Rapid Detection of A282S Mutation in the <i>RDL1</i> Gene of Rice Stem Borer via the Mutation-Specific LAMP Technique
Zijiao Song,
Sugui Zhang,
Wentao Man,
Yao Li,
Guanghua Luo,
Jinyao Yu,
Yilan Fu,
Junxi Yao,
Yiqu Chen,
Chunqing Zhao
Affiliations
Zijiao Song
Key Laboratory of Integrated Pest Management on Crops in East China, Ministry of Agriculture and Rural Affairs, Department of Entomology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China
Sugui Zhang
Key Laboratory of Integrated Pest Management on Crops in East China, Ministry of Agriculture and Rural Affairs, Department of Entomology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China
Wentao Man
Key Laboratory of Integrated Pest Management on Crops in East China, Ministry of Agriculture and Rural Affairs, Department of Entomology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China
Yao Li
Key Laboratory of Integrated Pest Management on Crops in East China, Ministry of Agriculture and Rural Affairs, Department of Entomology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China
Guanghua Luo
Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China
Jinyao Yu
Key Laboratory of Integrated Pest Management on Crops in East China, Ministry of Agriculture and Rural Affairs, Department of Entomology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China
Yilan Fu
Key Laboratory of Integrated Pest Management on Crops in East China, Ministry of Agriculture and Rural Affairs, Department of Entomology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China
Junxi Yao
Key Laboratory of Integrated Pest Management on Crops in East China, Ministry of Agriculture and Rural Affairs, Department of Entomology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China
Yiqu Chen
College of Plant Science, Tibet Agriculture and Animal Husbandry University, Nyingchi 860000, China
Chunqing Zhao
Key Laboratory of Integrated Pest Management on Crops in East China, Ministry of Agriculture and Rural Affairs, Department of Entomology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China
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/μL), dNTP (0.8 mM), Mg2+ (4 mM), betaine (0.6 M), forward inner primer and backward inner primer (1.6 μM), F3 and B3 (0.4 μM), and hydroxyl naphthol blue (150 mM), respectively, and the optimal reaction condition was 63 °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.
