupertino, Fernanda B. ..... 419Curlango-Rivera, G. ........... 541Czymmek, Kirk................... 142DDagdas, Yasin .................... 165Dagdas, Yasin F. ................ 492Dahlmann, Tim .................. 313Dahms, Tanya ..................... 88Dalmay, T. ......................... 426Dang Tat, Thanh ................ 325Darbyshir, Heather L. ... 687Darwiche, Sabrina ............. 701Dasgupta, Arko.................. 389Daskalov, Asen .......... 345, 637Davidson, Fordyce ............. 159Davies, Emma.................... 689Davis, Carol E. ..................... 63Davis, C. Britton ................ 540Davis, Meryl ...................... 365Davis, Meryl A. .................. 366Dawodu, Omolola ............... 84Dawson, Thomas ............... 350Day, Ryan ............................ 78Dean, Ralph ............... 166, 715Dean, Ralph A............ 476, 573de Beer, Z. Wilhelm ........... 206de Bekker, Charissa ........... 581Debets, Alfons J. M. .. 654, 734Debieu, Danièle ................... 15DeBlasio, Daniel ................ 679Deborde, C. ......................... 38de Bruijn, Irene ......... 275, 525Debuchy, R. ............... 321, 453de Castro, Patrícia A. ......... 691de Gracia, M. ..................... 665de Groot, P. W. J. .............. 122De Haro, J. P. ..................... 426de Jonge, Ronnie ............... 251Dekhang, Rigzin ................. 383de Koster, C. ...................... 122Delancy, E. ........................ 147de la Providencia, Ivan ...... 673Delaye-Arredondo, L. J. ..... 521DelBove, Claire .................... 84Delgado-Álvarez, Diego ..... 173Delgado-Ramos, L. ............ 714Deller, Siãn ........................ 322Delorey, Toni M. ............... 659deMello, A. J. .................... 741deMello, Andrew J. ........... 706Deng, Cecilia ............. 517, 526Deng, Shuang ...................... 13Denison, Steven H. .............. 81Denning, D. ....................... 690Derbyshire, Mark .............. 631de Sena-Tomas, Carmen ... 137Deshpande, Raamesh ....... 244Desmarini, Desmarini ........ 126Dettmann, Anne ....... 174, 176de Vries, Ronald P.694, 695, 696de Vries, R. P. ...................... 27de Wit, Pierre .................... 344de Wit, Pierre JGM ............ 336de Wit, P. J. G. M............... 272Dhillon, B. ................. 332, 666Dhingra, Sourabh ...... 356, 364Diaz, Sara ............................ 22Díaz-Choque, Rodrigo ....... 444Dickmanns, Achim ............ 132Didierjean, Claude .............. 45Dieguez-Uribeondo, Javier .. 21Diernfellner, Axel ................ 44Dieryck, Cindy ................... 495Dietrich, Bob ..................... 631Dijksterhuis, J. ... 156, 161, 207Dijksterhuis, Jan ........ 136, 151Dijkwel, Paul ....................... 48Dinh Thi My, Hang ............ 325Di Pietro, Antonio554, 555, 556Dirschnabel, Daniela ......... 404Divon, Hege H. .................. 318Dixelius, C. ........................ 453Dixelius, Christina ............. 402Dixit, Cheshil ....................... 78Djamei, Armin ... 494, 532, 619Djordjevic, Julianne ........... 126Dobinson, Katherine F. ..... 300Dodhia, Kejal ..................... 724Doehlemann, Gunther622, 624Doernte, Bastian ......... 64, 470Doktycz, Mitchel ....... 515, 606Dolling, Rick ...................... 647Donaldson, M. E. ............... 330Dong, Suomeng................. 531Donia, M. .......................... 604Donner, J. .......................... 291Doré, J. .............................. 327Doughan, Benjamin .......... 454Dowd, P. ........................... 710Downes, Damien ............... 365Downes, Damien J............. 366Downey, Kurtis.................. 396Downs, Bradley ................... 82Doyle, C. E. ........................ 730Drevet, C. .......................... 321Drews, Kelly ...................... 518Drews, Kelly C. .................. 483Driessen, Arnold ................. 55Druzhinina, Irina S. ...... 65, 341Dubey, Mukesh ................. 193Dubiel, Wolfgang ................ 94DuBois, Juwen C. ............... 420Dubos, Tiphaine ................ 671Du Fall, Lauren A. .............. 595Dumarçay, Stéphane .......... 45Dumas, B........................... 520Dumur, Jérôme ................. 586Duncan, Randall ................ 142Duncan, Shona M. .............. 52Dunlap, Jay ............... 383, 389Dunlap, Jay C. ..................... 73Dunstan, Kelly ................... 569Duplessis, Sebastien ......... 519Duplessis, Sébastien ......... 339Duran, Rocio ..................... 726Durel, C.-E. ........................ 665Duressa, Dechassa ............ 300Dutken, Nicolette ............. 187D. Walton, Jonathan ........... 51Dyer, Paul ......................... 202Dyer, Paul S. .............. 640, 687EEastlack, Steven .................. 78Eaton, Carla .............. 162, 612Eberle, M. ......................... 690Echauri-Espinosa, Ramón O.121Efrat, M. ............................ 168Egan, Martin ....................... 92Egan, M. J. ........................ 214Ehrlich, K. .......................... 309Ehrlich, K. C. ...................... 710Ekengren, Sophia .............. 614Elfstrand, Malin ................ 562El Ghalid, Mennat ............. 403Elliott, Candace ................... 56Elsenpeter, Ryan ............... 213Emerson, Jillian ................. 383Emily, Fox ......................... 377Emri, Tamás ........................ 91Ene, Iuliana V. ..................... 16Eng, Hayde ........................ 166Ennos, Richard .................. 650Entwistle, Ruth ................... 54Epstein, Lynn ...................... 43Erbs, Gitte ......................... 548Erdmann, Susann .............. 203Erika, Kothe ...................... 729Escudero, Nuria ................ 570Eslami, H, .......................... 474Espagne, E......................... 321Espeso, E. A....................... 484Espino, J. ................... 238, 407Esquivel-Naranjo, EU. ....... 435Evangelisti, Edouard ......... 477Eyzaguirre, Jaime .............. 236FFaça, Vitor M. ..................... 47Fagnan, Kirsten ................. 306Fahlgren, Noah ................. 415Faivre Talmey, Y. ............... 374Fajardo, R.......................... 117Fakhoury, Ahmad ............. 542Fan, Tao ............................ 299Fang, Weiguo .................... 592Farazkhorasani, Fatemeh ... 86Faris, Justin ....................... 524Faris, Justin D. ................... 627Farman, Mark ................... 256Farmer, Andrew D. ........... 631Favier, Frédérique .............. 45Faville, Marty J. ................. 608Feau, N. ............................ 332Feau, Nicolas .................... 336Fekete, Erzsébet ............... 232Feldbruegge, M. ............... 216Feldbrügge, M. ................. 713Feldman, D. ...................... 168Feldmann, Jörg ................. 500Felício, A. P. ........................ 42Feng, Mu .......................... 175Ferdous, Janna.................. 507Feretzaki, Marianna .. 125, 243Fernandes, Caroline M. .... 691Fernandes-Matos, Larissa . 639Ferrer, S. ........................... 530Ficner, Ralf .................. 79, 132Figueroa, Melania ............. 491Filichkin, Sergei ................. 491Filler, Scott.................. 76, 379Filler, Scott G. ....................... 3Filler, S. G.......................... 484Filley, Timothy R. ................ 52Fillinger, Sabine .......... 15, 329Finlayson, M. .................... 122Firat, Demet ..................... 545Fischbach, M..................... 604Fischer, Gregory J. ............ 486Fischer, R. ........................... 80Fischer, Reinhard ....... 8, 95, 98Fischer, Steve ................... 338Fisher, Jenny ..................... 679Fitak, Robert R. ................. 679Flaherty, J. ........................ 147Flajsman, M. ..................... 271Fleetwood, Damien .. 324, 610Fleetwood, Damien J. ....... 569Fleissner, André ........ 179, 370Fleissner, Andre ................ 174Flipphi, Michel .................. 232Florence, Cambon ............ 265Floudas, Dimitris ............... 306Floyd, Anna ....................... 246Fluhr, Robert .................... 277Foley, Anna ....................... 643Foley, Rhonda ................... 588Fontaine, Thierry .................. 3Forbes, G. ......................... 661Ford, Kathryn .................... 537Forester, Natasha ............. 559Forester, Natasha T. ......... 459Fortwendel, Jarrod R. ... 72, 74Foster, Gary D. ............ 57, 537Foulongne-Oriol, M. ......... 708Fountaine, James ..... 564, 641,......................................... 650Fourie, G. .......................... 260Fournier, E. ....................... 652Fournier, Elisabeth ........... 222Fowler, Thomas J. ............. 208Fox, K. ............................... 484Fraczek, Marcin ................ 689308

AUTHOR LISTFrancis, Martin ................. 560Franck, Bill ........................ 476Franck, William ................. 166Franck, William L. ............. 573Frandsen, Rasmus .............. 53Frandsen, Rasmus J. N. ....... 28Fränzel, Benjamin ...... 199, 210Fraser, James A. ................ 366Free, Stephen J. . 118, 128, 153Freihorst, Daniela ............. 203Freitag, Johannes ........62, 616Freitag, M. ........................ 384Freitag, Michael ........ 175, 242,.......................... 380, 383, 385Freitas, F. .....................42, 421Freitas, Fernanda Z. .......... 419Frey, Regula ...................... 731Fricker, Mark .................... 196Friedlander, Gilgi .............. 277Friedman, Steven ............. 380Friesen, Timothy ............... 524Friesen, Timothy L. ........... 627Friman, E. ......................... 602Frisvad, Jens ....................... 10Fry, W. E. .......................... 661Fu, Ci................................. 153Fudal, Isabelle................... 403Fujii, R. ................................ 12Fujimura, M. ..................... 183Fujimura, Makoto ..... 131, 191,......................................... 401Fukada, Fumi .................... 113Fuller, Kevin K. .................... 73Funck Jensen, Dan ............ 259Funt, Jason ....................... 502Furuki, Eiko ....................... 724GGadd, Geoffrey ................. 159Gagey, M.-J. ...................... 240Gajdeczka, M. ................... 682Galazka, Jonathan ............ 175Gao, L. .............................. 326García-Martínez, Jorge ..... 141Garcia-Pedrajas, Maria ..... 300Gardiner, Donald M. ......... 261Gardner, Richard .............. 610Gargas, Andrea .......... 352, 688Gargouri, Ali ..................... 412Garibaldi, Angelo .............. 544Garnek, Joshua A. ............. 416Garre, Victoriano .............. 436Garud, AMruta ................. 184Garvin, David .................... 491Gashgari, Rukaia M. .......... 599Gaulin, E. .......................... 520Gauthier, Gregory M. ....... 441Gautier, Angélique ........... 737Gawinska-Urbanowicz, Hanna543Gay, G. .............................. 327G-David, Biron .................. 265ge, Lingxiao ......................... 34Gebhart, Dana .................. 171Gehrke, Christopher ........... 74Gelhaye, Eric....................... 45Geller, Alexander...................3Gendrault, A. ..................... 652Geoffrion, N. ..................... 371Georg, Raphaela ................ 739Georgieva, Marina L. ......... 734Gérardin, Philippe ...............45Germain, Hugo .................. 519Gherbawy, Youssuf A. ....... 599Ghimire, Sita R................... 567Ghosh, Anupama ............... 619Giang, Anh ...........................78Gibeaux, R. ........................ 134Giesbert, Sabine ................ 496Gill, S. ................................ 484Gilmore, Sarah A. .............. 155Giotia, Anastasia ............... 350Giraldo, Martha ................. 165Giraldo, Martha C. ............. 492Girard, Vincent .................. 495Girlanda, M. ...................... 327Gladieux, Pierre ......... 660, 663Glass, L. ............................. 720Glass, Louise ...................... 192Glass, N. L. ......................... 303Glass, N. Louise………154, 185,…………198, 315, 418, 466, 721Gleason, Cynthia ............... 588Gleason, Frank .................. 712Glenn, Anthony ................. 538Glenn, Anthony E. ..... 539, 540Gloss, Andrew ................... 679Gobec, Stanislav ..................67Goebels, Carolin ................ 392Goehre, V. ......................... 216Goity, A. .................... 386, 387Gokce, Emine .................... 573Gold, Scott E. ............. 539, 540Goldman, Gustavo............. 361Goldman, Gustavo H. ........ 691Gomi, K. ...............................12Gomi, Katsuya ........... 104, 427Gonçalves, R. ....... 42, 384, 421Gong, Jinjun ...................... 458Gonoi, Tohru ..................... 456Gonzalez-Hilarion, Sara ..... 392González Roncero, M. I.552, 553Goodwin, S. B. ................... 332Goodwin, Stephen B.......... 336Gordon, Colin ......................94Gordon, Sean .................... 491Gordon, T. R. ..................... 264Gorka-Niec, Wioletta ......... 563Gorman, Jeffrey................. 724Gorsich, Steve ................... 194Goss, E. M. ........................ 661Gotanda, Yasutaka ..............59Gotfredsen, Charlotte H. .......9Gottfredsen, Charlotte H. ......6Gough, Kathleen M. ............86Gourgues, Mathieu ........... 477Gouvêa, Paula ................... 361Govers, Francine ....... 130, 220Govetto, Benjamin ............ 477Gow, N. ............................. 699Gow, Neil ........................... 159Gow, Neil A. R.16, 503, 565, 722Graczyk, Sebastian ............ 563Gradnigo, Julien S. ............. 732Grahl, Nora ............... 355, 485Grandaubert, Jonathan280, 618Granek, Josh ...................... 644Gras, Diana E. .................... 430Grashel, Tayler .................. 172Grassi, Paola ...................... 705Grava, S. ............................ 158Gravelat, Fabrice N. .............. 3Grayburn, Scott ................. 726Grebogi, Celso ................... 722Greenberg, E. Peter ........... 515Greenshields, David .......... 611Gregory, Hurst .................. 515Greimel, Katrin .................... 65Grell, M. ............................ 735Grell, Morten .................... 697Grell, Morten N. ................ 704Grell, Morten Nedergaard... 60Grenville-Briggs, Laura20, 119, 275Grice, C. M. ....................... 484Grigoriev, I. ............... 327, 332Grigoriev, Igor .. 227, 228, 281,……306, 590, 606, 678Grigoriev, Igor V.237, 336, 649Grisafi, Paula L. ................. 440Grognet, Pierre ......... 157, 733Gross, Nathan W. ................ 82Groth, Marco .................... 231Gruetzmann, Konrad ......... 231Grum-Grzhimaylo, Alexey A.734Grund, E. ........................... 240Gruntjes, Thijs ................... 695Grünwald, Niklaus J. .. 415, 683Gryganskyi, Andrii ..... 600, 606Gryganskyi, A. P. ............... 305Grzebyta, Jacek ................. 229Gsaller, Fabio ............ 357, 471Guazzone, Natalia ............. 493Gube, Matthias ................. 729Guebitz, Georg M. ............... 65Gueldener, U. .................... 407Guérin, C. .......................... 652Guerin, Cyprien ................. 222Guillemette, Thomas ......... 586Güldener, Ulrich 140, 263, 269Gullino, Maria Lodovica .... 544Gunatilaka, Leslie .............. 594Gunel, Aslihan ................... 299Günther, Alix ..................... 729Guo, Chun jun ..................... 54Guo, Jinhu ......................... 390Guo, L. ............................... 326Gupta, Yogesh K. ....... 165, 492Gurr, Sarah ........................ 196Guthke, Reinhard .............. 231Gutierrez, Matias .............. 468Gutiérrez, S. ...................... 669Gutiérrez, Sammy ............. 123Gutzeit, Jonathon .............. 187Guzman-Guzman, P. ......... 521Gyawali, Rachana127, 510, 700HHaar, C. ............................. 216Haas, Brian ........................ 223Haas, Hubertus1, 357, 358, 471Habel, Andreas ..................... 2Haβing, Berit ..................... 204Hacquard, Stéphane . 519, 603Hadar, Yitzhak ................... 711Hadeler, Birgit ... 545, 546, 547Haffner, Fernanda ............. 678Hagiwara, Daisuke ... 367, 368,.................................. 427, 456Hagiwara, Hiroko .............. 234Hahn, Matthias ................. 108Häkkinen, M. ..................... 432Hall, Bradley A..................... 26Hall, Charles ...................... 721Hall, Nathan ...................... 526Hall, Rebecca A. ................ 503Hallen-Adams, Heather…..266,................................... 501,703Hamelin, R. ............... 332, 666Hamelin, Richard C............ 336Hammel, Kenneth E. ........... 52Hammond Kosack, Kim ..... 262Hammond-Kosack, Kim ..... 229Hampel, Martin ................. 616Han, Dong-Min...... 85, 93, 233Han, J.-G.................... 276, 662Han, Kap-Hoon ... 93, 129, 233,.......................................... 234Hane, James ...................... 724Hane, James K. .......... 588, 632Hankemeier, Thomas .......... 55Hanlon, Regina .................. 250Hann-Soden, Christopher 660,.......................................... 663Hansberg, Wilhelm ........... 123Hanschen, Erik .................. 679Hansen, Guido .................. 505Hansen, Tilde J. ..................... 6Happel, Petra .................... 512Harb, Omar ....................... 338Hargarten, Jessica C. ......... 498Harland, Duane P. ............. 565Harris, Steven ..................... 96Harris, Steven D. ................. 82Harris-Coward, P. .............. 710Harry, Elizabeth ................ 507Hartmann, Thomas ............... 4Harvengt, Luc ...................... 45Harwood, Caroline ............ 515Hasim, S. ............................. 17Haslam, Stuart .................. 705Hassani-Pak, Keywan ........ 631Hastie, Alex ....................... 290Hastie, Marcus .................. 724Hatakeyama, Shin ..... 188, 472Hatakka, Annele ............ 32, 37Hauber, Joachim ............... 549Havel, Virginia E. ............... 508Havis, Neil ................. 564, 641Hawes, M. ......................... 541Hayes, Patrick M. ................ 57Hayes, Tristan ................... 52327th Fungal Genetics Conference | 309

AUTHOR LISTHayes, Tristan A. ............... 483Haynes, Ken ...................... 722Hays, Shan M. ................... 440He, Xiaoxiao Sean ................ 87Heck, Albert J. R. ............... 100Heck, Carolin ..................... 527Hecker, Arnaud ................. 519Heilig, Yvonne ... 174, 176, 177Heimel, Kai ........................ 616Heinek, Thorsten ................... 2Heinekamp, Thorsten358, 359,.......................................... 487Heitman, J. ........ 114, 305, 682Heitman, Joseph ...... 109, 125,... 197, 243, 246, 350, 416, 644.......................................... 701Held, Benjamin .................. 306Heller, Jens ................ 185, 496Hemetsberger, Christoph .. 624Hench, Virginia K. ...... 333, 353Henderson, A. ................... 604Hendrickson, Brittney ....... 355Hengartner, Michael ......... 705Henke, Catarina ................ 582Henley, Jordan .................... 84Henricot, Beatrice ............. 537Henrissat, Bernard ... 247, 306,.......................................... 336Herbert, Ben ..................... 507Hernandez-Rodriguez,Yainitza ............................... 83Herr, Andreas .................. 8, 98Herrera-Estrella, A. .. 433, 435,.......................................... 521Herrera-Estrella, Alfredo ... 124Herrero Acero, Enrique ....... 65Herrmann, Andrea ............ 457Herrmann, S. ..................... 327Hersh, M. H. ...................... 686Hertweck, Christian ....... 2, 725Herzog, Britta .................... 204Hess, Matthias .......... 649, 678Hesselbart, Ana ................. 317Hibbet, David .................... 656Hibbett, David ........... 247, 306Hihlal, ElKbir ........................ 68Hijri, Moahmed ................. 673Hildén, Kristiina ..... 32, 37, 328Hill, Robert ........................ 493Hill, Terry............................. 84Hillman, Bradley ........ 279, 674Hirayama, Hiroto ............... 193Hirsch, R. ........................... 568Hoebe, Peter ............. 641, 650Hoehenwarter, Wolfgang.. 623Hoff, Birgit ................... 49, 202Hoffmann, Lucien ...... 262, 671Hoffmeister, Dirk .......... 50, 56Högberg, Nils OS ............... 560Holland, Linda ................... 239Holloman, William K.......... 137Holm, Dorte K. ................ 9, 31Honda, Shinji ..................... 445Honda, Yoichi .................... 707Hong, C.............................. 386Hong, Seung-Bum ............. 234Hong, Sung-Yong ................. 51Hori, Chiaki ....................... 247Horn, Bruce W. ................. 668Horton, Stephen J. .............. 33Hortschansky, Peter .......... 357Horwitz, Benjamin A. 336, 579Hosomi, Akira ................... 193Houterman, Petra M. ........ 248Houterman, PM. ............... 672Howard, Brad .................... 523Howarth, Clinton............... 340Howlett, Barbara 56, 280, 618Howlett, Barbara J. ........... 653Hsiang, Tom ...................... 288Hsieh, Ming-Chen ............... 64Hsu, P. W.-C. ..................... 205Hsueh, Yen-Ping ................ 535Hu, Sufen .......................... 338Hu, Yang ............................ 562Hua, Chenlei.............. 130, 220Huang, Qianli .................... 287Huang, Xiaohua ................... 94Huang, Y. ........................... 735Huff, Yulon .......................... 78Hug, Josie ............................ 34Hughes, David ................... 581Hughes, T. ......................... 387Hughes-Hallet, James ........ 679Huh, Aram ................. 221, 715Huinink, Henk P. ............... 136Huinink, H. P. ............ 156, 161Hull, Christina M. .............. 395Hunter, Cameron .............. 365Hunter, Cameron C. .......... 366Hur, J.-S. .............................. 66Hurley, Jennifer ................. 389Huskey, D. ......................... 541Hutchison, Elizabeth ......... 721Hutchison1, Elizabeth A. ... 198Hynes, Michael ................. 423Hynes, Michael J. ...... 212, 366Hyun, M.-W. ............. 276, 662Hyun, S.-H. ........................ 276IIchiishi, Akihiko . 131, 191, 401Idnurm, Alexander ... 170, 211,.................................. 398, 399Igarashi, Kiyohiko .............. 247Ikeda, Y. ............................ 235Iliuk, Anton ....................... 578Illgen, Julia ........................ 174Imaru, N. ........................... 235Imhoff, Johannes ................ 68Inderbitzin, Patrik ............. 455Ingersoll, Tom ..................... 36Inglis, Diane O. .......... 224, 340Inman, Annie .................... 119Inoue, Hirokazu ................. 188Ip Cho, Vong shian Simon . 548Irniger, Stefan ..................... 79Isaac, Dervla ...................... 591Ishii, Akira ........................... 70Ishii, T................................ 282Ishii, Tomoko ...................... 13Ishitsuka, Yuji ...................... 95Ishiuchi, kan'ichiro .............. 59Islam, K. T. ........................ 542Issi, Luca ........................... 502Ivanov, Ivaylo P. ................ 467Ivanova, Christa .................. 61Iwashita, K. ............... 235, 450Iwashita, Kazuhiro ............ 223JJackson-Hayes, Loretta ....... 84Jacob, Tiago R. .......... 465, 617Jacobsen, I. D. ................... 482Jacobsen, Ilse D. 358, 359, 487Jacobsen, Mette ............... 722Jacquot, Jean-Pierre ........... 45Jahan, Sultana N. .............. 402Jahng, Kwang-Yeop ..... 85, 129Jahng, K.-Y. ....................... 439Jakse, J. ............................. 225Jamal, Paiman K. ................... 9James, Steven ..................... 78James, T. Y. ....................... 305Jamur, M. C. ...................... 504Jan, Stenlid ....................... 560Janbon, Guilhem ............... 392Jang, Y.-S. .......................... 727Jani, Niketa ....................... 109Jankowski, S. ..................... 713Janowska-Sedja, Elzbieta .. 229Jason, Arnold .................... 128Jaspersen, S. L. .................. 134Javornik, B. ............... 225, 271Jayasundera, Keerthi ........ 578Jelen, V. ............................ 225Jennifer, Chinnici .............. 128Jeon, Jongbum .................. 221Jeon, Junhyun .. 221, 297, 298,.................................. 634, 715Jeong, Haeyoung .............. 269Jeong, Y.-S. ........................ 727Jin, Cheng ............................. 4Jin, Hailing......................... 497Jo, Seong-mi ..................... 411Joechl, Christoph .............. 358Joensuu, Jussi ..................... 29Joergensen, Mikael S. ......... 31Johanesson, Hanna ........... 676Johannes, Raff .................. 729Johannesen, Pia F. .............. 31Johannesson, Hanna ......... 677Johansson, T. .................... 327Johansson, Tomas ..... 442, 704Jöhnk, B. ........................... 117Jöhnk, Bastian ................... 487Johns, A. E. ................ 488, 690Johnson, D. ....................... 615Johnson, Darryl ................... 83Johnson, Derek ................. 252Johnson, Linda J. 459, 559, 569Johnson, Richard D. .. 565, 608Johnson, Richard J. ........... 569Johnson, Shakira ............... 526Joly, David L. ............. 519, 628Joly, D. L. ........................... 331Jonathan, Binkley ............. 224Jones, Dan ........................ 249Jones, Daniel............. 517, 526Jones, Meriel G. ................ 363Jonkers, W. ....................... 326Jonkers, Wilfried ............... 154Jörg, Linde ........................ 729Jørgensen, Thomas ........... 102Jourdain, Isabelle .............. 612JP Brown, Alistair .............. 500Ju, K. ................................. 386Judelson, H. ...................... 462Judelson, Howard ............... 30Judelson, Howard S. .. 63, 112,......................................... 429Jun, Sang-Cheol ................ 129Jung, Boknam ................... 143Jung, E.-M. ........................ 209Jung, Kwang-Woo ..... 246, 393Jung, Seunho ...................... 97Juntunen, Kari .................... 14Jurgens, Joel A. ................... 52Juvvadi, Praveen R. ....... 72, 74KKabat, Anna M. ..................... 9Kaczmarek, Maciej .... 564, 641Kagan, Sarah ..................... 244Kagda, M. ......................... 462Kagda, Meenakshi .............. 30Kahmann, Regine..... 494, 532,.......... 619, 621, 623, 625, 626Kalchschmidt, J. ................ 484Kale, S. .............................. 530Kale, Shiv ........... 518, 523, 528Kale, Shiv D. .............. 295, 483Kallio, Pauli T. ................... 706Kaloriti, Despoina ............. 722Kamakura, T. ..................... 480Kamakura, Takashi.............. 70Kamei, M. ......................... 183Kamei, Masayuki .............. 131Kaminskyj, Susan .. 87, 88, 360,......................................... 611Kaminskyj, Susan GW . 86, 728Kämper, Jörg..................... 616Kamvar, Zhian N. .............. 683Kaneko, S. ......................... 437Kang, Ji Young ..................... 85Kang, Seogchan ....... 142, 221,.......................... 253, 254, 551Karaffa, Levente ............... 232Karagiosis, Sue .................... 13Karlovsky, Petr .................. 408Karlsson, Magnus ..... 193, 259Kasanen, R. ....................... 702Kastora, Stavroula .............. 16Katrin, Flemming .............. 729Katschorowski, Alexandra .. 49Katz, Margaret E. .............. 449Kaur, Jan Naseer ............... 394Kavanagh, Kevin ............... 636Kawaguchi, T. ........... 373, 424310

