Agricultural Microbes Genome 2 Conference

Poster: Microbial Genome Sequencing/Programs
P01_08.html

GENOME SEQUENCING AND GENE DISCOVERY IN THE RICE BLAST FUNGUS, MAGNAPORTHE GRISEA: THE OPENING ROUND

¹ Fungal Genomics Laboratory, NC State University, Raleigh, NC, 27695, USA

² 2 Department of Molecular Cellular and Developmental Biology, Yale University, New Haven, CT, 06502, USA

³ 3 Clemson University Genomics Institute, Clemson, SC, 29634, USA

⁴ 4 Department of Plant Pathology, University of Kentucky, Lexington, KY, 40546, USA

⁵ 5 Department of Plant Pathology and Microbiology, Texas A&M University, College Station, TX, 77843, USA

Magnaporthe grisea, the causal agent rice blast disease, is one of the most devastating threats to food security worldwide. The fungus is amenable to classical and molecular genetic manipulation and is a compelling experimental system for elucidating numerous aspects of pathogenesis, including infection-related morphogenesis, host species and cultivar specificity, and signaling pathways. Sequence information will be a rich resource for elucidating the molecular basis of pathogenicity and will be invaluable for designing novel environmentally sound control strategies. A 25X large insert (130 kb) HindIII BAC library was used to construct a physical map of the genome. BAC clones were fingerprinted and assembled into 188 contigs. These were aligned into a physical map by anchoring to mapped RFLP markers. Chromosome 7 (4.2 Mb) has been studied in the greatest detail and a set of 42 BAC clones representing a minimum tiling path covering >95% of the chromosome has been deduced. The sequence of one BAC clone (6J18) has been completed. The entire BAC library was end sequenced providing sequence tag connectors (STC) every 3-4 kb across the genome. A federated database integrating data from relational and object-oriented databases is being developed. We have initiated a draft sequence (~5X coverage) of chromosome 7 using the BAC by BAC approach coupled with information from our STC/fingerprint databases. A comprehensive EST program has been launched. 30,000 ESTs will be derived from 8 cDNA libraries prepared from different stages of growth and development as well as cells subjected to various stress conditions. A set of ~5,000 ESTs representing unique genes will be further sequenced. ESTs and consensus sequence data for individual BAC clones will be annotated, including BLAST and open reading frame analysis. A publicly available graphic web based interface will be created to search and view assembled and annotated data.