烟粉虱MED隐种TRP基因全长cDNA克隆及生物信息学分析

Molecular cloning and bioinformatics analysis of the Bemisia tabaci Mediterranean(MED) transient receptor potential(TRP) gene

【目的】近几年,昆虫温度适应的感知机理备受研究者的关注。由瞬时感受器离子通道(Transient receptor potential,TRP)介导的温度感知系统是影响昆虫温度感知的关键组成。研究温度感知机制关键调控因子的功能可为明确温度感知以及温度适应调控机理提供理论依据。TRP具有感知高温、低温以及在适温范围内温度选择等功能,对昆虫感知温度有重要的作用。【方法】本研究以烟粉虱Bemisia tabaci(Gennadius)Mediterranean(MED)隐种为研究对象,采用反转录PCR(Reverse transcription PCR,RT-PCR)和c DNA末端快速扩增PCR(Rapid amplification of c DNA ends PCR,RACE-PCR)技术。【结果】首次克隆获得烟粉虱MED隐种TRP基因的c DNA全长序列,命名为Bt TRP。结果表明,该基因含有一个3 501bp的开放阅读框,编码1 166个氨基酸,其编码产物有6次跨膜结构,属于瞬时感受器离子通道家族成员。同源性比较分析表明,Bt TRP与豌豆蚜Acyrthosiphon pisum和欧洲熊蜂Bombus terrestris的TRP编码的氨基酸序列的一致性较高,分别为75%和71%。【结论】该结果为进一步明确Bt TRP在烟粉虱温度感知中的作用奠定基础。

[Objectives] Recently, researchers have paid more attention to the insect temperature sensing mechanism of which the transient receptor potential （TRP） is one of key components. Studying the temperature sensing mechanism will provide a theoretical basis for understanding the regulatory mechanism involved in temperature adaptation. TRP plays important roles in perceiving temperature, such as avoiding extreme temperatures and choosing the optimum temperature. [Methods] In the present study, we used 3＇ RACE and 5＇ RACE to isolate the TRP gene eDNA sequence from the Bemisia tabaci （Gennadius） Mediterranean （MED） cryptic species, and bioinformatics to characterize the TRP genes molecular structure and analyze its phylogenetic relationships. [Results] The TRP gene of B. tabaci MED, named BtTRP, had an open reading frame of 3 501 bp, encoding 1 166 amino acids. The encoded products included 6 transmembrane structures, which are one of the features of transient receptor ion channels. Bemisia tabaci MED BtTRP displayed 75% and 71% similarity with that ofAcyrthosiphon pisum （Harris） and Bombus terrestris （Linnaeus）, respectively. IConelusion] These results provide basic information for further study of the BtTRP gene function underlying B. tabaci MED temperature perception, and provide a theoretical basis for understanding the temperature perception of insects and their adaptability.