AUTHOR LISTKawamoto, Susumu... 427, 456Kawamura, A. ................... 373Kawauchi, M. .................... 450Kazama, Yusuke ................ 472Kazan, Kemal .................... 261Keema, Gert ..................... 329Keller, M. .......................... 158Keller, Nancy ..................5, 54,.................................. 692, 726Keller, Nancy P. ........ 452, 486,.................................. 566, 633Kellner, Ronny .................. 664Kema, Gert HJ ................... 336Kema, G. H. J. ............ 272, 332Kempken, Frank.......... 68, 186,......... 283, 284, 285, 286, 302,.................................. 308, 692Kemuriyama, K. ................ 133Kennell, John .................... 187Kern, Kyle ......................... 194Kersey, Paul ...................... 334Ketelaar, Tijs .............. 130, 220Khalid, Abdul Nasir ........... 513Khorramizadeh, M. R. ....... 474Khorramizadeh, MR. ......... 434Kim, D.-H. .. 414, 439, 727, 736Kim, Hee-Kyoung ....... 269, 411Kim, Hye-Seon .................. 142Kim, J. A. ............................. 66Kim, J.-C. ............ 144, 145, 195Kim, J.-M.... 414, 439, 727, 736Kim, Jong-Guk................... 323Kim, Jong-Hwa ..... 93, 129, 234Kim, Jung-Eun ................... 142Kim, Kaeun ....................... 634Kim, Ki-Tae ....................... 297Kim, S.................................. 66Kim, Seongbeom ....... 221, 634Kim, S. H. .......................... 276Kim, Sun Chang................... 97Kim, W. ............................. 615Kim, Y.-T. .......................... 635Kirkpatrick, T. ................... 601Kissinger, Jessica............... 338Kistler, C. H. ...................... 326Kistler, H. Corby ................ 571Kita, Hirohito .................... 523Klammer, Veronika ........... 357Kleemann, Jochen ............ 603Klei, Karlijn ....................... 557Klejnstrup, Marie L. .............. 9Klimes, Anna ..................... 300Klinter, Stefan ..............20, 119Klis, F. M. .......................... 122Klitgaard, Andreas ................ 6Klocko, Andrew D. ............ 440Klosterman, Steven J. ....... 300Knabe, N. .......................... 209Kniemeyer, Olaf ................ 231Knogge, Wolfgang ............ 489Knop, Doriv ....................... 711Knopp, M. ......................... 713Knox, Benjamin .................. 54Knox, Benjamin P.............. 341Kobayashi, Tetsuo ............ 463Kobie, Julie ......................... 78Kobiler, Ilana .................... 479Koch, Laurel ....................... 685Kodama, Sayo ......................71Koepke, J. .......................... 216Koh, Y. J. ..............................66Kohler, A............................ 327Kohler, Annegret ....... 227, 560Köhler, Julia R. ................... 109Koike, H. .................... 282, 372Koike, Hideaki......................13Kokkelink, Leonie .............. 105Kollath-Leiβ, Krisztina ........ 302Kolláth-Leiβ, Krisztina ........ 186Kombrink, Anja .. 251, 505, 522Komel, Radovan .......... 67, 288Konieczka, Jay ................... 659Konopka, James B. ............ 155Koopmanschap, Bertha ..... 655Kopke, Katarina ...................49Kordbacheh, P. .................. 434Korosec, Branka ..................67Kosawang, Chatchai .......... 259Kothe, E. .................... 209, 400Kothe, Erika203, 582, 583, 585Kots, Kiki .................... 130, 220Kovács, Ákos T. .................. 694Kovalchuk, Andriy.............. 656Kowbel, David ... 315, 660, 721Koyama, Yasuji .................. 463Kozubowski, L. ................... 114Kramer, Annemarie .............68Krasevec, Nada ............ 67, 226Krause, K. .......................... 400Krause, Kartrin .................. 582Krause, Katrin ............ 583, 585Kreibich, Saskia ................. 625Kreplak, J. .......................... 652Kretschmer, Matthias ........ 620Krijgsheld, Pauline ............. 100Krisp, Christoph ................. 210Kröger, Cathrin .. 545, 546, 547Kroj, T. ............................... 652Krombach, Sina ................. 625Kronstad, Jim ..................... 620Kruszewska, J. ................... 362Kruszewska, Joanna S.543, 563Kruzel, Emilia K. ................. 395Krysan, Damian J. .............. 244Kubicek, Christian .......... 61, 65Kubicek, Christian P. .. 341, 464Kubo, Yasuyuki .... 71, 113, 593Kück, Ulrich ........ 49, 199, 202,.......................... 210, 313, 404Kües, Ursula 64, 218, 219, 470Küfner, Isabell ................... 532Kühn, Barbara ................... 725Kuhn, Marie-Line ............... 477Kuijt, Suzanne J. H. ............ 569Kuipers, Oscars P. .............. 694Kumar, Abhishek ...... 283, 284,.................. 285, 286, 302, 308Kumasaka, Mayu .................70Kunitake, E. ............... 373, 424Künzler, M. ........................ 741Künzler, Markus ........ 705, 706Kuo, Alan ........................... 227Kuo, H.-C. .......................... 205Kurashima, Kiminori .......... 188Kurata, C. .......................... 183Kurtenbach, Eleonora ....... 691Kuuskeri, Jaana ......... 289, 328Kuwata, S. ......................... 572Kuyper, Thomas W. ........... 655Kwan, Hoi Shan . 287, 307, 469Kwon, H.-W. ...................... 276Kwon, Min Jin .................... 102Kwon, Nak-Jung ............ 89, 97Kwon, Seomun .................. 221Kwon-Chung, Kyung J. ....... 640LLaarhoven, K. A., van ......... 156Labbé, Jessy ...................... 606Labes, Antje ........................ 68LaButti, Kurt ...................... 606Lackner, Gerald ................... 50Lagendijk, Ellen ................. 103Lah, Ljerka ................... 67, 288Lahrmann, U. .................... 327Laine, Pia ................... 289, 328Lalucque, Herve ................ 157Lalucque, Hervé ................ 733Lam, Ernest ....................... 290Lamacchia, Marina ............ 637Lamant, Tiphaine ................ 45Lambie, Scott .................... 620Lamers, Gerda ..................... 99Lamont, Iain ...................... 559Lamont, Iain L. ................... 459Lamoth, Frédéric ................. 74Lane, Geoffrey A. ...... 459, 559Lanen, Catherine ......... 15, 329Lang, C....................... 134, 158Lang, Elza .......................... 382Lang, Elza A. S.................... 617Lange, L. ............................ 735Lange, Lene ........ 60, 343, 697,.................................. 704, 712Langner, T. ........................ 216Lankhorst, Peter .................. 55Lanver, Daniel ................... 621Lapalu, Nicolas .................. 737Larkov, Olga ...................... 579Larrondo, L. ....................... 386Larrondo, L. F. ................... 387Larrondo, Luis F. ................ 444Larsen, Thomas ................... 10Larsen, Thomas O........ 6, 9, 31Larson, Jennifer R. ............. 115Lascoux, Martin ................. 676Lass-Flörl, Cornelia .... 481, 636Lastovetsky, Olga .............. 600Latge, Jean-Paul ............ 3, 485Latorse, Marie-Pascale ...... 495Lauinger, Linda .................... 44Laval, Valérie ..................... 737Lawrence, Christopher ...... 336Lawrence, Christopher B. .. 483Lawry, Robert .................... 493Lazzez, Houcine ................. 412Leach, Michelle ................. 110Leach, Michelle D. ............... 16Lebrun, Marc-Henri222, 322, 737Lebrun, M.-H. ............ 240, 652Le Cam, B. ......................... 665Le Cam, Bruno ................... 517Lechner, Beatrix E. ............ 357Lee, Chang-Soo ................. 323Lee, Dayoung .................... 221Lee, Gir-Won ............. 297, 715Lee, H. ............................... 386Lee, Im-Soon ....................... 89Lee, In H. ............................. 58Lee, J. ................ 145, 146, 195Lee, Jang-Won .................. 246Lee, Jin H. ............................ 58Lee, Jungkwan................... 143Lee, Jung-Youn .................. 138Lee, Mark J. ........................... 3Lee, Mi-Kyung ..................... 89Lee, Sang S. ......................... 58Lee, Seunghoon ........ 269, 411Lee, Soo Chan ........... 197, 416Lee, Theresa ...................... 269Lee, Yin-Won ............. 142, 143Lee, Y. J. ............................ 144Lee, Yong-Hwan 221, 296, 297,... 298, 349, 634, 656, 715, 716Lee, Yung Yung .................. 287Lee, Y.-W. .. 144, 145, 146, 195Leeder, Abigail C. .............. 154Lefebvre, F. ....................... 331Legoahec, L. ........................ 38Lehneck, Ronny ................. 132Leinonen, Taija .................... 14Leisge, Elisa ......................... 62Lemaire, C. ........................ 665Lendenmann, M. ............... 658Leng, Yueqiang .................. 319Lengeler, Klaus .......... 150, 200Lengeler, Klaus B. .............. 120Leroy, T. ............................ 665Lev, Sophie ................ 126, 579Levander, F. ...................... 602Levasseur, Anthony........... 306Levin, Ana M. .................... 100Levinson, Dana .................. 711Levitis, Dan ....................... 642Lewis, Sara ........................ 366Lewis, Zack ........................ 443Li, Alicia ............................. 197Li, Dan ............................... 528Li, Guotian......... 268, 438, 574Li, Lei ................................. 287Li, M. ................................. 635Li, Nuo ............................... 463Li, P. .................................. 309Li, Shengnan Jill ................... 87Li, Simeng .......................... 346Li, W. ................................. 682Li, W.-C.............................. 205Li, Wei ............................... 338Li, Wenjun ......................... 350Li, Yang ........................ 39, 578Li, Yiming............................. 95Liang, X. ............................ 635Liao, H.-L. .................. 305, 314Liaw, Edward..................... 338Liberti, D. .......................... 63527th Fungal Genetics Conference | 311

AUTHOR LISTLichius, Alex ...................... 464Lichtenzveig, Judith ........... 632Lim, Fang Yun ........................ 5Lin, Ching-Hsuan ............... 377Lin, Xiaorong ...... 77, 127, 391,………………. 509, 510, 700, 738Lin, Xiu .............................. 539Lind, Mårten ..................... 562Lindner, Daniel L. .............. 688Lindner, Herbert................ 471Link, Stephanie L. .............. 208Linne, Uwe .......................... 62Litvintseva, A. .................... 312Litvintseva, A. P. ................ 305Liu, Hong ................. 3, 76, 484Liu, Huiquan ...................... 267Liu, JinGe ........................... 490Liu, Meigang ...................... 268Liu, Peng............................ 164Liu, Yi ................................. 390Liu, Zhaohui............... 524, 684Löbach, Lars ...................... 525Long, Nanbiao ................... 369Lopes Gomes, S. ................ 723López-García, Sergio ......... 436Lopez-Llorca, Luis V. .......... 580Lopez-Llorca, Luis Vicente315, 570Lopez-Moya, Federico ....... 315Loprete, Darlene ................. 84Lorch, Jeffrey M. ............... 688Loros, Jennifer ................... 389Loros, Jennifer J. ................. 73Love, Brenda ..................... 679Lovingood, Rachel V. ........... 72Lowe, Rohan G. T. ............. 618Lu, Ling .............................. 369Ludovic, Bonhomme ......... 265Ludwig, Sarah .................... 174Lue, Neal F. ....................... 137Lugones, Luis G. ................ 237Lundell, Taina .................... 328Lundell, Taina K. ................ 289Luo, Hong ............................ 51Luo, Jerry........................... 452Luo, Jing ............................ 674Lutzoni, François ....... 278, 685Lyhne, Ellen ......................... 10Lynch, Denise .................... 239Lysenko, Artem ................. 229Lysøe, Erik ......................... 318MMa, Jiwen .......................... 267Ma, L. ................................ 326Ma, Liang .......................... 274Ma, Li-Jun .......................... 603Ma, Liqiu ........................... 472Ma, Lisong ......................... 248MacCallum, Donna ............ 500Macdonald, D. A................ 690Macek, Peter ..................... 226Machado Jr., Joel ........ 24, 639Macheleidt, Juliane ........... 359312Machida, M. .............. 282, 372Machida, Masayuki ............ 13,.................................. 233, 234Mack, B. ............................ 309Mackie, Roderick .............. 678MacLean, Kirstin ............... 493Maddi, Abhiram ................ 118Madhani, Hiten D. ............. 244Madhavan, S. .................... 400Mahanti, Parag ................. 535Maheswari, Uma ............... 334Mahoney, D. P. ................. 321Majer, A. ........................... 225Mäkelä, Miia ....... 37, 289, 328Mäkelä, Miia R. ................... 32Mäkinen, Susanna ............... 14Malagnac, Fabienne .. 157, 733Mallappa, Chandru ........... 389Mallet, L. ........................... 652Mallet, Ludovic ................. 222Malm, Erik ........................... 21Malmierca, M. G. .............. 669Mandel, M. Alejandra ...... 354,.......................... 594, 629, 679Mandelc, S. ............... 225, 271Manjrekar, J. ..................... 575Mann, Matthias ................ 625Manners, John .................. 261Manning, Gerard ............... 306Manning, V. A. .................. 561Manning, Viola .................. 460Manzoor, Nikhat ............... 698Mao, J. F. ........................... 666Marchegiani, Elisabetta .... 322Marcos, A. T. ..................... 714Marcroft, Steve J. .............. 653Mardones, Wladimir ......... 236Marold, Annemarie ............. 65Martha, Arnaud B. ............ 224Martin, F. .......................... 327Martin, Francis ......... 227, 278,.................. 306, 515, 606, 649Martin, Francis M.............. 301Martin, T. .......................... 453Martinez, A. I. ................... 122Martinez, Leonora ............ 189Martinez-Rocha, Ana Lilia . 549Martinez-Rossi, Nilce ........ 382Martinez-Rossi, Nilce M. .. 430,.................................. 465, 617Martino, E. ........................ 327Martins, Maíra P. .............. 617Martins, Natalia ................ 229Marty, Amber J. ................ 441Maruthachalam, K ............ 455Maruyama, Jun-Ichi ............ 79Masuo, Shunsuke ................ 83Mathieu, Yann .................... 45Mathiot, Sandrine ............... 45Matsumoto, Teruyuki ....... 707Mattanovich, Diethard ........ 11Mattos, Larissa F. ................ 24Maucourt, M. ...................... 38Maurer, Elisabeth ..... 481, 636May, Georgiana ........ 667, 685McCallum, Brent D. ........... 628McCluskey, Kevin .............. 304McCormick, S. P. ............... 669McDonald, B. A. ................ 658McDonald, Bruce .............. 643McDonald, Bruce A. .. 448, 657McDonald, Tami R. ........... 245McDowall, Mark ............... 334McDowell, John M. ........... 533McLaggan, Debbie ............ 514McLaggan, Debra .............. 275Mead, Matthew E. ............ 395Medeiros, Luciano N. ........ 691Medina, Edgar .................. 294Meerupati, T. .................... 602Mehrabi, R. ....................... 272Mehrotra, Pankaj .............. 503Meijer, Harold .......... 130, 220Melida, Hugo ...................... 21Mélida, Hugo .............. 20, 119Mendoza, Artemio ............ 493Mendoza-Mendoza, A ...... 124Meng, S. ............................ 330Meng, Xiangchun .............. 277Menke, Jon ....................... 571Menke, Jon R. ..................... 52Mennat, El Ghalid ............. 554Mentlak, Thomas A. .......... 492Mesarich, Carl ... 249, 517, 526Mesters, Jeroen R. ............ 505Mettälä, Aila ....................... 37Meux, Edgar........................ 45Meyer, Vera ........ 99, 101, 102Michael, Valinluck ............. 169Michkov, Alexander V. ...... 190Mideros, Santiago ............. 506Mideros, Santiago X. ......... 607Midorikawa, Yura ..... 367, 368Mieczkowski, Piotr ............ 416Miettinen, Otto ................. 648Migheli, Quirico ................ 262Miki, S. .............................. 577Milbredt, Sarah ................... 49Miller, Robert ................... 316Min, K. ...................... 145, 195Minami, A. .......................... 12Minet-Kebdani, Naima ...... 477Minnis, Andrew M. ........... 688Minz-Dub, Anna ................ 105Miralles-Duran, A ...... 211, 473Mirzadi Gohari, A. ............. 272Misener Bloom, Amanda L.396Misiek, Mathias .................. 50Mitchell, Aaron ................. 379Mitchell, T. ........................ 312Miyara, Itay....................... 479Miyasato, Stuart R. ........... 340Miyauchi, Shingo .............. 425Miyazaki, Y. ....................... 437Moazeni, M. .............. 434, 474Mockler, Todd .................. 491Moffat, Caroline ............... 647Mogannam, John ................ 43Mogg, C. ........................... 709Möhlman, Tim .................. 655Moing, A. ............................ 38Moktali, Venkatesh ... 253, 254Molnar, Thomas ............... 279Molzahn, L. ....................... 135Momany, Michelle.............. 83Mondo, Stephen ............... 600Mondo, Stephen J............. 255Monteiro, Valdirene . 739, 740Monteiro, Valdirene………….18Montenegro-Montero, A. . 387Montibus, M. .................... 410Montis, Valeria ................. 408Moolhuijzen, Paula ........... 261Moraga, Roger A. .............. 608Morel, Melanie ................... 45Moretti, A. ........................ 670Moretti, M. ....................... 630Moretti, Marino ................ 621Morin, E. ........................... 327Morin, Emmanuelle .. 278, 306Moriyama, E. N. ................ 732Morozov, Igor Y. ............... 363Morris, Paul ........................ 34Mortensen, Uffe H. 6, 9, 28, 31Mortensen, Uffe Hasbro ....... 7Mørtz, Ejvind .................... 614Motteram, Juliet ............... 631Mouriño-Pérez, R. ............ 173Mouriño-Pérez, Rosa R. .... 121Moxon, S. ......................... 426Moyrand, Frédérique ....... 392M. Sgambelluri, Robert ....... 51Muddiman, David ............. 166Muddiman, David C. ......... 573Mueller, Andre ................. 622Mueller, Sebastian ............ 231Muensterkoetter, M. ........ 407Mujic, Alija ........................ 335Müller, Christoph ............. 481Muller, Laura K. ................ 688Müller, Olaf ...................... 278Müller, Wally H. ................ 100Mulock, Barbara ............... 628Muñoz, Alberto .................. 75Munro, C. ......................... 699Munro, Carol .................... 110Munro, Carol A. .................. 16Munro, E. .......................... 371Münsterkötter, Martin .... 140,......................................... 263Muraguchi, H. ................... 133Murakami, Shigeyuki ........ 707Murat, Claude ................... 649Murray, Sandra ................. 423Mustalahti, Eero ................. 29Mwambutsa, Faustin .......... 78Mycorrhizal Genome InitiativeConsortium ....................... 301Mycorrhizal GenomicsInitiative (MGI) Consortium227Myers, Chad L. .................. 244

