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Acer Or167 m RNA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c DNA序列,命名为Acer Or167(Gen Bank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Acer Or167与西方蜜蜂Amel Or167的一致性最高,达94%,而与毕氏粗角蚁Cbir Or4-like的一致性最低,为49%;荧光定量结果显示,Acer Or167 m RNA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Acer Or167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。
In this study, the cDNA sequence of the odorant receptor gene Or161 from Apis cerana ceranawas 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 0rl61was obtained and named as Acer0rl61 （ GenBank accession number ; KF239369） , the open readingframe of Acer0r／61 is 1311 bp, encoding 436 amino acid residues, and homology analysis showed thatthe amino acid sequences of Acer0r／61 contain five-transmenbrane domains. The multiple alignment andphylogenetic tree analyses showed that Acer0r／61 shared the highest identity of 94% with Amel0r161, and shared the lowest identity of 49% with Cbir0r4-like. The results of qRT-PCR showed that Acer0r／61 was clearly detected in different tissues at different developmental stages of adult workers. The expression of AcerOrl61 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 oi AcerOr／61 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 AcerOrl61 in 2 0 th was significantly higher than other days. Acer0r／61 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.
出处
《环境昆虫学报》
CSCD
北大核心
2017年第1期39-47,共9页
Journal of Environmental Entomology
基金
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)
