Monoclonal Antibody Immunoblot Procedure for Detecting Neotyphodium Coenophialum in Seedling Tall Fescue

Abstract

In the United States, tall fescue (Festuca arundinacea Schreb.) turf and forage cultivars are being touted as “endophyte enhanced” or “endophyte-free”. Emerging technology suggests that knowledge of infection status of seed lots will become increasingly important for forage cultivars as endophyte-infected non-toxic germplasms are produced. Seed testing services routinely test for presence of endophyte by peeling inner epidermal and subtending cortical cells from the leaf sheath and histologically staining the tissue for microscopic analysis. The procedure is tedious and time consuming and requires a grow out of the seed for a period of 4 to 8 weeks. Hence, a rapid, inexpensive, and reproducible technique is needed to evaluate tall fescue as to the presence or absence of the endophytic fungus, Neotyphodium coenophialum (Morgan-Jones and Gams) Glenn, Bacon, and Hanlin, comb. nov. The purpose of this study was to develop a tissue immunoblot procedure using monoclonal antibodies developed against N. coenophialum mycelial protein that detected the presence of N. coenophialum in tall fescue plants and seeds.