ABC转运蛋白基因在草地贪夜蛾对茚虫威抗性形成中的功能分析  

Functional analysis of ABC transporter genes in the development of resistance to indoxacarb in Spodoptera frugiperda(Lepidoptera:Noctuidae)

作　　者：胡榕 陈慧淋 徐翔 吕佳鑫 彭安春 蒲建 杨继芝[1] 王学贵[1,3] HU Rong;CHEN Hui-Lin;XU Xiang;L Jia-Xin;PENG An-Chun;PU Jian;YANG Ji-Zhi;WANG Xue-Gui(College of Agronomy,Sichuan Agricultural University,Chengdu 611130,China;Plant Protection Station,Sichuan Provincial Department of Agriculture and Rural Affairs,Chengdu 610041,China;State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China,Sichuan Agricultural University,Chengdu 611130,China)

机构地区：[1]四川农业大学农学院,成都611130 [2]四川省农业农村厅植物保护站,成都610041 [3]四川农业大学,西南作物基因资源发掘与利用国家重点实验室,成都611130

出　　处：《昆虫学报》2024年第12期1597-1605,共9页Acta Entomologica Sinica

基　　金：四川省草地贪夜蛾抗药性监测及其抗药性基因分子检测(2022YFSY0034);全国区域草地贪夜蛾绿色防控技术集成与示范(2021YFD1400705-2)。

摘　　要：【目的】本研究旨在探讨ABC转运蛋白基因对草地贪夜蛾Spodoptera frugiperda茚虫威抗性形成的生物学功能,为该害虫的综合防治提供理论依据。【方法】采用点滴法以茚虫威单独处理及ABC转运蛋白抑制剂维拉帕米盐酸盐与茚虫威联合处理草地贪夜蛾茚虫威抗性种群DC-22和敏感品系WH的3龄幼虫,24 h时计算致死中浓度(median lethal concentration,LC_(50))和维拉帕米盐酸盐对茚虫威的增效比;利用RT-qPCR检测草地贪夜蛾茚虫威敏感品系WH及4个茚虫威抗性种群(DC-22,CX-22,MY-22和RH-22)3龄幼虫中7个ABC转运蛋白基因(SfABCG20,SfABCC2,SfABCF4,SfABCA1,SfABCA5,SfABCG23和SfABCG9)的表达量;通过对草地贪夜蛾DC-22和WH的3龄幼虫注射ds SfABCG 23,对响应茚虫威高表达的ABC转运蛋白基因SfABCG23进行RNAi,48 h后利用RT-qPCR检测SfABCG23的表达量,并测定RNAi后LC_(30)浓度茚虫威处理24 h时的死亡率。【结果】维拉帕米盐酸盐显著提高了草地贪夜蛾茚虫威抗性种群DC-22对茚虫威的敏感性,增效比达1.73。相对于敏感品系WH,SfABCG23在茚虫威抗性种群DC-22和CX-223龄幼虫中表达量分别上调2.56和4.05倍,且SfABCG23表达量与抗性倍数呈显著正相关,相关系数为0.941。注射ds SfABCG23后,茚虫威抗性种群DC-22和敏感品系WH中基因沉默效率分别为65.04%和39.55%;与注射ds GFP的对照组相比,注射ds SfABCG 23后茚虫威处理24 h时茚虫威抗性种群DC-22和敏感品系WH 3龄幼虫的死亡率分别增加30.55%和25.00%。【结论】本研究结果提示,ABC转运蛋白基因在草地贪夜蛾茚虫威抗性种群中对抗药性形成起重要调节作用,SfABCG23的过表达可能在草地贪夜蛾对茚虫威抗性形成中起重要作用。【Aim】The aim of this study is to investigate the biological function of ABC transporter genes in the development of resistance to indoxacarb in Spodoptera frugiperda,so as to provide a theoretical basis for the comprehensive control of this pest.【Methods】Indoxacarb alone and combined with ABC transporter inhibitor verapamil hydrochloride were used to treat the 3rd instar larvae of the indoxacarb-resistant population DC-22 and the indoxacarb-susceptible strain WH of S.frugiperda by the topical application method,and the median lethal concentration(LC_(50))and the synergistic ratio of verapamil hydrochloride to indoxacarb were calculated at 24 h after treatment.The expression levels of seven ABC transporter genes(SfABCG20,SfABCC2,SfABCF4,SfABCA1,SfABCA5,SfABCG23 and SfABCG9)in the 3rd instar larvae of the indoxacarb-susceptible strain WH and four indoxacarb-resistant populations,including DC-22,CX-22,MY-22 and RH-22,were examined.The highly expressed ABC transporter gene SfABCG23 in response to indoxacarb was silenced through RNAi by injecting ds SfABCG23 into the 3rd instar larvae of DC-22 and WH.The expression level of SfABCG23 was detected by RT-qPCR at 48 h after RNAi,and the mortality was detected at 24 h after exposure to LC_(30)of indoxacarb following RNAi.【Results】Verapamil hydrochloride significantly increased the susceptibility of the indoxacarb-resistant population DC-22 to indoxacarb,with the synergistic ratio of 1.73.The expression levels of SfABCG23 in the 3rd instar larvae of the indoxacarb-resistant populations DC-22 and CX-22 were up-regulated by 2.56-and 4.05-fold,respectively,as compared with that in the indoxacarb-susceptible strain WH,and the expression level of SfABCG23 was significantly positively correlated with the resistance ratio,with the correlation coefficient of 0.941.After ds SfABCG23 injection,the gene silencing efficiency was 65.04%and 39.55%,respectively,in the indoxacarb-resistant population DC-22 and the indoxacarb-susceptible strain WH,and compared with the ds GFP-in

关 键 词：草地贪夜蛾 抗药性 茚虫威 ABC转运蛋白 维拉帕米盐酸盐 增效作用 

分 类 号：Q966[生物学—昆虫学]