MONOCLONAL-ANTIBODIES FOR DETECTION OF NEOTYPHODIUM-COENOPHIALUM

A simple endophyte detection system is vital for breeding efforts desi gned to produce low alkaloid, endophyte-infected tall fescue (Festuca arundinacea Schreb,) cultivars, The objectives of this research were t o generate monoclonal antibodies specific to endophytes for immunochem ical detection purposes, Protein extracts from Neotyphodium coenophial um (Morgan-Jones and Gams) Glenn, Bacon, and Hanlin comb, nov, isolate EDN11 were injected into Balb/c mice to elicit an immune response. Hy bridoma cell lines were screened for antibody production to the Neotyp hodium extract and for cross recognition of protein extracts from N. u ncinatum Glen, Bacon, and Hanlin and N. starrii Glen, Bacon, and Hanli n, as well as other fungi in the genera Cladosporium, Penicillium, Fus arium, and Aspergillus. Three hybridoma cell lines produced antibodies that reacted to Neotyphodium spp, proteins, but not with proteins iso lated from the other fungi, Monoclonal antibody 5C7 was immunochemical ly tested for specificity to endophyte in leaf sheaths of plant genoty pe DN11. Fluorescent staining suggested the antibody preferentially bo und to the cell wall of the endophyte, Antibodies were tested for affi nity to protein extracts from 14 endophyte-infected and endophyte-free tall fescue genotypes by an enzyme-linked immunosorbent assay (ELISA) method. Mean absorbance values for the protein extracts from the endo phyte-infected genotypes differed and ranged from 0.317 to 0.583. Mean absorbance values for the endophyte-free forms did not differ (P grea ter than or equal to 0.05). All infected plants had higher absorbance values than non-infected plants regardless of antibody tested, Monoclo nal antibodies can be used in an ELISA to detect Neotyphodium spp, fun gal mycelia present in tall fescue.