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绿盲蝽气味受体基因AlucOR40的克隆及功能研究
  • ISSN号:0454-6296
  • 期刊名称:《昆虫学报》
  • 时间:0
  • 分类:Q966[生物学—昆虫学]
  • 作者机构:[1]山西师范大学生命科学学院,山西临汾041004, [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
  • 相关基金:国家自然科学基金项目(31471833,31301245)
中文摘要:

【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。

英文摘要:

【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.

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期刊信息
  • 《昆虫学报》
  • 北大核心期刊(2011版)
  • 主管单位:中国科学院
  • 主办单位:中国昆虫学会 中国科学院动物研究所
  • 主编:黄大卫
  • 地址:北京东中关村中国科学院动物研究所
  • 邮编:100101
  • 邮箱:kcxb@ioz.ac.cn
  • 电话:010-64807173 010-64807099
  • 国际标准刊号:ISSN:0454-6296
  • 国内统一刊号:ISSN:11-1832/Q
  • 邮发代号:2-153
  • 获奖情况:
  • 国家期刊方阵“双百期刊”
  • 国内外数据库收录:
  • 俄罗斯文摘杂志,美国化学文摘(网络版),英国农业与生物科学研究中心文摘,美国剑桥科学文摘,美国生物科学数据库,英国动物学记录,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:16190