A POLYMERASE CHAIN REACTION-BASED PROCEDURE FOR DETECTION OF ACREMONIUM-COENOPHIALUM IN TALL FESCUE

Inserts from clones of a genomic library constructed using DNA from th e fungal endophyte Acremonium coenophialum were screened using dot blo ts and Southern blots. Several inserts that hybridized to DNA from A, coenophialum and to DNA from endophyte-infected (E+) tall fescue (Fest uca arundinacea), but not to DNA from endophyte-free (E-) grass, were sequenced. Oligonucleotide primers were synthesized, and polymerase ch ain reaction (PCR) was carried out using DNA from E+ and E- tall fescu e genotypes. PCR with one set of 21-mer primers yielded a prominent 1- kb product with DNA from A. coenophialum-infected plants but not from uninfected plants. This PCR-based procedure provides an accurate, rapi d, and sensitive means of detecting A. coenophialum in tall fescue.