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Rapid in situ quantification of the strobilurin resistance mutation G143A in the wheat pathogen Blumeria graminis f. sp. tritici.
Dodhia, Kejal N; Cox, Belinda A; Oliver, Richard P; Lopez-Ruiz, Francisco J.
Afiliación
  • Dodhia KN; Centre for Crop and Disease Management, School of Molecular and Life Sciences, Curtin University, Bentley, Perth, WA, 6102, Australia.
  • Cox BA; Faculty of Science and Engineering, Curtin University, Perth, WA, 6102, Australia.
  • Oliver RP; Curtin University, Perth, WA, 6102, Australia.
  • Lopez-Ruiz FJ; Centre for Crop and Disease Management, School of Molecular and Life Sciences, Curtin University, Bentley, Perth, WA, 6102, Australia. fran.lopezruiz@curtin.edu.au.
Sci Rep ; 11(1): 4526, 2021 02 25.
Article en En | MEDLINE | ID: mdl-33633193
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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Ascomicetos / Farmacorresistencia Fúngica / Estrobilurinas / Fungicidas Industriales / Mutación Idioma: En Revista: Sci Rep Año: 2021 Tipo del documento: Article País de afiliación: Australia Pais de publicación: Reino Unido
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Ascomicetos / Farmacorresistencia Fúngica / Estrobilurinas / Fungicidas Industriales / Mutación Idioma: En Revista: Sci Rep Año: 2021 Tipo del documento: Article País de afiliación: Australia Pais de publicación: Reino Unido