| 杜亚丽*，潘建芳*，王树杰，杨爽，赵慧婷，姜玉锁**,2017,中华蜜蜂气味受体基因AcerOr167的克隆及表达分析[J].环境昆虫学报,39(1):39-47 |
| 中华蜜蜂气味受体基因AcerOr167的克隆及表达分析 |
| Cloning and expression analysis of odorant receptor gene AcerOr167 in the Chinese honeybee, Apis cerana cerana (Hymenoptera: Apidae) |
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| DOI： |
| 中文关键词: 中华蜜蜂 AcerOr167 基因克隆 荧光定量 表达谱 |
| 英文关键词:Apis cerana cerana AcerOr167 gene cloning qRT-PCR expression profile |
| 基金项目:国家自然科学基金项目（31502021，31272513）；山西农业大学科技创新基金（2014012） |
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| 中文摘要: |
| 本研究通过RT-PCR获得中华蜜蜂气味受体基因Or167的cDNA序列，并采用多种生物信息学软件对其结构特征进行预测分析；利用荧光定量PCR检测AcerOr167 mRNA在工蜂羽化后不同发育阶段（1、5、10、15、20、25和30日龄）和不同组织（触角、头、胸、腹、足、翅）中的相对表达量。克隆获得中华蜜蜂Or167的cDNA序列，命名为AcerOr167（GenBank登录号为KF239369），其全长1311 bp，编码436个氨基酸，预测有5个跨膜结构域；多序列比对结果显示，中华蜜蜂AcerOr167与西方蜜蜂AmelOr167的一致性最高，达94%，而与毕氏粗角蚁CbirOr4-like的一致性最低，为49%；荧光定量结果显示,AcerOr167 mRNA在成蜂不同发育期的各个组织中均有表达，且触角中的表达量极显著高于其它各个组织（P<0.01），在头、胸、腹、足、翅中有少量表达；从触角不同发育阶段的表达情况来看，1日龄表达量最低，5日龄表达量显著升高，20日龄表达量最高，且极显著高于其它各日龄（P<0.01）。推测AcerOr167可能在中华蜜蜂的嗅觉识别系统中起着重要的作用，与中华蜜蜂外出采集，识别花香气味有关。本研究为进一步深入研究中华蜜蜂传统气味受体的功能奠定理论基础。 |
| 英文摘要: |
| In this study, the cDNA sequence of the odorant receptor gene Or167 from Apis cerana cerana was cloned by RT-PCR，and the relative expression of this gene at different developmental stages (1, 5, 10, 15, 20, 25 and 30 days) and in different tissues (antennae, head (without antennae), thorax, abdomen, legs and wings) of A. c. cerana were profiled by qRT-PCR. The cDNA sequence of the Or167 was obtained and named as AcerOr167 (GenBank accession number: KF239369), the open reading frame of AcerOr167 is 1311 bp, encoding 436 amino acid residues, and homology analysis showed that the amino acid sequences of AcerOr167 contain five-transmenbrane domains. The multiple alignment and phylogenetic tree analyses showed that AcerOr167 shared the highest identity of 94% with AmelOr167, and shared the lowest identity of 49% with CbirOr4-like. The results of qRT-PCR showed that AcerOr167 was clearly detected in different tissues at different developmental stages of adult workers. The expression of AcerOr167 in the antennae was significantly higher than other tissues (P<0.01), and the slight expression was detected in head (without antennae), thorax, abdomen, legs and wings. From the expression level of AcerOr167 in every developmental stages, expression was lowest in the 1st, expression increased significantly in the 5th and reached the highest in the 20th. The expression of AcerOr167 in 20th was significantly higher than other days. AcerOr167 may play an important role in the olfactory system of A. c. cerana, and was closely related to the behavior that Chinese honeybee collect pollen and identify the fragrance of flowers. This findings will provide theoretical basis for further functional study of the traditional odorant receptor genes. |
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