Kit for detection of fungal endophytes of grasses yields inconsistent results

Summary 1. The Agrinostics field immunoblot assay is a colour-imprint assay originally developed for routinely and rapid detection of mycotoxin-producing fungi Neotyphodium spp./Epichloe spp. from samples of pasture, turf, cereals and seeds of agricultural grasses. 2. In addition the Agrinostics immunoblot assay has been used and recommended for exploring the presence and ecological role of Neotyphodium and the closely related Epichloe in several cool season grasses. 3. Here we demonstrate that this method is not limited to detection of only these two species, as several other fungal endophyte genera or species also yielded positive results when mycelia were loaded onto cards. Moreover, other closely related endophyte genera or species were found to yield negative result. Thus, the assay is neither specific to isolates of Neotyphodium/Epichloe nor is it universal for fungal endophyte detection. 4. Our study underscore the fact that the Agrinostics field immunoblot assay should not be used on its own for field research into the broader ecological significance of Neotyphodium/Epichloe. 5. We suggest that the assay may be best suited for use in agricultural fields or in controlled greenhouse experiments for hyphal load estimates, and should always be accompanied by other methods (e.g. microscopic visualization, DNA-sequencing) to confirm infection frequencies and certify that the endophyte present actually is Neotyphodium/Epichloe.