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首页> 外文期刊>Plant Disease >Simultaneous Detection of Multiple Benzimidazole-Resistant p-Tubulin Variants of Botrytis cinerea using Loop-Mediated Isothermal Amplification
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Simultaneous Detection of Multiple Benzimidazole-Resistant p-Tubulin Variants of Botrytis cinerea using Loop-Mediated Isothermal Amplification

机译：使用环介导的等温扩增同时检测肉豆蔻菌的多种苯并咪唑抗性P-管蛋白变体

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摘要

Optimal disease management depends on the ability to monitor the development of fungicide resistance in plant pathogen populations. Benzimid-azole resistance is caused by the point mutations of the p-tubulin gene in Botrytis cinerea, and three mutations (E198A, E198K, and E198V) at codon 198 account for more than 98% of all resistant strains. Although traditional methods remain a cornerstone in monitoring fungicide resistance, molecular methods that do not require the isolation of pathogens can detect resistance alleles present at low frequencies, and require less time and labor than traditional methods. In this study, we present an efficient, rapid, and highly specific method for detecting highlybenzimidazole-resistant B. cinerea isolates based on loop-mediated isothermal amplification (LAMP). By using specific primers, we could simultaneously detect all three resistance-conferring mutations at codon 198. The LAMP reaction components and conditions were optimized, and the best reaction temperatures and times were 60 to 62°C and 45 min, respectively. When B. cinerea field isolates were assessed for benz-imidazole resistance, similar results were obtained with LAMP, minimal inhibition concentration, and sequencing. The LAMP assay developed in the current study was highly suitable for detection of highly benzimidazole-resistant field isolates of B. cinerea.
机译：最佳疾病管理取决于监测植物病原体群中杀菌剂抗性的发展的能力。苯并咪唑抗性是由Botrytis Cinerea的P-Timulin基因的点突变引起的，并在CoCon 198处占所有抗性菌株的98％以上的三个突变（E198a，E198k和E198V）。虽然传统方法仍然是监测杀菌剂抗性的基石，但不需要分离病原体的分子方法可以检测在低频下存在的阻力等位基因，并且比传统方法需要更少的时间和劳动力。在这项研究中，我们提出了一种基于环介导的等温扩增（灯）的高效，快速，高度特异性的方法，用于检测高度低位咪唑抗性B.灰质分离物。通过使用特异性引物，我们可以同时检测密码子198处的所有三个电阻赋予突变。灯反应组分和条件优化，最佳的反应温度和时间分别为60至62℃和45分钟。当评估B.Cherea场分离株时，对苯甲酰咪唑抗性评估，用灯，最小抑制浓度和测序获得了类似的结果。目前研究中开发的灯测定非常适合于检测B.Careea的高苯并咪唑抗性场分离株。

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  • 来源
    《Plant Disease》 |2018年第10期|共9页
  • 作者

    Ya Bing Duan; Ying Yang; Jian Xin Wang; Chang Jun Chen; Gero Steinberg; Bart A Fraaije; Ming Guo Zhou;

  • 作者单位

    College of Plant Protection State &

    Local Joint Engineering Research Center of Green Pesticide Invention and Application Nanjing Agricultural University Nanjing Jiangsu Province 210095 China;

    College of Plant Protection State &

    Local Joint Engineering Research Center of Green Pesticide Invention and Application Nanjing Agricultural University Nanjing Jiangsu Province 210095 China;

    College of Plant Protection State &

    Local Joint Engineering Research Center of Green Pesticide Invention and Application Nanjing Agricultural University Nanjing Jiangsu Province 210095 China;

    College of Plant Protection State &

    Local Joint Engineering Research Center of Green Pesticide Invention and Application Nanjing Agricultural University Nanjing Jiangsu Province 210095 China;

    Biosciences University of Exeter Stacker Road Exeter EX4 4QD UK;

    Biointeractions &

    Crop Protection De-partment Rothamsted Research Harpenden Hertfordshire 'AL5 2JQ UK;

    College of Plant Protection State &

    Local Joint Engineering Research Center of Green Pesticide Invention and Application Nanjing Agricultural University;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 植物保护;
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