基于转录组的亚洲小车蝗气味结合蛋白基因的鉴定及其表达谱分析

发布时间：2018-06-02 05:50

本文选题：转录组 + 第二代高通量测序　；参考：《内蒙古农业大学》2015年硕士论文

【摘要】：嗅觉感受系统在昆虫取食、寻找寄主、躲避天敌、交配和产卵等一系列生命活动中发挥了重要作用；在昆虫嗅觉感受系统中,气味分子结合蛋白(odorant-binding protein, OBP)扮演着重要的角色。本研究利用第二代高通量测序技术在亚洲小车蝗(Oedaleus asiaticus)触角中首次鉴定出15个编码OBP的基因,命名为OBP1-15,并对其表达谱进行了测定与分析,此外,又对亚洲小车蝗OBP与包括东亚飞蝗在内的其它六种昆虫的OBP进行了系统发育分析。主要结果如下：1.亚洲小车蝗雌、雄成虫触角转录组测序、数据分析及气味分子结合蛋白基因的鉴定。通过对亚洲小车蝗成虫雌、雄触角分别进行转录组测序,获得总clean reads数分别为137,426,706和127,822,838;总clean碱基数分别为12,368,403,540 nt和11,504,055,420 nt,均超过10GB,经过软件组装后,共获得61,629条all-unigene,其中有26,064(42.30%)条all-unigene获得了功能注释信息。首次在亚洲小车蝗成虫触角中鉴定出15个OBP基因,分析发现它们均具有全长开放阅读编码框(open reading frame, ORF)。2.亚洲小车蝗气味分子结合蛋白基因表达谱测定。通过实时荧光定量PCR (quantitative real-time PCR, qRT-PCR)实验,发现只有OBP7在雌触角中的表达量要远大于在雄触角中的表达量；有9个OBP在雄触角中的表达量要大于在雌触角中的表达量。通过半定量PCR实验,发现OBP9和OBP10只特异性的表达于触角,而其它OBP则在触角、口器或翅膀等化学感受组织中表达。3.亚洲小车蝗OBP与其他昆虫OBP系统发育分析。通过对亚洲小车蝗15个OBP与其它来自于六个不同目的六种昆虫OBP氨基酸序列之间构建的进化树进行系统发育分析,发现亚洲小车蝗的大部分OBP都与东亚飞蝗(Locusta migratoria) OBP亲缘关系最近,但是亚洲小车蝗有些OBP并不是与东亚飞蝗亲缘关系最近。这表明二者的OBP基因最初由同一个祖先基因进化而来,但是为了适应不同的生态环境,各自进化出了独特的化学感受系统。
[Abstract]:Olfactory sensing system plays an important role in a series of life activities, such as feeding, finding hosts, avoiding natural enemies, mating and spawning, etc. In insect olfactory sensing system, odor molecule binding protein odorant-binding protein (OBPP) plays an important role. In this study, the second generation high-throughput sequencing technique was used to identify for the first time 15 genes encoding OBP in the antennae of Oedaleus asiaticus, named OBP1-15, and their expression profiles were determined and analyzed. The phylogenetic analysis of OBP and six other insects, including migratory locust in East Asia, was also carried out. The main results are as follows: 1. Transcriptome sequencing, data analysis and identification of odour molecular binding protein gene in female and male adults. By sequencing the female and male antennae, the total clean reads numbers were 137426706 and 127822838, and the total clean base numbers were 12368403540 NT and 11504055420 NT, respectively. A total of 61629 all-unigenetics were obtained, of which 26064 and 42.30) all-unigene obtained functional annotation information. For the first time, 15 OBP genes were identified from the antennae of Acridostoma asiatica. It was found that all of them had full-length open reading frame, ORF.2. Detection of molecular binding protein gene expression profile of Asiatic locust. By real-time quantitative PCR quantitative real-time PCR (qRT-PCR) experiment, it was found that only OBP7 was expressed in the female antennae and 9 OBP in the male antennae were higher than those in the female antennae. By semi-quantitative PCR assay, it was found that OBP9 and OBP10 expressed only in the antennae, while other OBP expressed .3in chemosensory tissues such as antennae, mouthparts or wings. Phylogenetic analysis of OBP and other insects OBP. Phylogenetic analysis of the phylogenetic tree constructed between 15 OBP and other OBP amino acid sequences of six insect species from six different purposes was carried out. It was found that most of the OBP was closely related to Locusta migratoria OBP, but some OBP were not closely related to the Asian migratory locust. This indicates that the OBP gene of the two genes was originally evolved from the same ancestor gene, but in order to adapt to different ecological environments, they have evolved a unique chemical sensing system.
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S433.2;Q78

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