中华蜜蜂气味受体基因AcerOR113的克隆与时空表达分析

发布时间：2018-07-09 13:28

本文选题：中华蜜蜂 + 工蜂　；参考：《昆虫学报》2017年05期

【摘要】：【目的】鉴定中华蜜蜂Apis cerana cerana气味受体基因并分析其结构特征,明确其在外界蜜粉源充足和缺乏时期不同发育阶段各组织中的表达差异,以期为该基因的功能研究提供理论依据。【方法】利用RT-PCR技术从中华蜜蜂采集蜂触角中扩增获得气味受体基因的cDNA序列;利用生物信息学软件预测分析其编码蛋白的结构特性;采用MEGA6.0邻接法(neighbor-joining)构建系统进化树;利用实时荧光定量PCR技术分析该基因在不同蜜粉源条件下在中华蜜蜂不同日龄(1,5,10,15,20,25和30日龄)工蜂的不同组织(触角、头(去除触角)、胸(去除翅)、腹、足和翅)中的表达差异。【结果】从中华蜜蜂采集蜂触角中克隆获得了中华蜜蜂气味受体基因Acer OR113的cDNA序列(Gen Bank登录号:KT252877.1)。该基因的开放阅读框(ORF)长1 035 bp,编码344个氨基酸,成熟蛋白分子量为40.13 kD,理论等电点(pI)7.05,无信号肽,含有6个跨膜结构且N端位于胞内,在第59-343位氨基酸之间存在一个昆虫气味受体家族7tm-6 superfamily保守结构域。Acer OR113与西方蜜蜂Apis mellifera的Amel OR113亲缘关系最近,氨基酸序列一致性高达94%。实时荧光定量PCR结果显示,无论外界环境中蜜粉源是否充足,Acer OR113在不同日龄工蜂触角中的表达量均极显著高于其他组织(P0.01),腹部中的表达量最低;外界蜜粉源充足时各日龄工蜂触角中Acer OR113的表达量均极显著低于蜜粉源缺乏时相应日龄工蜂触角中的表达量(P0.01)。【结论】Acer OR113具有昆虫气味受体的典型特征,其基因特异性高表达于中华蜜蜂成虫触角中,且表达量在外界蜜粉源缺乏时期高于在蜜粉源充足时期,推测其主要与识别外界蜜粉源的花香气味有关。
[Abstract]:[objective] to identify the odor receptor gene of honeybee Apis cerana and analyze its structural characteristics, and to determine the difference of its expression in different developmental stages of external nectar source. [methods] the cDNA sequence of the odour receptor gene was amplified from the antennae of honeybee bee by RT-PCR, and the structural characteristics of the encoded protein were predicted by bioinformatics software. The phylogenetic tree was constructed by MEGA 6.0 contiguous method (neighbor-joining), and the antennae of different tissues (antennae) of worker bees were analyzed by real-time fluorescence quantitative PCR under different nectar source conditions. [results] the cDNA sequence of the odor receptor gene Acer OR113 was cloned from the antennae of honeybees (GenBank accession number: KT252877.1). The open reading frame (ORF) of the gene is 1 035 BP long, encodes 344 amino acids, the molecular weight of mature protein is 40.13 KD, the theoretical isoelectric point (Pi) is 7.05, it contains 6 transmembrane structures and the N terminal is located in the cell. There is a conserved domain of insect odor receptor family 7tm-6 superfamily between the 59-343 amino acids. The conserved domain of Acer OR113 is closely related to the Amel OR113 of western honeybee Apis mellifera. The results of real-time fluorescence quantitative PCR showed that the expression of Acer OR113 in the antennae of worker bees at different ages was significantly higher than that in other tissues (P0.01), and the lowest in abdomen, regardless of whether the nectar source was sufficient or not. The expression of Acer OR113 in the antennae of worker bees was significantly lower than that in workers' antennae in the absence of nectar powder (P0.01). [conclusion] Acer OR113 has the typical characteristics of insect odor receptors. Its gene specificity was highly expressed in the antennae of adult honeybee, and the expression level was higher in the period of lack of external honey powder than in the sufficient period of pollen source, which was probably related to the identification of the smell of flowers from the external pollen source.
【作者单位】：山西农业大学动物科技学院;山西农业大学生命科学学院;云南省农业科学院蚕桑蜜蜂研究所;
【基金】：国家自然科学基金项目(31502021,31272513)
【分类号】：Q78;S891

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