A PCR-based technique for detection of Neotyphodium endophytes in diverse accessions of tall fescue

Abstract

A previously described polymerase chain reaction (PCR)-based method used for detection of Neotyphodium coenophialum in tall fescue detected Neotyphodium endophytes in some, but not all, infected plants from a geographically diverse sample. In the study reported here, a different set of primers, based on intervening sequences of the tubulin 2 gene, were prepared and used for PCR. PCR with these primers yielded the expected 444 base pair amplification product with DNA from 104 of the 106 infected accessions tested. In addition, one accession originally scored as endophyte-free on the basis of a tissue culture test was correctly rated as endophyte-infected using the PCR procedure. Results suggest that primers based on intervening sequences of the tubulin 2 gene can be used for PCR-based detection of Neotyphodium endophytes in tall fescue accessions of diverse origin.