摘要
【目的】从绿盲蝽Apolygus lucorum触角中克隆气味受体基因Aluc OR40,研究该气味受体基因在绿盲蝽不同组织中的表达水平,然后对该基因的功能进行研究,为理解绿盲蝽的嗅觉识别机制提供理论基础。【方法】在绿盲蝽成虫触角转录组测序与分析的基础上,通过PCR技术克隆得到气味受体基因Aluc OR40的全长序列。用半定量RT-PCR研究该基因在雌雄虫不同组织中的表达水平。然后通过爪蟾卵母细胞体外表达该基因,并结合双电极电压钳检测了该气味受体对60种气味分子包括绿盲蝽性信息素组分和植物挥发物的反应。然后,利用触角电位技术测定羽化后3 d的绿盲蝽成虫对Aluc OR40的气味配体的触角电位反应。【结果】克隆了Aluc OR40(Gen Bank登录号:KU886190)。Aluc OR40编码398个氨基酸,蛋白具有7个跨膜结构域,N末端位于胞内,C端位于细胞外,符合昆虫气味受体的典型特征。半定量RT-PCR的结果显示,Aluc OR40在触角中特异表达,且在雄虫触角中的表达水平高于雌虫。双电极电压钳记录结果显示,Aluc OR40只对反-2-己烯醇一种气味分子具有特异反应。EAG实验结果表明,羽化后3 d的绿盲蝽雌雄虫都对反-2-己烯醇有反应,且雄虫触角的EAG反应显著高于雌虫。【结论】结果提示Aluc OR40参与了绿盲蝽对反-2-己烯醇的识别过程,推测其在绿盲蝽交配过程中雄虫对雌虫的识别中发挥作用。
【Aim】This study aims to clone an odorant receptor gene,Aluc OR40,from Apolygus lucorum,to characterize its expression profiles in different tissues and to explore its function,so as to provide a theoretical basis for understanding the mechanisms of odor reception in A. lucorum. 【Methods 】The antennal transcriptome sequencing and RT-PCR amplification was used to clone the full-length c DNA of Aluc OR40. The relative expression levels of this gene in different tissues of male and female adults of A.lucorum were detected using semi-quantitative RT-PCR. Through the combination of heterologous expression in Xenopus oocytes and two-electrode voltage clamp electrophysiological recording technique,the response of this odorant receptor to 60 odorants including sex pheromone of A. lucorum and plant volatiles was determined. The EAG responses of the 3-d-old adults of A. lucorum to the odorant ligand of Aluc OR40 were further measured. 【Results】Aluc OR40（ Gen Bank accession number： KU886190） was cloned. Aluc OR40 encodes 398 amino acids and the deduced protein has 7 transmembrane domains with an intracellular N-terminus and an extracellular C-terminus,which conforms to the typical structure of insect odorant receptors. The results of semi-quantitative RT-PCR showed that Aluc OR40 was specifically expressed in the antenna of adults and the expression level was obviously higher in males than females.Two-electrode voltage clamp electrophysiological recording showed that Aluc OR40 specifically responded to（ E）-2-hexenol among the tested odorants. The EAG experiments showed that the 3-d-old adults had EAG response to（ E）-2-hexenol,but the responses of males were significantly higher than those of females. 【Conclusion】Our results suggest that the Aluc OR40 is involved in identification of（ E）-2-hexenol and may play an important role in identification of female by male during mating process.
出处
《昆虫学报》
CAS
CSCD
北大核心
2016年第6期592-601,共10页
Acta Entomologica Sinica
基金
国家自然科学基金项目(31471833,31301245)
关键词
绿盲蝽
气味受体
半定量RT-PCR
双电极电压钳
触角电位
Apolygus lucorum
odorant receptor
semi-quantitative RT-PCR
two-electrode voltage clamp
electroantennography
