| 棉铃虫烟碱类乙酰胆碱受体α7亚基基因的鉴定及功能分析 |
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| 引用本文：刘少凯,谢丙堂,魏纪珍,梁革梅.棉铃虫烟碱类乙酰胆碱受体α7亚基基因的鉴定及功能分析.植物保护学报,2021,48(5):1016-1025 |
| DOI：10.13802/j.cnki.zwbhxb.2021.2021853 |
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| 中文摘要:为探究和挖掘更多杀虫剂靶标受体，采用PCR方法结合RACE技术克隆了棉铃虫烟碱类乙酰胆碱受体（nicotinic acetylcholine receptor，nAChR）的α7亚基（nAChR-α7）基因，通过荧光定量PCR技术进行时空表达分析，并采用电生理检测以及RNAi技术研究该基因对棉铃虫生长发育的调控作用。结果表明，棉铃虫nAChR-α7基因（GenBank登录号：KM884875）全长3 632 bp，包含编码496个氨基酸的1 491 bp开放阅读框。该基因具有nAChR典型结构，与其他鳞翅目昆虫nAChR-α7基因同源性较高，且包括胞外N端配体结合区、半胱氨酸环、Loop A~Loop F、跨膜区（TM1~TM4）和1个短的C端胞外尾。nAChR-α7基因在棉铃虫幼虫头部及成虫期高表达；通过将nAChR-α7基因在非洲爪蟾Xenopus oocytes卵母细胞中表达，并检测电生理信号，确定该基因是nAChR受体基因，但不能持续响应ACh信号。干扰试验结果显示nAChR-α7基因对棉铃虫幼虫生长发育无显著影响。推测该基因可能不是主要的nAChR基因，而是与其他亚基基因共同起作用。 |
| 中文关键词:棉铃虫 烟碱型乙酰胆碱受体 时空表达 功能 电生理学 |
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| Identification and functional studies of nicotinic acetylcholine receptor α7 subunit in cotton bollworm Helicoverpa armigera |
| Author Name | Affiliation | E-mail | | Liu Shaokai | State key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China
College of Plant Protection, Henan Agricultural University, Zhengzhou 450002, Henan Province, China | | | Xie Bingtang | State key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China | | | Wei Jizhen | College of Plant Protection, Henan Agricultural University, Zhengzhou 450002, Henan Province, China | | | Liang Gemei | State key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China | gmliang@ippcaas.cn |
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| Abstract:In order to explore more targets for insecticides, the full-length cDNA sequence of nicotinic acetylcholine receptor (nAChR) α7 subunit (nAChR-α7) gene was cloned from cotton bollworm Helicoverpa armigera by PCR and RACE (rapid amplification of cDNA ends) technologies in this study. Meanwhile, the expression patterns of nAChR-α7 gene were also assayed by qRT-PCR. The function of nAChR α7 subunit as a nicotinic acetylcholine receptor was confirmed by being heterologously expressed in Xenopus oocytes to record the changing currents using the two-microelectrode voltage-clamp technique. In addition, the regulatory function of nAChR-α7 gene in the development of cotton bollworm was investigated by using RNAi technology. The results showed that the full-length cDNA sequence of nAChR-α7 gene (GenBank accession no. KM884875) was 3 632 bp in length, which contained an open reading frame of 1 491 bp encoding 496 amino acids. It had the common characters of the cysteine ring receptor family:N- and C-terminal ex-cytosolic tail, Loop A-Loop F, the four transmembrane domains TM1, TM2, TM3 and TM4, and the big cytosolic ring. The relatively high expression of nAChR-α7 was detected in larval heads and adults. The pharmacological property analysis by constructing the X. oocytes cell expression confirmed that it was a typical nicotinic acetylcholine receptor, though the electrophysiological experiment showed that the current did not sustain as the doses of Ach increased. The down-regulation of nAChR-α7 gene had no significant effect on the development of H. armigera. The results indicated that nAChR-α7 gene was not an important nicotinic acetylcholine receptor and might serve a function with other nAchRs. |
| keywords:Helicoverpa armigera nicotinic acetylcholine receptors (nAChR) spatio-temporal expression function electrophysiology |
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