• Ralph A. Dean, Laboratory of Fungal Genomics, Department of Plant Pathology,
  North Carolina State University. (PI). http://www.cals.ncsu.edu/fungal_genomics
• Daniel Ebbole, Department of Plant Pathology and Microbiology,
  Texas A&M University. (co-PI). http://plantpathology.tamu.edu/people/bios/ebbole.html

Rice blast disease, caused by Magnaporthe grisea, is recognized as one of the main pathological threats to world food supplies.  Moreover, few fungal plant pathogens rival rice blast for the sophistication of molecular and classical genetic tools that have been developed, nor the breadth and the depth of the rice blast research community both in the US and elsewhere. The rice blast fungus is a compelling experimental system for elucidating numerous aspects of pathogenesis, including infection-related morphogenesis, host species and cultivar specificity and associated signaling pathways.

The specific objectives of this proposal are:

1.      Generate a total of 35,000 ESTs from a minimum of 8 cDNA libraries prepared from different stages of growth and development as well as cells subjected to various stress conditions.  Each library will be 5’end sequenced until ~70% redundancy or until a total of 30,000 lanes have been run.  From these, a set of ~5,000 ESTs representing unique genes will be sequenced from the 3’ end.

2.      Obtain ~5 X shotgun sequence of 41 BAC clones covering chromosome 7.  BAC clones will be sheared to ~2 kb fragments and cloned in a plasmid vector.  Sequence will be obtained from both ends of ~815 clones per BAC.  [41 X 1,625 = 66,625 sequence lanes]

3.      Provide basic sequence analysis and integration into physical map of chromosome 7.  Sequence data will be processed using Phred/Phrap.  Assemblies will be made first for each BAC clone, then the consensus sequence of ESTs and  BACs will be aligned and merged with BAC end sequence data.   Consensus sequence data for individual BAC clones and ESTs will be deposited at GenBank immediately.  A publicly available BLAST server will be created.  FASTA files will be publicly available.

The project provides new opportunities for integrating research and education of graduate students.  Availability of sequence information is essential for elucidating the molecular basis of pathogenicity and will be crucial for designing novel environmentally sound control strategies.