Location: Horticultural Crops ResearchTitle: Development of a quantitative loop-mediated isothermal amplification (qLAMP) assay for the detection of Magnaporthe oryzae airborne inoculum in turf ecosystems
|VILLARI, CATERINA - The Ohio State University|
|Mahaffee, Walter - Walt|
|MITCHELL, THOMAS - The Ohio State University|
|PEDUTO HAND, FRANCESCA - The Ohio State University|
Submitted to: American Phytopathological Society Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 5/25/2015
Publication Date: 8/1/2015
Citation: Villari, C., Mahaffee, W.F., Mitchell, T.K., Pedley, K.F., Pieck, M.L., Peduto Hand, F. 2015. Development of a quantitative loop-mediated isothermal amplification (qLAMP) assay for the detection of Magnaporthe oryzae airborne inoculum in turf ecosystems. In: Poster sessions of the American Phytopathological Society Annual Meeting. August 1-5, 2015, Pasadena, California. 443-P.
Technical Abstract: Grey Leaf Spot (GLS) is a detrimental disease of perennial ryegrass caused by a host-specialized form of Magnaporthe oryzae (Mot). In order to improve turf management, a quantitative loop-mediated isothermal amplification (LAMP) assay coupled with a simple spore trap is being developed to monitor GLS airborne inoculum. LAMP has been shown to be suitable for implementation by practitioners to initiate and time fungicide applications in vineyards. Since the most common conserved regions such as ITS, LSU and Calmodulin lack the needed heterogeneity among the different host-specialized forms of M. oryzae, LAMP primers were designed to a non-annotated genomic DNA region unique to Mot. Reactions were performed on a Real-Time PCR Detection System and optimized to provide a response in approximately 30 minutes. Quantification of the inoculum was obtained by comparison with a standard curve developed using 6 independent serial spore dilutions. The LAMP assay was specific to the host-specialized forms of M. oryzae and was able to detect greater than 500 spores, indicating that it is capable of detecting the pathogen at the very beginning of the disease epidemic. The implementation of this assay could be a useful decision support tool to guide initiation and timing of fungicide applications for GLS management.