Rapid in situ quantification of the strobilurin resistance mutation G143A in the wheat pathogen Blumeria graminis f. sp. tritici

Abstract As the incidence of fungicide resistance in plant pathogens continues to increase, control of diseases and the management of resistance would be greatly aided by rapid diagnostic methods. Quantitative allele-specific PCR (ASqPCR) is an ideal technique for the in-field analysis of fungicide...

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Autores principales:	Kejal N. Dodhia, Belinda A. Cox, Richard P. Oliver, Francisco J. Lopez-Ruiz
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/5bdf8b1ebbd7487aaa1a818cccc2d5d3
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spelling	oai:doaj.org-article:5bdf8b1ebbd7487aaa1a818cccc2d5d32021-12-02T15:53:03ZRapid in situ quantification of the strobilurin resistance mutation G143A in the wheat pathogen Blumeria graminis f. sp. tritici10.1038/s41598-021-83981-92045-2322https://doaj.org/article/5bdf8b1ebbd7487aaa1a818cccc2d5d32021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-83981-9https://doaj.org/toc/2045-2322Abstract As the incidence of fungicide resistance in plant pathogens continues to increase, control of diseases and the management of resistance would be greatly aided by rapid diagnostic methods. Quantitative allele-specific PCR (ASqPCR) is an ideal technique for the in-field analysis of fungicide resistance as it can quantify the frequency of mutations in fungicide targets. We have applied this technique to the fungal pathogen Blumeria graminis f. sp. tritici (Bgt), the causal agent of wheat powdery mildew. In Australia, strobilurin-resistant Bgt was first discovered in 2016. Molecular analysis revealed a nucleotide transversion in the cytochrome b (cytb) gene in the cytochrome bc1 enzyme complex, resulting in a substitution of alanine for glycine at position 143 (G143A). We have developed an in-field ASqPCR assay that can quantify both the resistant (A143) and sensitive (G143) cytb alleles down to 1.67% in host and Bgt DNA mixtures, within 90 min of sample collection. The in situ analysis of samples collected during a survey in Tasmania revealed A143 frequencies ranging between 9–100%. Validation of the analysis with a newly developed laboratory based digital PCR assay found no significant differences between the two methods. We have successfully developed an in-field quantification method, for a strobilurin-resistant allele, by pairing the ASqPCR assay on a lightweight qPCR instrument with a quick DNA extraction method. The deployment of these type of methodologies in the field can contribute to the effective in-season management of fungicide resistance.Kejal N. DodhiaBelinda A. CoxRichard P. OliverFrancisco J. Lopez-RuizNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-15 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Kejal N. Dodhia Belinda A. Cox Richard P. Oliver Francisco J. Lopez-Ruiz Rapid in situ quantification of the strobilurin resistance mutation G143A in the wheat pathogen Blumeria graminis f. sp. tritici
description	Abstract As the incidence of fungicide resistance in plant pathogens continues to increase, control of diseases and the management of resistance would be greatly aided by rapid diagnostic methods. Quantitative allele-specific PCR (ASqPCR) is an ideal technique for the in-field analysis of fungicide resistance as it can quantify the frequency of mutations in fungicide targets. We have applied this technique to the fungal pathogen Blumeria graminis f. sp. tritici (Bgt), the causal agent of wheat powdery mildew. In Australia, strobilurin-resistant Bgt was first discovered in 2016. Molecular analysis revealed a nucleotide transversion in the cytochrome b (cytb) gene in the cytochrome bc1 enzyme complex, resulting in a substitution of alanine for glycine at position 143 (G143A). We have developed an in-field ASqPCR assay that can quantify both the resistant (A143) and sensitive (G143) cytb alleles down to 1.67% in host and Bgt DNA mixtures, within 90 min of sample collection. The in situ analysis of samples collected during a survey in Tasmania revealed A143 frequencies ranging between 9–100%. Validation of the analysis with a newly developed laboratory based digital PCR assay found no significant differences between the two methods. We have successfully developed an in-field quantification method, for a strobilurin-resistant allele, by pairing the ASqPCR assay on a lightweight qPCR instrument with a quick DNA extraction method. The deployment of these type of methodologies in the field can contribute to the effective in-season management of fungicide resistance.
format	article
author	Kejal N. Dodhia Belinda A. Cox Richard P. Oliver Francisco J. Lopez-Ruiz
author_facet	Kejal N. Dodhia Belinda A. Cox Richard P. Oliver Francisco J. Lopez-Ruiz
author_sort	Kejal N. Dodhia
title	Rapid in situ quantification of the strobilurin resistance mutation G143A in the wheat pathogen Blumeria graminis f. sp. tritici
title_short	Rapid in situ quantification of the strobilurin resistance mutation G143A in the wheat pathogen Blumeria graminis f. sp. tritici
title_full	Rapid in situ quantification of the strobilurin resistance mutation G143A in the wheat pathogen Blumeria graminis f. sp. tritici
title_fullStr	Rapid in situ quantification of the strobilurin resistance mutation G143A in the wheat pathogen Blumeria graminis f. sp. tritici
title_full_unstemmed	Rapid in situ quantification of the strobilurin resistance mutation G143A in the wheat pathogen Blumeria graminis f. sp. tritici
title_sort	rapid in situ quantification of the strobilurin resistance mutation g143a in the wheat pathogen blumeria graminis f. sp. tritici
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/5bdf8b1ebbd7487aaa1a818cccc2d5d3
work_keys_str_mv	AT kejalndodhia rapidinsituquantificationofthestrobilurinresistancemutationg143ainthewheatpathogenblumeriagraminisfsptritici AT belindaacox rapidinsituquantificationofthestrobilurinresistancemutationg143ainthewheatpathogenblumeriagraminisfsptritici AT richardpoliver rapidinsituquantificationofthestrobilurinresistancemutationg143ainthewheatpathogenblumeriagraminisfsptritici AT franciscojlopezruiz rapidinsituquantificationofthestrobilurinresistancemutationg143ainthewheatpathogenblumeriagraminisfsptritici
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Rapid in situ quantification of the strobilurin resistance mutation G143A in the wheat pathogen Blumeria graminis f. sp. tritici

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