AUTHOR LISTNNadimi, Maryam ............... 673Naesby, Michael ................. 28Nagoshi, T. ........................ 133Nagy, Laszlo ...................... 306Naik, Vikram ..................... 623Naito, Mizue ..................... 609Nakamura, M.................... 437Nakashima, Jin .................. 567Nakayama, Mayumi... 367, 456Nakazawa, Takehito ........... 59Nantel, A. ......................... 378Narra, Hema P. ................. 594Narukawa, M. ................... 480Narukawa, Megumi ............ 70Naseem, Shamoon ........... 155Natorff, R. ......................... 362Natorff, Renata ................. 543Navarathna, Dhammika H. M.L. P. ................................... 499Navarro, Eusebio .............. 436Navarro-Gonzaléz, Monica 218Navarro Velasco, Gesabel Y.555Nawrath, Thorben ............ 179Ndonwi, Maze .................. 187Nelson, Paul ..................... 667Nelson, Rebecca ............... 506Nelson, Rebecca J. ............ 607Ness, Frédérique .............. 637Nett, Markus ...................... 56Neumann, Piotr ................ 132Nevalainen, Helena .......... 425Newman, Mari-Anne ........ 548Newton, Adrian ................ 564Nguyen, Andy ................... 290Nguyen, Mimi ..................... 78Nguyen, N.-L. .................... 727Nguyen, Thuat Van .... 546, 547Nguyen Thanh, Thuy ........ 325Ni, Peixiang ....................... 676Nickerson, K. ...................... 17Nickerson, Kenneth .......... 703Nickerson, Kenneth W. .... 498,......................................... 499Nicolas-Molina, F. E. ......... 426Nielsen, Jakob B. ..... 6, 7, 9, 31Nielsen, Kirsten ................ 245Nielsen, Kristian F. ...........6, 31Nielsen, Michael L. ............... 9Nielsen, Morten T. .............. 31Niemi, Merja ...................... 14Nienhaus, Ulrich ................. 95Nierman, William C. ............. 3Nieto-Jacobo, Maria Fernanda124, 493Nieuwenhuis, Bart ............ 655Nijland, Jeroen ................... 55Nitsche, Benjamin M. . 99, 100,......................................... 102Nogami, Yuhei .................. 191Noguchi, Hiroshi ................. 59Nogueira, F. ...................... 699Nolan, Matt ...................... 606Nong, Wenyan .................. 287Noorbakhsh, F. ................. 434Nordzieke, Steffen ............ 199Noronha, Eliane F. ............. 316Novak, Metka ......................67Novikova, Olga .......... 256, 257Nowrousian, Minou .. 258, 313Nürnberger, Thorsten ....... 532Nuss, Donald ..................... 273Nygren, Kristiina ................ 259Nystedt, Björn ................... 350OOakley, Berl .........................54Oakley, Liz ...........................54Ochoa Tufiño, Valeria ........ 344O'Connell, Richard J........... 603Odds, Frank C. .....................16Oeser, B. ............................ 238Ogawa, Masahiro .............. 463O'Gorman, Céline .............. 202O'Gorman, Céline M. ......... 640Oh, Dong-Soon .. 129, 233, 234Oh, J. ......................... 276, 662Oh, Yeonyee ...... 166, 476, 573Oh, YounLee ...................... 323Ohashi, S. .......................... 372Ohba, Ayumi ..................... 427Ohkura, Mana ........... 347, 679Ohm, R. ............................. 332Ohm, Robin ....... 590, 606, 678Ohm, Robin A. ... 237, 336, 649Ohsato, S. .......................... 572Ohtsuka, K. ........................ 577Oikawa, H. ...........................12Oksanen, Ilona .................. 289Olarte, Rodrigo A. .............. 668Olivares-Yañez, C. .............. 387Oliveira, Fenix.................... 739Oliveira, Vanderci .............. 382Oliver, Brian ...................... 502Oliver, C. ............................ 504Oliver, Richard ........... 647, 724Oliver, Richard P. ............... 632Olmedo-Monfil, V. ............. 521Olson, Ake ......................... 562Olsson, Lisbeth .................. 325Olsson, Stefan ................... 548Omrane, Selim................... 329Oome, Stan ................. 34, 596Oono, Ryoko ...................... 685Orbach, Marc ............ 347, 629Orbach, Marc J. . 354, 594, 679Orlando, Ron .......................83Ortega, R. .......................... 305Ortega, Rebecca ................ 606Ortega-Abboud, E.............. 652Orzechowski, Amanda .........78Ose, T. ............................... 576Oses- Ruiz, Miriam ............ 478Oskarsson, Therése ........... 150Osmani, Stephen A. ........... 115Other members of theDothideomycetes community336other PMI researchers(http://pmi.ornl.gov) ......... 515Otillar, Robert ................... 306Ouedraogo, Jean-Paul ....... 101Ouyang, Shouqiang ........... 178Owensby, C. Alisha ............ 320PPadhi, Suhasini .................... 33Padula, Matthew............... 507Pakula, T. ........................... 432Palamarczyk, Grazyna ....... 563Palma-Guerrero, Javier ..... 185Palmer, Jonathan M. ......... 726Paloheimo, Marja .............. 428Pan, Juan ........................... 511Panabieres, Franck ............ 477Panaccione, Daniel G. ....... 511Pandelova, I. ..................... 561Pandelova, Iovanna ........... 460Panepinto, J. ....................... 23Panepinto, John ................ 394Panepinto, John C. ... 396, 397,.......................................... 508Paoletti, Mathieu .............. 637Pappas, Georgios J. ........... 316Pareja-Jaime, Yolanda ....... 550Park, A. R. .......................... 145Park, Bongsoo ................... 253Park, Heesoo ....................... 90Park, Hee Soo ...................... 97Park, J.-A. .......................... 414Park, Jaejin ........................ 716Park, Joohae .............. 102, 103Park, Sang-Wook ............... 523Park, S.-M. ......................... 414Park, Sook-Young ..... 221, 297,.......................... 298, 634, 715Park, Young-Jin .................. 323Pascon, Renata .................. 639Pasquali, Matias 262, 408, 671Paszewski, A. ..................... 362Pathirana, Ruvini U. .......... 499Patterson, Miranda ........... 722Patyshakuliyeva, A. ............. 27Paul, Biplab ......................... 88Paulin, L............................. 702Paulin, Lars ................ 289, 328Paun, Linda ......................... 68Pawlowska, Teresa .... 600, 609Pawlowska, Teresa E. ........ 255Payne, Gary ......................... 69Payne, Michael J. .............. 597Peay, K. G. ......................... 314Peccoud, Jean ................... 250Pedersoli, Wellington .......... 47Pedersoli, Wellington R. .... 413Pedro, Heldér .................... 334Pelletier, Dale ............ 515, 606Peñate, X. .......................... 714Penselin, Daniel................. 489Penttilä, M. ....................... 432Peraza-Reyes, L. ................ 321Peres, Nalu ........................ 382Peres, Nalu TA ................... 465Peres, Nalu T. A. ........ 430, 617Perez-Martin, Jose ............ 137Perez-Martín, José ............ 215Pérez-Nadales, Elena ........ 556Perfect, J. .......................... 312Perfect, John R. ................. 485Perlin, Michael .................. 339Perlin, Michael H. ................ 35Perlinska-Lenart, Urszula .. 563Perotto, S. ......................... 327Persinoti, Gabriela ............ 382Persinoti, Gabriela F. . 430, 465Petersen, Lene .................... 10Petersen, Lene M. ........... 6, 31Petre, Benjamin ................ 519Petro, Thomas M. ............. 498Petrs-Silva, Hilda ............... 691Pfannenstiel, Brandon ...... 365Pfender, William ............... 491Pfiffner, Jenna M............... 659Pflugmacher, M. ............... 638philippe, Lecomte ............. 265Philippsen, P. ........... 122, 134,.................................. 135, 158Piao, Hailan ....................... 678Piehler, Sebastian ............. 140Pilgaard, Bo ....................... 712Pilsyk, Sebastian........ 543, 563Pinney, Deborah ............... 338Pinson-Gadais, L. ....... 447, 708Piotrowska, Marta .... 641, 650Plamann, Michael ..... 213, 217Plaza, D. F.......................... 741Plummer, Kim ........... 249, 517Plummer, Kim M. .............. 526Pócsi, István ........................ 91Podobnik, Barbara .............. 67Poeggeler, Stefanie ........... 139Pöggeler, Stefanie .... 132, 202,.................................. 204, 258Pogoda, Friederike ............ 671Pohlmann, T. ..................... 216Poidevin, Laetitia .............. 429Polacheck, Itzhack ............. 244Polaino Orts, Silvia ............ 211Poland, Jesse ..................... 506Poli, Anna .......................... 544Politi, A. Z. ......................... 134Ponts, N. ..... 38, 410, 447, 708Poppe, Stephan......... 512, 664Porter, T. M. ...................... 305Post, Harm ........................ 100Potrykus, Joanna ............... 500Poullard, Ashley .................. 84Pourmand, M. R. ............... 474Poussereau, N. .................. 240Poussereau, Nathalie ........ 495Praseuth, Mike .................... 54Prenzler, K. ........................ 638Presley, Gerald N. ............... 52Priegnitz, Bert-Ewald ........ 370Priegnitz, Ewald ................ 179Priest, Henry ..................... 491Pringle, Anne .............. 40, 642Proctor, R. H.............. 669, 670Proctor, Robert H. ............. 318Prosper, Pascalita ............... 45Prusinkiewicz, Martin ......... 86Prusky, Dov ....................... 27727th Fungal Genetics Conference | 313

AUTHOR LISTPrusky, Dov B. ................... 479Pukkila, Patricia J. ..... 333, 337,.......................................... 353Pupelis, Linas ..................... 704Puranen, Terhi .................... 14Puttikamonkul, Srisombat . 485QQi, Xiaodong ........................ 39Qin, Jing ............................ 287Qin, Xiaotian ..................... 600Quesneville, Hadi .............. 737Quester, Katrin .................. 719Quinn, Janet ...................... 722RRaab, Andrea .................... 500Raaymakers, Tom .............. 596Rabe, Franziska ................. 494Radisek, S. ......................... 225Rahnama, Mostafa ............ 610Ram, Arthur .............. 101, 103Ram, Arthur F. J. .. 99, 100, 102Ramada, Marcelo HS ........... 46Ramada, Marcelo H. S. ...... 316Ramamoorthy, Vellaisamy 364Ramamurthy, Raghuraman338Ramegowda, Y. B. ............. 613Ramesh, Marilee A. ........... 348Ramírez-del Villar, Arianne 121Ramirez-Garces, D. ............ 520Ramos-Guelfo, M. S. ......... 714Rampitsch, Christof ........... 299Rand, Dustin P. .................... 39Rangel, Pablo .................... 123Rao, Reeta ......................... 502Rasmussen, Tim ................ 525Ravasio, D. ........................ 451Read, Nick ........... 75, 182, 564Read, Nick D. ..................... 565Reck-Peterson, Samara ....... 92Reck-Peterson, S. L. ........... 214Redkar, Amey .................... 624Regev, Aviv ................ 502, 659Regulin, Annika ................. 692Reilly, M. ........................... 720Reissmann, Stefanie .......... 625Remme, Nicole ...................... 2Rems, Ana ........................... 53Rep, M....................... 270, 672Rep, Martijn ............. 148, 149,.................................. 248, 557Repas, Tim......................... 611Requena, Natalia ............... 527Reschka, Eva ..................... 204Restrepo, S. ....................... 661Reuveni, Eli ....................... 277Rey, Patrice ....................... 737Reynolds, Hannah ............... 36Reza, Md Hashim .............. 575Rezaie, S. ........................... 434Rezaie, S, ........................... 474Rezonja, Sasa .................... 226R. Gonçalves, I. .................. 374Rhind, Nicholas R. ............. 290Rhodes, J. .......................... 147Ribitsch, Doris ..................... 65Richard, Bennett ............... 377Richard Albert, J. ............... 371Richard-Forget, F. ...... 38, 410,.................................. 447, 708Ridenour, J. B. ................... 601Riedel, Sabine ................... 204Ries, Marco ......................... 55Rigden, Daniel ................... 363Riley, Robert ............. 228, 306Rineau, Francois ................ 442Ringel, Till ......................... 625Ringleberg, Carol S. ............. 73Riquelme, M. ............ 117, 163Riquelme, Meritxell .......... 189Ritland, K. .......................... 666Rivera, Lorena ................... 241Rizzoli, Silvio........................ 79Robb, S. ............................. 305Robb, Sofia ........................ 606Roberson, R. ..................... 117Roberts, A. J. ..................... 214Roberts, Anthony ................ 92Roberts, David D. .............. 499Robledo Briones, AngélicaMariana ............................ 381Rodolphe, F. ...................... 652Rodriguez, Gabriela .......... 673Rodriguez-Carres, M. ........ 305Rollins, J. ........................... 635Rollins, Jeffrey ................... 454Roncero, M. Isabel G. ........ 550Ronen, Mordechai ............ 579Roos, David ....................... 338Roque-Barreira, Maria Cristina18Rosikiewicz, Pawel ............ 584Rosling, Anna .................... 560Rossi, Antonio382, 430, 465, 617Rossi, M. ................... 167, 630Rot, G. ............................... 225Roth, Martin ......................... 2Rouhier, Nicolas ................ 519Rountree, Michael R. ........ 440Rouvinen, Juha .................... 14Rouxel, Thierry .. 280, 403, 618Roy, S. ............................... 462Roy, Sourav ....................... 429Roy, Sushmita ................... 659Rudd, Jason ....................... 522Rudd, Jason J. .................... 631Rugbjerg, Peter ................... 28Ruger-Herreros, Carmen ... 446Ruiz Herrera, José ............. 381Ruiz Roldan, C. .................. 553Ruiz Roldán, C. .................. 552Ruiz-Roldán, Carmen ........ 550Ruiz-Vázquez, R. M. .......... 426Ruiz-Vazquez, Rosa ........... 416Rumore, Amanda .............. 523Runa, Farhana ..................... 69Ruoff, Peter ......................... 41Ruprich-Robert, Gwenäel . 219Rytioja, Johanna ............ 32, 37Ryu, Jae San ........................ 52Rzeszutek, Elzbieta ............. 22SSachs, Matthew 383, 390, 738Sachs, Matthew S. ............ 467Saha, Surya ....................... 607Sain, Divya ........................ 241Saitoh, Hiromasa .............. 492Sakaguchi, Ayumu ...... 71, 113Sakalidis, M. ...................... 332Sakamoto, Y. ..................... 437Sakulkoo, Wasin ............... 116Salamat, Khalid ................. 345Salamat, Muhammad Khalid637Salame, Tomer .................. 711Salamov, Asaf ................... 306Salamov, Asaf A. ............... 228Salavirta, Heikki ................ 289Saleem-Batcha, Raspudin . 505Saloheimo, M. ................... 432Saloheimo, Markku ............. 29Samalova, Marketa ........... 196Samejima, Masahiro ......... 247Samwel, Simon ................. 596Sancar, Gencer.................. 446Sanches, Pablo R. .............. 617Sanchez, E. ........................ 163Sanchez, James ................... 54Sánchez-Barrionuevo, L. ... 714Sánchez-Vallet, Andrea ..... 505San Clemente, H. .............. 520Sandai, Doblin ..................... 16Sanders, I. ......................... 675Sanders, Ian ...................... 584Sandoval-Sierra, J. Vladimir 21Sandrock, Björn .................. 62Sano, H. ............................ 437Sano, M............................. 372Sanyal, K. .......................... 114Saqi, Mansoor ........... 229, 631Sarabia, Miguel-Ángel ....... 123Saraiva, Marcia ................. 275Sarikaya Bayram, Oezlem79, 93Sarkari, P. .......................... 713Sarowar, Mohammad N.... 514Sarwar, Samina ................. 513Sasaki, K. ........................... 480Sato, Michio ........................ 59Sato, Y. .............................. 577Satoh, Y. ............................ 576Sauer, Michael .................... 11Saunders, Charles ............. 350Saupe, Sven ...................... 637Saupe, Sven J. ........... 293, 345Sauter, T. .......................... 638Saville, Barry J. .................. 628Saville, B. J. ............... 330, 730Savytskyi, O. V. ................. 305Scazzocchio, C. .................. 693Scazzocchio, Claudio ......... 232Schackwitz, Wendy ........... 337Schadt, Christopher .. 515, 606Schaefer, Amy ........... 515, 606Schaefer, Katja .................. 556Schäfer, Wilhelm ..... 140, 545,.......................... 546, 547, 549Schäfers, Christian ............ 404Schafferer, Lukas .............. 358Schardl, Chris .................... 324Schardl, Christopher L. ...... 26,................................. 490, 511Scharf, Daniel H. ................... 2Scharfenstein, L. ............... 309Scherlach, Kirstin .................. 2Scherm, Barbara ............... 262Scheyniuse, Annika ........... 350Schilling, Jonathan S. .......... 52Schink, Kay Oliver ..... 152, 160Schinke, Josua .................... 94Schipper, K. ....................... 713Schipper, Kerstin .............. 625Schirawski, J. ..................... 291Schlunk, Ines ..................... 583Schmaler, Tilo ..................... 94Schmidt, A. ....................... 135Schmidt, Irina L. ................ 621Schmidt, Sarah Maria ....... 248Schmidt, S. M.................... 270Schmidt-Heck, Wolfgang .. 359Schmieder, S. S. ................ 741Schmieder, Stefanie.......... 705Schmoll, Monika ....... 201, 431Schnaβ, Nicole .................. 210Schnittker, Robert ... 213, 217,......................................... 304Schoch, Conrad ................. 280Schoch, Conrad L. ............. 336Scholten, Stefan ............... 140Schotanus, Klaas ............... 242Schott, Veronika ............... 664Schreiver, Ines .................. 248Schrettl, Markus ....... 358, 471Schroeder, Frank .............. 535Schuerg, Timo ................... 192Schuler, Herwig ................ 614Schultz, David ................... 339Schulz, Stefan ................... 179Schumacher, Julia ..... 106, 495Schürmann, Janine ........... 457Schwarzer, A. .................... 638Schwenk, Daniel ................. 56Scott, Barry .. 48, 417, 587, 612Scott, Eileen ...................... 731Scott, James A. ................. 336S. Druzhinina, Irina ........... 558Secombes, Christopher..... 525See, Pao Theen ................. 647Seger, S. ............................ 158Segers, F. J. J. .................... 161Segers, Frank J. J. .............. 136Séguin, Armand ................ 519Seiboth, Bernhard .............. 61Seidl-Seiboth, Verena ....... 464Seiler, S. ............................ 173Seiler, Stephan .. 174, 176, 177Sekimoto, S. ...................... 305Selker, Eric ................ 380, 445Selker, Eric U. ................... 440Sellam, A. .......................... 378Selway, Laura...................... 16314

AUTHOR LISTSenftleben, Dominik ......... 585Senoo, F. ........................... 235Seo, J.-A. ........................... 145Sepcic, Kristina ................. 226Servin, Jacqueline ............. 162Severing, Edouard ............ 344Sexton, M. ........................ 147Sghyer, Hind ..................... 329Shah, Firoz ................. 442, 704Shah, Prachi ............... 224, 340Shahi, Shermineh ............. 148Shail, Kabrawala ............... 377Shakya, Viplendra ............. 211Shakya, Viplendra P. S. ..... 170Shalaby, Samer ................. 579Shantappa, S. .................... 710Shantappa, Sourabha ....... 364Shapiro, Justin .................... 84Sharma Khatiwada, S. ....... 601Sharon, Amir .................... 105Sharpee, William C. .......... 476Shbat, Yousef.................... 308Shelest, Ekaterina ............. 725Shelver, Weilin ................. 524Shen, Gui .......................... 458Shen, Qirong ..................... 558Shepardson, Kelly M. ........ 485Sheppard, Donald C. ............. 3Sherlock, Gavin .. 223, 224, 340Shertz, Cecelia .................. 416Shiller, Jason .............. 249, 526Shimizu, Kiminori .............. 427Shin, Miho ........................ 221Shin, Pyung-Gyun ............. 323Shingler, Kristin .................. 78Shintani, T. ......................... 12Shintani, Takahiro...... 104, 427Shjerve, Rachel A. ............. 627Shnaiderman, Chen .......... 479Shoji, Jun-ya ..................... 567Shrestha, B. ............... 276, 662Shubitz, Lisa ...................... 629Shubitz, Lisa F. .................. 594Sibthorp, Christopher ....... 223Sicault-Sabourin, Martine . 637Sieber, Christian ............... 263Siebert, Kendra ................. 365Siebert, Kendra S. ............. 366Siegmund, Ulrike .............. 496Sienko, M. ........................ 362Sienko, Marzena ............... 543Sierotzki, Helge ................. 731Sietiö, Outi-Maaria ............. 32Sil, A.................................. 604Sil, Anita ............ 155, 171, 468Silar, P............................... 321Silar, Philippe ............. 157, 733Sillaots, S. ......................... 371Silva, Larissa ..................... 382Silva, Larissa G. .......... 465, 617Silva, Roberto .. .... 18, 47, 413,……………………………….422, 740Simison, Matt ................... 340Simon, A. .......................... 374Simon, Adeline .......... 106, 495Simoneau, Phillippe .......... 586Simonin, Anna .................. 507Simpson, Wayne R. ... 569, 608Singh, Kanwar ........... 649, 678Singh, Karam B. ......... 588, 632Singh, Rakhi ....................... 668Skoneczny, M. ................... 362Skovlund, Dominique A. ......31Skrzypek, Marek S. .... 224, 340Slinski, S. L. ........................ 264Sluga, Nina ........................ 226Smet, F. ............................. 714Smith, Deborah A. ............. 722Smith, J. E. ......................... 613Smith, Kristina ................... 383Smith, Kristina M. ...... 242, 385Smulian, A. George ............ 420Snook, Maurice E. ............. 540So, K.-K. ............................. 727Soanes, Darren M. ............. 478Solis, Norma ...................... 379Solomon, Peter S. .............. 595Solomons, J. T. Graham ..... 397Somai, Benesh M. ............. 461Somerville, C. R. ................ 303SON, H. .............................. 146Son, H. ............... 144, 145, 195Son, Hokyoung .......... 142, 143Sone, T. ............. 480, 576, 577Song, Min-Hee................... 246Sophianopoulou, V. ........... 693Sorrell, Tania ..................... 126Souibguy, Eytham.............. 495Soukup, Alexandra ...... 54, 452Sova, Matej .........................67Soyer, Jessica..................... 403Spanu, Francesca............... 262Spatafora, J........................ 305Spatafora, Joseph ...... 252, 335Spatafora, Joseph W. ....... 311,.................................. 320, 336Sperschneider, Jana .......... 261Spraker, Joe E. ................... 566Srivastava, Akhil ................ 590Srivastava, Vaibhav .............21S. Scott Craig, John ..............51Stahl, D. ............................. 638Staines, Daniel M. ............. 334Stajich, Jason .... 164, 241, 258,.................................. 294, 338Stajich, Jason E. ................. 475Stajich, J. E. ........................ 305Stanley, A. M. .................... 669Stanley, C. E. ...................... 741Stanley, Claire E. ................ 706Starr, Trevor ...................... 192Stassen, Joost ............ 107, 375Stea, G. .............................. 670Stead, David ................ 16, 500Steenkamp, Emma T. ........ 206Steenkamp, E. T. ................ 260Steffens, Eva ...................... 210Steidemann, Michelle ....... 194Steiger, Matthias G. ............11Steinbach, William J. ..... 72, 74Steindorff, Andrei S. .... 46, 316Stenlid, Jan ........................ 562Stephenson, Karen ............ 159Sternberg, Paul .................. 535Stevens-Lagos, A. .............. 387Stewart, Alison .......... 124, 493Stewart, E. ......................... 658Steyaert, Johanna ............. 493Stock, J. ............................. 713Stöckli, Martina A. ............. 706Stojan, Jure ......................... 67Stokes, Tricia ..................... 194Stoll, Thomas .................... 724Storms, R. .......................... 371Stukenbrock, Eva............... 512Stukenbrock, Eva H. . 242, 448,.................................. 657, 664Stumpf, Sina ........................ 94Subbarao, Krishna ............. 455Subramaniam, G. .............. 709Sugimoto, Satoru ................ 59Sugui, Janyce A. ................. 640Suhr, Mallory .................... 501Suhre, Karsten ................... 263Sukno, Serenella ............... 277Suleman, Essa ................... 461Sumitani, J. ................ 373, 424Sun, Furong ....................... 528Sun, H. ............................... 702Sun, Jianping ..................... 466Sun, Lei .................................. 4Sun, Sheng ........................ 644Sun, Shun-Kuo ..................... 25Sundelin, Thomas .............. 548Sung, G.-H. ................ 276, 662Sung, Ki-Ho ........................ 323S. Urgast, Dagmar ............. 500Suriani, Marcela ................ 739Susca, A. ............................ 670Suzuki, Tadashi .................. 193Swilaiman, Sameira S. ....... 640Swoboda, Ginger A.............. 26Syme, Robert .................... 724TTabima, J. F. ...................... 661Tad, Woraratanadharm .... 169Takach, J. E. ....................... 651Takach, Johanna E. .............. 26Takahashi, Hiroki ............... 456Takahashi, Masakazu ........ 131Takahashi, Tsukasa............ 401Takeda, I. ........................... 282Takeshita, N. ....................... 80Takeshita, Norio .................. 95Takeuchi, S. ....................... 577Takken, Frank L. W. ........... 248Talbo, Gert ........................ 526Talbot, J. M. ...................... 314Talbot, Nicholas J. .... 116, 165,.................................. 478, 492Tamano, K. ................ 282, 372Tamano, Koichi ................... 13Tan, K. ............................... 214Tan, Kaeling ......................... 92Tan, Kar-Chun ................... 724Tanaka, Kaoru ................... 593Tanaka, M. .......................... 12Tanaka, Mizuki .................. 104Tanaka, Shigeyuki ............. 619Tanaka, Shuuitsu ....... 188, 472Tang, Dingzhong ............... 230Tanguay, P. ....................... 332Tani, S. ...................... 373, 424Tao, Andy .......................... 578Taranto, Adam .................. 526Tarrka, M. ......................... 327Taylor, Jen ......................... 261Taylor, John....... 660, 663, 681Taylor, John W. ................. 292Taylor, J. W. ...................... 314Taylor, Samantha ...... 250, 518Tebbji, F. ........................... 378Tefsen, Boris ..................... 103Tegelaar, Martin ............... 237Teichert, Ines .... 199, 210, 404Teimoori-Toolabi, L. .......... 434Temme, N. ................ 238, 638Templeton, M. .................. 665Templeton, Matthew ....... 249,.................................. 517, 526Tenor, J. ............................ 312Tenorio-Rammer, Liliana E. . 65Te'o, Junior ....................... 425Terado, S. .......................... 235Terai, Goro ........................ 234Terauchi, R. ....................... 576Terauchi, Ryohei ............... 492Terhonen, E....................... 702Terpitz, Ulrich ................... 141Tharreau, D. ...................... 652Tharreau, Didier ................ 222Theisen, Jeffrey ................. 726Thierry, Langin .................. 265Thomas, A. ........................ 601Thomma, Bart ... 251, 522, 526Thomma, Bart P. H. J. 505, 516Thompkins, A. ................... 147Thompson, Dawn .............. 502Thompson, Dawn A. .......... 659Thomson, Darren .............. 111Thon, Michael ........... 277, 603Thornton, Christopher R. .. 570Tian, Chengming ............... 346Tian, Xiuyun ...................... 510Tian, Xunyun ..................... 127Tillmann, Anna .................. 722Tillmann, Britta ................. 457Tillmann, Britta A. M. ........ 160Timonen, Sari ...................... 32Timpner, Christian .... 536, 589Tipton, J. ........................... 601Tisch, Doris ............... 201, 431Tisserant, E. ...................... 327Titz, Alexander .................. 705Toda, T. ............................. 372Todd, Richard .................... 365Todd, Richard B. ................ 366Toffaletti, D. ...................... 312Toh, Su San ....................... 339Tollot, Marie ..................... 626Toloczko, Aleksandra ........ 525Tong, Xiaoxue ................... 697Toquin, V. .......................... 240Torres-Martínez, S. ........... 42627th Fungal Genetics Conference | 315

