Program Book - 27th Fungal Genetics Conference

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