Program Book - 27th Fungal Genetics Conference

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