AUTHOR LISTTorres-Martinez, Santiago 416Torres-Martínez, Santiago 436Torriani, Stefano F. F. 448, 657Traeger, Stefanie ............... 258Tran, Van Tuan .......... 536, 589Treitschke, Steffi ............... 622Tremblay, Julien ........ 649, 678Trengove, Robert D. .......... 632Treves, David..................... 339Trien, Hien ........................ 629Tringe, Susannah ............... 678Tringe, Susannah G. .......... 649Trinh, Hien ........................ 594Tripathy, Sucheta .............. 338Trivitt, Gloria ..................... 518Tschaplinski, Timothy ........ 515Tsui, C................................ 666Tsukagoshi, Yuko ............... 131Tudzinsky, Paul.................. 495Tudzynski, B. ............. 260, 407Tudzynski, Bettina ..... 405, 409Tudzynski, P. ..................... 238Tudzynski, Paul......... 105, 106,.................................. 457, 496Tumlinson, James .............. 551Tung, S.-Y. ......................... 205Tunlid, A. ................... 327, 602Tunlid, Anders ........... 442, 704Turgeon, B. Gillian ............. 336Turgeon, Gillian ......... 506, 534Turina, M. ................. 167, 630Turk, Samo .......................... 67Turner, Ashley ................... 645Turra, David ...................... 554Tuskan, Gerald .......... 515, 606Tyler, B. ............................. 530Tyler, Brett 250, 338, 518, 523Tyler, Brett M. .......... 295, 415,.......................... 483, 528, 533Tymon, L. .......................... 615Tzelepis, Georgios ............. 193UUberbacher, Edward ......... 515Ulhoa, Cirano ............ 739, 740Ulhoa, Cirano J. ........... 46, 316Umemura, M. ............ 282, 372Umemura, Myco ................. 13Upadhyay, Srijana ....... 77, 509Urban, Martin ................... 229VVaillancourt, Lisa ............... 603Valent, Barbara . 138, 165, 492Valerius, Oliver ............ 79, 487Valiante, V. ........................ 484Valiante, Vito .................... 231Valim, C. X. R. .................... 504Valkenburg, Dirk-Jan . 505, 522Valkonen, Jari P. T. ............ 349Valkonen, M. ..................... 432Vallance, Jessica ................ 737Vallet, Julie ........................ 322Vallino, M. ......................... 167van Dam, P. ....................... 270van Dam, Peter ................. 248Van den Ackerveken, Guido596van den Berg, A. Herbert .. 514van den Berg, Grardy ........ 251van den Esker, Marielle H. 694van der Burgt, Ate ............. 344van der Does, Charlotte .... 557van der Holst, Kyle W. ....... 461van der Lee, T.................... 670vanderLee, Theo ............... 230van der Merwe, N. A. ........ 260VanderSluis, Benjamin ...... 244van de Vondervoort, Peter JI.687Van de Wouw, Angela ....... 618van de Wouw, Angela ....... 249Van de Wouw, Angela P. ... 653van Die, Irma ..................... 103van Doorn, Tineke M. ....... 151Van Etten, H. ..................... 541van Gemert, Jerom ........... 557van het Hoog, M. .............. 378Van Hove, F. ...................... 670van Kan, Jan ...... 107, 108, 375van Laarhoven, K. A. ......... 161van Laarhoven, Karel A. .... 136van Leeuwen, Martin Richard151van Leeuwen, M. R. .......... 207van Peer, Arend ................ 323van Veluw, Jerre ............... 695Van Werven, Folkert J. ...... 109van West, Pieter ...... 275, 514,.................................. 525, 532Vargas, Itzel ...................... 123Vehmaanperä, Jari .............. 14Veneault-Fourrey, C. ......... 327Ver Loren van Themaat, Emiel603Verma, Surbhi ........... 398, 399Vetukuri, Ramesh R. ......... 402Viaud, M. .......................... 374Viaud, Muriel ............ 106, 495Viefhues, A. ....................... 238Vila, A. ............................... 426Vilaplana, Francisco .......... 119Vilchis-Nestor, Alfredo Rafael719Vilgalys, R. ......... 305, 314, 686Vilgalys, Rytas ........... 515, 606Villalobos-Escobedo, JM. .. 435Villalobos-Escobedo, J. M. 433Villalta, Christopher F. ...... 171Vinck, Arman..................... 695Vintila, Simina ................... 614Virgilio, S. .......................... 388Virgilio, Stela ..................... 419Visentin, Ivan .................... 408Vitikainen, M..................... 432Vitikainen, Marika ............... 29Vlaardingerbroek, Ido 149, 557Voegeli, S. ................. 122, 158Vogel, John ....................... 491Voigt, Christian A. ............. 529Voigt, Oliver ...................... 139Voisey, Christine R. ... 565, 569Voorhies, Mark ................. 468Vreeken, Rob ...................... 55Vu Nguyen, Thanh ............ 325WWaalwijk, C. ...................... 670Waalwijk, Cees .................. 230Wagener, Jeanette ........... 503Walbot, Virginia ................ 624Walker, Anne-Sophie .. 15, 329Walker, Janet ...................... 16Walker, L. .......................... 699Walker, Louise .................. 110Walsh, Christopher ............... 5Walther, A. ....................... 451Walther, Andrea 120, 200, 317Wang, Bin ......................... 389Wang, Chenfang ............... 267Wang, Chen fang .............. 268Wang, Chengshu ............... 274Wang, C.-L. ....................... 205Wang, Clay .......................... 54Wang, Guanghui ....... 438, 574Wang, Linqi ...... 127, 391, 509,.................................. 510, 738Wang, Ming ...................... 497Wang, Ping........................ 458Wang, Q. ........................... 530Wang, Sha ......................... 369Wang, T.-F. ....................... 205Wang, Xu .......................... 138Wang, Xuying .... 125, 243, 701Wang, Yanli ....................... 458Wang, Yizhou .................... 475Wang, Yonglin ................... 346Wang, Yuanchao ............... 531Wang, Yun ........................ 605Ward, T. J. ......................... 670Warn, P. A. ........................ 488Wasserstroem, Lisa ........... 200Wasserström, Lisa............. 120Watanabe, Kenji ................. 59Watters, Michael K. .......... 172Wawra, Stephan ....... 525, 532Weber, Jakob .................... 571Weerasinghe, Harshini C. .. 597Wehmeier, Silvia ............... 111Wehr, Michaela ................ 152Wei, Jiajie.......................... 467Weiberg, Arne .................. 497Weichert, Martin .............. 179Weimer, L. ........................ 638Weirauch, M. .................... 387Welch, Juliet ..................... 721Wender, Carly ..................... 33Wendland, J. ..................... 451Wendland, Jürgen .... 120, 150,.......................................... 317Wendland, Jurgen W. ....... 200Werner, Ernst R. ........... 1, 357Westerholm-Parvinen, A. . 432Whisson, Stephen C. . 402, 532Whiston, Emily A. ............. 292White, David ..................... 598White, Theodore............... 502Whiteway, M. ................... 378Whittington, Amy ............. 458Whittle, Carrie .................. 677Wickramage, Amritha ....... 629Wiemann, Phillipp ............ 409Wiest, Aric ........................ 304Wigen, Lauren J. ............... 376Wilken, P. Markus............. 206Williams, Angela ............... 724Williams, Angela H. ........... 632Willingmann, Peter ........... 549Wilson, Mandy ................. 250Wilson, R. ......................... 635Wilson, R. A. ..................... 732Wingfied, Mike J. .............. 206Wingfield, B. D. ......... 260, 264Wingfield, Brenda D. ........ 206Wingfield, M. J. ................. 260Wise, Hua ......................... 483Wise, Hua Z....................... 533Wisecaver, Jennifer H. ...... 679Wit, Pierre ........................ 230Wohlschlager, Therese ..... 705Wolfers, Simon ................. 202Wollenberg, T. .................. 291Wolters, Dirk .................... 199Wolters, Dirk A. ................ 210Wong, Kin Sing.................. 287Wong, Koon Ho ........ 366, 423Wong, Man Chun .............. 287Wool, Nathan K. ............... 396Woolford, Carol ................ 379Woriedh, Mayada ............. 549Worley, Jeremy................. 679Wormley, Floyd ................ 509Worthington, Carolyn J. .... 668Wortman, Jennifer ........... 223Wortman, Jennifer Russo . 340Wösten, H. A. B. ............... 207Wösten, Han A. B.100, 237, 695Wozniak, Karen ................ 509Wu, Bo-Ming .................... 455Wu, Cheng ........................ 738Wu, Dongliang .................. 534Wu, Jiayao ........................ 349Wyatt, Timon T. ........ 151, 207Wymore, Farrell........ 224, 340XXiang, Meichun ................. 598Xiang, Qijun ...................... 389Xiong, Yi .................... 418, 466Xiong, Z. ............................ 541Xu, J. ................................. 326Xu, Jin-Rong ............. 267, 268,................................. 438, 574Xu, Jinrong ........................ 578Xu, Jun .............................. 350Xu, Liangsheng .................. 598Xu, Wenjie ........................ 379Y316

AUTHOR LISTYablonowski, Jacob .......... 172Yadav, V. ........................... 114Yadeta, Koste ................... 251Yamaguchi, M. .................. 114Yamakawa, Yohei ............. 463Yamane, N. ....................... 372Yang, A. ............................ 387Yang, Chun-Hsiang.............. 25Yang, Cui ........................... 268Yang, Dong-Hoon ............. 246Yang, Fei ........................... 383Yang, M.-S. ....................... 736Yang, Shuang ........................ 7Yarden, O.......................... 168Yarden, Oded ................... 711Yeadon, P. Jane ......... 717, 718Yeh, Hsu-Hua ...................... 54Yerra, Lakshmi Preethi ....... 96Yi, Mihwa ................... 138, 492Yin, Q. Y. ........................... 122Yin, Weixiao ...................... 531Yin, Zhikang .................16, 722Yoo, Soyeon ...................... 296Yoo, Young-Bok ................. 323Yoshimi, Akira ........... 367, 368You, Tao ............................ 722Young, C. A. ....................... 651Young, Carolyn ............ 48, 324Young, Carolyn A. ........ 26, 511Young, Mark ...................... 514Younomaru, T. ................... 572Yu, Eun .............................. 445Yu, Eun Young ................... 137Yu, Jae-Hyuk ............ 89, 91, 97Yu, JJae-Hyuk .......................90Yu, Yidong ................. 218, 219Yun, S.-H. ................... 439, 736Yun, Sung-Hwan ........ 269, 411Yuzon, J. ..............................27ZZala, M. ............................. 658Zala, Marcello .................... 448Zamborszky, J. ................... 386Zehraoui, E. ............... 410, 447Zembek, Patrycja .............. 563Zeraati, H. ......................... 434Zhai, Bing .................. 509, 738Zhang, Dong-Xiu ................ 273Zhang, Hui ............................. 7Zhang, Jian ........................ 558Zhang, Juanyu ................... 267Zhang, Lei .......................... 274Zhang, Lisha ...................... 375Zhang, Ning ....................... 674Zhang, Ruifu ...................... 558Zhang, Shijie ...................... 267Zhang, Shizhu .................... 369Zhang, Shuping.................. 509Zhang, Ying .................. 98, 467Zhao, Chunzhao ................ 230Zhao, G. ............................. 326Zhao, H. ............................. 735Zhao, Hai ............................. 60Zhao, Jiuhai ....................... 721Zhao, Shuang .................... 674Zhao, X. ............................. 326Zheng, Dawei .................... 267Zheng, Hailin ..................... 369Zheng, He .......................... 605Zheng, Qian ....................... 268Zheng, Y. ........................... 371Zhong, Shaobin ......... 319, 336Zhou, Mian ........................ 390Zhou, Miaomiao ................ 694Zhou, Tian ......................... 518Zhou, Xiaoying .......... 438, 578Zhou, Zhihua ..................... 274Zickler, D. .......................... 321Ziemann, Sebastian ........... 622Zimmerman, Kolea .............. 40Zimmerman, Kolea C. K. .... 642Ziv, C. ................................ 168Zou, Gen ........................... 274Zuccaro, A. ........................ 327Zupan, B. ........................... 225Zupanec, Neja ..................... 6727th Fungal Genetics Conference | 317

LIST OF PARTICIPANTSDuur K. AanenWageningen UniversityPhone: 0031-317-483144duur.aanen@wur.nlMikael R AndersenDepartment of Systems BiologyPhone: 4545252675mr@bio.dtu.dkNina K AroVTT Technical Research CentrePhone: 3.5850356463e+011nina.aro@vtt.fiKerrie W BarryDOE Joint Genome InstitutePhone: (925) 296-5672kwbarry@lbl.govSalma AbdeljalilCentre of Biotechnology of SfaxPhone: 21622731918salma.abdeljalil@gmail.comJames B AndersonUniv TorontoPhone: (905) 828-5362janderso@utm.utoronto.caFelipe R ArredondoOregon State UniversityPhone: (541)737-3679felipe.arredondo@oregonstate.eduSalomon Bartnicki-GarciaCICESEPhone: 52 646-175-0590bart@citrus.ucr.eduGerard C Adams JrUniv of Nebraska-LincolnPhone: 402-472-2858gadams3@unl.eduJennifer L AndersonUppsala University, EBCPhone: +46 018-471 2827jennifer.anderson@ebc.uu.seFred O AsiegbuUniversity of HelsinkiPhone: +358 9 191 58109Fred.Asiegbu@Helsinki.fiEvelina Y. BasenkoUniversity of GeorgiaPhone: (706) 542-1550ebasenko@uga.eduKate Juliette AffeldtUniversity of Wisconsin-MadisonPhone: (608) 262-1958kjschmitt2@wisc.eduJonathan P AndersonCSIROPhone: +61 8 93336103jonathan.anderson@csiro.auLea AtanasovaVienna University of TechnologyPhone: 4.3676583722e+011lea.atanasova@tuwien.ac.atSebastian BaumannInstitute for MicrobiologyPhone: 0049 (0)211 8114834baumann@hhu.deOleg AgafonovUniversity of StavangerPhone: (47)40340499oleg.agafonov@uis.noMatthew Z AndersonUniversity of MinnesotaPhone: 408-394-0489mzanders@umn.eduSara J AtkinsonCoker CollegePhone: (843) 340-3255sara.atkinson@coker.eduOezguer BayramInst Microbiology & GeneticsPhone: 49 551 39 37 72obayram@gwdg.dePablo S AguilarInstitut Pasteur de MontevideoPhone: +598 2522 0910aguilar@pasteur.edu.uyKarl-Magnus AnderssonLund UniversityPhone: 46730377338karl-magnus.andersson@biol.lu.seJin-Ho BaekChonbuk National Univ.Phone: 82 10 2728 3600sadasoul@nate.comHoda BazafkanAIT Austrian Institute of TechnoPhone: 0043 664 88390594karin.schlaudoschich@ait.ac.atJesus L AguirreI de Fisiologia Celular-UNAMPhone: 5255 5622-5651jaguirre@ifc.unam.mxAlex AndrianopoulosUniversity of MelbournePhone: 61 3-9344-5164alex.a@unimelb.edu.auYong-Sun BahnYonsei UniversityPhone: -7601ysbahn@yonsei.ac.krLasse BechAalborg UniversityPhone: 4522431410lab@bio.aau.dkAudrey M V Ah-FongUniv California, RiversidePhone: (951) 827-3932audreya@ucr.eduCaroline AngelardUniversity of LausannePhone: 41786714773caroline.angelard@unil.chScott E BakerPacific Northwest Natl LabPhone: (509) 372-4759scott.baker@pnnl.govNicola BeckmannBiocenter, Innsbruck Med. Uni.Phone: 4.3650525009e+011nicola.beckmann@i-med.ac.atSteven AhrendtUniversity of California, RiversidePhone: (951) 827-2363sahre001@ucr.eduMd. Kausar AlamUniversity of SaskatchewanPhone: 306 341 2765kausar.alam@usask.caSabine E AlbermannWWU MuensterPhone: 4.9251832499e+011sabine.albermann@wwu.deNoam AlkanWeizmann Institute of SciencePhone: 972-8-9342739noam.alkan@weizmann.ac.ilFausto AlmeidaUniversity of São PauloPhone: +55(16)36023066fbralmeida@usp.brJoelle AmselemINRAPhone: 33 1-30-833395joelle.amselem@versailles.inra.frGeorge E AnasontzisChalmers University of TechnologyPhone: 46733603904george.anasontzis@chalmers.seSarah L AnglinMillsaps ColPhone: (601) 974-1414mcguisl@millsaps.eduAmanda C.C. AntoniêtoUniversity of São PauloPhone: (16) 81004410amandaantonieto@yahoo.com.brDiana AnyaoguTechnical University of DenmarkPhone: +45 45252500dca@bio.dtu.dkKahandawa AriyawansaMassey UniversityPhone: 6421 1435681sameera.ariyawansa@agresearch.co.nzJose ArnauNovozymesPhone: +45 44461218joau@novozymes.comMartha B ArnaudStanford University School of MedicinePhone: (415) 602-7488arnaudm@stanford.eduA. Elizabeth ArnoldThe University of ArizonaPhone: 5206217212arnold@ag.arizona.eduClara BaldinHans-Knoell-InstitutePhone: +49 (0)3641 532-1382clara.baldin@hki-jena.deThomas BaldwinUniv of GeorgiaPhone: 518-322-9435tbaldwin@uga.eduDithi BanerjeeUniversity at Buffalo, SUNYPhone: 7168292091dithiban@buffalo.eduNabaraj BanjaraUniversity of Nebraska-LincolnPhone: 402-472-2825nabaraj.banjara@huskers.unl.eduFlora BanuettCalifornia State UniversityPhone: (562)-985-5535fbanuett@csulb.eduSharief BarendsDuPont IBPhone: 31715686159sharief.barends@dupont.comBridget M BarkerMontana State UnivPhone: 406-994-7467bridget.barker@montana.eduMaria-Soledad BenitezDuke UniversityPhone: 9196607362maria.benitez.ponce@duke.eduJoan W BennettRutgers UniversityPhone: 848-932-6223profmycogirl@yahoo.comRichard BennettBrown UniversityPhone: (401) 863-3121richard_bennett@brown.eduCalmes BenoitUniversity of AngersPhone: 689424176calmes.b@gmail.comIsabelle BenoitCBS-KNAWPhone: +31 (0)30 2122621i.benoit@cbs.knaw.nlJohan Philipp BenzUC BerkeleyPhone: (510) 666-2556ph.benz@berkeley.eduMary L BerbeeUniv British ColumbiaPhone: (604) 822-3780mary.berbee@gmail.com318

LIST OF PARTICIPANTSLeslie G BeresfordNovozymes Inc.Phone: (530) 757-8100 4627LSBF@novozymes.comBurton H BluhmUniversity of ArkansasPhone: (479)575-2677bbluhm@uark.eduAxel A BrakhageLeibniz Institute Infection BiolPhone: 49 3641-5321000axel.brakhage@hki-jena.deSteve BrownNovozymes IncPhone: (530) 757-8104shbr@novozymes.comRandy M BerkaNovozymes, IncPhone: (530) 757-4974ramb@novozymes.comMarike J. BoenischBiocenter Flein FlottbekPhone: -43105marike.boenisch@uni-hamburg.deSara BrancoUniversity of CaliforniaPhone: 510 688 8583sara.mayer.branco@gmail.comMichael BrunnerUniv HeidelbergPhone: 49 6221-544207michael.brunner@bzh.uni-heidelberg.deJudith G BermanTel Aviv UniversityPhone: 03-640-7633jberman@post.tau.ac.ilDaniela BoettgerHans-Knöll-InstitutePhone: 1.1491765511e+014daniela.boettger@hki-jena.deAlexandra C BrandUniv AberdeenPhone: 44 1224-555878a.brand@abdn.ac.ukTom D BrunsUniv California, BerkeleyPhone: (510) 642-7987pogon@berkeley.eduDaniel BerryMassey UniversityPhone: 6463505543d.berry@massey.ac.nzMichael BolkerPhilipps UniversityPhone: 49 6421-2821536boelker@staff.uni-marburg.deChristoph BraunBayer SAS - Bayer CropSciencePhone: 33 472 85 23 43christophandreas.braun@bayer.comJerzy BrzywczyInstitute of Biochemistry &BiophPhone: -5921291jurek@ibb.waw.plMaria Célia BertoliniInstitute of ChemistryPhone: 55-16-33019675mcbertol@iq.unesp.brStephanie R BollmannOregon State UniversityPhone: (541)737-3679stephanie.bollmann@oregonstate.eduGerhard H BrausGeorg-August UnivPhone: 49 551-39-3771gbraus@gwdg.deJoanna A BuecheUC BerkeleyPhone: 510-643-2546jbueche@berkeley.eduMargherita BertuzziImperial College LondonPhone: 020 7594 7409margherita.bertuzzi06@imperial.ac.ukMelvin D BoltonUSDA - ARSPhone: 701 239 1373melvin.bolton@ars.usda.govSusanna A Braus-StromeyerGeorg-August-Univ GoettiPhone: (0551) 393817sbraus@gwdg.deHayley E BugejaUniversity of MelbournePhone: +61 411 409 949h.bugeja@unimelb.edu.auSinem BeyhanUniversity of California, San FranciscoPhone: (415) 502-4810sinem.beyhan@ucsf.eduGregory BonitoDuke UniversityPhone: 9196607363gmb2@duke.eduGustavo Adolfo Bravo RuizUniversity of CordobaPhone: 670472051b12brrug@uco.esVincent BuloneRoyal Inst Biotech (KTH)Phone: 46 70 389 1122bulone@kth.seMonica BhaskarHelsinki UniversityPhone: 3.5844530354e+011monica.bhaskar@helsinki.fiKatherine A BorkovichUniversity of CaliforniaPhone: (951) 827-2753katherine.borkovich@ucr.eduErin L BredewegOregon State UnivPhone: (541) 737-4399erin.bredeweg@gmail.comKathleen M BurchhardtNorth Carolina State UniversityPhone: 919-513-4840kmpitche@ncsu.eduMichael J BidochkaBrock UniversityPhone: 905-688-5550 x3392bidochka@brocku.caJoerg BormannUniversity HamburgPhone: 4.9404281631e+011bormannj@botanik.uni-hamburg.deMarin BrewerUniversity of GeorgiaPhone: 7065421254mtbrewer@uga.eduMaria A. BurggraafInstitute of Biology LeidenPhone: +31(0)71 5274745m.a.van.welzen@biology.leidenuniv.nlElaine M BignellImperial College LondonPhone: 00 44 207 594 2074e.bignell@imperial.ac.ukIndrani BoseWestern Carolina UniversityPhone: (828) 227-3658ibose@email.wcu.eduSusan M BrewerDuPont Industrial BiosciencesPhone: 650-846-4632susan.brewer@dupont.comClaire BurnsWashington and Jefferson CollegePhone: (812) 219-6499burnsc@indiana.eduRobert Blake BillmyreDuke UniversityPhone: 4438040705bbillmyre@gmail.comUlrike BinderMedical University InnsbruckPhone: -80220ulrike.binder@i-med.ac.atVasileios BitasPennsylvania State UnivPhone: (814) 441-0690vzb107@psu.eduSandra BloemendalRuhr-University BochumPhone: 4.9234322247e+011sandra.bloemendal@rub.deSara J BlosserMontana State UniversityPhone: (406) 994-7468sara.wezensky@msu.montana.eduJordi C BoshovenWageningen UniversityPhone: 31613970288jordi.boshoven@wur.nlOurdia BouzidUtrecht UnivPhone: 31 030 2122600o.bouzid@uu.nlJoanna K BowenPlant & Food ResearchPhone: +64 9 925 7154joanna.bowen@plantandfood.co.nzBarry J BowmanUniversity of CaliforniaPhone: (831) 459-2245bbowman@ucsc.eduEmma-Jean BowmanUniv CaliforniaPhone: (831) 459-3448rbowman@biology.ucsc.eduMatthias BrockFriedrich Schiller University and HansKnoell InstPhone: 4.9364153217e+012matthias.brock@hki-jena.deStuart BrodyUniv California, San DiegoPhone: (858) 534-2619sbrody@ucsd.eduAlistair J P BrownUniversity of AberdeenPhone: +44 1224 437482al.brown@abdn.ac.ukDaren W BrownUSDA/ARSPhone: (309) 681-6230browndw@ncaur.usda.govJessica C BrownU. California, San FranciscoPhone: (617) 258-5190jessica.brown@ucsf.eduKathryn E BushleyOregon State UnivPhone: (541) 908-0116bushleyk@science.oregonstate.eduGeraldine ButlerUniv Col DublinPhone: 353 1-7166885gbutler@ucd.ieNicole BühlerKarlsruhe Institut of TechnologyPhone: 0721-60844633Nicole.Buehler@kit.eduJulia BöhmRuhr-University BochumPhone: 4.9234322566e+011julia.boehm@rub.deIlva Esther CabreraUniv of California, RiversidePhone: (951) 827-3190ilva.cabrera@email.ucr.edu27th Fungal Genetics Conference | 319

