Fungal genetic tools in Magnaporthe oryzae for reactive oxygen species detection and identification of host basal immunity components
N. DONOFRIO (1), K. Huang (1), T. Mitchell (2), R. Wilson (3), J. Sweigard (4), J. Caplan (5), K. Czymmek (6) (1) University of Delaware, Newark, DE, U.S.A.; (2) The Ohio State University, Columbus, OH, U.S.A.; (3) Univerrsity of Nebraska-Lincoln, Lincoln, U.S.A.; (4) DuPont, U.S.A.; (5) University of Delaware, U.S.A.; (6) Zeiss, U.S.A.

Bolstering plant basal immunity holds promise for controlling plant diseases, however its deployment first requires a better understanding of the genetic components contributed by both organisms, and how they interact. The interaction between the devastating rice blast fungus Magnaporthe oryzae and its hosts is well-studied, but the fungal factors that modulate basal immunity- particularly reactive oxygen species (ROS)- and the host components they impact, are poorly understood. We propose to fill this knowledge deficit using our available fungal mutant strains coupled with identification of new fungal genes via forward genetic screens. We will take advantage of a genetically-encoded ROS-sensing fungal strain recently generated in our lab to generate a random insertion library. Our ROS sensing line can be used with either confocal microscopy or in a 96-well plate reader format, making high-throughput screening, feasible, and can detect ROS during invasive growth in rice and barley up to 72 hours post-inoculation. While our mutant screening is not yet underway as we have just begun the project, our ultimate goal is to test candidate mutants on a panel of differentially susceptible rice lines using pathogenicity assays, confocal ROS imaging, transcriptomics and metabolic profiling to determine which fungal genes trigger basal immunity. The ROS-sensing line is freely available to the M. oryzae community, and we will present on current results, as well as immediate next steps.

Abstract Number: S2-6
Session Type: Special Session