柞蚕与家蚕前胸腺的比较转录组分析

发布时间：2018-06-16 20:52

  本文选题：前胸腺 + 转录组　； 参考：《沈阳农业大学》2017年硕士论文

【摘要】：前胸腺是昆虫体内最重要的内分泌器官之一,合成和分泌的蜕皮激素在调控昆虫生长发育过程中起着关键作用。然而,我们对这一重要内分泌器官在分子水平的认识还相当有限,原因在于前胸腺基因组信息的匮乏。为了填补这一缺憾,本论文以两种重要的经济昆虫(家蚕和柞蚕)为材料,利用高通量测序技术,对这两种昆虫前胸腺的转录组进行了比较。同时,基于转录组数据,在前胸腺中鉴定出了一整套的生物钟基因。此外,对柞蚕转录组数据中的SSR位点信息进行了分析。本论文是首次进行昆虫前胸腺的比较转录组分析,为深入理解这一重要内分泌器官的特征和功能提供非常有价值的信息资源。主要结果如下:1.柞蚕和家蚕前胸腺的转录组测序和比较分析。拼接组装后,柞蚕得到64,301个transcripts(平均长度 657 bp)和 44,067 个 Unigenes。家蚕得到 49,287 个 transcripts(平均长度1,064 bp)和32,302个Unigenes。柞蚕和家蚕数据均已储存在NCBI Sequence Read Archive中,登录号分别为SRR5119598和SRR5119600。柞蚕和家蚕中分别有22,402个(50.84%)和 23,157 个(71.69%)Unigenes 具有匹配的 homologous hits。比较分析发现,这两个前胸腺在GO terms、COG category和KO assignments的功能注释结果是高度相似的。另外,两个前胸腺转录组数据中,共鉴定出6,357对直系同源基因。2.生物钟基因的鉴定和RT-PCR检测。生物钟在昆虫的行为和生理过程中起着重要调控作用。从柞蚕和家蚕前胸腺转录组数据中,我们鉴定出了 13个生物钟基因,包括cryptochrome 1、cryptochrome 2、period、timeless、clock、cycle、shaggy、double-time、vrille、PAR-domainprotein 1、slimb、casein kinase Ⅱalpha、casein kinase Ⅱbetaa。RT-PCR检测结果表明,这些基因在前胸腺中均有表达。这些结果表明,在昆虫前胸腺中也存在一个外围生物钟。3.柞蚕SSR位点信息分析。在柞蚕转录组数据中,共鉴定出2,322个SSR位点,分布于1,615条Unigenes中,发生频率为37.60%,平均分布距离为4.76 kb。柞蚕SSR中,以单昔酸重复占主导地位。获得的转录组数据为SSR引物设计提供了信息依据,进一步为柞蚕遗传多样性分析、功能基因发掘、遗传图谱构建等的奠定了基础。
[Abstract]:Prothymus is one of the most important endocrine organs in insects. Ecdysone synthesized and secreted plays a key role in the regulation of insect growth and development. However, our understanding of this important endocrine organ at the molecular level is rather limited, due to the lack of information on the genome of the prethymus. In order to fill this shortcoming, two important economic insects (Bombyx mori and Antheraea pernyi) were used in this paper to compare the transcriptome of prethymus using high-throughput sequencing technique. At the same time, a whole set of biological clock genes were identified in the prethymus based on transcriptional data. In addition, SSR site information in transcriptional data of tussah was analyzed. This paper is the first time to carry out comparative transcriptome analysis of insect prethymus to provide valuable information resources for further understanding the characteristics and functions of this important endocrine organ. The main results are as follows: 1. Transcriptome sequencing and comparative analysis of prethymus of tussah and Bombyx mori. After stitching and assembling, 641 transcripts (average length 657 BP) and 44067 Unigenes were obtained from tussah silkworm (Antheraea pernyi). Bombyx mori obtained 49287 transcripts (average length 1064 BP) and 32302 Unigenes. The data of Antheraea pernyi and Bombyx mori have been stored in NCBI sequence read Archive, and the login numbers are SRR5119598 and SRR5119600respectively. In tussah silkworm and Bombyx mori, there are 22402 and 71.69 Unigenes, respectively. A comparative analysis showed that the functional annotation results of the two prothymocytes in go termscog category and KO assignments were highly similar. In addition, 6357 pairs of homologous genes. 2. 2 were identified from two pre thymus transcriptional data. Identification of biological clock gene and RT-PCR detection. Biological clock plays an important role in insect behavior and physiological processes. From transcriptional data of prethymus of Antheraea pernyi and Bombyx mori, we have identified 13 clock genes, including cryptochrome 1 Cryptochrome 2 (cryptochrome 1) Cryptochrome 2 (cryptochrome 2) and clock. The results of double-time detection of PAR-domainprotein 1 and kinase 鈪，

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