LIST OF PARTICIPANTSMark X CaddickUniv LiverpoolPhone: 44 151-795-4566caddick@liv.ac.ukMARTA CASTRILLO JIMÉNEZUniversity of SevillePhone: +34 954 555 948martacastrillo@us.esXuanjin ChengThe Chinese University of Hong KongPhone: (852)39431331jeanch12345@gmail.comSamuel T CoradettiUniversity of CaliforniaPhone: 510-666-2559scorad.register@gmail.comMarie-Cecile CaillaudThe Sainsbury LaboratoryPhone: +44(0)1603450430marie-cecile.caillaud@tsl.ac.ukErnestina Castro-LongoriaCICESEPhone: 646 1750500ecastro@cicese.mxHelene ChiapelloCR INRA Toulouse Midi-PyreneesPhone: 33 5 61 28 52 87helene.chiapello@toulouse.inra.frPadraic G CorcoranUppsala UniversityPhone: 735853162padraic.corcoran@ebc.uu.seSilvia Calo VarelaDUKE University Medical CenterPhone: 919.684.2809silvia.calovarela@duke.eduDavid E A CatchesideFlinders UnivPhone: 618 8201-2335david.catcheside@flinders.edu.auYangrae ChoUniversity of Hawaii at ManoaPhone: 808-956-5305yangrae@hawaii.eduBen J.C. CornelissenUniversity of AmsterdamPhone: 0031 20 5257707b.j.c.cornelissen@uva.nlAna M Calvo-ByrdNorthern Illinois UniversityPhone: (815) 753-0451amcalvo@niu.eduGustavo C CerqueiraBroad InstitutePhone: 2405155696gustavo@broadinstitute.orgJaehyuk ChoiSeoul National UniversityPhone: -5484jaehyukc@gmail.comNicolas CorradiUniversity of OttawaPhone: +1 6132529432ncorradi@uottawa.caLeona CampbellUniv SydneyPhone: 61293513758leona.campbell@sydney.edu.auFrançois CesbronHeidelberg Biochemistry CentrePhone: -8412261cesbron.francois@gmail.comJaeyoung ChoiSeoul National UniversityPhone: -5746nizam84@snu.ac.krCristina Corral RamosUniversity of CordobaPhone: 34665882311z12corac@uco.esThiago S CandidoUNESP - IQ - AraraquaraPhone: (55) (14) 96057215thiago.s.candido@gmail.comNadia ChackoTexas A&M UniversityPhone: (979) 845-7259nchacko@bio.tamu.eduTetsuya ChujoMassey UniversityPhone: -9397t.chujo@massey.ac.nzLuis M CorrochanoUniversity of SevillePhone: 34 954 550 919corrochano@us.esPaulo Francisco CanessaUniversidad Catolica de ChilePhone: 5626862348paulo.canessa@gmail.comJimmy ChanDuPont Industrial BiosciencesPhone: 6166179146jimmy.chan@dupont.comHyunjung ChungSeoul National UniversityPhone: -5484comcato@snu.ac.krPeter J CottyUniversity ArizonaPhone: (520) 626-5049pjcotty@email.arizona.eduNallely Cano-DominguezCICESEPhone: 01 (646) 175 05 00ncanodominguez@gmail.comChia-Chen ChangUniversity of EdinburghPhone: +44(0)131 651 3337jeanwhale@gmail.comAlice C L ChurchillCornell UnivPhone: (607) 255-0872acc7@cornell.eduMikael S CourbotSyngenta Crop Protection MunchwilenPhone: 41628660368mikael.courbot@syngenta.comDavid CanovasUniv de SevillaPhone: 34 954 55 59 47davidc@us.esJinhui ChangThe Chinese University of Hong KongPhone: (852)39431331tjchang@cuhk.edu.hkLynda M CiuffettiOregon State UnivPhone: (541) 737-2188ciuffetl@science.oregonstate.eduSarah F CovertUniv GeorgiaPhone: (706) 542-1385covert@uga.eduIgnazio CarboneNorth Carolina State UniversityPhone: (919) 513-4866ignazio_carbone@ncsu.eduNohemi Carreras-VillasenorLANGEBIO-CinvestavPhone: 52 462 1527287ncarreras@ira.cinvestav.mxDee A CarterUniv SydneyPhone: 61 29351-5383dee.carter@sydney.edu.auGemma M CartwrightMassey UnivPhone: 64 63505515g.m.cartwright@massey.ac.nzJeffrey W CaryARS-SRRCPhone: 504-286-4264Jeff.Cary@ars.usda.govSonia M CastanheiraCNB-CSICPhone: +34 923294912smcastanheira@cnb.csic.esPranatchareeya ChankhamjonHKIPhone: +49 (0)3641 532 1392pranatchareeya.chankhamjon@hkijena.deNikki D CharltonThe Samuel Roberts Noble FoundationPhone: 580-224-6964ndcharlton@noble.orgBiju V ChellappanUniversity of Amsterdam, SILSPhone: 31687098694b.chellappan@uva.nlJulian J-L ChenArizona State UniversityPhone: 480-965-3650JLChen@asu.eduWeidong ChenUSDA ARSPhone: 5093359178w-chen@wsu.eduYuan ChenDuke University Medical CenterPhone: 9193217870yuan.chen@duke.eduPierre-Henri J ClergeotStockholms UniversitetPhone: 46 8 16 37 56clergeot@botan.su.seJeffrey J ColemanRhode Island Hosp/Alpert MedicalSchool of Brown UniversityPhone: (401) 444-7309jjcoleman@partners.orgJérôme CollemareWageningen UniversityPhone: (+31) 0317-483096jerome.collemare@wur.nlBradford J CondonCornell UnivPhone: (607) 255-3200bjc225@cornell.eduMichael R CooperUniversity of LouisvillePhone: 5023381405michael.cooper@louisville.eduEvelyne CoppinUniv Paris - SudPhone: 33 169 157012evelyne.coppin@igmors.u-psud.frRobert A CramerGeisel School of Medicine at DartmouthPhone: (603) 650-1040robert.a.cramer.jr@dartmouth.eduRhaisa CrespoUniv of Puerto Rico MayaguezPhone: 787-614-7278rhaisa.crespo@upr.eduJuliana CrestaniUSPPhone: 5.5129730841e+011julicrestani@gmail.comDaniel CrollETH ZurichPhone: +41 44 632 3387daniel.croll@me.comMichael CsukaiSyngentaPhone: +44 1344 414094michael.csukai@syngenta.comFernanda B CupertinoInstituto de Quimica,UNESPPhone: 55-16-33019804fernanda_cupertino@yahoo.com.br320

LIST OF PARTICIPANTSTim A DahlmannRuhr-University BochumPhone: 4.9234322221e+011tim.dahlmann@rub.deSourabh DhingraNorthern Illinois UniversityPhone: 815 753 1468dhingra@niu.eduKelly DrewsVirginia TechPhone: 7037281190kcdrews@gmail.comEDOUARD EVANGELISTIINRA/CNRS/Université de NicePhone: 492386588edouard.evangelisti@sophia.inra.frPascale Daran-LapujadeDelft Univ. of TechnologyPhone: +31 15 2789965p.a.s.daran-lapujade@tudelft.nlAntonio Di PietroUniv de CordobaPhone: 34 957 2189 81ge2dipia@uco.esBernard DumasUMR5546 CNRS Univ P SabatierPhone: 33 562193503dumas@lrsv.ups-tlse.frJaime EyzaguirreUniversidad Andres BelloPhone: (562) 26618070jeyzaguirre@unab.clHeather L DarbyshirUniversity of NottinghamPhone: +44 115 951 3203sbxhd4@nottingham.ac.ukGeorge DiallinasUniversity of AthensPhone: 3.0210727465e+011diallina@biol.uoa.grJay C DunlapGeisel Schl Medicine@DartmouthPhone: (603) 650-1108jay.c.dunlap@dartmouth.eduDavid EzraARO The Volcani CtrPhone: 972-3-9683555dezra@volcani.agri.gov.ilAsen I DaskalovIBGC UMR5095Phone: 629669841asen.daskalov@ibgc.cnrs.frMarty DickmanPlant Genomics and BiotechnoloPhone: (979) 862-4788mbdickman@tamu.eduSebastien DuplessisINRA - Tree-Microbe InteractionsPhone: 33 383 394013duplessi@nancy.inra.frYohann Faivre TalmeyUniversity Claude BernardPhone: 04 72 85 23 78yohann.faivre@bayer.comCarol E DavisUCRPhone: (951) 827 3932cdavis@ucr.eduAxel CR DiernfellnerBZHPhone: +49 6221 544778axel.diernfellner@bzh.uni-heidelberg.deDannie DurandCarnegie Mellon UniversityPhone: (412) 268-6036durand@cmu.eduRosa FajardoCICESEPhone: 646 175 05 00rfajardo@cicese.mxCharissa de BekkerPennsylvania State UniversityPhone: 814-321 2226amd32@psu.eduFred S DietrichDuke UniversityPhone: 919 684 2857fred.dietrich@duke.eduPaul S DyerUniversity of NottinghamPhone: 44 115-9513203paul.dyer@nottingham.ac.ukAhmad M FakhourySouthern Illinois UnivPhone: (618) 453-1782amfakhou@siu.eduRonnie de JongeVIB - PSBPhone: 3293313537ronnie.dejonge@psb.ugent.beDaniela DirschnabelRuhr-University BochumPhone: 4.923432277e+011daniela.dirschnabel@rub.deCarla J EatonMassey UniversityPhone: 64 63569099c.j.eaton@massey.ac.nzWeiguo FANGZhejiang UniversityPhone: 86-571-88206668wfang1@zju.edu.cnRonald P de VriesCBS-KNAW Fungal Biodiversity CentrePhone: 31 302-122600r.devries@cbs.knaw.nlPeter DoddsCSIROPhone: +61 2 62465039peter.dodds@csiro.auDaniel J EbboleTexas A&M UnivPhone: (979) 845-4831d-ebbole@tamu.eduYufeng FangVirginia TechPhone: 5408086725yufengf1@vt.eduRalph A DeanNorth Carolina State UnivPhone: (919) 513-0020ralph_dean@ncsu.eduGunther DoehlemannMax Planck InstitutePhone: -7152doehlemann@mpi-marburg.mpg.deMartin John EganHarvard Medical SchoolPhone: (617) 432-7177martin_egan@hms.harvard.eduErzsébet FeketeUniversity of DebrecenPhone: +3652512900 62488kicsizsoka@yahoo.comDaniele DebieuINRAPhone: 33130814564debieu@versailles.inra.frBastian DoernteUniversity of GoettingenPhone: 4.9551391409e+011bdoernt@gwdg.deSophia K EkengrenStockholm UnivPhone: 46 8 16 37 54ekengren@botan.su.seMichael FeldbruggeHeinrich-Heine UniversityPhone: 49 211 15475feldbrue@hhu.deRobert DebuchyUniv Paris-SUDPhone: 33 1-69157012robert.debuchy@igmors.u-psud.frMichael E DonaldsonTrent UniversityPhone: 705 748 1011michaeldonald2@trentu.caCandace E ElliottUniversity of MelbournePhone: (613)8344 5056ce@unimelb.edu.auDaria FeldmanThe Hebrew University of JerusalemPhone: 97289489315daria.feldman@mail.huji.ac.ilDiego Delgado-ÁlvarezCICESEPhone: +52 646 1265163ddelgado@cicese.edu.mxNicole M DonofrioUniv DelawarePhone: (302) 831-1372ndonof@udel.eduLynn EpsteinUniversity of CaliforniaPhone: 530-754-7916lepstein@ucdavis.eduCarl R FellbaumSouth Dakota State UniveristyPhone: 6052167854carl.fellbaum@sdstate.eduToni M DeloreyBroad InstitutePhone: 617-714-8225delorey@broadinstitute.orgTammy DotyNovozymes, Inc.Phone: 530 757-8100tmmd@novozymes.comNuria EscuderoUniversity of AlicantePhone: (+34)965903400 x 328nuria.escudero@ua.esMarianna FeretzakiDuke UnivPhone: (919) 684-2809mf55@duke.eduSteven H DenisonEckerd CollegePhone: (727) 864-8456denisosh@eckerd.eduBenjamin DoughanUniversity of FloridaPhone: 319-504-9168bdoughan@ufl.eduJose EspinoUniversity of MuensterPhone: 4.9251832481e+011espino@uni-muenster.deCaroline M FernandesFederal UniversityPhone: 55-21-25626573cmota@biof.ufrj.brAnne DettmannMolecular Plant PhysiologyPhone: 0049-0761203266adettma@gwdg.deDamien DownesKansas State UniversityPhone: 785-532-1348djdownes@ksu.eduJessie FernandezUniversity of Nebraska-LincolnPhone: (402) 613-6223jfernandez99@huskers.unl.edu27th Fungal Genetics Conference | 321

LIST OF PARTICIPANTSMelania FigueroaOregon State UniversityPhone: (541) 737-2234figuerme@science.oregonstate.eduGerda FourieFABIPhone: +2712 420 3938gerda1.fourie@fabi.up.ac.zaFumi FukadaKyoto Prefectural UniversityPhone: 81-75-703-5613ddswb383@yahoo.co.jpMARTHA C GIRALDOKANSAS STATE UNIVERSITYPhone: (785) 532-2337mgiraldo@ksu.eduScott G FillerLos Angeles Biomedical ResInstPhone: (310) 222-3813sfiller@ucla.eduElisabeth FournierINRAPhone: 33 499624863elisabeth.fournier@supagro.inra.frKevin K FullerGeisel Sch Medicine at DartmouthPhone: (603) 650-1120kevin.fuller@dartmouth.eduTatiana GiraudCNRS- Univ Paris 11Phone: 33 1 6915-5669tatiana.giraud@u-psud.frSabine FillingerINRAPhone: 33 1-3081 4565sabine.fillinger@versailles.inra.frThomas J FowlerSIUEPhone: (618) 650-5231tfowler@siue.eduJames GalaganBoston UniversityPhone: 617 875 9874jgalag@bu.eduPierre GladieuxUC BerkeleyPhone: 510-642-8441pgladieux@berkeley.eduAmanda J FischerNovozymes IncPhone: (530) 757-8100amfs@novozymes.comWilliam L FranckNC State UniversityPhone: 919-513-0167wlfranck@ncsu.eduJonathan M GalazkaOregon State UniversityPhone: 541-737-4399galazkaj@onid.orst.eduLouise GlassUniv CaliforniaPhone: (510) 643-2399lglass@berkeley.eduGregory J FischerUniv of Wisconsin-MadisonPhone: 715-574-8857gfischer2@wisc.eduRasmus J N FrandsenTechnical University of DenmarkPhone: 45 4525 2708rasf@bio.dtu.dkJORGE GARCÍA-MARTÍNEZUniversity of SevillePhone: +34 954 555 948jorgegarcia@us.esScott E GoldUSDA-ARSPhone: (706) 542-1259sgold@uga.eduReinhard FischerKarlsruhe Inst. of TechnologyPhone: 49 721-608-4630reinhard.fischer@kit.eduStephen J FreeSUNY Univ, BuffaloPhone: (716) 645-4935free@acsu.buffalo.eduDonald M GardinerCSIROPhone: 61-7-32142370donald.gardiner@csiro.auKatsuya GomiTohoku UnivPhone: 81 22-717-8901gomi@biochem.tohoku.ac.jpJoseph E FlahertyCoker CollegePhone: (843) 383-8079jflaherty@coker.eduStanley FreemanVolcani CtrPhone: 972 3-9683537freeman@volcani.agri.gov.ilAndrea GargasSymbiology LLCPhone: (608) 827-5164andreagargas@symbiology.comRodrigo D Gonçalvesinstituto de Química, UNESPPhone: 55 (16) 82007360rdgoncalves@gmail.comMarko FlajsmanUniv of LjubljanaPhone: 386 1 3203295marko.flajsman@bf.uni-lj.slDaniela FreihorstFriedrich-Schiller-UniversitätPhone: +49 3641 949293danielafreihorst@aol.comVictoriano GarreUniversity of MurciaPhone: 34 868887148vgarre@um.esStephen B GoodwinUSDA-ARS/Purdue UnivPhone: (765) 494-4635sgoodwin@purdue.eduDamien FleetwoodAgResearchPhone: -7917damien.fleetwood@agresearch.co.nzJohannes FreitagPhilipps-UniversitätPhone: 4.9642128271e+012JohannesFreitag@gmx.netAmruta GarudUC, RiversidePhone: 858-231-7773amruta.garud@email.ucr.eduMathieu GourguesUniversité de NicePhone: 33492386588mathieu.gourgues@sophia.inra.frAndre FleissnerTU BraunschweigPhone: 49 531-3915795a.fleissner@tu-bs.deMichael FreitagOregon State UnivPhone: (541) 737-4845freitagm@onid.orst.eduAudrey P GaschUniv Wisconsin, MadisonPhone: (608) 265-0859agasch@wisc.eduFrancine GoversWageningen UnivPhone: 31 317-483-138francine.govers@wur.nlDimitrios FloudasClark UniversityPhone: (508) 847-5892dfloudas@clarku.eduFernanda Z FreitasIQ-UNESPPhone: 55-16-33019804fernanda.zanolli@gmail.comElodie GaulinLRSV UMR5546 CNRS/UPSPhone: +33 534323803gaulin@lrsv.ups-tlse.frNeil A R GowUniv AberdeenPhone: 44 1224-437483n.gow@abdn.ac.ukKathryn FordUniversity of BristolPhone: 7738247075Kathryn.Ford@bristol.ac.ukSteven FriedmanOregon State UniversityPhone: (410) 967-0974stevenfriedman8@gmail.comGregory M. GauthierUniversity of WisconsinPhone: (608) 262-7703gmg@medicine.wisc.eduJonathan GrandaubertINRAPhone: 33130814569jonathan.grandaubert@versailles.inra.frNatasha (Tash) T ForesterAgResearch / University of OtagoPhone: 6463518315natasha.forester@agresearch.co.nzTimothy L FriesenUSDA-ARSPhone: 701-239-1337timothy.friesen@ars.usda.govAnupama GhoshMPI MarburgPhone: 0049-6421-178571anupama.ghosh@mpi-marburg.mpg.deFabrice N GravelatMcGill UniversityPhone: 1 (514) 398-4434fabrice.gravelat@mcgill.caJarrod R FortwendelUniversity of South AlabamaPhone: 251-460-6681jfortwendel@southalabama.eduCi FuUniversity at BuffaloPhone: (716)645-4936cifu@buffalo.eduArit GhoshUniversity of California, RiversidePhone: 951-827-3190aghos003@ucr.eduDave GreenshieldsNovozymes BioAg LtdPhone: 3066578269dvgs@novozymes.comJames M FountaineScotland's Rural CollegePhone: 4.4131535437e+011james.fountaine@sruc.ac.ukMakoto FujimuraToyo UnivPhone: 81 276-82-9216mfujimura@toyo.jpSarah A GilmoreUCSFPhone: 4155024810sarah.gilmore@ucsf.eduLaura J Grenville-BriggsRoyal Institute of TechnologyPhone: +46 (0)5537 8890lauragb@kth.se322

LIST OF PARTICIPANTSIgor V GrigorievJoint Genome InstPhone: (925) 296-5860ivgrigoriev@lbl.govUlrich GüldenerHelmholtz-Zentrum MünchenPhone: +49 89 3187 3582u.gueldener@helmholtz-muenchen.deJessica HargartenUniversity of Nebraska- LincolnPhone: 925-300-7007jessica.c.teixeira@gmail.comYainitza Hernandez-RodriguezUniv GeorgiaPhone: (706) 542-6026yrodriguez@plantbio.uga.eduPierre GrognetUniversité Paris SudPhone: 33169154657pierre.grognet@igmors.u-psud.frThomas HaarmannAB Enzymes GmbHPhone: 4.9615136804e+012thomas.haarmann@abenzymes.comSteven D HarrisUniv NebraskaPhone: (402) 472-2938sharris2@unl.eduAndreas HerrKarlsruhe Inst of TechnologPhone: 4.9721608446e+012andreas.herr@kit.eduGuido GrossmannUniversity of HeidelbergPhone: -551832grossmann@stanford.eduHubertus HaasInnsbruck Medical UniversityPhone: -11363969hubertus.haas@i-med.ac.atThomas HartmannChinese Academy of SciencesPhone: 0086 1860 237 384hartmann.tom@gmail.comAlfredo H Herrera-EstrellaCINVESTAV-IPN, IrapuatoPhone: 52-462-1663041aherrera@langebio.cinvestav.mxAlexey A. Grum-GrzhimayloWageningen UniversityPhone: 31684940430alexey.grumgrzhimaylo@wur.nlYitzhak HadarHebrew UnversityPhone: 972-8-9489935hadar@agri.huji.ac.ilSahar HasimUnivesity of Nebraska LincolnPhone: 402-2022326saharm@huskers.unl.eduAndrea HerrmannWWU MuensterPhone: 4.9251832499e+011andrea.herrmann@uni-muenster.deElisabeth GrundBayer CropSciencePhone: 33649116097elisabeth.grund@bayer.comStefan A HaefnerBASF SEPhone: +49 621 60-42902stefan.haefner@basf.comShin HatakeyamaSaitama UnivPhone: 81-48-858-3414shinh@mail.saitama-u.ac.jpChristian HertweckHKIPhone: +49 3641 5321100christian.hertweck@hki-jena.deAndrii GryganskyiDuke UniversityPhone: 9196607363apg10@duke.eduDaisuke HagiwaraChiba universityPhone: -2983dhagi@chiba-u.jpVirginia E HavelUniversity at BuffaloPhone: 7168292091vehavel@buffalo.eduBritta HerzogGeorg-August-UniversityPhone: 4.9551391967e+011bherzog@gwdg.deFabio GsallerMedical Univ of InnsbruckPhone: 0043 650 6617957Fabio.Gsaller@i-med.ac.atCharles R HallDupont Industrial BiosciencesPhone: (510) 350-7426charles.hall@dupont.comPatrick M HayesUniversity of BristolPhone: 44117 928 7475patrick.hayes@bristol.ac.ukJaqueline HessHarvard UniversityPhone: 617-496-9741jhess@fas.harvard.eduMatthias GubeFSU JenaPhone: 4.936419494e+011matthias.gube@uni-jena.deHeather E Hallen-AdamsUniversity of Nebraska-LincolnPhone: (402) 472-2825hhallen-adams2@unl.eduKim Hee-KyoungSoonchunhyang UniversityPhone: -1777beargn@naver.comAna G Hesselbart MarquezCarlsberg LaboratoryPhone: 4533275235yonosoyana@gmail.comJANBON GuilhemInstitut PasteurPhone: 33 1 45688356janbon@pasteur.frRichard C HamelinUniversity of British ColumbiaPhone: 604-827-4441richard.hamelin@ubc.caYvonne HeiligMolecular Plant PhysiologyPhone: 4.9761203267e+011yheilig@gwdg.deDavid S HibbettClark UniversityPhone: 508-793-7332dhibbett@clarku.eduThomas GuillemetteUniversity of AngersPhone: 33241735260thomas.guillemette@univ-angers.frAmy M GundersenNovozymes, Inc.Phone: 559-706-0062aygm@novozymes.comYogesh K GuptaUniversity of ExeterPhone: -7404494002ykg201@exeter.ac.uksarah j gurrUniv oxfordPhone: 4.4186527581e+011sarah.gurr@plants.ox.ac.ukPaulina Guzman-GuzmanUniversity of GuanajuatoPhone: 473 1090556paulina.gzmn2@gmail.comRachana GyawaliTexas A&M UniversityPhone: (979) 845-7259rgyawali@bio.tamu.eduKim E Hammond-KosackRothamsted ResearchPhone: 44-1582-763133kim.hammondkosack@rothamsted.ac.ukJae-Gu HanRural Development AdministrationPhone: -4631180jaeguhan@gmail.comKap-Hoon HanWoosuk UnivPhone: 82 63-290-1427khhan@woosuk.ac.krChristopher T Hann-SodenUC BerkeleyPhone: 9258722716channsoden@berkeley.eduWilhelm HansbergInst. Fisiología Celular/UNAMPhone: 5255 5622 5655whansberg@ifc.unam.mxOmar HarbUniv of PennsylvaniaPhone: 215-746-7019oharb@pcbi.upenn.eduKai HeimelGeorg-August-UniversityPhone: 49551-393815kheimel@gwdg.deJoseph HeitmanDuke UnivPhone: 919-684-2824heitm001@duke.eduJens HellerUC-BerkeleyPhone: 510-501-3152jens.heller@berkeley.eduGinnie K HenchUNC Chapel HillPhone: (919) 923-2309ghench@med.unc.eduCatarina HenkeFriedrich-Schiller Univ - MPIPhone: 49 03461 949299chenke@ice.mpg.deBernard HenrissatCNRSPhone: 33 49182 55 87Bernard.Henrissat@afmb.univ-mrs.frYujiro HiguchiUniversity of ExeterPhone: +44 (0) 1392 269178Y.Higuchi@exeter.ac.ukKristiina HildenUniv HelsinkiPhone: 35891911kristiina.s.hilden@helsinki.fiTerry W HillRhodes CollegePhone: 901-843-3559hill@rhodes.eduRobert Louis HirschUniversity of ArkansasPhone: 479-575-4952rhirsch@uark.eduAndrea HlubekBASFPhone: +49 621 60-59338andrea.a.hlubek@basf.comDirk HoffmeisterFriedrich-Schiller-UniversityPhone: 01149-3641-949850dirk.hoffmeister@hki-jena.de27th Fungal Genetics Conference | 323

