Detection of endophytic fungi in Festuca spp. by means of tissue print immunoassay

Many wild and cultivated grasses live in mutualistic symbiosis with endophytic fungi of the genera Neotyphodium and Epichloe¨. These associations are of agronomic importance because endophytes may induce a range of beneficial effects for the host plant but also produce alkaloids detrimental to livestock. Conventional detection of endophytes by means of histological staining is time-consuming and not suited for large numbers of samples. Therefore, in order to simplify the detection of endophytic fungi the utility of tissue print immunoassay (TPIA) was studied and compared with the commonly used microscopic analysis. Ecotypes collected from natural grassland habitats and plants from field experiments were analyzed for endophyte infection. Both methods provided similar results. Based on stained or non-stained mycelium in tissue prints, endophyte-infected (E+) and endophyte-free (E–) tillers and inflorescences of Festuca pratensis, F. arundinacea and F. rubra were clearly distinguishable. Prints of cross sections of tillers allow the precise localization of endophyte infection within the plant tissues. Because TPIA allows the examination of endophytes in individual branches and segments of inflorescences it is a useful method for dissemination studies. Tissue print immunoassay appears to be a reliable method suited for routine work in research, practical grassland management and selection of defined E+ or E– material for breeding.