Rapid detection of benzimidazole resistance in Botrytis cinerea by loop-mediated isothermal amplification View Full Text


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Article Info

DATE

2019-12

AUTHORS

Fei Fan, Matthias Hahn, Guo-Qing Li, Yang Lin, Chao-Xi Luo

ABSTRACT

Benzimidazole fungicides (MBCs) have been widely used in agriculture since 1970s, and resistance to this class of fungicides in Botrytis cinerea is reported worldwide. Resistance to MBCs in B. cinerea is related to mutations in the target β-tubulin gene (TUB2). Compared with the mutation at codon 200, the substitutions from glutamic acid to alanine (E198A), valine (E198V), or lysine (E198K) at codon 198 are currently the predominant mutations in MBC resistant populations of B. cinerea. In this study, a loop-mediated isothermal amplification (LAMP) method was established for rapid detection of benzimidazole resistance in B. cinerea. On the basis of the three mutations at TUB2 codon 198, three sets of LAMP primers were designed, and each of these primer sets was able to specifically amplify the DNA containing its corresponding mutation, while no amplification was detected with other mutated or the wild type DNA. After optimization, the sensitivity and specificity tests illustrated that this LAMP assay had good sensitivity and specificity to detect specific resistant genotypes in B. cinerea. Result also showed that the LAMP assay had good reproducibility. Above all, boiled mycelia could be used as templates, which simplified the process and increased the efficiency of the assay. Considering its rapidity, simplicity, and high efficiency, the LAMP assay developed in this study is a promising tool for the diagnosis of benzimidazole resistance in B. cinerea, and will contribute to the monitoring of resistance development to MBCs in the future. More... »

PAGES

10

References to SciGraph publications

  • 2014-10. The rising threat of fungicide resistance in plant pathogenic fungi: Botrytis as a case study in CHEMISTRY CENTRAL JOURNAL
  • 2012-07. Development and evaluation of a loop-mediated isothermal amplification assay for detection of Erwinia amylovora based on chromosomal DNA in EUROPEAN JOURNAL OF PLANT PATHOLOGY
  • 2016-11. One-step reverse transcription loop-mediated isothermal amplification for the detection of Capsicum chlorosis virus in Phalaenopsis orchids in JOURNAL OF GENERAL PLANT PATHOLOGY
  • 2015-05. Establishment and application of a loop-mediated isothermal amplification (LAMP) system for detection of cry1Ac transgenic sugarcane in SCIENTIFIC REPORTS
  • 2009-09. Identification of a novel point mutation in the β-tubulin gene of Botrytis cinerea and detection of benzimidazole resistance by a diagnostic PCR-RFLP assay in EUROPEAN JOURNAL OF PLANT PATHOLOGY
  • 2015-12. Development and application of loop-mediated isothermal amplification for detecting the highly benzimidazole-resistant isolates in Sclerotinia sclerotiorum in SCIENTIFIC REPORTS
  • 2010-04. Detection and characterization of benzimidazole resistance of Botrytis cinerea in greenhouse vegetables in EUROPEAN JOURNAL OF PLANT PATHOLOGY
  • 2013-06. Loop-mediated isothermal amplification (LAMP): recent progress in research and development in JOURNAL OF INFECTION AND CHEMOTHERAPY
  • 2008-05. Loop-mediated isothermal amplification (LAMP) of gene sequences and simple visual detection of products in NATURE PROTOCOLS
  • 2015-05. Development and application of loop-mediated isothermal amplification for detection of the F167Y mutation of carbendazim-resistant isolates in Fusarium graminearum in SCIENTIFIC REPORTS
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    URI

    http://scigraph.springernature.com/pub.10.1186/s42483-019-0016-8

    DOI

    http://dx.doi.org/10.1186/s42483-019-0016-8

    DIMENSIONS

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