薇甘菊颈盲蝽谷胱甘肽S-转移酶PmGSTd1基因克隆及表达分析  

Cloning and Expression Analysis of Glutathione S-transferase PmGSTd1 Gene in Pachypeltis micranthus(Hemiptera:Miridae)

作　　者：张传光 泽桑梓[3] 朱家颖 季梅[2] 刘凌[2] 张乃明[1] ZHANG Chuanguang;ZE Sangzi;ZHU Jiaying;JI Mei;LIU Ling;ZHANG Naiming(College of Plant protection,Yunnan Agricultural University,Kunming 650201 China;Bureau of Forestry Pest Control and Quarantine of Yunnan Province,Kunming 6500519 China;Yunnan Academy of Forestry,Kunming 650204 China;Southwest Forestry University,Kunming 650224 China)

机构地区：[1]云南农业大学植物保护学院,云南昆明650201 [2]云南省林业科学院,云南昆明650204 [3]云南省林业有害生物防治检疫局,云南昆明650051 [4]西南林业大学,云南昆明650224

出　　处：《生态环境学报》2018年第12期2179-2184,共6页Ecology and Environmental Sciences

基　　金：国家自然科学基金项目(31360154).

摘　　要：薇甘菊颈盲蝽(Pachypeltis micranthus)是入侵植物——薇甘菊(Mikania micrantha)的重要天敌昆虫,专食薇甘菊。谷胱甘肽S-转移酶(GSTs)是一种多功能蛋白,在代谢有毒化合物的过程中起着重要作用,探明其在不同部位的表达情况,可为深入研究其在薇甘菊颈盲蝽与寄主互作中的作用奠定基础。采用RACE技术克隆薇甘菊颈盲蝽GST基因,实时荧光定量PCR检测其在不同部位的表达情况。结果表明,Pm GSTd1基因全长977 bp,共编码240个氨基酸,理论分子量为27.28 kDa,N端1-18号氨基酸为信号肽,与温带臭虫(Cimex lectularius)GST基因的亲缘关系最近。该基因在雌、雄虫的各部位均有表达,且皆表现为足部表达量明显高于其他部位;雄虫各部位的表达量(残体除外)均显著高于雌虫,雄虫翅膀、触角、足部、残体中的表达量分别是雌虫相应部位的4.49倍、2.11倍、1.68、1.33倍。雌虫触角和残体中PmGSTd1的表达量基本相同,翅膀中的表达量显著低于其他部位;雄虫触角与翅膀中的表达水平基本相同,残体中的表达水平显著低于其他部位。薇甘菊颈盲蝽PmGSTd1是属于GST Delta家族的分泌蛋白,此基因功能可能包括代谢食物源毒素、降解气味分子及飞翔肌的结构蛋白。The mirid bug,Pachypeltis micranthus Mu et Liu(Hemiptera:Miridae),is a natural enemy of the widespread invasive weed,Mikania micrantha.Glutathione S-transferase(GSTs),a multifunctional protein,plays an important role in metabolizing toxic compounds.Proving its expression in different parts provides valuable information to further exploration of the role of GSTs involved in the interaction between P.micranthus and its host.A cDNA encoding GST was cloned using rapid amplification of cDNA ends(RACE)strategy,and fluorescence quantitative real time PCR(qPCR)was used to detect the expression of GST gene in different parts.Results showed that PmGSTd1 gene was 977 bp in full length,which encoded 240 amino acids,and its predicted molecular weight was 27.28 kDa.N-terminal amino acids 1-18 were signal peptides.The homology of PmGSTd1 from P.micranthus had the closest relationship with GST of Cimex lectularius.PmGSTd1 was expressed in all parts of the female and male,and the expression level of the foot was significantly higher than that of other parts.The expression of male in all parts(except the residue body)was significantly higher than that of female.The expression of male in their wings,antennae,feet and residue body was 4.49,2.11,1.68 and 1.33 times respectively of female in corresponding parts.The expression of PmGSTd1 in antennae and residue body of female was basically the same,and the expression in wings was significantly lower than other parts.The expression level of antennae and wings of male was basically the same,and the expression level of residue body was significantly lower than other parts.PmCYP4C1 was a secretory protein which belongs to delta family.The function of this gene may include the metabolism of toxic compounds from food,degradation of odor molecules and the structural protein of the flying muscle.

关 键 词：薇甘菊颈盲蝽 谷胱甘肽S-转移酶 基因克隆 表达分析 

分 类 号：Q965.9[生物学—昆虫学] X17[环境科学与工程—环境科学]