LIST OF PARTICIPANTSLinda HollandUniversity College DublinPhone: 3.5387664498e+011linda.holland@ucd.ieDavid HuskeyUniversity of ArizonaPhone: 520-621-9390dahuskey@email.arizona.eduJernej JakseUniv. of LjubljanaPhone: 386 1 3203280jernej.jakse@bf.uni-lj.slRob JoostenDyadic Nederland BVPhone: +31 317 465456rjoosten@dyadic.nlWilliam K HollomanCornell Univ Weill Med ColPhone: (212) 746-6510wkhollo@med.cornell.eduMichael J HynesUniv MelbournePhone: 61 03-83446239mjhynes@unimelb.edu.auBranka JavornikBiotechnical FacultyPhone: +386 1 3203260branka.javornik@bf.uni-lj.siHoward S JudelsonUniv CaliforniaPhone: (951) 827-4199howard.judelson@ucr.eduShinji HondaUniversity of FukuiPhone: -9456SHINJI@u-fukui.ac.jpChristian HongUniversity of CincinnatiPhone: 5135583289christian.hong@uc.eduSung-Yong HongMichigan State UniversityPhone: 517-353-2270hongsun7@msu.eduChiaki HoriUniversity of TokyoPhone: -11020chori718@jcom.home.ne.jpBenjamin A HorwitzTechnion-ITTPhone: 972 4 8293976horwitz@tx.technion.ac.ilSara HosseiniSLUPhone: 0046-18-672379sara.hosseini@slu.sePetra HoutermanUniversity of AmsterdamPhone: +31 20 525 7706P.M.Houterman@uva.nlBarbara J HowlettUniv MelbournePhone: 61 3-8344-5062bhowlett@unimelb.edu.auYen-Ping HsuehCalifornia Inst of TechnologyPhone: (626) 395-2669yph@caltech.eduYuhong HuangAalborg UniversityPhone: +45 99402596huyu@bio.aau.dkRichard HungRutgers, The State University of NewJerseyPhone: (732) 850-6220Richard.Hung@rutgers.eduCameron HunterKansas State UniversityPhone: 913-907-8289cameron.hunter15@gmail.comJae-Seoun HurSunchon National UniversityPhone: -4112jshur1@sunchon.ac.krNils OS HögbergBioCenter, SLUPhone: 4618671875nils.hogberg@slu.seAkihiko IchiishiToyo UniversityPhone: 81 276-82-9203akihiko@toyo.jpAlexander IdnurmUniv Missouri-Kansas CityPhone: (816) 235-2265idnurma@umkc.eduKiyohiko IgarashiUniversity of TokyoPhone: 81-3-5841-5258aquarius@mail.ecc.u-tokyo.ac.jpPatrik InderbitzinUC DavisPhone: 916 668 9078patrik.inderbitzin@gmail.comDiane O InglisStanford UniversityPhone: (415) 713-1408dianeoinglis@gmail.comVong shian Simon Ip ChoUniversity of CopenhagenPhone: 4535332627sich@life.ku.dkAkira IshiiTokyo Univ. of SciencePhone: -16194j6412701@ed.noda.tus.ac.jpKazi Tariqul IslamSouthern Illinois UniversityPhone: 6182033089tariq@siu.eduChrista IvanovaTechnical University ViennaPhone: +43 6801127145christa.ivanova@tuwien.ac.atKazuhiro IwashitaNational Research Institute of BrewingPhone: -2688iwashitact@nrib.go.jpSultana N JahanSveriges lantbruksuniversitetPhone: +46 18-673238sultana.jahan@slu.seKwang-Yeop JahngChonbuk Natl Univ Col Nat SciPhone: 82 63-270-3358goodear@chonbuk.ac.krGregory JeddPhone: 65 93380462gregory@tll.org.sgJunhyun JeonSeoul National UniversityPhone: 82-2-880-4684plantdr1@snu.ac.krQiming JinNovozymes IncPhone: (530) 757-0827qmji@novozymes.comSeongmi JoSoonchunhyang Univ.Phone: -1777kinosjo@gmail.comHanna JohannessonUppsala UnivPhone: 46 18 4716662hanna.johannesson@ebc.uu.seAnnette H JohansenNovozymes A/SPhone: (0045)44460736ahjo@novozymes.comTomas JohanssonLund UniversityPhone: 46 46-222-4549tomas.johansson@biol.lu.seAnna JohnsUniv. of ManchesterPhone: 447 7205 62237anna.johns@postgrad.manchester.ac.ukDerek JohnsonOregon State UniversityPhone: 541-737-5302johnsde4@science.oregonstate.eduLinda J JohnsonAgResearch LimitedPhone: 6463518189linda.johnson@agresearch.co.nzRichard D JohnsonAgresearch GrasslandsPhone: +64 6 3518090richard.johnson@agresearch.co.nzDavid L JolyAgriculutre and Agri-Food CanadaPhone: (250) 404-3318David.Joly@agr.gc.caWilfried JonkersUC BerkeleyPhone: 510-643-2546wjonkers@berkeley.eduBoknam JungDong-A UniversityPhone: 82-51-200-5670wjdqhrska89@naver.comElke-Martina JungFriedrich Schiller UniversityPhone: 1.1493641949e+013elke-martina.jung@uni-jena.deKWANG-WOO JUNGYonsei UniversityPhone: 82-70-7578-1596jkwoo1983@hanmail.netPraveen R JuvvadiDuke University Medical CenterPhone: 919-681-2613praveen.juvvadi@duke.eduBastian JöhnkGeorg August UniversityPhone: +49 551 393821bjoehnk@gwdg.deMaciej KaczmarekScotland's Rural CollegePhone: 7706957947maciej.kaczmarek@sruc.ac.ukJoerg T KaemperKITPhone: 49 721 609 45670joerg.kaemper@kit.eduMeenakshi KagdaUC, RiversidePhone: 951-756-6400mkagd001@ucr.eduRegine KahmannMPI Terrestrial MicrobiologyPhone: 49 6421-178501kahmann@mpi-marburg.mpg.deShiv D KaleVirginia Tech.Phone: (540)-231-3784sdkale@vt.eduVincent KamNovozymes, Inc.Phone: 530-757-8100 x 4624vck@novozymes.comHisae KamakuraTokyo Univ. of SciencePhone: -16194rheingold@i.softbank.jpTakashi KamakuraTokyo Univ. of SciencePhone: -16194kamakura@rs.noda.tus.ac.jp324

LIST OF PARTICIPANTSMasayuki KameiToyo UniversityPhone: -9493dx110002x@toyo.jpJung-Eun KimPenn State UniversityPhone: 814-863-2074juk30@psu.eduAnja KombrinkWageningen UniversityPhone: 0031-317485325anja.kombrink@wur.nlEmi KunitakeOsaka Prefecture UniversityPhone: 81-72-254-9466kunitake@biochem.osakafu-u.ac.jpSusan G W KaminskyjUniversity of SaskatchewanPhone: +1 306 966 4422susan.kaminskyj@usask.caJung-Mi KimWonkwang UniversityPhone: 82-63-850-6676micro@wku.ac.krMichael KoonceNational Science FoundationPhone: 702-292-7144mkoonce@nsf.govMarkus KunzlerETH ZurichPhone: 41 446324925markus.kuenzler@micro.biol.ethz.chZhian N. KamvarOregon State UniversityPhone: (541) 738-4074kamvarz@science.oregonstate.eduSeongbeom KimSeoul National UniversityPhone: -5434ssungbum@snu.ac.krBranka KorosecNational Institute of ChemistryPhone: 38614760262branka.korosec@ki.siAlan KuoDOE Joint Genome InstPhone: 925-899-1364akuo@lbl.govSeogchan KangPennsylvania State UnivPhone: (814) 863-3846sxk55@psu.eduCorby KistlerCereal Disease LaboratoryPhone: (612) 625-9774hckist@umn.eduErika KotheFriedrich-Schiller UniversityPhone: 49 3641-949291erika.kothe@uni-jena.deKiminori KurashimaSaitama UnivercityPhone: 81-48-858-3414s09db004@mail.saitama-u.ac.jpLevente KaraffaUniversity of DebrecenPhone: +3652512900 62488karaffa.levente@science.unideb.huSylvia KlaubaufCBS-KNAW Fungal Biodiversity CentrePhone: 31302122600s.klaubauf@cbs.knaw.nlLukasz KozubowskiDuke Universtiy Medical CenterPhone: (919) 308-5974lukasz.kozubowski@duke.eduJaana T KuuskeriUniversity of HelsinkiPhone: 3.5891915928e+011jaana.kuuskeri@helsinki.fiMargaret E KatzUniv New EnglandPhone: 61 2-6773-3016mkatz@une.edu.auStefan KlinterRoyal Institute of TechnologyPhone: 46735570644klinter@kth.seNada KrasevecNational Institute of ChemistryPhone: +386(1)4760-262nada.krasevec@ki.siHoi S KwanChinese Univ, Hong KongPhone: 85239436251hoishankwan@cuhk.edu.hkJan Naseer KaurUniversity at BuffaloPhone: 716-829-2091jannasee@buffalo.eduAndrew D. KlockoUniversity of OregonPhone: 541-346-5197klocko@uoregon.eduKatrin KrauseFriedrich Schiller UniversityPhone: +49 (0)3641-949399Katrin.Krause@uni-jena.deGerald LacknerUniversity JenaPhone: +49 17623132017gerald.lackner@hki-jena.deMoriyuki KawauchiHiroshima UniversityPhone: -1245m-kawauchi@nrib.go.jpSteve J KlostermanUSDA-ARSPhone: (831) 755-2845Steve.Klosterman@ars.usda.govPauline KrijgsheldUtrecht UniversityPhone: 31302533041p.krijgsheld@uu.nlLjerka LahNational Inst. of ChemistryPhone: 38641384557Iljerka.lah@ki.siNancy P KellerUniv Wisconsin, MadisonPhone: (608) 262-9795npkeller@wisc.eduRonny KellnerMax Planck Institute MarburgPhone: 49 642 1178 632ronny.kellner@mpi-marburg.mpg.deFrank KempkenChristian-Albrechts UniversityPhone: 49 431-880-4274fkempken@bot.uni-kiel.deJohn C KennellSt Louis UnivPhone: (314) 977-3905kennellj@slu.eduNemat O KeyhaniUniv FloridaPhone: (352) 392-2488keyhani@ufl.eduDae-Hyuk KimChonbuk National UnivPhone: 82 63 270 3440dhkim@chonbuk.ac.krHye-Seon KimUniv of DelawarePhone: 302-831-3429hyeseon@udel.eduOlaf KniemeyerLeibniz Institute for Natural ProductResearch and Infection Biology (HKI)Phone: (0049) 3641-5321071olaf.kniemeyer@hki-jena.deTetsuo KobayashiNagoya UniversityPhone: 81527894085koba@agr.nagoya-u.ac.jpSayo KodamaKyoto Prefectural UniversityPhone: 81-75-703-5613ze14skodama@gmail.comAnnegret KohlerINRA Centre de NancyPhone: +33 383394072kohler@nancy.inra.frLinda M KohnUniv Toronto MississaugaPhone: 905-277-1321linda.kohn@utoronto.caHideaki KoikeAISTPhone: 81 29-861-6679hi-koike@aist.go.jpKaihei KojimaNovozymes Japan Ltd.Phone: -7028kka@novozymes.comBrent A KronmillerOregon State UniversityPhone: 510-825-8153bkronmiller@gmail.comJames W KronstadUniv British ColumbiaPhone: (604) 822-4732kronstad@interchange.ubc.caYasuyuki KuboKyoto Prefectural UniversityPhone: 81 75-703-5613y_kubo@kpu.ac.jpUlrich KueckRuhr Univ BochumPhone: 49 234-3226212ulrich.kueck@rub.deUrsula KuesGeorg-August UnivPhone: 49 551-397024ukuees@gwdg.deGretchen KuldauPenn StatePhone: 814-863-7232kuldau@psu.eduAbhishek KumarInstitute of Botany, CAU KielPhone: 4.9179759067e+011akumar@bot.uni-kiel.deHerve LalucqueUniversité Paris SudPhone: 33 1-69154668herve.lalucque@igmors.u-psud.frScott LambieUniversity of British ColumbiaPhone: 604 822 2217scott.lambie@gmail.comElza A. S. LangUniversity of Sao PauloPhone: 5516 3602-3078elzalang@usp.brThorsten LangnerHeinrich-Heine UniversityPhone: 0049 (0)211 81 11529thorsten.langner@hhu.deNicolas NL LAPALUINRA- URGIPhone: 33130833810nicolas.lapalu@versailles.inra.frLuis F LarrondoPontificia Universidad Católica de ChilePhone: (56)-2-23541916llarrondo@bio.puc.clJennifer R LarsonOhio State UnivPhone: (614) 247-6873larson.315@osu.edu27th Fungal Genetics Conference | 325

LIST OF PARTICIPANTSOlga LastovetskyCornell UniversityPhone: 607-220-7351ol57@cornell.eduSophie LevUniversity of SydneyPhone: 61-2-98457046levsophie@gmail.comYi LiuUT Southwestern Medical CtrPhone: (214) 645-6033yi.liu@utsouthwestern.eduJuliane MacheleidtHans-Knoell-InstitutePhone: 4.9364153213e+012Juliane.Macheleidt@hki-jena.deLinda LauingerBZHPhone: +49 6221 544345linda.lauinger@bzh.uni-heidelberg.deStuart LevitzUniversity of MassachusettsPhone: 508-856-1525stuart.levitz@umassmed.eduZhaohui LiuNorth Dakota State UniversityPhone: (701) 231-7454zhh.liu@ndsu.eduMasayuki MachidaNatl Inst Advanced Sci TechPhone: 81 298-61-9447m.machida@aist.go.jpMichelle LeachUniversity of TorontoPhone: (647) 405-4437michelle.leach@abdn.ac.ukZachary A LewisUniversity of GeorgiaPhone: (541) 346-5197zlewis@uga.eduLars LobachSchool of Biological SciencesPhone: 4.475346927e+011lars.loebach@abdn.ac.ukSoumya MadhavanFriedrich Schiller UniversityPhone: 4.936419494e+011soumya.madhavan@uni-jena.deMarc-Henri LebrunINRAPhone: (33) 130814551marc-henri.lebrun@versailles.inra.frIn H LeeKookmin UnivPhone: 82 2-910-4771leei@kookmin.ac.krJUNGKWAN LEEDong-A UniversityPhone: 82-51-200-7554jungle@dau.ac.krMi Kyung LeeUniversity of Wisconsin MadisonPhone: 6082636830mlee234@wisc.eduSooChan LeeDuke University Medical CenterPhone: 919-684-3036soochan.lee@duke.eduYong-Hwan LeeSeoul National UnivPhone: 82 2-880-4674yonglee@snu.ac.krYoon Ji LeeSeoul National UnivPhone: -5481damnkool@snu.ac.krFrançois LefebvreUniversite LavalPhone: (418) 977-0963francois.lefebvre.3@ulaval.caRonny LehneckGeorg-August-UniversityPhone: 49551393579rlehnec@gwdg.deTaija LeinonenRoal OyPhone: 3.5892904216e+011taija.leinonen@roal.fiYueqiang LengNorth Dakota State UniversityPhone: 6122290411yueqiang.leng@ndsu.eduKlaus B LengelerCarlsberg LaboratoryPhone: +45 3327 5236KlausB.Lengeler@carlsberglab.dk326Guotian LiPhone: 765-237-2617leeguotian@163.comSimeng LiBeijing Forestry UniversityPhone: +86 18611426618lisimeng00@gmail.comXiangqian LiMonsantoPhone: 63017xiangqian.li@monsanto.comXiaofei LiangPhone: (352)-328-9853xiaofeiliang@ufl.eduHui-Ling LiaoDuke UnivPhone: 9196607362sunny.liao@duke.eduEdward Yi-Hong LiawUniv of California, RiversidePhone: (949)682-8289edward.liaw@ucr.eduAlex LichiusVienna Univ of TechnologyPhone: +43(1)58801-166524alexlichius@gmail.comFang Yun LimUniversity of Wisconsin-MadisonPhone: 608-262-1958fylim@wisc.eduChing-Hsuan LinNational Taiwan UniversityPhone: 886-2-33664449chinghsuanlin@ntu.edu.twXiaorong LinTexas A&M UniversityPhone: (979) 845-7274xlin@bio.tamu.eduBjörn D LindahlSLUPhone: +46 18 672725Bjorn.Lindahl@slu.seHaoping LiuUniv California Col MedicinePhone: (949) 824-1137h4liu@uci.eduHuiquan LiuNorthwest A&F UniversityPhone: 86-29-87081270liuhuiquan@nwsuaf.edu.cnJennifer K LodgeWashington Univ Sch MedPhone: 314-747-0515lodgejk@wustl.eduLuis V Lopez-LlorcaUniveristy AlicantePhone: 34-965903400lv.lopez@ua.esFederico Lopez-MoyaUniversity of AlicantePhone: 0034965903400 x3280federicolopezmoya@gmail.comJennifer LorangOregon State UniversityPhone: 541-737-5282lorangj@science.oregonstate.eduJennifer J LorosGeisel Sch Medicine@DartmouthPhone: (603) 650-1154jennifer.loros@dartmouth.eduEdward J LouisUniv NottinghamPhone: 44 115-823-0375ed.louis@nottingham.ac.ukRohan G T LoweUniversity of MelbournePhone: 61383445056rohan.lowe@unimelb.edu.auLing LuNanjing Normal UniveristyPhone: 86-25-85891791linglu6465@hotmail.comTaina K LundellUniversity of HelsinkiPhone: +358 9 191 59316taina.lundell@helsinki.fiHong LuoMichigan State UnivPhone: (517) 353-4886hongluo@msu.eduLiang MaSIBS,CASPhone: 86-21-54924049liangm2008@yahoo.comDarel Anne MacdonaldUniversity of ManchesterPhone: 4.4759566856e+011darel.macdonald@postgrad.manchester.ac.ukLori A Maggio-HallEI Dupont de Nemours & CoPhone: (302) 695-1480lori.a.maggio-hall@usa.dupont.comUma MaheswariEMBL-EBIPhone: 4.4122349468e+011uma@ebi.ac.ukAlejandra MandelUniversity of ArizonaPhone: (520) 621-9419mandel@ag.arizona.eduViola A ManningOregon State UnivPhone: (541) 737-2234manniv@science.oregonstate.eduElisabetta MarchegianiINRAPhone: 033-130814560elisabetta.marchegiani@versailles.inra.frMark R MartenUniv Maryland,Baltimore CountyPhone: (410) 455-3439marten@umbc.eduFrancis M MartinINRAPhone: 33 383-394080fmartin@nancy.inra.frLeonora MartinezCICESEPhone: 175 0500lotoleias@gmail.comAna Lilia Martinez-RochaUniversity of HamburgPhone: +49 40 428 16 309analiliamartinezrocha@googlemail.comNilce M Martinez-RossiSao Paulo UniversityPhone: 55 16-36023150nmmrossi@usp.brEmma MasterUniversity of TorontoPhone: 416-946-7861emma.master@utoronto.caElisabeth MaurerMedical University InnsbruckPhone: -80204elisabeth.maurer@i-med.ac.at

LIST OF PARTICIPANTSGeorgiana MayUniv MinnesotaPhone: (612) 624-6737gmay@umn.eduAlejandro Miralles-DuranFacultad de BiologiaPhone: +34 954556473amiralles@us.esDonna L MoyerNovozymes IncPhone: (530) 757-8115dlm@novozymes.comGesabel Y Navarro VelascoUniversidad de CordobaPhone: 0034 957212421z72naveg@uco.esKevin McCluskeyUniv Missouri, Kansas CityPhone: (816) 235-6484mccluskeyk@umkc.eduAmir Mirzadi GohariPlant Research International (PRPhone: 686323223amir.mirzadigohari@wur.nlAndre MuellerMPI for Terrestrial MicrobiologyPhone: 49 642 1178 612andre.mueller@mpi-marburg.mpg.deAttila NemethCodexisPhone: -6921attila.nemeth@codexis.comBruce A McDonaldETH-Zentrum, LFW B16Phone: 41 44-632-3848bruce.mcdonald@usys.ethz.chAmanda L Misener BloomSUNY University at BuffaloPhone: 716-829-2091amisener@buffalo.eduAlija Bajro MujicOregon State UniversityPhone: (541) 737-5302mujica@science.oregonstate.eduHelena KM NevalainenMacquarie UniversityPhone: 61403162038helena.nevalainen@mq.edu.auTami R McDonaldUniversity of MinnesotaPhone: (612) 625-4975mcdo0264@umn.eduAaron P MitchellCarnegie Mellon UnivPhone: (412) 268-5844apm1@andrew.cmu.eduAlberto MuñozUniversity of EdinburghPhone: (+44)-(0)1316513337alberto.munoz@ed.ac.ukConnie B NicholsDuke Medical CtrPhone: (919) 684-5054connie.nichols@duke.eduMatt MeadUniversity of Wisconsin-MadisonPhone: 608-265-5689mmead@wisc.eduPankaj MehrotraAberdeen Fungal GroupPhone: 0044-01224 4374p.mehrotra@abdn.ac.ukHarold MeijerWageningen UniversityPhone: 31 317-483138harold.meijer@wur.nlHugo MelidaRoyal Inst of Technology (KTH)Phone: -13869hmelida@kth.seArtemio MendozaLincoln UnivPhone: +64 3321 8472artemio.mendoza@lincoln.ac.nzEl Ghalid MennatUniversidad de CordobaPhone: 0034 682280250m.elghalid@gmail.comMartha W MerrowLudwig-Maximilians-Universität-MünchenPhone: 4.9171381266e+011merrow@lmu.deYasumasa MiyazakiFFPRIPhone: -9056ymiyazak@ffpri.affrc.go.jpCaroline MoffatCurtin UniversityPhone: 61892669186Caroline.Moffat@curtin.edu.auVenkatesh MoktaliPenn State UnivPhone: (814) 321-5762vpm104@psu.eduMichelle C MomanyUniv GeorgiaPhone: (706) 542-2014momany@plantbio.uga.eduStephen J MondoCornell UniversityPhone: (607) 342-8087sjm284@cornell.eduValdirene MonteiroUniversidade Estadual de GoiásPhone: +55(62)33281160valdirene.neves@ueg.brMelanie MorelINRAPhone: 999 0383684228Melanie.Morel@scbiol.uhp-nancy.frHajime MuraguchiAkita Prefectural UnivPhone: 81 18-872-1580muraguchi@akita-pu.ac.jpGloria Muzzi ErichsenNovozymes Inc.Phone: 15307578100GMEr@novozymes.comMiia R MäkeläUniv HelsinkiPhone: 3.5891915931e+011miia.r.makela@helsinki.fiMaryam NadimiUniveristy of MontrealPhone: (514)746-2624maryam.nadimi@umontreal.caVikram NaikMPI for Terrestrial MicrobiologyPhone: 49 6421 178 522naik@mpi-marburg.mpg.deMizue NaitoCornell UniversityPhone: 607-280-3774mn389@cornell.eduMasaya NakamuraFFPRIPhone: -9059nmasaya@ffpri.affrc.go.jpFrancisco E Nicolas-MolinaUniv de MurciaPhone: 34648104533fnicolas@um.esEva-Maria NiehausUniversity of MuensterPhone: 4.9251832499e+011e.niehaus@uni-muenster.deJakob B NielsenDTU Systems BiologyPhone: 45 45252657jbn@bio.dtu.dkMichael L NielsenNovozymesPhone: +45 26805119mlxn@novozymes.comMaria Fernanda Nieto-JacoboLincoln UniversityPhone: +64 3 321 8113maria.nieto-jacobo@lincoln.ac.nzBart NieuwenhuisLab of Genetics, WURPhone: 31641125774bpsnieuw@gmail.comMarie NishimuraNatl Inst Agrobiol SciencesPhone: 81 2-838-8461marie@affrc.go.jpAlexander V MichkovUniversity of California, RiversidePhone: (951)8273190alexander.michkov@email.ucr.eduMarino MorettiMPI for Terrestrial MicrobiologyPhone: 49 0 6421 178 521marino.moretti@mpi-marburg.mpg.deMayumi NakayamaTohoku UniversityPhone: -9437mayumi-n@niche.tohoku.ac.jpClarissa Jane NobileUCSFPhone: (415) 476-8097Clarissa.Nobile@ucsf.eduSantiago Xavier MiderosCornell UniversityPhone: (607)339-8711sxm2@cornell.eduPaul F MorrisBowling Green State UnivPhone: (419) 372-0481pmorris@bgsu.eduTakehito NakazawaUniversity of ShizuokaPhone: 81 54 264-5662gp1595@u-shizuoka-ken.ac.jpYuhei NogamiToyo UniversityPhone: -9493s39101200242@toyo.jpKyunghun MinSeoul National UnivPhone: 82 2-8804681mymin117@snu.ac.krUffe H MortensenDTU Systems BiologyPhone: 45 4525 2701um@bio.dtu.dkAndre NantelNational Research CouncilPhone: 1 (514) 496-6370Andre.Nantel@nrc-cnrc.gc.caFilomena A NogueiraUniversity of AberdeenPhone: 4.4758376676e+011r01mfd0@abdn.ac.ukAnna Minz-DubTel-Aviv UniversityPhone: 9.725447883e+011annamin1@post.tau.ac.ilRosa R Mourino-PerezCICESEPhone: 52 646-1750500rmourino@cicese.mxNaweed I NaqviTemasek Life Sciences LaboratoryPhone: 65-68727493naweed@tll.org.sgSteffen NordziekeRuhr-University BochumPhone: 4.9234322566e+011steffen.nordzieke@rub.de27th Fungal Genetics Conference | 327

