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Development and application of loop-mediated isothermal amplification for detection of the F167Y mutation of carbendazim-resistant isolates in Fusarium graminearum

Resistance of Fusarium graminearum to carbendazim is caused by point mutations in the β(2)-tubulin gene. The point mutation at codon 167 (TTT → TAT, F167Y) occurs in more than 90% of field resistant isolates in China. To establish a suitable method for rapid detection of the F167Y mutation in F. gra...

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Autores principales: Duan, Yabing, Zhang, Xiaoke, Ge, Changyan, Wang, Yong, Cao, Junhong, Jia, Xiaojing, Wang, Jianxin, Zhou, Mingguo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4235284/
https://www.ncbi.nlm.nih.gov/pubmed/25403277
http://dx.doi.org/10.1038/srep07094
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author Duan, Yabing
Zhang, Xiaoke
Ge, Changyan
Wang, Yong
Cao, Junhong
Jia, Xiaojing
Wang, Jianxin
Zhou, Mingguo
author_facet Duan, Yabing
Zhang, Xiaoke
Ge, Changyan
Wang, Yong
Cao, Junhong
Jia, Xiaojing
Wang, Jianxin
Zhou, Mingguo
author_sort Duan, Yabing
collection PubMed
description Resistance of Fusarium graminearum to carbendazim is caused by point mutations in the β(2)-tubulin gene. The point mutation at codon 167 (TTT → TAT, F167Y) occurs in more than 90% of field resistant isolates in China. To establish a suitable method for rapid detection of the F167Y mutation in F. graminearum, an efficient and simple method with high specificity was developed based on loop-mediated isothermal amplification (LAMP). A set of four primers was designed and optimized to specially distinguish the F167Y mutation genotype. The LAMP reaction was optimal at 63°C for 60 min. When hydroxynaphthol blue dye (HNB) was added prior to amplification, samples with DNA of the F167Y mutation developed a characteristic sky blue color after the reaction but those without DNA or with different DNA did not. Results of HNB staining method were reconfirmed by gel electrophoresis. The developed LAMP had good specificity, stability and repeatability and was suitable for monitoring carbendazim-resistance populations of F. graminearum in agricultural production.
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spelling pubmed-42352842014-11-25 Development and application of loop-mediated isothermal amplification for detection of the F167Y mutation of carbendazim-resistant isolates in Fusarium graminearum Duan, Yabing Zhang, Xiaoke Ge, Changyan Wang, Yong Cao, Junhong Jia, Xiaojing Wang, Jianxin Zhou, Mingguo Sci Rep Article Resistance of Fusarium graminearum to carbendazim is caused by point mutations in the β(2)-tubulin gene. The point mutation at codon 167 (TTT → TAT, F167Y) occurs in more than 90% of field resistant isolates in China. To establish a suitable method for rapid detection of the F167Y mutation in F. graminearum, an efficient and simple method with high specificity was developed based on loop-mediated isothermal amplification (LAMP). A set of four primers was designed and optimized to specially distinguish the F167Y mutation genotype. The LAMP reaction was optimal at 63°C for 60 min. When hydroxynaphthol blue dye (HNB) was added prior to amplification, samples with DNA of the F167Y mutation developed a characteristic sky blue color after the reaction but those without DNA or with different DNA did not. Results of HNB staining method were reconfirmed by gel electrophoresis. The developed LAMP had good specificity, stability and repeatability and was suitable for monitoring carbendazim-resistance populations of F. graminearum in agricultural production. Nature Publishing Group 2014-11-18 /pmc/articles/PMC4235284/ /pubmed/25403277 http://dx.doi.org/10.1038/srep07094 Text en Copyright © 2014, Macmillan Publishers Limited. All rights reserved http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder in order to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Duan, Yabing
Zhang, Xiaoke
Ge, Changyan
Wang, Yong
Cao, Junhong
Jia, Xiaojing
Wang, Jianxin
Zhou, Mingguo
Development and application of loop-mediated isothermal amplification for detection of the F167Y mutation of carbendazim-resistant isolates in Fusarium graminearum
title Development and application of loop-mediated isothermal amplification for detection of the F167Y mutation of carbendazim-resistant isolates in Fusarium graminearum
title_full Development and application of loop-mediated isothermal amplification for detection of the F167Y mutation of carbendazim-resistant isolates in Fusarium graminearum
title_fullStr Development and application of loop-mediated isothermal amplification for detection of the F167Y mutation of carbendazim-resistant isolates in Fusarium graminearum
title_full_unstemmed Development and application of loop-mediated isothermal amplification for detection of the F167Y mutation of carbendazim-resistant isolates in Fusarium graminearum
title_short Development and application of loop-mediated isothermal amplification for detection of the F167Y mutation of carbendazim-resistant isolates in Fusarium graminearum
title_sort development and application of loop-mediated isothermal amplification for detection of the f167y mutation of carbendazim-resistant isolates in fusarium graminearum
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4235284/
https://www.ncbi.nlm.nih.gov/pubmed/25403277
http://dx.doi.org/10.1038/srep07094
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