摘要
细胞色素P450是昆虫体内一类代谢外源物质重要的酶系。为了解梨小食心虫细胞色素P450基因的分子特性及在阿维菌素代谢中的作用,对转录组数据库检索细胞色素P450基因、全长克隆,并对其进行生物信息学分析;并采用RT-qRCR测定目的基因在梨小食心虫不同发育阶段、不同组织及不同浓度阿维菌素处理成虫后的表达量。结果表明,通过克隆结果获得1个梨小食心虫细胞色素P450基因CYP6AE123的全长cDNA序列,开放阅读框(ORF)长1524 bp,编码507个氨基酸;其二级结构表明,该基因编码的蛋白主要由α螺旋和无规则卷曲组成;系统发育分析表明,此基因与苹果蠹蛾、小线角木蠹蛾、棉铃虫的氨基酸序列一致性分别高达83.67%、57.39%、52.61%。RT-qPCR分析结果表明,CYP6AE123在成虫期和5龄幼虫期的表达量较高,且在成虫脂肪体和中肠中有较高的表达。高浓度阿维菌素(179.66μg/mL)处理梨小食心虫后基因表达量显著增加,表明该基因可能在成虫对阿维菌素的解毒代谢中发挥重要作用。研究结果可为进一步通过RNAi进行功能验证提供理论依据。
Cytochrome P450 is one of the most important detoxification enzymes in insects and plays an important role in the metabolism of endogenous and exogenous compounds,including chemical insecticides.To identify the structure and function of cytochrome P450 gene in Grapholita molesta,the full-length cDNA sequence of cytochrome P450 gene,namely CYP6AE123,was blasted based on Grapholita molesta transcriptome data.Its structure was analyzed by bioinformation software,and the gene expression pattern of CYP6AE123 was further analyzed in different developmental stages,different tissues and different concentrations of abamectin using real-time quantitative RCR(RT-qPCR).The results showed that a full-length cDNA sequence of CYP6AE123 was 1524 bp,encoded 507 amino acids.The secondary structure of CYP6AE123 was composed of alpha helix and random coil.The phylogeny analysis indicated that CYP6AE123 protein had 83.67%,57.39%and 52.61%of similarity with Cydia pomonella,Streltzoviella insularis and Helicoverpa armigera,respectively,and Grapholita molesta was the most closely related to Mythimna separata.The RT-qPCR analysis showed that CYP6AE123 had a higher expression level in the adult stage and 5th instar larvae,and in fat body and midgut during the adult stage.The CYP6AE123 gene expression increased significantly after treated with high concentration(179.66μg/mL)of abamectin,indicating that it might play a crucial role in the detoxification metabolism of abamectin in adults.This study result will provide a theoretical basis for further functional verification using RNAi technology.
作者
韩慧
王彪龙
李恩
胡军
马瑞燕
高玲玲
郭艳琼
HAN Hui;WANG Biaolong;LI En;HU Jun;MA Ruiyan;GAO Lingling;GUO Yanqiong(College of Plant Protection,Shanxi Agricultural University,Taigu 030801,China;CSIRO,Agriculture and Food,Private Bag 5,Wembley,WA 6913,Austral)
出处
《山西农业科学》
2021年第5期539-544,共6页
Journal of Shanxi Agricultural Sciences
基金
山西省研究生教育创新项目(2020SY217)
山西省重点研发计划(农业)项目(201803D221004-7)
山西省留学人员科技活动择优资助项目(20200018)。
