Program Book - 27th Fungal Genetics Conference

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