苹果蠹蛾瞬时感受器离子通道基因的克隆及温度胁迫表达分析  

Cloning and expression analysis of transient receptor potential gene in Cydia pomonella under temperature stress

作　　者：梁林 吕志创[2] 武强[2] 刘怀[1] 刘万学[2] 万方浩[2] LIANG Lin;Lü Zhichuang;WU Qiang;LIU Huai;LIU Wanxue;WAN Fanghao(College of Plant Protection,Southwest University,Chongqing 400715,China;State Key Laboratory for Biology of Plant Diseases and Insect Pests,Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193,China)

机构地区：[1]西南大学植物保护学院,重庆400715 [2]中国农业科学院植物保护研究所,植物病虫害生物学国家重点实验室,北京100193

出　　处：《植物保护》2021年第1期48-54,60,共8页Plant Protection

基　　金：国家自然科学基金(31672088);国家重点研发计划(2016YFC1200603)。

摘　　要：瞬时感受器离子通道(transient receptor potential,TRP)是影响昆虫温度感知系统的关键组成。为探讨苹果蠹蛾温度感知和温度适应的机理,本研究以苹果蠹蛾Cydia pomonella为研究对象,通过分子克隆获得TRPA家族中的CpPainless和CpWater_witch基因,进行生物信息学分析,并利用实时荧光定量PCR技术分析靶基因在高低温胁迫后的表达。结果表明,CpPainless基因编码938个氨基酸,N端有8个锚蛋白重复序列。CpWater_witch基因编码980个氨基酸,N端有10个锚蛋白重复序列,二者编码产物均有6个跨膜结构,具瞬时感受器离子通道家族成员结构典型特征。表达分析结果显示,和对照26℃相比,高低温胁迫1 h后,CpPainless在5龄幼虫雌虫体内的表达量显著下调,而5龄幼虫雄虫经低温胁迫1 h后CpPainless表达量显著上调,但高温胁迫1 h后表达差异不显著;CpWater_witch在雌雄虫体内均没有显著变化。研究结果为研究瞬时感受器离子通道在苹果蠹蛾温度感知中的作用奠定基础。TRP(transient receptor potential)is a key component of the temperature sensing system in insects.To understand the mechanism of temperature perception and temperature adaptation in Cydia pomonella,CpPainless and CpWater_witch,which belong to TRPA subfamily,were cloned by RT-PCR(reverse transcription PCR).After conducting bioinformatics analysis,quantitative real-time PCR was applied to characterize the relative expression levels of two genes at different temperature stresses.CpPainless and CpWater_witch were successfully predicted and cloned,which encoded 938 amino acids and 980 amino acids,respectively.Protein structure analysis indicated that CpPainless and CpWater_witch had 8 ankyrin repeats and 10 ankyrin repeats,respectively.Both of the genes had 6 transmembrane domains,which is the structural signature of TRP.The RT-PCR analysis showed that the expression level of CpPainless was significantly decreased in female 5th-instar larvae 1 h after temperature stress,but significantly increased in male 5th-instar larvae 1 h after low-temperature stress,whereas there was no significant difference in male larvae 1 h after high-temperature stress;besides,CpWater_witch did not show significant difference in both male and female 5th-instar larvae after temperature stress.The results laid a foundation for further understanding of the role of transient receptor channels in temperature perception of C.pomonella.

关 键 词：苹果蠹蛾 瞬时感受器离子通道 温度感知 温度胁迫 表达量 

分 类 号：S436.611.29[农业科学—农业昆虫与害虫防治]