薇甘菊颈盲蝽细胞色素P450 PmCYP4基因克隆及表达分析  

Cloning and expression analysis of a cytochrome P450 PmCYP4 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, Yunnan 650201, China;Yunnan Academy of Forestry,Kunming, Yunnan 650204, China;Bureau of Forestry Pest Control and Quarantine of Yunnan Province, Kunming,Yunnan 650051, China;Southwest Forestry University, Kunming, Yunnan 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：《生物安全学报》2019年第1期13-18,共6页Journal of biosafety

基　　金：云南省重点研发项目(2018BB009);中央财政林业科技推广项目(云[2017]TG 07号)

摘　　要：【目的】薇甘菊颈盲蝽是入侵植物薇甘菊的天敌昆虫。CYP4家族基因在专食性昆虫与宿主植物的相互作用中发挥着极其重要的作用,探明其在不同部位的表达情况,可为薇甘菊生物控制提供科学依据。【方法】采用RACE技术克隆薇甘菊颈盲蝽CYP基因,实时荧光定量PCR检测其在不同部位的表达情况。【结果】PmCYP4C1基因全长1713 bp,其中ORF长1500 bp,共编码500个氨基酸,理论分子质量为57.44 ku,无信号肽;与其他昆虫CYP4家族基因的同源性大于40%,与温带臭虫CYP的亲缘关系最近。该基因在雌、雄虫各部位均有表达,且都是足部的表达量明显地高于其他部位;雌、雄虫的表达差异在于雄虫翅膀中的表达量明显地高于触角和残体,但在雌虫中这3个部位的表达量无显著差异,且雄虫翅膀中的表达量显著地高于雌虫,是其2.37倍。【结论】薇甘菊颈盲蝽PmCYP4基因除参与代谢有毒物质外,其主要功能可能是编码与薇甘菊颈盲蝽运动相关的酶。【Aim】 Pachypeltis micranthus is a specialist herbivore feeding on the invasive plant Mikania micrantha . CYP4 family genes play a very important role in the interaction between insects and host plants.【Method】 A cDNA encoding CYP was cloned using rapid amplification of cDNA ends (RACE) strategy. Fluorescence quantitative real time PCR (qPCR) was used to detect the expression of CYP gene in different body parts.【Result】 PmCYP4 gene was 1713 bp in full length, and 1500 bp in ORF, which encoded 500 amino acids with a predicted molecular weight of 57.44 ku. There was no signal peptide in this deduced amino acid sequence. The homology of PmCYP4 of P. micranthus with CYP4 family of other insects was more than 40%, with the closest relationship with CYP of Cimex lectularius . PmCYP4 gene was expressed in different body parts with the highest expression in leg. The expression in wings of males was significantly higher than in antennae and residue and 2.37 times higher than females, but there were no significant differences in these body parts for females.【Conclusion】 In addition to metabolic toxins, the main function of PmCYP4 gene may be to encode for enzymes related to the movement of P. micranthus .

关 键 词：薇甘菊颈盲蝽 细胞色素P450 基因克隆 表达分析 

分 类 号：S451[农业科学—植物保护] S476