梨小食心虫ABC转运蛋白基因的克隆及其分子特性分析  

Cloning and molecular characterization of ATP-binding cassette transporter genes in oriental fruit moth Grapholita molesta

作　　者：杨燕玉 贺文杰 李朗程 高玲玲 马瑞燕[1] 郭艳琼[1] Yang Yanyu;He Wenjie;Li Langcheng;Gao Lingling;Ma Ruiyan;Guo Yanqiong(College of Plant Protection,Shanxi Agricultural University,Taigu 030801,Shanxi Province,China;Agriculture and Food,Commonwealth Scientific and Industrial Research Organization(CSIRO),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]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4期981-990,共10页Journal of Plant Protection

基　　金：国家现代农业产业技术体系(CARS-28-19);山西省自然科学研究面上项目(202103021224153);山西省研究生教育创新项目(2022Y345)。

摘　　要：为明确梨小食心虫Grapholita molesta三磷酸腺苷结合盒(ATP-binding cassette,ABC)转运蛋白基因的分子特性,采用反转录PCR(reverse transcription-PCR,RT-PCR)技术对筛选出的ABC转运蛋白基因进行克隆,对克隆所得序列进行生物信息学分析,并利用实时荧光定量PCR(real-time quantitative PCR,RT-qPCR)技术对其时空表达模式以及3种杀虫剂处理后的表达特性进行分析。结果表明,克隆得到3条梨小食心虫ABC转运蛋白基因,分别命名为GmABCB7、GmABCG4和GmABCG4X1,其开放阅读框分别为2151、1932和2106 bp,且均具有ABC转运蛋白家族基因的典型结构特征。GmABCB7和GmABCG4X1基因在梨小食心虫成虫期的相对表达量均最高,而GmABCG4基因在3龄幼虫期和成虫期显著高表达;GmABCB7基因在马氏管中显著高表达,其他2个基因则在脂肪体中显著高表达。与对照相比,在高浓度(LC50)的吡虫啉和高效氯氟氰菊酯以及低浓度(LC10)的吡虫啉和阿维菌素处理24 h后,梨小食心虫成虫中GmABCB7、GmABCG4和GmABCG4X1基因被显著诱导高表达。表明ABC转运蛋白可能参与梨小食心虫对吡虫啉、阿维菌素和高效氯氟氰菊酯的解毒代谢。In order to understand the molecular characteristics of the ATP-binding cassette(ABC)transporter genes of oriental fruit moth Grapholita molesta,the ABC transporter gene sequences were obtained by reverse transcription PCR(RT-PCR),and the cloned sequences were analyzed with bioinformatics.Real-time quantitative PCR(RT-qPCR)technology was used to analyze their spatiotemporal expression patterns and expression profiles after treatment with three insecticides.Three ABC transporter genes were cloned and named GmABCB7,GmABCG4,and GmABCG4X1,respectively.Their open reading frames were 2151,1932,and 2106 bp,respectively,and all contained the typical structural features of ABC transporter family genes.The RT-qPCR results showed that the relative expression levels of GmABCB7 and GmABCG4X1 were the highest in G.molesta adults;and GmABCG4 was significantly overexpressed in the 3rd instar larvae and adults.GmABCB7 was highly expressed in the Malpighian tubules and the other two genes were highly expressed in the fat body.After treatment for 24 hours,compared with the control group,the expressions of the three genes were significantly induced in G.molesta adults under high concentrations(LC50)of imidacloprid and L-cyhalothrin as well as low concentrations(LC10)of imidacloprid and avermectin.The results suggested that ABC transporter might be involved in the detoxification metabolism of imidacloprid,avermectin and L-cyhalothrin in G.molesta.

关 键 词：ABC转运蛋白 梨小食心虫 分子特性 表达分析 杀虫剂 

分 类 号：S433[农业科学—农业昆虫与害虫防治]