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ARS Home » Midwest Area » West Lafayette, Indiana » Crop Production and Pest Control Research » Research » Publications at this Location » Publication #357635
Research Project: Molecular Mechanisms of Host-Fungal Pathogen Interactions in Cereal Crops

Location: Crop Production and Pest Control Research

Title: Development of a multiplex RT-PCR assay for simultaneous detection of 10 major viral pathogens of wheat

Author
item DEB, MAHUA - Purdue University
item ANDERSON, JOSEPH - Purdue University
item Scofield, Steven - Steve

Submitted to: Agronomy Journal
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/9/2023
Publication Date: 3/16/2023
Citation: Deb, M., Anderson, J.M., Scofield, S.R. 2023. Development of a multiplex RT-PCR assay for simultaneous detection of 10 major viral pathogens of wheat. Agronomy Journal. 13:833. https://doi.org/10.3390/agronomy13030833.
DOI: https://doi.org/10.3390/agronomy13030833

Interpretive Summary: Wheat and other small grain cereals are vulnerable to a large number of viruses that can cause major yield loss. Infection by these diverse viruses cause very similar symptons, making it difficult to identify the specific agent causing disease. Additionally, more than one virus may be present at the same time. This work reports the development of a PCR-based assay that can simultaneously detect and identify 10 major viruses affecting wheat production. This rapid and cost-effective test will be an important new tool for plant disease diagnosticians, extension specialists, and plant pathologists.

Technical Abstract: Wheat production can be limited by a wide range of viruses. The similar symptoms resulting from these diverse viruses makes it difficult to identify the specific disease causing agent. This paper describes a robust multplex PCR assay that is able to simultaneously detect 10 of the major viral pathogens of wheat. The method uses a set of primers that detect the target viruses: Triticum mosaic virus (TriMV), High plains virus (HPV), Barley/Cereal yellow dwarf viruses -PAV, -MAV, -SGV, -RPV, -RMV, Wheat spindle streak mosaic virus (WSSMV), Soil-borne wheat mosaic virus SBWMV and Wheat streak mosaic virus (WSMV). Each forward primer is fluorescently tagged so that each unique viral PCR product can be detected via capillary electrophoresis (CE). By constructing mixes, it demonstrated that all 10 viruses are detected simultaneously.