Program Book - 27th Fungal Genetics Conference

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