白背飞虱细胞色素P450还原酶基因的分子特征与表达模式分析  

Molecular characterization and expression profiles of cytochrome P450 reductase gene in Sogatella furcifera(Hemiptera:Delphacidae)

作　　者：余航[1] 刘苏[1] 朱晴子[1] 周文武[1] 梁庆梅[1] 史肖肖[1] 祝梓杰 祝增荣[1] 

机构地区：[1]浙江大学昆虫科学研究所/水稻生物学国家重点实验室/农业部农业昆虫学重点实验室,杭州310058

出　　处：《浙江大学学报（农业与生命科学版）》2016年第4期391-400,共10页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：农业部公益性行业(农业)科研专项(201403030);国家自然科学基金(31371935);国家重点基础研究发展计划(973计划)(2010CB126200);亚洲发展银行-国际水稻研究所(ADB-IRRI)稻飞虱项目(RETA13;RETA14;RETA15)

摘　　要：从水稻重要害虫白背飞虱(Sogatella furcifera)中克隆了一条编码细胞色素P450还原酶(cytochrome P450reductase,CPR)的全长cDNA,命名为SfCPR,其开放阅读框全长为2 034bp,编码一个含有677个氨基酸残基的蛋白质.SfCPR蛋白质的2级结构具有CPR的典型特征,例如N端跨膜区、黄素单核苷酸结合域、黄素腺嘌呤二核苷酸结合域和烟酰胺腺嘌呤二核苷酸磷酸结合域,以及保守的催化残基.系统进化分析结果显示,SfCPR与灰飞虱和褐飞虱的CPRs聚在同一分支.荧光定量聚合酶链式反应结果显示:SfCPR基因在白背飞虱各龄期均有表达,其中以成虫表达量最高;在白背飞虱成虫各组织中均能够检测到SfCPR基因的表达,其中,以腹部表达量最高.使用亚致死剂量的溴氰菊酯、吡虫啉和噻嗪酮处理白背飞虱3龄若虫后,试虫的SfCPR表达水平均显著上升.上述结果为进一步研究该基因的生理功能奠定了基础.Summary Nicotinamide adenine dinucleotide phosphate （NADPH）-cytochrome P450 reductase （CPR） is an electron supplier for various cytochrome P450 monooxygenases. Most P450-mediated catalytic reactions in insects require involvement of CPR, such as detoxification of insecticides and plant secondary metabolites. So far, CPR genes have been identified and characterized from many model and non-model insect species. Since insect P450 system is one of the most important metabolic adaptive traits involved in the degradation of xenobiotics and regulation of endogenous substrates. CPR, as the indispensable electron donor of P450 system, has attracted increasing attention as a potential candidate to develop novel chemical inhibitor to manage target insect pests. Rice planthoppers, such as Nilaparvata lugens, Laodelphaa- striatellus and Sogatella furcifera, are considered to be the most serious pests of rice. Previously, some studies on L. striatellus and N. lugens found that silencing the CPR gene by RNAi technology could increase their sensitivity to insecticides, but little was known about S. furcifera. This work firstly reports the identification of CPR gene in S. furcifera and up-regulation of the CPR transcription by chemical insecticides. The research will facilitate further study on the function of CPR in S. furcifera. In this study, a full-length eDNA encoding CPR was cloned from S. furcifera. The phylogenetic relationships of SfCPR with other insect CPRs were estimated by neighbor-joining method, and its distribution in various tissues and different developmental stages were analyzed by reM-time quantitative polymerase chain reaction （qPCR）. Finally, after exposure of deltamethrin, buprofezin and imidacloprid at sublethal concentrations for 6, 12 and 24 h, the relative expression levels of SfCPR in the third instar nymphs were investigated. By searching the transeriptome data sets of S. furcifera, a eDNA fragment encoding putative CPR （named SfCPR） was identified. This eDNA fragment was then amplif

关 键 词：细胞色素P450还原酶 白背飞虱 杀虫剂 表达模式 

分 类 号：S435.112.3[农业科学—农业昆虫与害虫防治] Q963[农业科学—植物保护]