我国草地贪夜蛾田间种群有机磷和氨基甲酸酯类杀虫剂靶标基因ace-1的基因型和突变频率  

Genotype and mutation frequency of ace-1,the target gene of organophosphorus and carbamate insecticides,in field populations of Spodoptera frugiperda(Lepidoptera:Noctuidae)in China

作　　者：李妍 龚丽凤 王欢欢 李曦 孙歌 谷少华 梁沛[2] 高希武[2] LI Yan;GONG Li-Feng;WANG Huan-Huan;LI Xi;SUN Ge;GU Shao-Hua;LIANG Pei;GAO Xi-Wu(State Key Laboratory for Biology of Plant Diseases and Insect Pests,Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193,China;Department of Entomology,China Agricultural University,Beijing 100193,China)

机构地区：[1]中国农业科学院植物保护研究所,植物病虫害生物学国家重点实验室,北京100193 [2]中国农业大学昆虫学系,北京100193

出　　处：《昆虫学报》2020年第5期574-581,共8页Acta Entomologica Sinica

基　　金：国家重点研发计划项目“草地贪夜蛾防控关键技术研究与集成示范”(2019YFD0300103);科技基础资源调查专项“重要作物害虫抗药性基因和敏感基因资源调查”(2018FY101100);国家自然科学基金项目(31772164)。

摘　　要：【目的】对入侵我国的草地贪夜蛾Spodoptera frugiperda有机磷和氨基甲酸酯类杀虫剂靶标基因ace-1的基因型进行分子检测,明确抗性基因频率,进而指导田间科学用药。【方法】采集中国12省份的草地贪夜蛾田间种群幼虫样本,提取单头样本的基因组DNA,利用特异性引物进行PCR扩增,获得ace-1基因片段。根据碱基、氨基酸序列比对和测序峰图分析,明确与有机磷和氨基甲酸酯类杀虫剂抗性相关的3个氨基酸突变位点A201S, G227A和F290V的基因型和抗性基因频率。【结果】通过DNA检测分析中国12省份草地贪夜蛾田间种群589头个体ace-1基因的基因型和突变频率发现,在A201S位点检测到137头个体为抗性杂合基因型,抗性基因频率为11.6%,未发现抗性纯合基因型个体;G227A位点589头个体均为敏感纯合基因型;F290V位点的抗性基因频率最高,达到57.1%,携带抗性基因的个体数量达到523头(占样本总数的88.8%)。【结论】结果表明入侵我国的草地贪夜蛾种群携带高频率的对有机磷和氨基甲酸酯类杀虫剂抗性基因。田间防治建议不用或少用有机磷和氨基甲酸酯类杀虫剂,同时进一步加强田间抗性监测工作。【Aim】The objective of this study is to detect the genotype of ace-1,the target gene of organophosphorus and carbamate insecticides,in field populations of Spodoptera frugiperda in China,and to identify the frequency of resistance genes,so as to provide guidance for the scientific use of insecticides in the field.【Methods】Samples of S.frugiperda larvae were collected from maize fields in 12 provinces in China,and the genomic DNA was extracted individually.Then the ace-1 gene fragment was cloned using gene specific primers,and the genotype and resistance gene frequency of three amino acid mutation sites,A201S,G227A and F290V,were analyzed through base and amino acid sequence alignment,DNA sequencing and peakTrace Basecaller analysis.【Results】The genotype and mutation frequency of the ace-1 gene of 589 individuals of field populations of S.frugiperda from 12 provinces in China analyzed through DNA detection method revealed that 137 individuals exhibited resistant heterozygous genotype at locus A201S,and the resistance gene frequency reached 11.6%.Meanwhile,no resistant homozygous genotype was found.At locus G227A,all the 589 individuals displayed susceptible homozygous genotypes.The resistance gene frequency at locus F290V was the highest(57.1%),and there were 523 individuals carrying homozygous or heterozygous resistance genes,accounting for 88.8%of the total samples.【Conclusion】The results indicate that S.frugiperda invading in China carries a high-level frequency of resistance genes to organophosphorus and carbamate insecticides.Organophosphorus and carbamate insecticides should not or less be used in the field control of S.frugiperda,and resistance monitoring in fields should be strengthened.

关 键 词：草地贪夜蛾 ace-1 基因型 抗性基因频率 有机磷杀虫剂 氨基甲酸酯类杀虫剂 抗药性 

分 类 号：Q965.9[生物学—昆虫学]