Program Book - 27th Fungal Genetics Conference

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