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意大利蜜蜂α1亚型烟碱型乙酰胆碱受体功能表达和分析

Functional expression and analysis of Apis mellifera α1 nicotinic acetylcholine receptor

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【作者】 耿俊杰王昱权张坤吴少英

【Author】 GENG Junjie;WANG Yuquan;ZHANG Kun;WU Shaoying;School of Breeding and Multiplication (Sanya Institute of Breeding and Multiplication), Hainan University;School of Tropical Agriculture and Forestry, Hainan University;

【通讯作者】 张坤;吴少英;

【机构】 海南大学南繁研究院(三亚南繁研究院)海南大学热带农林学院

【摘要】 尽管作用于昆虫烟碱型乙酰胆碱受体(nicotinic acetylcholine receptors,nAChRs)的新烟碱类杀虫剂是农作物害虫防治的常用工具,但它们对意大利蜜蜂等传粉昆虫产生严重的负面影响。本研究克隆意大利蜜蜂nAChR α1亚基基因,对其进行系统发育分析,并利用双电极电压钳技术研究α1亚型nAChR与新烟碱类杀虫剂(吡虫啉、噻虫胺和呋虫胺)的相互作用机制。序列比对和系统进化树分析显示,意大利蜜蜂nAChR α1亚基具有典型的n AChRs亚基家族基因结构域特征,与半翅目昆虫的α1亚基关系最远,与同属膜翅目蜜蜂的α1亚基关系最近。将意大利蜜蜂nAChR α1亚基与大鼠nAChR β2亚基在非洲爪蟾卵母细胞中共表达,电生理结果显示吡虫啉对Amα1/rβ2 nAChR的激动效率最高,噻虫胺最低,而Amα1/rβ2 nAChR对呋虫胺的激动亲和力最高,对吡虫啉最低。这表明意大利蜜蜂nAChR α1亚基是新烟碱类杀虫剂的作用靶标之一,并且Amα1/rβ2nAChR对不同新烟碱类杀虫剂敏感性存在差异。本研究结果对于开发作用于害虫的新型高选择性杀虫剂具有重要理论指导意义。

【Abstract】 Neonicotinoids that act on insect nicotinic acetylcholine receptors(nAChRs) are common tools for crop pest control, but they have serious negative effects on pollinators such as Apis mellifera. In this study, the nAChR α1 subunit gene was cloned from A. mellifera and systematically analyzed. The interaction mechanism between the nAChR α1 subtype and neonicotinoids(imidacloprid, clothianidin, and dinotefuran) was investigated by using the two-electrode voltage clamp technique. Sequence alignment and phylogenetic tree analysis showed that the nAChR α1 subunit of A. mellifera had typical domain characteristics in the nAChR subunit family genes, and was most distantly related to the α1 subunit of Hemiptera insects and most closely related to the α1 subunit of bees from Hymenoptera. The combination of A. mellifera nAChR α1 subunit and Rattus norvegicus nAChR β2 subunit was co-expressed in Xenopus laevis oocytes. Electrophysiological results showed that imidacloprid had the highest agonistic efficiency and lowest agonistic affinity, that clothianidin had the lowest agonistic efficiency and that dinotefuran had the highest agonistic affinity for Amα1/rβ2 nAChR, indicating that the α1 subunit of A. mellifera nAChRs is one of the targets of neonicotinoids and the sensitivity of Amα1/rβ2 nAChR to different neonicotinoids is different. All these results have important theoretical significance for the development of novel highly selective insecticides for target insects.

【基金】 海南省重大科技计划项目(ZDKJ2021007);国家自然科学基金地区科学基金项目(32360663,32260666)
  • 【文献出处】 热带生物学报 ,Journal of Tropical Biology , 编辑部邮箱 ,2024年05期
  • 【分类号】TQ453
  • 【网络出版时间】2024-03-12 17:57:00
  • 【下载频次】235
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