甘蓝夜蛾CYP9A90基因的克隆及溴氰菊酯对其诱导表达  

Cloning of CYP9 A90 and effects of deltamethrin on its mRNA expression in Mamestra brassicae

作　　者：刘月庆[1] 樊星[2] 周夏[1] 钱金[1] 杨文庆[1] 杨静[1] 樊东[1] Liu Yueqing Fan Xing Zhou Xia Qian Jin Yang Wenqing Yang Jing Fan Dong(College of Agriculture,Northeast Agricultural University, Harbin 150030, China College of Resources and Environment, Northeast Agricultural University, Harbin 150030, China)

机构地区：[1]东北农业大学农学院,哈尔滨150030 [2]东北农业大学资源与环境学院,哈尔滨150030

出　　处：《植物保护》2017年第1期54-60,82,共8页Plant Protection

基　　金：黑龙江省教育厅科学技术研究项目(12531045)

摘　　要：P450 CYP9A家族基因与昆虫对拟除虫菊酯类杀虫剂的抗性相关。为证实溴氰菊酯对甘蓝夜蛾CYP9A基因的诱导效果,本研究采用RT-PCR和RACE技术克隆获得甘蓝夜蛾CYP9A基因,real-time PCR检测该基因在甘蓝夜蛾不同组织中的表达差异及溴氰菊酯处理甘蓝夜蛾5龄幼虫不同时间后该基因相对表达量变化,研究结果可为甘蓝夜蛾对溴氰菊酯的抗性治理提供依据。结果表明:克隆得到甘蓝夜蛾CYP9A基因cDNA全长序列,该序列包含1 828bp,包括1个125bp的5′非编码区,1个104bp的3′非编码区和1个1 599bp的开放阅读框,编码532个氨基酸,分子量约为61.1kDa,等电点为8.84,GenBank登录号为KR676343,被国际P450命名委员会命名为CYP9A90。Real-time PCR分析结果表明,该基因在甘蓝夜蛾5龄幼虫6个组织中表达情况不同,其中在脂肪体中表达量最高。低剂量溴氰菊酯作用后不同时间点CYP9A90mRNA总体呈现诱导表达趋势。Genes in CYP9A subfamily of P450 are associated with the resistance of insects to pyrethroids. In order to verify the induction effect of deltamethrin to CYP9A in Mamestra brassicae, RT-PCR and RACE were used to clone the full-length cDNA sequence of CYP9A gene. Its mRNA expression profiles in different tissues at fifth-instar larvae and the effect of deltamethrin were detected by real-time PCR. The results would give us a basis for resolving the management problem of the resistance of M. brassicae to deltamethrin. The results showed that the cDNA sequence was 1 828 bp in length, which contained a 125 bp 5＇-untranslated region, a 104 bp 3＇ untranslated region and a 1 599 bp open reading frame, and coded a polypeptide of 532 amino acid residues with a predicted molecular weight of 61.1 kDa and pl 8.84 （GenBank accession no. KR676343）. It was named CYP9A90 by the P450 Nomenclature Committee. The real-time quantitative RT-PCR assay showed that CYP9A90 gene expression levels were different in six tissues of the fifth-instar larvae. The expression level was at the highest point in fat body. Low dosage of deltamethrin treatment could induce CYP9A90 mRNA expression at different time points after treatment.

关 键 词：甘蓝夜蛾 P450 基因克隆 溴氰菊酯 表达 

分 类 号：Q785[生物学—分子生物学] S436.35[农业科学—农业昆虫与害虫防治]