Location: Foreign Disease-Weed Science
Project Number: 1920-22000-041-08
Start Date: Sep 01, 2012
End Date: Sep 30, 2014
Whole genome sequencing and post-sequencing comparisons will be used to rapidly identify unique loci between the M. oryzae pathotypes that infect wheat and the pathotypes that infect related grass species. This approach is made feasible by virtue of “next generation” sequencing platforms that enable fast and cost-effective whole genome sequencing projects. We have previously utilized this technology to sequence an isolate of M. oryzae obtained from a 2011 outbreak of wheat blast in Bolivia. As part of this agreement, we will collect and prepare isolates of M. oryzae from North and South America collected from wheat and other grasses. Although the virulent pathotype of M. grisea (i.e. "South American wheat blast") has not been found outside of South America, it is important to sequence isolates collected from both continents in order to design a robust detection assay that is not prone to signaling false-positives. Additionally, RNA from wheat plants infected with M. oryzae will be used for deep sequencing to ascertain the genes involved in pathogenicity/resistance, and we will perform genotyping by sequencing of wheat varieties that have been tested for wheat blast resistance / susceptibility. Preparation of the M. oryzae isolates and isolation of high-quality genomic DNA and RNA will be performed at the FDWSRU at Ft. Detrick in the necessary containment laboratory/greenhouse facilities, and the sequencing will be performed by collaborators at Kansas State University. Post-sequencing analysis will be performed jointly between ARS and KSU, and final testing of the molecular diagnostic assays will be performed by ARS staff at Ft. Detrick.