LIST OF PARTICIPANTSOlga NovikovaUniversity at AlbanyPhone: 859-940-5160novikova@albany.eduCarsten L OlsenNovozymes A/SPhone: +45 30776013cols@novozymes.comHEESOO PARKUniversity of Wisconsin MadisonPhone: 608-263-6830hpark48@wisc.eduJose C PerezUCSFPhone: 415-476-8097jperez@ucsf.eduMinou NowrousianRuhr University BochumPhone: 49 234-3224588minou.nowrousian@ruhr-unibochum.deDonald L NussUniversity of MarylandPhone: (240) 314-6218dnuss@umd.eduKristiina NygrenSwedish Univesity of AgriculturalSciencesPhone: (46) 734-226900kristiina.nygren@gmail.comRichard J O'ConnellINRA-BIOGERPhone: 49 2215062311oconnel@mpipz.mpg.deCéline M O'GormanUniversity of NottinghamPhone: -8232112celine.ogorman@nottingham.ac.ukBerl R OakleyUniv KansasPhone: (785) 864-8170boakley@ku.eduKen OdaUniv GeorgiaPhone: (706) 542-6026koda@plantbio.uga.eduYeon Y OhNCSUPhone: (919) 513-0167yoh2@ncsu.eduAyumi OhbaTohoku universityPhone: -9561ayumiohba@gmail.comMana OhkuraUniversity of AZPhone: 520-342-4794manabanana@email.arizona.eduRobin A OhmDOE Joint Genome InstitutePhone: 925- 927-2546RAOhm@lbl.govShuichi OhsatoMeiji UniversityPhone: -8710ohsato@gmail.comRodrigo A OlarteNCSUPhone: 919 513-4867raolarte@ncsu.eduVianey G Olmedo-MonfilUniversidad de GuanajuatoPhone: 52 4737320006vianey_olmedo@yahoo.com.mxAke OlsonSW Univ Agricultural SciPhone: +46 18 671876ake.olson@slu.seRyoko OonoDuke Univ. & NC State Univ.Phone: 5073696394ryoko.oono@duke.eduMarc J OrbachUniv ArizonaPhone: (520) 621-3764orbachmj@ag.arizona.eduMiriam Oses-RuizUniversity of ExeterPhone: 440-1392264689mo256@exeter.ac.ukTherése OskarssonCarlsberg LaboratoryPhone: 0046 70 8898397tos@crc.dkStephen A OsmaniOhio State UnivPhone: (614) 247-6791osmani.2@osu.eduJean-Paul J OuedraogoInstitut of BiologyPhone: 003171 5274914jpaul_oued@yahoo.frShouqiang OuyangUniversity of California, RiversidePhone: 951-827-3190souyang@ucr.eduCatherine Alisha OwensbyOregon State UniversityPhone: 5417375284owensbyc@science.oregonstate.eduJavier Palma-GuereroUC BerkeleyPhone: 510-643-2546jpalma@berkeley.eduMarja PaloheimoRoal OyPhone: 358 9-29042-128marja.paloheimo@roal.fiFranck P PanabieresINRA-ISAPhone: (+33) 4 92 38 65 18franck.panabieres@sophia.inra.frIovanna PandelovaOregon State UniversityPhone: (541)7372234pandeloi@science.oregonstate.eduAe Ran ParkSeoul National UnivPhone: 82 2-8804681arpark@snu.ac.krJaejin ParkSeoul National UniversityPhone: -5484asdfppp2@gmail.comJoohae ParkInstitute of Biology LeidenPhone: 31715275072joohae@gmail.comSeung-Moon ParkChonbuk Natl UnivPhone: 82 63-270-4311smpark@moak.chonbuk.ac.krSook-Young ParkSeoul National UniversityPhone: 82-2-880-4635sookyp@gmail.comMatias PasqualiCRP GABRIEL LIPPMANNPhone: 352 470261436matias.pasquali@gmail.comRuvini U. PathiranaUniversity of Nebraska - LincolnPhone: 4028407086ruvini@huskers.unl.eduAleksandrina PatyshakuliyevaCBS-KNAWPhone: + 31 (0)30 2122600a.patyshakuliyeva@cbs.knaw.nlLinda PaunChristian-Albrechts-UniversityPhone: 4.9431880551e+011lpaun@bot.uni-kiel.deTeresa E PawlowskaCornell UnivPhone: (607) 254-8726tep8@cornell.eduKabir G PeayStanford UniversityPhone: 650-723-0552kpeay@stanford.eduMiguel PenalvaCSICPhone: 3491837 3112penalva@cib.csic.esGervette M PennySt. George's UniversityPhone: 14734564404gpenny001@hotmail.comDaniel PenselinInstitute of Plant BiochemistryPhone: +49 (0) 345 55821431Daniel.Penselin@ipb-halle.deNalu PeresUniv of Sao PauloPhone: 55 16 3602 3224nalu@usp.brJose Perez-MartinCentro Natl de BiotechnologyPhone: 34 91-585-4704jose.perez@csic.esMike PerlinUniv LouisvillePhone: (502) 852-5939mhperl01@louisville.eduPeter PhilippsenUniv BaselPhone: 41 61-2671480peter.philippsen@unibas.chBirgit PiechullaUniversity of RostockPhone: 0049 381 4986130birgit.piechulla@uni-rostock.deSebastian PilsykInst of Biochem & Biophysics PASPhone: 48 22-592-1209Seba@ibb.waw.plMarta J PiotrowskaScotland's Rural CollegePhone: 4.413153542e+011marta.piotrowska@sruc.ac.ukJohn W PitkinMonsantoPhone: (636) 737-5959john.w.pitkin@monsanto.comMichael PlamannUniv Missouri, Kansas CityPhone: (816) 235-2593plamannm@umkc.eduDavid F PlazaETH ZurichPhone: +41 44 632 49 24david.plaza@micro.biol.ethz.chClemence PlissonneauINRAPhone: 130814573clemence.plissonneau@versailles.inra.frKim M PlummerLa Trobe UnivPhone: 61 3-9479-2223k.plummer@latrobe.edu.auIstvan PocsiUniv of DebrecenPhone: 3652 512900ipocsi@gmail.comStefanie PoeggelerGeorg-August UniversityPhone: 49 551-3913930spoegge@gwdg.deLaetitia PoidevinUniv. of California, RiversidePhone: (951) 522-2025laetitia@ucr.edu328

LIST OF PARTICIPANTSNadia PontsINRAPhone: -57122466nadia.ponts@bordeaux.inra.frFranziska RabeMPI MarburgPhone: 49 64211 78533franziska.rabe@mpi-marburg.mpg.deStefanie G ReissmannMax-Planck InstPhone: 4.9642117185e+012reissmas@mpi-marburg.mpg.deMarika RossiNational Research CouncilPhone: 3.9011397792e+011m.rossi@ivv.cnr.itStephan PoppeMax Planck Institute MarburgPhone: 49 642 1178632poppeste@mpi-marburg.mpg.deRoman RabinovichDuPont Industrial BiosciencesPhone: 650-245-9059Roman.Rabinovich@dupont.comAna RemsTechnical University of DenmarkPhone: 4542570887anare@bio.dtu.dkThierry RouxelINRAPhone: 33 1 30 81 45 73rouxel@versailles.inra.frNathan PortierUnion Biometrica, Inc.Phone: 508-893-3115kbarnhart@unionbio.comMostafa RahnamaUniversity of AucklandPhone: 64220863333mrah233@aucklanduni.ac.nzMartijn RepUniversity of AmsterdamPhone: 31 20-525-7764m.rep@uva.nlJohn C RoyerDSMPhone: 617 913-5167John.royer@dsm.comJoanna PotrykusUniversity of AberdeenPhone: +44 1224 437 478j.potrykus@abdn.ac.ukMarcelo HS RamadaBrasilia UniversityPhone: 5.5629980492e+011marceloramada@gmail.comTimothy RepasUniversity of SaskatchewanPhone: 3062410474tsr852@mail.usask.caJason J RuddRothamsted ResearchPhone: +44 (0)1582 763133jason.rudd@rothamsted.ac.ukNathalie PoussereauUniversité Lyon IPhone: 33683184278nathalie.poussereau@univ-lyon1.frMarilee A RameshRoanoke CollegePhone: (540) 375-2464ramesh@roanoke.eduNatalia RequenaKarlsruhe Institute of TechnologyPhone: 49-721-60844626natalia.requena@kit.eduCarmen Ruger-HerrerosUniv de SevillaPhone: 34954556473carmenruger@us.esRobert J PrattDuPont Industrial BiosciencesPhone: (650) 846-7500robert.pratt@dupont.comChristof RampitschAgriculture & Agrifood CanadaPhone: 204 983-2385chris.rampitsch@agr.gc.caHannah T ReynoldsUniversity of AkronPhone: 919-698-2846hannah.t.reynolds@gmail.comFarhana RunaNorth Carolina State UniversityPhone: 626-325-4067fruna@ncsu.eduBert-Ewald PriegnitzTU BraunschweigPhone: 0531 391 5787B-E.Priegnitz@tu-bs.deDustin Parker RandArizona State UniversityPhone: (480) 965-1928dustin.rand@asu.eduHashim H RezaGenome Research CentrePhone: 9.1960139119e+011dreamzhashim908@gmail.comJesper H RungNovozymesPhone: 0045 44460349rung@novozymes.comAnne PringleHarvard UniversityPhone: (617) 496-9707pringle@oeb.harvard.eduCarlene A RaperUniversity of VermontPhone: 802-660-4809carlene.raper@uvm.eduRobert W RileyLawrence Berkeley National LabPhone: 925-296-5797rwriley@lbl.govLauren S RyderUniv ExeterPhone: 44 0-1392 264689l.s.ryder@ex.ac.ukRobert H ProctorNCAUR/ARS/USDAPhone: (309) 681-6380proctorh@ars.usda.govChad A RappleyeOhio State UniversityPhone: (614) 247-2718rappleye.1@osu.eduMeritxell RiquelmeCICESEPhone: 52 646-1750500riquelme@cicese.mxJohanna RytiojaUniversity of HelsinkiPhone: 3.5891915932e+011johanna.rytioja@helsinki.fiDov B PruskyAgricultural Research OrgPhone: 972-3-9683880dovprusk@volcani.agri.gov.ilDavide RavasioCarlsberg LaboratoryPhone: +45 33275235davide.ravasio@gmail.comBarbara RobbertseNCBIPhone: (301) 594-5068robberts@ncbi.nlm.nih.govElzbieta RzeszutekRoyal Institute of TechnologyPhone: +46-(0)8-5537-8448elrz@kth.sePatricia J PukkilaUniv North CarolinaPhone: (919) 966-5576pukkila@unc.eduNick ReadUniversity of EdinburghPhone: 44 131-650-5335Nick.Read@ed.ac.ukAngélica Mariana Robledo BrionesCinvestav Unidad IrapuatoPhone: +52 (462) 623 96 00jruiz@cinvestav.mxTeppo - RämäUniversity of Tromso, NorwayPhone: 5419744703teppo.rama@uit.noPeter J PuntTNOPhone: 31 88-8661728peter.punt@tno.nlSamara L Reck-PetersonHarvard Med SchPhone: 617-432-7178reck-peterson@hms.harvard.eduRusty J RodriguezSymbiogenicsPhone: (206) 661-8065rjrodriguez@symbiogenics.orgMatthew S SachsTexas A&M UnivPhone: (979) 845-5930msachs@mail.bio.tamu.eduTerhi J PuranenRoal OyPhone: 358 9 2904 2127terhi.puranen@roal.fiAmey RedkarMPI for Terrestrial MicrobiologyPhone: 49 64211 78610amey.redkar@mpi-marburg.mpg.deAntonis RokasVanderbilt UniversityPhone: (615) 936-3892antonis.rokas@vanderbilt.eduDivya SainUniv California, RiversidePhone: (951) 756-5449dsain001@ucr.eduXiaodong QiArizona State UniversityPhone: 480-965-1928xiaodong@asu.eduRegina S RedmanAdaptive Symbiotic TechnologiesPhone: (206) 661-8064ast-reginaredman@comcast.netM. Isabel G RonceroUniversity of CordobaPhone: +34 957218981roncero@uco.esWasin SakulkooUniversity of ExeterPhone: -7884157152ws254@exeter.ac.ukKatrin QuesterCICESEPhone: +52 646 1750500quester@cicese.edu.mxMorgann C ReillyUniversity of California, BerkeleyPhone: 510-666-2559mcreilly@berkeley.eduAntonio RossiFMRP-USPPhone: 55 16-36023112anrossi@usp.br27th Fungal Genetics Conference | 329

LIST OF PARTICIPANTSAsaf A SalamovDOE Joint Genome InstPhone: (925) 296-5782aasalamov@lbl.govGabriel ScallietSyngentaPhone: +41 62 8660122gabriel.scalliet@syngenta.comKlaas SchotanusMax-Planck Inst.Phone: +49 6421 178630klaas.schotanus@mpg-marburg.mpg.deDanyu ShenOregon State UniversityPhone: 541-602-9583shendanyu9@gmail.comMarketa SamalovaUniversity of OxfordPhone: 4.401865275e+012marketa.samalova@plants.ox.ac.ukPatrick C SchachtUniv California, RiversidePhone: (951) 823-0017pscha001@ucr.eduTimo SchuergUC BerkeleyPhone: 5105026980tschuerg@berkeley.eduKuwata ShigeruMeiji UniversityPhone: -7933kuwata@meiji.ac.jpEddy F Sanchez-LeonCICESEPhone: 52 646 1750500eddy.san.leon@gmail.comChristopher W SchadtOak Ridge National LaboratoryPhone: 865.576.3982schadtcw@ornl.govJulia SchumacherPurdue UniversityPhone: 4.9178600024e+011julia.schumacher@wwu.deJason ShillerLa Trobe UniversityPhone: 61426248488jshiller@students.latrobe.edu.auAndrea Sanchez-ValletWageningen UniversityPhone: 0031 (0)317485325andrea.sanchezvallet@wur.nlKatja SchaeferUniversity of CordobaPhone: 34957212421k_scha11@hotmail.deBarry ScottMassey UnivPhone: 6463505543d.b.scott@massey.ac.nzPatrick ShiuUniversity of MissouriPhone: 573-884-0020shiup@missouri.eduBjörn SandrockPhilipps-University MarburgPhone: +49 6421 2827080sandrock@staff.uni-marburg.deWilhelm SchaeferUniversity HamburgPhone: 0049-4042816266wilhelm.schaefer@googlemail.comFrank J.J. SegersCBS-KNAW Fungal Biodiversity CentrePhone: +31(0)302122669f.segers@cbs.knaw.nlJun-ya ShojiThe Samuel Roberts Noble FoundationPhone: 5802246968jyshoji@yahoo.co.jpDominique SanglardUniv of Lausanne and Hosp CtrPhone: 41213144883Dominique.Sanglard@chuv.chLes L ScharfensteinSouthern Regional Research CenterPhone: 504-286-4374les.scharfenstein@ars.usda.govStephan S SeilerUniversity FreiburgPhone: 49 551-393777sseiler@gwdg.deBhushan ShresthaRural Development AdministrationPhone: -11361bhushan.shrestha@gmail.comHiroaki SanoForestry and Forest Products ResearchInstitutePhone: +81 29 829 8279sanoh@affrc.go.jpMarcia R M SaraivaUniversity of AberdeenPhone: 4.4787902256e+011m.saraiva@abdn.ac.ukOezlem Sarikaya BayramInst. of Microbiology and GenetPhone: 49551393821osarika@gwdg.deMohammad N SAROWARInstitute of Medical SciencesPhone: 7741511109r01mns11@abdn.ac.ukKengo SasakiTokyo Univ. of SciencePhone: -16194j6412624@ed.tus.ac.jpYuki SATOHHokkaido Univ.Phone: -3138yuki-s@chem.agr.hokudai.ac.jpCharles W SaundersProcter & GamblePhone: (513) 622-4982saunders.cw@pg.comSven J SaupeCNRSPhone: 33 5-56999027sven.saupe@ibgc.u-bordeaux2.frBarry J SavilleTrent UniversityPhone: (705) 748-1011barrysaville@trentu.caKirstin ScherlachLeibniz Institute, HKIPhone: + 49 3641 5321034kirstin.scherlach@hki-jena.deJosua SchinkeGeorg-August-UniversityPhone: +49 17623974760jschink2@gwdg.deKerstin SchipperUniversity DüsseldorfPhone: 0049-211-8110451kerstin.schipper@hhu.deJan SchirawskiRWTH Aachen UniversityPhone: 49-241-8026616jan.schirawski@rwth-aachen.deMorten SchiøttUniversity of CopenhagenPhone: +45 35321255mschiott@bio.ku.dkGeorg SchmidtVTTPhone: +358 40 723 1815georg.schmidt@vtt.fiSarah Maria SchmidtUniversity of AmsterdamPhone: 31623236526s.m.schmidt@uva.nlStefanie S SchmiederETH ZurichPhone: +41 44 632 49 24sschmied@ethz.chMonika SchmollAustrian Institute of TechnologyPhone: 43 664 88390594monika.schmoll@ait.ac.atMarc-André SelosseCentre d'Ecologie Fonctionelle etEvolutive (CNRS)Phone: +33 607123418ma.selosse@wanadoo.frDominik SenftlebenFriedrich-Schiller-UniversitatPhone: 49 3641 949399dominik.senftleben@uni-jena.deAne SesmaCBGP/ Univ Politécnica de MadridPhone: +34 91 336 4593ane.sesma@upm.esFiroz ShahLund UniversityPhone: 46462223757Firoz.Shah@biol.lu.seShermineh ShahiUniversity of Amsterdam - SILSPhone: (0031) 20-525-8415s.shahi@uva.nlViplendra P.S. ShakyaUMKCPhone: (201)993-7038vps7y4@mail.umkc.eduAmir SharonTel Aviv UniversityPhone: 972 36406741amirsh@ex.tau.ac.ilSharpee C SharpeeNC State UniversityPhone: 608-576-8890wcsharpe@ncsu.eduBrian D ShawTexas A&M UnivPhone: (979) 862-7518bdshaw@tamu.eduChristian SieberHelmholtz-Zentrum MünchenPhone: +49 (0)89 3187 3577christian.sieber@helmholtzmuenchen.deUlrike SiegmundUniversity of MuensterPhone: 4.9521832499e+011ulrike.siegmund@uni-muenster.deAnita SilUniv California, San FranciscoPhone: (415) 502-1805sil@cgl.ucsf.eduLarissa SilvaUniversity of Sao PauloPhone: 55 (16) 3602-3078larissa.gomes@usp.brRoberto N. SilvaUniversity of São PauloPhone: 5.5163602326e+011rsilva@fmrp.usp.brPaul SkatrudMonsantoPhone: 636-737-7082paul.l.skatrud@monsanto.comDominique A SkovlundNovozymes A/SPhone: +45 44 465892dau@novozymes.comStephanie L SlinskiUniversity of PretoriaPhone: 27124206937stephanie.slinski@fabi.up.ac.zaJason SlotVanderbilt UniversityPhone: (615) 266-1433jason.c.slot@vanderbilt.edu330

