Rapid Detection of A282S Mutation in the RDL1 Gene of Rice Stem Borer via the Mutation-Specific LAMP Technique

文献类型： 外文期刊

第一作者： Song, Zijiao

作者： Song, Zijiao;Zhang, Sugui;Man, Wentao;Li, Yao;Yu, Jinyao;Fu, Yilan;Yao, Junxi;Zhao, Chunqing;Luo, Guanghua;Chen, Yiqu

作者机构： Nanjing Agr Univ, Coll Plant Protect, Dept Entomol, Key Lab Integrated Pest Management Crops East Chin, Nanjing 210095, Peoples R China;Jiangsu Acad Agr Sci, Inst Plant Protect, Nanjing 210014, Peoples R China;Tibet Agr & Anim Husb Univ, Coll Plant Sci, Nyingchi 860000, Peoples R China

关键词： Chilo suppressalis; LAMP; RDL; resistance monitoring

期刊名称：AGRONOMY-BASEL （ 2022影响因子：3.7； 五年影响因子：4.0 ）

ISSN：

年卷期： 2023 年 13 卷 2 期

页码：

收录情况： SCI

摘要： Rice stem borer Chilo suppressalis (Walker) is one of the most serious pests on rice and is distributed worldwide. With the long-term and continuous usage of insecticides, C. suppressalis has developed high levels of resistance to various kinds of insecticides, including phenylpyrazole insecticides. As is well known, the resistance of C. suppressalis to phenylpyrazole insecticides is determined by the A282S mutation of the GABA receptor RDL subunit. In order to efficiently detect the resistance of C. suppressalis, a rapid and sensitive loop-mediated isothermal amplification (LAMP) technique was established and optimized in this study. The optimal concentration of components was Bst DNA polymerase (0.24 U/mu L), dNTP (0.8 mM), Mg2+ (4 mM), betaine (0.6 M), forward inner primer and backward inner primer (1.6 mu M), F3 and B3 (0.4 mu M), and hydroxyl naphthol blue (150 mM), respectively, and the optimal reaction condition was 63 degrees C for 60 min, which could reduce the cost and time of detection. In addition, the accuracy of the optimized LAMP reaction system and parameters was verified in the field strains of C. suppressalis from different regions, including Jiangsu, Jiangxi, and Hu'nan provinces. The mutation (A2'S) was successfully detected in the field strains. As far as we know, this is the first report of the LAMP technique applied in the resistance monitoring of C. suppressalis to phenylpyrazole insecticides. According to our results, the optimized LAMP reaction system is feasible and easy to operate and to efficiently detect resistance-related mutation in a short time, as directly judged by the naked eye. Our results provide a new tool for detection of resistance of C. suppressalis, which is a very useful tool for comprehensive management of C. suppressalis.

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