Fungal endophyte detection in pasture grass seed utilising the infection layer and comparison to other detection techniques

A new detection technique, which exploited the biology surrounding the colonisation of pasture grass embryos by the endophytic fungus Neotyphodium, was developed to assess the proportion of endophyte infected seeds within selected accessions. A generalized linear mixed model followed by one-sided pairwise comparisons were conducted to compare the new infection layer (IL) technique and a common seed squash (SS) to the tissue print-immunoblot (TPIB). The latter is a practice used within the seed industry to test endophyte viability within accessions of Lolium arundinaceum and Lolium perenne. The IL was found to be a simple and accurate technique for predicting the proportion of viable endophyte in uncompromised seed accessions and was more accurate than the SS, which frequently recorded an over estimation. However, neither the SS nor the IL were a substitute for TPIB in detecting viable endophyte in compromised seed accessions such as those affected by certain systemic fungicides.