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ARS Home » Midwest Area » West Lafayette, Indiana » Crop Production and Pest Control Research » Research » Publications at this Location » Publication #373122
Research Project: Molecular Mechanisms of Host-Fungal Pathogen Interactions in Cereal Crops

Location: Crop Production and Pest Control Research

Title: Real-time loop-mediated isothermal amplification assay for rapid and accurate detection of fungal pathogens of wheat

Author
item GOMEZ, SANDRA - PURDUE UNIVERSITY
item Goodwin, Stephen - Steve

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 3/15/2020
Publication Date: 3/15/2020
Citation: Gomez, S.V., Goodwin, S.B. 2020. Real-time loop-mediated isothermal amplification assay for rapid and accurate detection of fungal pathogens of wheat. In: Proceedings of Plant Health 2020. August 8-12, 2020. Denver, Colorado.

Interpretive Summary:

Technical Abstract: Wheat is an important cereal crop grown on more than 240 million ha. This crop is affected by the Septoria complex caused by Zymoseptoria tritici and Parastagonospora nodorum, as well as other pathogens like Pyrenophora tritici-repentis (tan spot), Cochliobolus sativus (spot blotch) and Magnaporthe oryzae (wheat blast). It is difficult to visually distinguish between these pathogens at the onset of infection. Accurate identification of the causal agent is decisive to make the right decisions about disease management. LAMP is a new isothermal nucleic acid amplification method that does not use a thermal cycler. It is emerging as a simple and highly sensitive diagnostic tool for early detection of plant pathogens. In this study we wanted to standarize a LAMP-based diagnostic assay for effective, quick and specific detection of pathogens in wheat. A set of six primers for each pathogen was designed by using Primer Explorer V4 software. Genomic DNA was extracted from Z. tritici and P. nodorum. The LAMP reaction was heated to 65 °C for 60 min, 80 min and 120 min to standardize the reaction. The reaction at 65 °C for 60 min was optimum and DNA amplification was obtained. Amplified bands of 200 bp were detected by electrophoresis for Z. tritici and P. nodorum and no amplification was detected for the positive or negative controls. The correct identification of the species that are affecting a crop of interest is the first approach for phytopathology studies, making possible to study its biological features and conduct research to find an effective way to control the disease.