中华蜜蜂4个气味受体基因AcerOrs的时空差异性表达分析

发布时间：2018-03-03 17:31

本文选题：中华蜜蜂　切入点：气味受体　出处：《山西农业大学》2016年硕士论文　论文类型：学位论文

【摘要】：昆虫为了适应外界多变的环境,进化出了复杂且灵敏的嗅觉系统,通过嗅觉系统来感知外界变化,从而获取信息,趋利避害,维持个体发育和种群的繁荣。随着生物信息学的发展,学者们对昆虫嗅觉分子机理的研究也日渐深入,更好的解释了昆虫嗅觉的发生机理,以及昆虫与寄主植物协同进化的关系。中华蜜蜂是我国传统的蜂种,由于长期的适应与进化,形成了其独有的生物学特性,已成为我国山林地区不可替代的优良蜂种,它具有嗅觉灵敏、采集勤奋、善于利用零星蜜粉源、抗逆性和抗病力强等西方蜜蜂无可比拟的优点。本研究以中华蜜蜂为研究对象,克隆了中华蜜蜂气味受体基因AcerOr28和AcerOr113,分析了两基因以及基于转录组数据得到的AcerOr35和AcerOr96序列,并对4个基因在外界蜜粉源充足时和缺乏时的中华蜜蜂工蜂不同发育阶段、不同组织的差异性表达分别进行了检测分析，，结果如下：(1) AcerOr28、AcerOr35、AcerOr96和AcerOr113的CDS区全长分别为1218、1239、1206和1035 bp,各编码405、413、402和344个氨基酸。GenBank登录号分别为：KT454837.1、KX371646、KX371647和KT252877.1。(2)生物信息学分析表明,AcerOr28、AcerOr35、AcerOr67和AcerOr113氨基酸序列中存在着数量不等的潜在糖基化和磷酸化位点,4个蛋白分子均无信号肽,除AcerOr35具有7个跨膜结构域之外,AcerOr28, AcerOr96和AcerOr113均为6个跨膜结构域,均与气味受体典型的结构特征相符。(3)通过real-time PCR技术,对不同发育阶段和不同组织的中华蜜蜂AcerOr28, AcerOr35, AcerOr96和AcerOr113基因的表达情况进行了研究。结果表明,所有基因在工蜂不同发育阶段均有表达,而且触角的表达量极显著高于其它组织。分析4个基因在触角中的表达量,除AcerOr28的表达量是在蜜粉源充足时期高于蜜粉源不足时期外，AcerOr35, AcerOr96和AcerOr113的表达量均是在蜜粉源充足时期低于蜜粉源不足时期。AcerOr28不论在蜜粉源充足还是蜜粉源缺乏下,其发育期表达量的峰值也与其它3个基因不同,并且AcerOr28与西方蜜蜂假基因AmelOr28p的相似性较高,推测AcerOr28也可能为假基因,但这一推论尚待进一步证实。
[Abstract]:Insects in order to adapt to a changing environment, the evolution of the olfactory system is complex and sensitive, the olfactory system to perceive changes in the outside world, so as to obtain information, while avoiding disadvantages, the maintenance of individual development and population prosperity. With the development of bioinformatics, the scholars study on the molecular mechanism of insect olfaction is deeper, better explanation the occurrence mechanism of insect olfaction, as well as the coevolutionary relationship between insects and their host plants. The bee bee is a kind of Chinese traditional, because of long-term adaptation and evolution, formed its unique biological characteristics, has become the mountain area of our country can not be replaced by excellent bee species, it has a sensitive sense of smell, good at collecting diligence the use of scattered nectar, advantages of resistance and strong stress resistance There is nothing comparable to this western bees. In this study, the bees as the research object, the cloning of Chinese bee odor receptor gene AcerOr2 8 and AcerOr113, analyzed two genes and based on AcerOr35 and AcerOr96 sequences of transcriptome data is obtained, and the APIs ceranaceranafabricius 4 genes in the outside world and the lack of sufficient nectar at different developmental stages, detection and analysis, were differentially expressed in different tissues. The results are as follows: (1) AcerOr28, AcerOr35. AcerOr96 and AcerOr113 CDS sequences were 121812391206 and 1035 BP respectively, encoding 405413402 and 344 amino acids.GenBank accession number were KT454837.1, KX371646, KX371647 and KT252877.1. (2) bioinformatics analysis showed that AcerOr28, AcerOr35, there is a number of potential glycosylation and phosphorylation sites of AcerOr67 and AcerOr113 amino acids. In the sequence, 4 proteins had no signal peptide, except AcerOr35 has 7 transmembrane domains, AcerOr28, AcerOr96 and AcerOr113 are 6 transmembrane domains, with gas With structural characteristics of typical taste receptor. (3) by real-time PCR, of different developmental stages and different tissues of the honeybee AcerOr28, AcerOr35, AcerOr96 and AcerOr113 gene expression were studied. The results showed that all the genes in different developmental stages of bees were expressed, and the expression was significantly higher than that of the antenna analysis of other tissues. Expression of 4 genes in the tentacles, in addition to the expression of AcerOr28 is higher than in nectar foraging period sufficient period of shortage, AcerOr35, expression of AcerOr96 and AcerOr113 are less than.AcerOr28 in the period of shortage of nectar nectar foraging in sufficient or lack of adequate nectar period. The developmental expression of the peak and the other 3 genes, and AcerOr28 and Western bees AmelOr28p pseudogenes high similarity, we speculate that AcerOr28 may be a false base However, this inference has yet to be further confirmed.

【学位授予单位】：山西农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S891

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