湖北四个地区草地贪夜蛾田间种群的杀虫剂敏感性及靶标突变检测  

Detection of insecticide sensitivity and target site mutations in field populations of Spodoptera frugiperda(Lepidoptera:Noctuidae)in four regions of Hubei,central China

作　　者：郭志敏 邓晓倩 李静 袁茂钧 万虎 李建洪 马康生 GUO Zhi-Min;DENG Xiao-Qian;LI Jing;YUAN Mao-Jun;WAN Hu;LI Jian-Hong;MA Kang-Sheng(Hubei Insect Resources Utilization and Sustainable Pest Management Key Laboratory,College of Plant Science and Technology,Huazhong Agricultural University,Wuhan 430070,China)

机构地区：[1]华中农业大学植物科学技术学院,昆虫资源利用与害虫可持续治理湖北省重点实验室,武汉430070

出　　处：《昆虫学报》2020年第5期582-589,共8页Acta Entomologica Sinica

基　　金：中央高校基本科研业务费专项基金(2662019QD052);湖北省农业科技创新行动项目(NYKJ2019011)。

摘　　要：【目的】测定湖北草地贪夜蛾田间种群对4种杀虫剂的敏感性,并对杀虫剂靶标Ace-1,VGSC和RyR的基因型和突变频率进行检测,以明确湖北草地贪夜蛾田间种群药剂敏感性现状,进而指导田间科学用药。【方法】采集湖北黄冈、武穴、咸宁和荆州4个地区玉米田中的草地贪夜蛾幼虫,采用浸叶法测定其对氯虫苯甲酰胺、乙基多杀菌素、茚虫威和甲维盐4种杀虫剂的敏感性;提取4个种群80头个体的单头总RNA合成cDNA,利用特异性引物进行PCR扩增,获得目的基因片段,根据序列比对和测序峰图分析,确定Ace-1,VGSC和RyR的基因型和靶标突变位点的突变频率。【结果】生测结果表明,湖北这4个田间种群的草地贪夜蛾幼虫对氯虫苯甲酰胺、乙基多杀菌素、茚虫威和甲维盐均比较敏感,敏感性从高到低依次为:甲维盐>乙基多杀菌素>氯虫苯甲酰胺>茚虫威。分子检测结果表明,湖北草地贪夜蛾这4个田间种群Ace-1基因在A201S, G227A和F290V位点均存在抗性杂合突变,且在F290V位点检测到抗性纯合突变,而VGSC和RyR两个基因均没有检测到靶标位点突变。【结论】湖北草地贪夜蛾4个田间种群对氯虫苯甲酰胺、乙基多杀菌素、茚虫威和甲维盐这4种杀虫剂均较敏感,但这4个草地贪夜蛾田间种群均携带有对有机磷和氨基甲酸酯类杀虫剂的抗性基因,存在潜在的抗性风险,田间防治建议少用或不用该类杀虫剂,同时需要进一步加强抗性监测工作。【Aim】The aim of this study is to evaluate the sensitivity of field populations of the fall worm,Spodoptera frugiperda,in Hubei,central China to four insecticides,and to detect the genotype and mutation frequency of the insecticide target genes Ace-1,VGSC and RyR,so as to ascertain the insecticide sensitivity of field populations of S.frugiperda in Hubei and to provide guidance for scientific application of pesticides.【Methods】The sensitivity of S.frugiperda larvae collected from corn fields in four regions,Huanggang,Wuxue,Xianning and Jingzhou of Hubei to four insecticides,chlorantraniliprole,spinetoram,indoxacarb and methylamino abamectin benzoate,was investigated using a standard leaf-dip bioassay.Eighty larvae from 4 regions were used for total RNA isolation and cDNA synthesis,and the target fragments were amplified by PCR using specific primers.Besides,the genotypes and target site mutation frequencies of Ace-1,VGSC,and RyR were determined by sequence alignment and sequencing chromatogram peak analysis.【Results】Toxicity assays showed that the four field populations of S.frugiperda larvae in Hubei were susceptible to all the tested insecticides,and the sensitivity of these populations to four insecticides was as follows:methylamino abamectin benzoate>spinetoram>chlorantraniliprole>indoxacarb.The sequencing results demonstrated that in the four field populations of S.frugiperda the resistant heterozygote of all three mutations of Ace-1(A201S,G227A,and F290V)was detected,while the resistant homozygote of F290V was detected.However,no site mutation in VGSC and RyR was detected in all the tested field populations of S.frugiperda.【Conclusion】Though the four field populations of S.frugiperda in Hubei are susceptible to the four tested insecticides(chlorantraniliprole,spinetoram,indoxacarb and methylamino abamectin benzoate),these field populations have already carried resistant genes conferring resistance to organophosphorus and carbamate insecticides and possess high resistance risk to these insecti

关 键 词：草地贪夜蛾 药剂敏感性 Ace-1 VGSC RYR 基因型 突变频率 

分 类 号：Q965.9[生物学—昆虫学]