Q型烟粉虱UDP-糖基转移酶UGT354A1基因克隆及其在噻虫嗪抗性中的作用  

Cloning of the Bemisia tabaci Q biotype UDP-glycosyltransferase UGT354A1 gene and its role in thiamethoxam resistance

作　　者：杜田华 危学高 殷城 杨静 黄明娇 桂连友 杨鑫[2] 张友军 DU Tian-Hua;WEI Xue-Gao;YIN Cheng;YANG Jing;HUANG Ming-Jiao;GUI Lian-You;YANG Xin;ZHANG You-Jun(Institute of Insect,College of Agriculture,Yangtze University,Jingzhou 434025,China;Institute of Vegetables and Flowers,Chinese Academy of Agriculture Sciences,Beijing 100081,China)

机构地区：[1]长江大学农学院昆虫研究所,荆州434025 [2]中国农业科学院蔬菜花卉研究所,北京100081

出　　处：《应用昆虫学报》2022年第2期257-267,共11页Chinese Journal of Applied Entomology

基　　金：国家重点研发计划(2019YFD1002100);北京市自然科学基金(6212031)。

摘　　要：【目的】UDP-糖基转移酶(UGTs)是昆虫主要的Ⅱ期解毒酶,可能参与昆虫抗药性的形成。本研究旨在探究Q型烟粉虱中UGT基因是否参与其对噻虫嗪的抗药性。【方法】依据烟粉虱基因组数据库设计引物,克隆其UGT354A1基因的全长序列;采用qRT-PCR技术检测UGT354A1基因在烟粉虱不同发育阶段、不同组织部位以及抗敏品系中的表达量;通过RNA干扰试验,验证UGT354A1基因在烟粉虱噻虫嗪抗性中的作用。【结果】序列生物信息学分析发现,UGT354A1基因属于典型的昆虫UGTs,包括两个糖基供体(DBR1和DBR2)和一个保守特征性基序。qRT-PCR结果显示,UGT354A1基因在噻虫嗪抗性品系(THQR)中的表达量为敏感品系(THQS)的2.60倍,且噻虫嗪可显著诱导该基因的表达。在不同发育阶段和组织部位中,发现UGT354A1基因在烟粉虱若虫和成虫阶段及头部和胸部表达丰富。RNAi后,沉默UGT354A1基因能够显著增加噻虫嗪对Q型烟粉虱的毒杀作用。【结论】UGT354A1基因在Q型烟粉虱中对噻虫嗪具有重要的解毒作用,可能参与了抗性的形成。[Objectives]To investigate whether the Bemisia tabaci(Q-biotype)UDP-glycosyltransferase(UGT)gene is involved in resistance to thiamethoxam.[Methods]PCR primers were designed according to the genome database and the full-length sequence of the UGT354A1 gene was cloned.Expression levels of the UGT354A1 gene in different developmental stages and body parts,and in a thiamethoxam-resistant(THQR)and susceptible strain(THQS)were detected by q-PCR.The function of UGT354A1 gene in the resistance of B.tabaci to thiamethoxam was verified by RNAi.[Results]Sequence analysis shows that the UGT354A1 gene has a conserved UGTs domain that is typical of insect UGTs.Compared with the susceptible strain,the UGT354A1 gene was overexpressed(2.60-fold)in the thiamethoxam resistant strain.In addition,thiamethoxam could significantly induce the expression of the UGT354A1 gene.The experiments of the expression levels of the UGT354A1 gene in different developmental stages and body parts indicate that it is most highly expressed in nymphs and adults,and in the head and thorax.RNAi silencing of UGT354A1 gene expression significantly reduced the resistance of B.tabaci to thiamethoxam.[Conclusion]The UGT354A1 gene is involved in the formation of thiamethoxam resistance in B.tabaci.The results improve the understanding of the role of UGTs in the evolution of insecticide resistance,and provide a theoretical basis for the comprehensive management of B.tabaci.

关 键 词：Q型烟粉虱 UDP-糖基转移酶(UGTs) UGT354A1基因 基因克隆 RNAI 噻虫嗪抗性 

分 类 号：S433.3[农业科学—农业昆虫与害虫防治]