LIST OF PARTICIPANTSFrancis SmetUnion BiometricaPhone: 3214570628fsmet@unionbio.comClaire StanleyInstitute of Chemical and BioengineeringPhone: 41774262541claire.stanley@chem.ethz.chJohanna TakachThe Noble FoundationPhone: (580) 224-6852jetakach@noble.orgEeva TerhonenHelsinki UniversityPhone: +35 40 7510773eeva.terhonen@helsinki.fiKristina M SmithOregon State UnivPhone: (541) 737-4399smitkris@science.oregonstate.eduTrevor StarrUC BerkeleyPhone: 5108470789t_starr@berkeley.eduTsukasa TakahashiToyo UniversityPhone: -9581dx0900031@toyo.jpBart P ThommaWageningen UniversityPhone: 0031-317-484536bart.thomma@wur.nlKaren M SnetselaarSt Joseph's UnivPhone: (610) 660-1826ksnetsel@sju.eduEmma T SteenkampUniversity of PretoriaPhone: 27 12 420 3262emma.steenkamp@up.ac.zaNorio TakeshitaKarlsruhe Institute of TechnologyPhone: -45913norio.takeshita@kit.eduAshley ThompkinsCoker CollegePhone: (843) 241-4107ashley.thompkins@coker.eduPeter S SolomonAustralian National UniversityPhone: -1391peter.solomon@anu.edu.auMatthias G SteigerACIB GmbH / BOKUPhone: 4313180078matthias.steiger@boku.ac.atJennifer M TalbotStanford UniversityPhone: 6178693715jmtalbot@stanford.eduDawn ThompsonBroad InstitutePhone: 6177147763dawnt@broadinstitute.orgJames TG SolomonsSUNY BuffaloPhone: 716 829 2091jgsolomo@buffalo.eduGero SteinbergUniv ExeterPhone: 0044-7717-243650G.Steinberg@exeter.ac.ukNick J TalbotUniv ExeterPhone: 44 1392-72-3006n.j.talbot@exeter.ac.ukDarren ThomsonAbderdeen UniversityPhone: 7980405654d.thomson@abdn.ac.ukBenesh M SomaiNelson Mandela Metropolitan UniversityPhone: 27-71-0211-090benesh.somai@nmmu.ac.zaAndrei S SteindorffBrasilia UniversityPhone: 55-6291037954andreistecca@gmail.comKoichi TamanoAISTPhone: -10246tamano-k@aist.go.jpBritta A. M. TillmannPhilipps-University MarburgPhone: 4.9642128271e+012britta-tillmann@gmx.deHOKYOUNG SONSeoul National UnivPhone: 82 2-8804681hogongi7@snu.ac.krKaren StephensonUniversity of DundeePhone: 4.413823863e+011k.stephenson@dundee.ac.ukKaeling TanHarvard Medical SchoolPhone: (617) 432-7177kaeling_tan@hms.harvard.eduRichard B ToddKansas State UniversityPhone: (785) 532-0962rbtodd@ksu.eduTeruo SoneHokkaido UnivPhone: 81 11-706-2502sonet@chem.agr.hokudai.ac.jpIoannis StergiopoulosUC DavisPhone: (530) 400-9802istergiopoulos@ucdavis.eduKar-Chun TanCurtin UniversityPhone: 61892669916Kar-Chun.Tan@curtin.edu.auSu San TohUniversity of LouisvillePhone: 5028525939susan.toh@louisville.eduVicky SophianopoulouBiosciences and applicationsPhone: 30 210 6503602vicky@bio.demokritos.grEva H StukenbrockMax Planck InstitutePhone: 0049 6421 178 630eva.stukenbrock@mpi-marburg.mpg.deMizuki TanakaTohoku UniversityPhone: -9561mizu-t@biochem.tohoku.ac.jpMarie TollotMPI MarburgPhone: 49 64211 78571marie.tollot@mpl-marburg.deAlexandra SoukupUW-MadisonPhone: 6082621957soukup@wisc.eduJoy E SturtevantLSUHSCPhone: (504) 568-6116jsturt@lsuhsc.eduShuji TaniOsaka Prefecture UnivPhone: 81-72-254-9466shuji@biochem.osakafu-u.ac.jpXiaoxue TongAalborg UniversityPhone: (+45) 50359928xiat@bio.aau.dkJessica SoyerINRAPhone: 33-130814593jessica.soyer@versailles.inra.frMartina A StöckliETH ZurichPhone: 0041 44 632 45 53smartina@micro.biol.ethz.chJohn W TaylorUniv California, BerkeleyPhone: (510) 642-5366jtaylor@berkeley.eduStefano F TorrianiETH ZurichPhone: 41 (0)44 632 38 48stefano.torriani@agrl.ethz.chJoseph W SpataforaOregon State UnivPhone: (541) 737-5304spatafoj@bcc.orst.eduGopal SubramaniamAgriculture CanadaPhone: (613) 759-7619subramaniamra@agr.gc.caSamantha TaylorOregon State UniversityPhone: 304-942-6007samantha.taylor@oregonstate.eduFrances TrailMichigan State UnivPhone: 517-432-2939trail@msu.eduRichard SplivalloUniversity of GoettingenPhone: 4.9176649314e+012richard.splivallo@a3.epfl.chPeter E SudberySheffield UnivPhone: 114 2226186p.sudbery@shef.ac.ukInes TeichertRuhr University BochumPhone: 4.9234322497e+011ines.teichert@rub.deAdrian TsangConcordia UnivPhone: (514)848-2424 x3405adrian.tsang@concordia.caJoseph SprakerUW-MadisonPhone: (608)262-1958jspraker@wisc.eduMallory SuhrUniversity of Nebraska-LincolnPhone: 402-641-5999msuhr@huskers.unl.eduNora TemmeKWS SAAT AGPhone: 49 5561 311 1141nora.temme@kws.comBettina TudzynskiWestf Wilhelms Univ MunsterPhone: 49 251-832-4801bettina.tudzynski@uni-muenster.deJason StajichUniv California, RiversidePhone: (951) 827-2363jason.stajich@ucr.eduSheng SunDuke University Medical CenterPhone: (919) 684-3036sheng.sun@duke.eduHiroshi TeramotoNovozymes Japan Ltd.Phone: +81 43-296-6770hite@novozymes.comPaul TudzynskiWest F Wilhelms Univ MuensterPhone: 49 251-832-4998tudzyns@uni-muenster.de27th Fungal Genetics Conference | 331

LIST OF PARTICIPANTSAnders P V TunlidEcologyPhone: 46 46-2223757anders.tunlid@mbioekol.lu.seCharlotte van der DoesUniversity of AmsterdamPhone: 31205257764H.C.vanderdoes@uva.nlSusanne VoigtUniv. of HamburgPhone: 4940 4281 6331voigt@botanik.uni-hamburg.deXuying WangDuke UnivPhone: (205) 482-0213xuying.wang@duke.eduGillian TurgeonCornell UnivPhone: (607) 254-7458bgt1@cornell.eduJan van KanWageningen UnivPhone: 31 317 483126jan.vankan@wur.nlChristine R VoiseyAgResearchPhone: 61 6-3518080christine.voisey@agresearch.co.nzYizhou WangUniv. of CA, RiversidePhone: 951-275-6883ywang039@ucr.eduMassimo TurinaCNRPhone: 3.9011397792e+011m.turina@ivv.cnr.itDavid TurraUniversity of CordobaPhone: 0034 957 218981ge3tutud@uco.esBrett M TylerOregon State UniversityPhone: 541-737-3347brett.tyler@oregonstate.eduGeorgios TzelepisSLUPhone: 4618671835georgios.tzelepis@slu.seHiroaki UdagawaNovozymes JapanPhone: -432966689huda@novozymes.comJessie UehlingDuke UniversityPhone: 2088712701jku@duke.eduSrijana UpadhyayTexas A&M UniversityPhone: (979) 845-7259srijanaupadhyay@tamu.eduMartin UrbanRothamsted ResearchPhone: (44) 01582-763133martin.urban@bbsrc.ac.ukBarbara ValentKansas State UnivPhone: (785) 532-2336bvalent@ksu.eduClarissa Xavier Res ValimFMRP-USPPhone: 55-16-3602-3143clarissavalim@hotmail.comMari ValkonenVTTPhone: 358 20 722-5825mari.valkonen@vtt.fiPeter van DamUniversity of Amsterdam - SILSPhone: 31205258412p.vandam@uva.nlGuido Van den AckervekenUtrecht UniversityPhone: 31302533013g.vandenackerveken@uu.nlMartin van LeeuwenCBS-KNAW Fungal Biodiversity CentrePhone: 0031 (0)30 621666r.vanleeuwen@cbs.knaw.nlHans D VanEttenUniv ArizonaPhone: (520) 621-9355vanetten@ag.arizona.eduElizabeth A VealNewcastle UnivPhone: 44 191-222-7596e.a.veal@ncl.ac.ukSudhagar Veerabadran BalasundaramUniversity of OsloPhone: 4794824403sudhagar@bio.uio.noClaire Veneault-FourreyLorraine University / INRAPhone: 33 (0)3 83 39 40 81claire.fourrey@univ-lorraine.frSurbhi VermaUniversity of Missouri-Kansas CityPhone: 1-816-235-1553svb66@mail.umkc.eduPaul VerweijUMC ST RADBOUDPhone: -3614349P.verweij@mmb.umcn.nlMuriel ViaudINRAPhone: 33 1 30 81 45 68viaud@versailles.inra.frRytas VilgalysDuke UniversityPhone: (919) 660-7361fungi@duke.eduStela VirgilioInstituto de Quimica, UNESPPhone: 55-16-33019804stelav5@yahoo.com.brHans VisserDyadic Nederland BVPhone: 31 317 465457hvisser@dyadic.nlIdo VlaardingerbroekUniversity of AmsterdamPhone: 31205258414i.vlaardingerbroek@uva.nlChristian A VoigtBiocenter Klein FlottbekPhone: -42816322voigt@botanik.uni-hamburg.deJohannes WagenerUniversity of MunichPhone: -6708778johannes.wagener@med.unimuenchen.deAndrea WaltherCarlsberg LabPhone: +45 3327 5391andrea.walther@carlsberglab.dkJonathan WaltonMichigan State UniversityPhone: 517-353-4885walton@msu.eduChen fang WangNorthwest A&F UniversityPhone: 86-029-87081270wangchenfang@nwsuaf.edu.cnChengshu WangShanghai Institutes for BiologicalSciences, CASPhone: -54924092cswang@sibs.ac.cnClay WangUniv Southern CaliforniaPhone: (323) 442-1670clayw@usc.eduFengfeng WangUtrecht UniversityPhone: +31 30 2533632F.wang@uu.nlGuanghui WangNorthwest A & F UniversityPhone: 765-418-6246wgh2891458@163.comLinqi WangTexas A&M UniversityPhone: (979) 845-7259lqwang@bio.tamu.eduMING WANGUC, RiversidePhone: 951-961-1183mwang012@ucr.eduPing WangRes Inst Children/LSUPhone: (504) 896-2700pwang@chnola-research.orgQunqing WangOREGON STATE UNIVERSITYPhone: 541-737-3679qunqing.wang@oregonstate.eduTing-Fang WangAcademia SinicaPhone: 886-2-27899188tfwang@gate.sinica.edu.twYuanchao WangNanjing Agricultral UniversityPhone: 86-25-84399071wangyc@njau.edu.cnMichael WardDuPont Industrial BiosciencesPhone: (650) 846-5850mward@genencor.comTodd J WardUSDA ,ARS,NCAURPhone: (309) 681-6394wardtj@ncaur.usda.govStephan WawraUniv AberdeenPhone: 44 1224 437329s.wawra@abdn.ac.ukRoland Wedlich-SoldnerMPI of BiochemistryPhone: +49 89 8578 3410wedlich@biochem.mpg.deHarshini C WeerasingheThe University of MelbournePhone: 61411362559h.weerasinghe@student.unimelb.edu.auMichaela WehrPhilipps UniversityPhone: 64212827078Michaela_Wehr@web.deArne WeibergUCRPhone: 9512757333aweiberg@ucr.eduMartin WeichertTU BraunschweigPhone: -6660m.weichert@tu-bs.deJurgen W WendlandCarlsberg LaboratoryPhone: 45 3327 5230jww@crc.dkEmily A WhistonU.C. BerkeleyPhone: (510) 642-8441whiston@berkeley.eduGraham WhytesideDSMPhone: 31152792118graham.whyteside@dsm.comPhillipp WiemannUniv. MünsterPhone: 49 251 8324806philipp.wiemann@uni-muenster.de332

LIST OF PARTICIPANTSAric E WiestUniv Missouri, KCPhone: (816) 235-6485wiesta@umkc.eduRichard A WilsonUniversity of Nebraska-LincolnPhone: 4024722156rwilson10@unl.eduBrenda D WingfieldUniversity of PretoriaPhone: 27 12 420 3946brenda.wingfield@up.ac.zaHua Z WiseOregon State UniversityPhone: (512)9213055zhanghua72@gmail.comThomas J. WolpertOregon State UniversityPhone: 541-737-5293wolpertt@science.oregonstate.eduKoon Ho WongHarvard Medical SchoolPhone: 6174322105koonho_wong@hms.harvard.eduJennifer R WortmanBroad InstitutePhone: 240-441-9094jwortman@broadinstitute.orgDongliang WuCornell UniversityPhone: 607-257-2683dongliangwu@yahoo.comJiayao WuHelsinki UniversityPhone: 3.580417058e+012jiayao.wu@helsinki.fiTimon WyattCBS Fungal Biodiversity CentrePhone: 31302122600timonwyatt@gmail.comXin XiangUSUHSPhone: (301) 295-0000xxiang@usuhs.milYi XiongUniv California, BerkeleyPhone: 510-666-2559yi-xiong@berkeley.eduJin-Rong XuPurdue UnivPhone: (765) 496-6918jinrong@purdue.eduJun XuP&GPhone: 5136220290xu.j.1@pg.comWenjie XuCarnegie Mellon UniversityPhone: 8587228372wxu7206@gmail.comChun-Hsiang YangNational Ilan UniversityPhone: 8.8692870579e+011philosophyroy@hotmail.comDong-Hoon YangYonsei UniversityPhone: 82-70-7578-1596garaman99@nate.comMoon S YangChonbuk National UnivPhone: 82 63-270-3339mskyang@jbnu.ac.krOded YardenHebrew University of JerusalemPhone: 972 8 9489298Oded.Yarden@huji.ac.ilWenwu YeNanjing Agricultral UniversityPhone: 86-25-843990712010202018@njau.edu.cnLakshmi Preethi YerraUNLPhone: 201-925-8484s-lyerra1@unl.eduMihwa YiKansas State UniversityPhone: 785-532-2337mihwa@ksu.eduWang YonglinBeijing Forestry UniversityPhone: 13488765326ylwang@bjfu.edu.cnCarolyn A YoungNoble FoundationPhone: (580) 224-6860cayoung@noble.orgJaehyuk YuUniv Wisconsin, MadisonPhone: (608) 262-4696jyu1@wisc.eduSung-Hwan YunSoonchunhyang UnivPhone: 82 41-5301288sy14@sch.ac.krIvo ZadraSANDOZ GmbHPhone: +43 (0)5338 200 3509ivo.zadra@sandoz.comDonald R ZakUniversity of MichiganPhone: 734763-4991drzak@umich.eduDongxiu ZhangUM IBBRPhone: 240-314-6497dzhan5@umd.eduJian ZhangNanjing Agricultural UniversityPhone: (86) 15850560269zhangjian106@hotmail.comNing ZhangRutgers UnivPhone: 8145743939zhang@aesop.rutgers.eduShizhu ZhangNanjing Normal UniversityPhone: 86 (25) 85891895shizhuz@126.comYing ZhangKarlsruheInstitute of TechnologyPhone: 17655321539fionazhangying@hotmail.comZhenyu ZhangUT Southwestern Medical CenterPhone: 2149301249zhenyuzhang86@gmail.comJiuhai ZhaoUC berkeleyPhone: 650-215-3209jhzhao@berkeley.eduYun ZhengConcordia UniversityPhone: 514-848-2424 ext4025zhyun627@gmail.comShaobin ZhongNorth Dakota State UnivPhone: (701) 231-7427shaobin.zhong@ndsu.eduMian ZhouUT Southwestern Medical CenterPhone: 9729788908mian.zhou@utsouthwestern.eduXiaoying ZhouPurdue UniversityPhone: 765-418-6157maidou2010@gmail.comKolea ZimmermanHarvard UniversityPhone: 8089898031kzimmerman@fas.harvard.eduMiriam E ZolanIndiana UnivPhone: (812) 855-6694mzolan@indiana.eduAlga ZuccaroMax Planck InstitutePhone: 4.964211786e+011zuccaro.alga@mpi-marburg.mpg.de27th Fungal Genetics Conference | 333

STUDENT POSTER LISTAbdeljalil, salma .................... 412Addicott, Ethan ....................... 40Affeldt, Katharyn ................... 633Agafonov, Oleg ........................ 41Ahmad, Aijaz ......................... 698Ahrendt, Steven .................... 294Alam, Md. Kausar .................. 360Albermann, Sabine ................ 405Almeida, Fausto....................... 18Andersson, Karl-Magnus ....... 602Antoniêto, Amanda ............... 413Anyaogu, Diana .........................7Arazoe, Takayuki ................... 572Ariyawansa, Kahandawa ....... 565Baek, Jin-Ho ........................... 414Baldin, Clara .......................... 231Baldwin, Thomas ................... 538Banerjee, Dithi ........................ 23Banjara, Nabaraj .................... 703Bazafkan, Hoda ..................... 201Bech, Lasse .............................. 60Beckmann, Nicola ......................1Berry, Daniel............................ 48Billmyre, Robert .................... 243Bitas, Vasileios ....................... 551Blosser, Sara .......................... 355Boenisch, Marike ................... 140Boettger, Daniela .................. 725Boshoven, Jordi ..................... 516Bravo Ruiz, Gustavo Adolfo ... 552Bredeweg, Erin ...................... 383Burchhardt, Kathleen ............ 646Burggraaf, Maria ..................... 99Bühler, Nicole .......................... 80Böhm, Julia ............................ 202Cabrera, Ilva .......................... 162Candido, Thiago ...................... 42Carreras-Villaseñor, Nohemi . 435Cartwright, Gemma ............... 587Castanheira, Sónia ................. 215Castrillo Jimenez, Marta ........ 406Chang, Chia-Chen .................. 182Chang, Jinhui ......................... 307Chankhamjon,Pranatchareeya .........................2Chellappan, Biju .................... 672Cheng, Xuanjin ...................... 469Cherif, Chetouhi .................... 265Choi, Jaeyoung ...................... 297Chung, Hyunjung ................... 298Coradetti, Samuel .................. 418Corcoran, Padraic .................. 676Corral Ramos, Cristina ........... 553Dahlmann, Tim ...................... 313Darbyshir, Heather ................ 687Daskalov, Asen ...................... 345Delgado-Álvarez, Diego ......... 173Delorey, Toni ......................... 659Dhingra, Sourabh .................. 356Díaz-Choque, Rodrigo ............ 444Dirschnabel, Daniela.............. 404Doernte, Bastian ............. 64, 470Donaldson, Michael............... 330Doughan, Benjamin ............... 454Downes, Damien ................... 365Drews, Kelly ........................... 518DuBois, Juwen ....................... 420English, Bevin ........................ 591Evangelisti, Edouard .............. 477FAIVRE TALMEY, YOHANN ..... 374Fajardo, Rosa ........................ 117Feldman, Daria ..................... 168Feretzaki, Marianna .............. 125Fernandes, Caroline .............. 691Fischer, Gregory.................... 486Flajsman, Marko ................... 271Ford, Kathryn ........................ 537Forester, Natasha ................. 459Fourie, Gerda ........................ 260Freihorst, Daniela ................. 203Friedman, Steven .................. 380Fu, Ci ..................................... 153Fukada, Fumi ........................ 113Gajdeczka, Michael ............... 682GARCÍA-MARTÍNEZ, JORGE ... 141Garud, AMruta...................... 184Gomes, Eriston ..................... 422Gradnigo, Julien .................... 732Grandaubert, Jonathan ......... 280Grognet, Pierre ..................... 733Gross, Nathan ......................... 82Grum-Grzhimaylo, Alexey ..... 734Grund, Elisabeth ................... 240Gsaller, Fabio ................ 357, 471Gupta, Yogesh ...................... 165Guzman-Guzman, Paulina .... 521Gyawali, Rachana ................. 700Hann-Soden, Christopher ..... 663Hargarten, Jessica ................. 498Hasim, Sahar ........................... 17Havel, Virginia ....................... 508Hayes, Patrick ......................... 57Heilig, Yvonne ............... 176, 177Henke, Catarina .................... 582Herrmann, Andrea ................ 457Hirsch, Robert ....................... 568Huang, Yuhong ..................... 735Hunter, Cameron .................. 366Huskey, David ....................... 541Ishii, Akira ............................... 70Islam, Kazi ............................. 542Ivanova, Christa ...................... 61Jöhnk, Bastian ....................... 487Jahan, Sultana ....................... 402Johns, Anna .......................... 488Johnson, Derek ..................... 252Jung, Elke-Martina ................ 209Jung, Kwang-Woo ................. 393Kaczmarek, Maciej ................ 564Kagda, Meenakshi .................. 30Kamei, Masayuki ................... 131Kamvar, Zhian ....................... 683Kaur, Jan Naseer ................... 394Kawauchi, Moriyuki .............. 450Kim, Seongbeom ................... 634Kim, Wonyong ...................... 615Klinter, Stefan ......................... 20Kodama, Sayo ......................... 71Kombrink, Anja ..................... 522Krijgsheld, Pauline ................ 100Kuo, Alan .............................. 227Kurashima, Kiminori ............. 188Kuuskeri, Jaana ..................... 328Lambie, Scott ........................ 620Langner, Thorsten ................. 216Lastovetsky, Olga .................. 600Lauinger, Linda ....................... 44Lee, Yoon Ji ........................... 144Lefebvre, François ................ 331Lehneck, Ronny .................... 132Li, Guotian ............................. 574Li, Simeng .............................. 346liang, xiaofei .......................... 635Lim, Fang Yun ............................ 5Lobach, Lars .......................... 525Ma, Liang .............................. 274Ma, Liqiu ............................... 472Macdonald, Darel .................. 690Macheleidt, Juliane ............... 359Madhavan, Soumya .............. 400marchegiani, elisabetta ......... 322Martinez, Leonora ................. 189Maurer, Elisabeth.................. 636Mead, Matt ........................... 395Mehrotra, Pankaj .................. 503Mennat, El Ghalid ................. 554Michkov, Alexander .............. 190Min, Kyunghun ...................... 195Minz-Dub, Anna .................... 105Miralles-Duran, Alejandro ..... 473Mirzadi Gohari, Amir ............. 272Misener Bloom, Amanda ...... 396Moktali, Venkatesh ....... 253, 254Mondo, Stephen ................... 255Mueller, Andre ...................... 622Mujic, Alija ............................ 335Nadimi, Maryam ................... 673Naik, Vikram .......................... 623Naito, Mizue ......................... 609Navarro Velasco, Gesabel ..... 555Nogami, Yuhei ....................... 191Nogueira, Filomena ............... 699Nordzieke, Steffen ................ 199Ohba, Ayumi ......................... 427Ohkura, Mana ............... 347, 679Olarte, Rodrigo ...................... 668Oses- Ruiz, Miriam ................ 478Oskarsson, Therése ............... 150Owensby, Catherine .............. 320Park, Ae Ran .......................... 145Park, Heesoo ........................... 90Park, Jaejin ............................ 716Park, Joohae .................. 102, 103Pathirana, Ruvini ................... 499Patyshakuliyeva,Aleksandrina ........................... 27Paun, Linda ............................. 68Penselin, Daniel..................... 489Pilgaard, Bo ........................... 712Piotrowska, Marta ................ 650Plaza, David ........................... 741Poppe, Stephan ..................... 512Priegnitz, Bert-Ewald ............. 370Quester, Katrin ...................... 719Rabe, Franziska ..................... 494Rahnama, Mostafa ................ 610Ramada, Marcelo .................... 46Ravasio, Davide ..................... 451Redkar, Amey ........................ 624Rems, Ana ............................... 53Repas, Timothy ..................... 611Reza, Md ............................... 575Robledo Briones, AngélicaMariana................................. 381Rosikiewicz, Pawel ................ 584Rossi, Marika ......................... 167Ruger-Herreros, Carmen ....... 446Runa, Farhana ......................... 69Rytioja, Johanna ...................... 37Rzeszutek, Elzbieta .................. 22Sain, Divya ............................ 241Sakulkoo, Wasin ................... 116Salamov, Asaf ....................... 228Sanchez, Eddy ....................... 163Saraiva, Marcia ..................... 275Sarikaya Bayram, Oezlem ....... 93Sarowar, Mohammad ........... 514Sarwar, Samina ..................... 513Sasaki, Kengo ........................ 480SATOH, Yuki .......................... 576Schaefer, Katja ..................... 556Schinke, Josua ........................ 94Schotanus, Klaas ................... 242Segers, Frank ........................ 136Senftleben, Dominik ............. 585Shah, Firoz ............................ 442Shahi, Shermineh ................. 148Shakya, Viplendra ................. 170Sharma Khatiwada, Sandeep 601Sharpee, William .................. 476Shiller, Jason ......................... 249Sieber, Christian ................... 263Siegmund, Ulrike .................. 496Silva, Larissa ......................... 382SON, HOKYOUNG ................. 146Soukup, Alexandra................ 452Soyer, Jessica ........................ 403Steindorff, Andrei ................. 316Stöckli, Martina .................... 706Suhr, Mallory ........................ 501Takahashi, Tsukasa ............... 401Terhonen, Eeva .................... 702Thompkins, Ashley ............... 147thomson, darren .................. 111Tillmann, Britta ..................... 160Toh, Su San ........................... 339Tong, Xiaoxue ....................... 697Tzelepis, Georgios ................ 193Valim, Clarissa ...................... 504van Dam, Peter ..................... 270van Laarhoven, Karel ............ 161Verma, Surbhi ................ 398, 399Villalobos-Escobedo, JoseManuel ................................. 433Virgilio, Stela ........................ 388Vlaardingerbroek, Ido ........... 149Wang, Guanghui ................... 438Wang, Yizhou........................ 475Weerasinghe, Harshini ......... 597Wehr, Michaela .................... 152Weichert, Martin .................. 179Whiston, Emily ..................... 292Williams, Angela ................... 632Wu, Jiayao ............................ 349Wyatt, Timon........................ 207Yang, Chun-Hsiang .................. 25Yang, Dong-Hoon ................. 246Yerra, Lakshmi ........................ 96Zhang, Jian ............................ 558Zhang, Lisha .......................... 375Zhang, Ying ............................. 98Zhou, Mian ........................... 390Zimmerman, Kolea ............... 642334

Program Book - 27th Fungal Genetics Conference

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