Simple and Rapid Detection of Endophytic Fungi from Perennial Ryegrass seeds by Vise Blotting ELISA Procedure

Abstract

A simple method for estimatingAcremoniumendophyte (EP) infestation levels of perennial ryegrass (Lolium perenne, PR) seed lots was developed. To blot the PR seed sap onto nitrocellulose membranes (NCM), the seed were allowed to imbibe water for two days and pressed on NCM with a vise (Vise blotting ELISA procedure (VBE) ) . EP infestation levels of the seeds using the VBE were almost equal to the levels of the microscopic staining method. The antigen of EP, which was adhered to NCM by VBE, could be stored for 3 months in a desiccator and could be mailed without reduction of the reaction color intensity. The feeding damage among 27 cultivar by bluegrass webworm (Pediasia teterrellus) varied inversely with endophyte infestation levels using the VBE and there was a distinct negative correlation between them (r=-0.94) . Therefore, we believe that endophyte infestation levels using VBE will indicate resistance to feeding damage by bluegrass webworm.