Alteration of alkaloid profiles of forage and turf grasses by genetic manipulation of endophytic fungi

Objectives: Introduce modified endophyte strains (provided by the Panaccione laboratory at WVU) into endophytefree perennial ryegrass, and provide infected plant material for ergot alkaloid analyses. Knock out EASl cluster in strain of Neotyphodium coenophialum in which dmaW2 is already knocked out. Knock out EASl cluster in wild-type N. coenophialum. Knock out lolC in wild-type N. coenophialum. Introduce modified emdophyte strains into endophyte-free tall fescue, and, for the EASl knockout strains, provide infected plant material for ergot alkaloid analyses. Perform loline alkaloid analyses on the 10lC knockout strain. Introduce selected loline cluster genes into Neotyphodium lolii x Epichloe typhina strain Lp1. Test cultures of modified fungi for loline alkaloids. Introduce modified strains into endophyte-free perennial ryegrass, and analyze loline alkaloids. Help coordinate tests of anti-insect and anti-nematode activity by cooperating with collaborators at University of Kentucky and USDA ARS.