番茄潜叶蛾谷胱甘肽-S-转移酶基因TabsGSTs2的克隆及与α-番茄碱的分子对接分析  

Gene cloning of glutathione-S-transferase(TabsGSTs2)in Tuta absoluta and its molecular docking analysis withα-tomatine

作　　者：周昕雨 李金萍 黄聪 许博 崔建臣 万方浩[2] 张毅波[2] 桂富荣[1] 张桂芬[2] ZHOU Xin-Yu;LI Jin-Ping;HUANG Cong;XU Bo;CUI Jian-Chen;WAN Fang-Hao;ZHANG Yi-Bo;GUI Fu-Rong;ZHANG Gui-Fen(State Key Laboratory for Conservation and Utilization of Bioresources in Yunnan,Plant Protection College of Yunnan Agricultural University,Kunming 650201,China;State Key Laboratory for Biology of Plant Diseases and Insect Pests,Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193,China;Beijing Plant Protection Station,Beijing 100029,China)

机构地区：[1]云南农业大学植物保护学院,云南生物资源保护与利用国家重点实验室,昆明650201 [2]中国农业科学院植物保护研究所,植物病虫害综合治理全国重点实验室,北京100193 [3]北京市植物保护站,北京100029

出　　处：《环境昆虫学报》2025年第1期56-65,共10页Journal of Environmental Entomology

基　　金：国家重点研发计划项目(2021YFD1400200);中央级公益性科研院所基本科研业务费专项(S2023XM27);云南省科学技术厅基础研究专项(202301AT070485)。

摘　　要：谷胱甘肽-S-转移酶(Glutathione-S-transferases,GSTs)在昆虫适应植物次生代谢物质中发挥重要作用,为明确番茄潜叶蛾Tuta absoluta的GSTs基因在其响应α-番茄碱胁迫中的作用。本研究利用PCR技术克隆了番茄潜叶蛾TabsGSTs2基因全长,通过生物信息学方法分析了TabsGSTs2基因的序列特征、理化特性、保守结构域、基因结构和进化关系,通过RT-qPCR技术测定了α-番茄碱胁迫下TabsGSTs2基因的表达情况,利用同源建模和分子对接研究了TabsGSTs2和α-番茄碱的结合能力与结合模式。结果表明,TabsGSTs2基因的CDS全长为609 bp,编码203个氨基酸,理论等电点为8.47,分子量为23.895 kDa;TabsGSTs2具有4个β-折叠和9个α-螺旋,具有典型的GSTs保守结构域,包括GSH结合位点(G-site)和底物结合位点(H-site);TabsGSTs2属于sigma亚家族成员,与苹果蠹蛾Cydia pomonella的CpGSTs2亲缘关系较近;α-番茄碱胁迫下,TabsGSTs2基因在72 h的表达量显著高于对照;TabsGSTs2与α-番茄碱的结合能力较强,主要以氢键、疏水作用力和盐桥等相互作用维持稳定的结合。研究结果可为后续研究番茄潜叶蛾适应α-番茄碱的分子机制提供参考,为进一步挖掘番茄潜叶蛾防控新靶标提供基础。Glutathione-S-transferases(GSTs)play an important role in insect adaptation to plant secondary metabolites.To clarify the role of the GST gene in the detoxification of Tuta absoluta toα-tomatine,we first cloned the full-length sequence of TabsGSTs2 gene.Bioinformatic analysis was employed to analyze the sequence characteristics,physicochemical properties,conserved domains,gene structure,and evolutionary relationships of the TabsGSTs2 gene.The expression levels of TabsGSTs2 underα-tomatine stress were measured using RT-qPCR,and the binding capacity and mode of TabsGSTs2 toα-tomatine were investigated using homology modeling and molecular docking.The results showed that the CDS of TabsGSTs2 was 609 bp,encoding 203 amino acids,with a theoretical isoelectric point of 8.47 and a molecular weight of 23.895 kDa.TabsGSTs2 contains fourβ-sheets and nineα-helices,and had the typical conserved domains of GST genes,including the GSH binding site(G-site)and the substrate binding site(Hsite).Phylogenetic analysis indicated that TabsGSTs2 belongs to the sigma subfamily and was closely related to CpGSTs2 of Cydia pomonella.Underα-tomatine stress,the expression level of TabsGSTs2 was significantly higher than that of the control at 72 h.Molecular docking results suggested that TabsGSTs2 bind strongly toα-tomatine,with the interaction being stabilized primarily by hydrogen bonds,hydrophobic forces,and salt bridges.Our study provides reference for subsequent studies of the molecular mechanisms of T.absoluta adaptation toα-tomatine and provides a basis for further exploration of new targets for the control of T.absoluta.

关 键 词：番茄潜叶蛾 α-番茄碱 谷胱甘肽-S-转移酶 解毒代谢 分子对接 

分 类 号：Q963[生物学—昆虫学]