梨小食心虫谷胱甘肽S-转移酶基因序列分析及其与吡虫啉的相关性  

Sequence analysis of the glutathione S-transferases gene of oriental fruit moth Grapholita molesta and its correlation with imidacloprid

作　　者：庞钦玮 郭灵 张学尧[2] 高玲玲 马瑞燕[1] 郭艳琼[1] Pang Qinwei;Guo Ling;Zhang Xueyao;Gao Lingling;Ma Ruiyan;Guo Yanqiong(College of Plant Protection,Shanxi Agricultural University,Taigu 030801,Shanxi Province,China;Institute of Applied Biology,Shanxi University,Taiyuan 030006,Shanxi Province,China;Commonwealth Scientific and Industrial Research Organisation(CSIRO),Agriculture and Food,Wembley 6913,WA,Australia)

机构地区：[1]山西农业大学植物保护学院,太谷030801 [2]山西大学应用生物学研究所,太原030006 [3]澳大利亚联邦科学与工业研究组织农业与食品部,文布利6913

出　　处：《植物保护学报》2023年第2期459-467,共9页Journal of Plant Protection

基　　金：山西省自然科学研究面上项目(202103021224153);国家现代农业产业技术体系(CARS-28-19);山西农业大学植物保护学院“植保先锋”项目(2021ZBXF012)。

摘　　要：为明确梨小食心虫Grapholita molesta谷胱甘肽S-转移酶(glutathione S-transferase,GST)对吡虫啉的代谢作用,基于不同亚致死浓度吡虫啉处理的梨小食心虫转录组数据库进行筛选,并通过生物信息学分析、系统发育树构建、保守位点序列比对、分子对接和表达谱分析来确定梨小食心虫代谢吡虫啉过程中的关键GST。结果显示,共筛选到梨小食心虫的17个GST基因,其中GmGSTS1、GmGSTD2、GmGSTE5和GmGSTE4基因编码的蛋白与苹果蠹蛾Cydia pomonella的GST蛋白有较高的相似性,相似度达69.12%~89.66%。梨小食心虫GmGSTD2蛋白G位点和H位点的保守氨基酸与褐飞虱Nilaparvata lugens及烟粉虱Bemisia tabaci代谢吡虫啉相关GST氨基酸序列完全一致。吡虫啉与梨小食心虫GmGSTD2蛋白G位点的ARG-66和H位点的TYR-112可以形成稳定的氢键;与GmGSTE3蛋白G位点的LEU-45和H位点的LYS-123可以形成稳定的氢键。在吡虫啉处理后,梨小食心虫体内GmGSTD2和GmGSTE3基因的表达量显著高于对照。表明GmGSTD2和GmGSTE3为梨小食心虫代谢吡虫啉的关键GST基因。In order to understand the metabolism of imidacloprid by the glutathione S-transferase(GST)of oriental fruit moth Grapholita molesta,key GST genes were screened with bioinformatics analysis,phylogenetic tree construction,sequence alignment,molecular docking and expression analysis based on the transcriptome database established by treating G.molesta with different sublethal concentrations of imidacloprid.The results showed that 17 GSTs of G.molesta were identified,among which GmGSTS1,GmGSTD2,GmGSTE5 and GmGSTE4 showed high similarity with those of Cydia pomonella,with a similarity of 69.12%-89.66%.According to sequence alignment,the conserved amino acids at the G-site and H-site of GmGSTD2 were identical to the GSTs related to imidacloprid metabolism in Nilaparvata lugens and Bemisia tabaci.Imidacloprid could form stable hydrogen bonds with ARG-66 at the G-site and TYR-112 at the H-site of GmGSTD2,and with LEU-45 at the G-site and LYS-123 at the H-site of GmGSTE3.The expression of GmGSTD2 and GmGSTE3 in G.molesta was significantly higher than that of the control after imidacloprid treatment.It indicated that GmGSTD2 and GmGSTE3 were the key GST genes for metabolizing imidacloprid in G.molesta.

关 键 词：梨小食心虫 谷胱甘肽S-转移酶 吡虫啉 分子对接 实时荧光定量PCR 

分 类 号：S433.4[农业科学—农业昆虫与害虫防治]