Detection of Pyricularia grisea Causing Gray Leaf Spot of Perennial Ryegrass Turf by a Rapid Immuno-Recognition Assay

Abstract

An immuno-recognition assay using a monoclonal antibody was developed to detect Pyricularia grisea, the causal agent of gray leaf spot of perennial ryegrass (Lolium perenne). In vitro assays with isolates of P. grisea from perennial ryegrass, tall fescue (Festuca arundinacea), St. Augustinegrass (Stenotaphrum secundatum), crabgrass (Digitaria sanguinalis), finger millet (Eleusine coracana), wheat (Triticum aestivum), triticale (× Triticosecale rimpaui), and rice (Oryza sativa) showed positive reactions; however, the strength of the reactions differed among isolates. Reactions were more intense with isolates from perennial ryegrass, wheat, and triticale. All P. grisea isolates from perennial ryegrass collected from various regions of the United States showed positive reactions. P. grisea was detected at antigen dilution rates of 0.5×, 0.25×, 0.13×, 0.06×, and 0.03×. Dot-blot assays with Bipolaris sorokiniana, Colletotrichum graminicola, Curvularia lunata, Microdochium nivale, Pythium aphanidermatum, Rhizoctonia solani, or Sclerotinia homoeocarpa isolated from turfgrasses were negative. In vivo assays of symptomatic leaves of perennial ryegrass plants inoculated with P. grisea also showed positive reactions, and those inoculated with B. sorokiniana, P. aphanidermatum, R. solani, or S. homoeocarpa were negative. Intensity of reaction between the monoclonal antibody and P. grisea was proportional to disease severity in perennial ryegrass inoculated with P. grisea; however, there was no reaction in dot blots of leaf tissue collected during the latent period. P. grisea was detected in perennial ryegrass samples from golf course fairways affected by gray leaf spot in Connecticut, Massachusetts, Maine, New Jersey, Pennsylvania, and Rhode Island using this procedure. The monoclonal antibody recognition system is highly sensitive to P. grisea and can be used effectively for the rapid diagnosis of gray leaf spot of perennial ryegrass turf.