A PCR, qPCR, and LAMP Toolkit for the Detection of the Wheat Blast Pathogen in Seeds
Wheat blast is a devastating disease caused by the pathogenic fungus <i>Pyricularia oryzae.</i> 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 occu...
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doaj-e366695e873446fb8e0747e1dab5cc5f2020-11-25T02:36:59ZengMDPI AGPlants2223-77472020-02-019227710.3390/plants9020277plants9020277A PCR, qPCR, and LAMP Toolkit for the Detection of the Wheat Blast Pathogen in SeedsMaud Thierry0Axel Chatet1Elisabeth Fournier2Didier Tharreau3Renaud Ioos4ANSES Plant Health Laboratory, Mycology Unit, Domaine de Pixérécourt, Bâtiment E, F-54220 Malzeville, FranceANSES Plant Health Laboratory, Mycology Unit, Domaine de Pixérécourt, Bâtiment E, F-54220 Malzeville, FranceUMR BGPI, Montpellier University, INRAE, CIRAD, Montpellier SupAgro, 34398 Montpellier, FranceUMR BGPI, Montpellier University, INRAE, CIRAD, Montpellier SupAgro, 34398 Montpellier, FranceANSES Plant Health Laboratory, Mycology Unit, Domaine de Pixérécourt, Bâtiment E, F-54220 Malzeville, FranceWheat blast is a devastating disease caused by the pathogenic fungus <i>Pyricularia oryzae.</i> 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 <i>Triticum</i> lineage, only differing from the other <i>P. oryzae</i> 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 <i>P. oryzae</i> <i>Triticum</i> lineage. In this study, we adopted a comparative genomics approach to identify new loci specific to the <i>P. oryzae</i> <i>Triticum</i> 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.https://www.mdpi.com/2223-7747/9/2/277seed testingdiagnostic<i>pyricularia oryzae</i> |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Maud Thierry Axel Chatet Elisabeth Fournier Didier Tharreau Renaud Ioos |
spellingShingle |
Maud Thierry Axel Chatet Elisabeth Fournier Didier Tharreau Renaud Ioos A PCR, qPCR, and LAMP Toolkit for the Detection of the Wheat Blast Pathogen in Seeds Plants seed testing diagnostic <i>pyricularia oryzae</i> |
author_facet |
Maud Thierry Axel Chatet Elisabeth Fournier Didier Tharreau Renaud Ioos |
author_sort |
Maud Thierry |
title |
A PCR, qPCR, and LAMP Toolkit for the Detection of the Wheat Blast Pathogen in Seeds |
title_short |
A PCR, qPCR, and LAMP Toolkit for the Detection of the Wheat Blast Pathogen in Seeds |
title_full |
A PCR, qPCR, and LAMP Toolkit for the Detection of the Wheat Blast Pathogen in Seeds |
title_fullStr |
A PCR, qPCR, and LAMP Toolkit for the Detection of the Wheat Blast Pathogen in Seeds |
title_full_unstemmed |
A PCR, qPCR, and LAMP Toolkit for the Detection of the Wheat Blast Pathogen in Seeds |
title_sort |
pcr, qpcr, and lamp toolkit for the detection of the wheat blast pathogen in seeds |
publisher |
MDPI AG |
series |
Plants |
issn |
2223-7747 |
publishDate |
2020-02-01 |
description |
Wheat blast is a devastating disease caused by the pathogenic fungus <i>Pyricularia oryzae.</i> 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 <i>Triticum</i> lineage, only differing from the other <i>P. oryzae</i> 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 <i>P. oryzae</i> <i>Triticum</i> lineage. In this study, we adopted a comparative genomics approach to identify new loci specific to the <i>P. oryzae</i> <i>Triticum</i> 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. |
topic |
seed testing diagnostic <i>pyricularia oryzae</i> |
url |
https://www.mdpi.com/2223-7747/9/2/277 |
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