A Quantitative Real-Time PCR Method to Detect the Quinoa Downy Mildew Pathogen, Peronospora variabilis .

Autor:	Testen AL; USDA-ARS Application Technology Research Unit, Wooster, OH., Puri P; Department of Plant Pathology, Washington State University, Northwestern Washington Research and Extension Center, Mount Vernon, WA., Shaw RS; USDA-ARS Application Technology Research Unit, Wooster, OH., Domsic EC; Department of Crop and Soil Sciences, Washington State University, Pullman, WA., Griffin-LaHue D; Department of Crop and Soil Sciences, Washington State University, Northwestern Washington Research and Extension Center, Mount Vernon, WA., Murphy KM; Department of Crop and Soil Sciences, Washington State University, Pullman, WA., Mattupalli C; Department of Plant Pathology, Washington State University, Northwestern Washington Research and Extension Center, Mount Vernon, WA.
Jazyk:	angličtina
Zdroj:	Plant disease [Plant Dis] 2024 Sep; Vol. 108 (9), pp. 2887-2893. Date of Electronic Publication: 2024 Sep 03.
DOI:	10.1094/PDIS-11-23-2308-RE
Abstrakt:	Quinoa downy mildew, caused by Peronospora variabilis , is the most devastating disease of quinoa globally. Rapid, sensitive diagnostic methods are needed to detect and quantify this pathogen in seeds and plant tissue. A hydrolysis probe-based quantitative real-time PCR (qPCR) assay including a competitive internal control was developed for P. variabilis detection. This assay could detect as low as 20 ag of DNA or approximately 25 internal transcribed spacer (ITS) copies per reaction with efficiencies ranging from 93.9 to 98.2%. No nontarget amplification was observed when tested against DNA from other downy mildew pathogens and related oomycetes. P . variabilis strains from multiple countries were detected using this assay. The assay was successfully applied to quantify the pathogen in quinoa seeds from a field trial conducted in the state of Washington. Downy mildew disease was recorded on all 14 genotypes, with the genotypes 104.88 and 106.49 recording the highest area under the disease progress curve values (mean ± SE; 3,236 ± 303 and 2,851 ± 198, respectively) and J6 and Dutchess recording the lowest (441 ± 107 and 409 ± 129, respectively). Seed washes obtained from field samples were subjected to the qPCR assay, and the pathogen was detected in all samples. The highest pathogen ITS copy number was recorded with 106.49 (194,934 ± 38,171), and the lowest was observed in Pasto (5,971 ± 1,435) and Riobamba (9,954 ± 4,243). This qPCR assay could lead to improved detection and quantification of P. variabilis as well as increased understanding of quinoa- P. variabilis interactions and epidemiology. Competing Interests: The author(s) declare no conflict of interest.
Databáze:	MEDLINE
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