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Open AccessArticle

A PCR, qPCR, and LAMP Toolkit for the Detection of the Wheat Blast Pathogen in Seeds

1
ANSES Plant Health Laboratory, Mycology Unit, Domaine de Pixérécourt, Bâtiment E, F-54220 Malzeville, France
2
UMR BGPI, Montpellier University, INRAE, CIRAD, Montpellier SupAgro, 34398 Montpellier, France
3
CIRAD, UMR BGPI, F-34398 Montpellier, France
*
Author to whom correspondence should be addressed.
These authors contribute equally to this work.
Plants 2020, 9(2), 277; https://doi.org/10.3390/plants9020277
Received: 7 February 2020 / Revised: 17 February 2020 / Accepted: 18 February 2020 / Published: 21 February 2020
(This article belongs to the Special Issue Detection and Diagnostics of Fungal and Oomycete Plant Pathogens)
Wheat blast is a devastating disease caused by the pathogenic fungus Pyricularia oryzae. Wheat blast first emerged in South America before more recently reaching Bangladesh. Even though the pathogen can spread locally by air-dispersed spores, long-distance spread is likely to occur via infected wheat seed or grain. Wheat blast epidemics are caused by a genetic lineage of the fungus, called the Triticum lineage, only differing from the other P. oryzae lineages by less than 1% genetic divergence. In order to prevent further spread of this pathogen to other wheat-growing areas in the world, sensitive and specific detection tools are needed to test for contamination of traded seed lots by the P. oryzae Triticum lineage. In this study, we adopted a comparative genomics approach to identify new loci specific to the P. oryzae Triticum lineage and used them to design a set of new markers that can be used in conventional polymerase chain reaction (PCR), real-time PCR, or loop-mediated isothermal amplification (LAMP) for the detection of the pathogen, with improved inclusivity and specificity compared to currently available tests. A preliminary biological enrichment step of the seeds was shown to improve the sensitivity of the tests, which enabled the detection of the target at an infection rate as low as 0.25%. Combined with others, this new toolkit may be particularly beneficial in preventing the trade of contaminated seeds and in limiting the spread of the disease. View Full-Text
Keywords: seed testing; diagnostic; Pyricularia oryzae seed testing; diagnostic; Pyricularia oryzae
MDPI and ACS Style

Thierry, M.; Chatet, A.; Fournier, E.; Tharreau, D.; Ioos, R. A PCR, qPCR, and LAMP Toolkit for the Detection of the Wheat Blast Pathogen in Seeds. Plants 2020, 9, 277. https://doi.org/10.3390/plants9020277

AMA Style

Thierry M, Chatet A, Fournier E, Tharreau D, Ioos R. A PCR, qPCR, and LAMP Toolkit for the Detection of the Wheat Blast Pathogen in Seeds. Plants. 2020; 9(2):277. https://doi.org/10.3390/plants9020277

Chicago/Turabian Style

Thierry, Maud; Chatet, Axel; Fournier, Elisabeth; Tharreau, Didier; Ioos, Renaud. 2020. "A PCR, qPCR, and LAMP Toolkit for the Detection of the Wheat Blast Pathogen in Seeds" Plants 9, no. 2: 277. https://doi.org/10.3390/plants9020277

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