Q型烟粉虱细胞色素P450 CYP6DV5基因克隆及其在烟粉虱对噻虫嗪抗性中的作用  

Cloning and expression profiles of cytochrome P450 CYP6DV5 in Bemisia tabaci and its role in resistance of B.tabaci to thiamethoxam

作　　者：杨峰山[1] 殷城 杨静 危学高 杜田华 杨鑫[2] 王少丽[2] 张友军[2] YANG Fengshan;YIN Cheng;YANG Jing;WEI Xuegao;DU Tianhua;YANG Xin;WANG Shaoli;ZHANG Youjun(College of Life Science,Heilongjiang University,Heilongjiang Provincial Key Laboratory of Ecological Restoration and Resource Utilization for Cold Region,Engineering Research Center of Agricultral Microbiology Technology,Ministry of Education,Key Laboratory of Microbiology,College of Heilongjiang Province,Harbin 150030,China;Institute of Vegetables and Flowers,Chinese Academy of Agricultural Science,Beijing 100081,China)

机构地区：[1]黑龙江大学生命科学学院,黑龙江省寒地生态修复与资源利用重点实验室,农业微生物技术教育部工程研究中心,黑龙江省普通高校分子生物学重点实验室,哈尔滨150030 [2]中国农业科学院蔬菜花卉研究所,北京100081

出　　处：《农药学学报》2021年第1期83-89,共7页Chinese Journal of Pesticide Science

基　　金：国家重点研发计划(2018YFD0201205,2016YFD0200500);现代农业产业技术体系专项资金资助(CARS-24-C-02);北京市重点实验室.

摘　　要：烟粉虱Bemisia tabaci已严重危害农作物的正常生长,而新烟碱类杀虫剂噻虫嗪已被广泛用于烟粉虱的防治,但由于常年应用,致使烟粉虱对噻虫嗪产生了严重的抗性,而目前其抗药性分子机制尚不明确。本研究通过荧光定量PCR比较分析烟粉虱噻虫嗪抗性及敏感种群,发现细胞色素P450基因CYP6DV5在噻虫嗪抗性品系中的表达量为敏感品系的2.95倍,依据基因组数据库克隆该基因的全长序列,序列分析结果表明,该基因具有细胞色素P450基因的保守结构域,属于典型的昆虫P450基因。进一步分析其在不同发育阶段和部位中的表达量,发现细胞色素P450基因CYP6DV5在烟粉虱成虫阶段和头部表达丰富,同时噻虫嗪能够显著诱导该基因的表达。通过RNA干扰试验,发现敲低CYP6DV5基因表达能够显著提高烟粉虱对噻虫嗪的敏感性。结果表明,CYP6DV5基因参与了Q型烟粉虱对噻虫嗪抗性的形成。研究结果将有助于揭示烟粉虱对噻虫嗪的抗性形成机制,并为烟粉虱的综合治理提供理论基础。Bemisia tabaci seriously damages the normal growth of agriculture crops.Thiamethoxam is a secondary product of neonicotinoid insecticide that is widely used in the control of B.tabaci.However,due to the increasing usage of thiamethoxam,B.tabaci has developed severe resistance to thiamethoxam in the fields.The mechanism of B.tabaci resistance to thiamethoxam is still unknown.In this study,we found that cytochrome P450 CYP6DV5 gene was overexpressed(2.95-fold)in a thiamethoxam resistant strain by comparing with the susceptible strain(Q-biotype).The full-length sequence of the gene was cloned according to the genome database.Sequence analysis results showed that the gene had conserved domains of cytochrome P450 gene,which was a typical insect P450 gene.Further analysis of the expression of CYP6DV5 in different developmental stages and body parts showed that CYP6DV5 gene was abundant in the adult stage and head of B.tabaci.We also found that thiamethoxam could significantly induce the expression of this gene.Finally,the knockdown of CYP6DV5 gene could significantly increase the sensitivity of B.tabaci to thiamethoxam.Results demonstrated that CYP6DV5 was involved in thiamethoxam resistance in B.tabaci.The results will contribute to reveal the mechanism of resistance formation of B.tabaci to thiamethoxam,and will provide a theoretical basis for the management of B.tabaci.

关 键 词：烟粉虱 细胞色素P450 CYP6DV5基因 基因克隆 RNA干扰 噻虫嗪抗性 

分 类 号：Q78[生物学—分子生物学] S481.4[农业科学—农药学]