不同发育阶段斯氏副柔线虫比较转录组学分析

发布时间：2018-05-27 16:03

  本文选题：斯氏副柔线虫 + 转录组　； 参考：《中国农业科学》2017年23期

【摘要】：【目的】探明不同发育阶段骆驼斯氏副柔线虫的转录组差异,了解不同发育阶段虫体在功能分类和代谢通路等方面的生物学特征,挖掘生长发育相关功能基因,丰富寄生性线虫转录组学信息。【方法】采用Illumina Hi Seq2000TM高通量测序技术对斯氏副柔线虫虫卵、第三期幼虫和雌虫进行转录组测序,构建3个样本的c DNA文库,评估建库质量;利用Trinity软件对所得序列进行De novo组装及组装效率评估;之后对获得的有效序列进行功能注释及相关生物信息学分析。【结果】测序及组装后虫卵、第三期幼虫和雌虫分别获得47 717、76 342和54 624个Unigenes,在虫卵和第三期幼虫比对中共有33 579个差异表达基因,其中表达上调的基因有20 477个,表达下调的基因有13 102个;在雌虫和第三期幼虫比对中共有32 199个差异表达基因,其中表达上调的基因有9 293个,表达下调的基因有22 906个。将成对比较的差异表达基因分别进行GO功能分类,其中虫卵和第三期幼虫比对中有6 617、3 891和8 755个Unigenes,雌虫和第三期幼虫比对中有7 043、3 686和10 177个Unigenes分别注释到生物过程、细胞组成和分子功能三大类中;通过KEGG pathway数据库分析,虫卵和第三期幼虫比对中有6 521个差异表达基因参与到251条通路中,显著富集MAPK信号通路、Wnt信号通路和氧化磷酸化等通路,雌虫和第三期幼虫比对中有6 528个参与到251条通路中,显著富集新陈代谢通路、DNA复制和细胞周期等通路。差异表达基因功能聚类分析中,雌虫和虫卵在调控生长速率和生殖发育等方面高表达;第三期幼虫在防御机制和糖类代谢等方面高表达;且3个发育阶段均在胚胎发育,胚后发育中富集高表达,其中在胚胎发育和胚后发育中分别有242和202个相关功能基因在3个发育阶段都表达,这些基因可能在胚胎(后)发育过程中起核心作用。此外,获得了9种异时性基因(如:LIN-28、LIN-14和RHEB-1等)、48个核激素受体(NHRs),包括NHR-49,NHR-48,NHR-40,NHR-1等以及36个锌金属蛋白酶(NAS),包括NAS-36、NAS-33和NAS-14等,并对其在斯氏副柔线虫虫卵、第三期幼虫和雌虫中的富集程度进行分析,发现这些基因在维持线虫正常生长发育过程中发挥关键作用,【结论】利用RNA-seq技术对3个发育阶段的斯氏副柔线虫进行测序和生物信息学分析,研究了不同发育阶段虫体的差异表达基因在GO功能分类、KEGG代谢通路和基因功能聚类等方面的生物学特性,筛选出多种异时性基因和发育相关重要基因,为后续开展斯氏副柔线虫功能基因组学研究,虫体与宿主互作、致病机制、免疫逃避等研究提供了理论依据。
[Abstract]:[Objective] to explore the differences in the transcriptional groups of camel's parasitic nematode in different developmental stages, to understand the biological characteristics of different developmental stages in functional classification and metabolic pathways, to excavate growth and development related genes and to enrich the transcriptional information of parasitic nematodes. [Methods] high throughput sequencing of Illumina Hi Seq2000TM was used. Technology was used to sequence the egg, third stage larvae and females of the Coriolis slovaki, construct the C DNA Library of 3 samples, evaluate the quality of the building, and evaluate the assembly and assembly efficiency of the De novo using Trinity software; and then the effective sequence was annotated and the related bioinformatics analysis. The third stage larvae and females obtained 47 717,76 342 and 54624 Unigenes respectively. There were 33579 differentially expressed genes in the egg and third stage larvae, of which 20477 genes were up-regulated, and 13102 expressed down regulated genes, and 32199 differences were expressed in the female and third stage larvae. In the gene, there were 9293 up-regulated genes and 22906 down-regulated genes. The differentially expressed genes were divided into GO functional classifications, including 6 617,3 891 and 8755 Unigenes for insect eggs and third stage larvae, and 7 043,3 686 and 10177 Unigenes respectively annotated by 7 043,3 686 and 10177 Unigenes respectively. In the three major categories of biological processes, cell composition and molecular function, through the analysis of KEGG pathway database, 6521 differentially expressed genes in the egg and third stage larvae were involved in the 251 pathways, which significantly enriched the MAPK signaling pathway, the Wnt signaling pathway and the oxidative phosphorylation pathway, and 6528 of the third and third stage larvae were involved. In the 251 pathways, the metabolic pathways, DNA replication and cell cycle pathways were significantly enriched. In the functional cluster analysis of differentially expressed genes, the female and the eggs were highly expressed in the regulation of growth rate and reproductive development, and the third stage larvae were highly expressed in the defense mechanism and carbohydrate metabolism, and the 3 developmental stages were in the embryonic development and after the embryo. There are 242 and 202 related functional genes in the embryonic development and postembryonic development, respectively, in which 242 and 202 related genes are expressed in 3 developmental stages. These genes may play a core role in the development of the embryo. In addition, 9 kinds of isochronic genes (such as LIN-28, LIN-14 and RHEB-1, etc.), and 48 nuclear hormone receptors (NHRs), including NHR-, are obtained. 49, NHR-48, NHR-40, NHR-1 and 36 zinc metalloproteinases (NAS), including NAS-36, NAS-33 and NAS-14, and their enrichment in the eggs of the Coriolis slovaki, third stage larvae and females, and found that these genes play a key role in maintaining normal growth and development of the nematodes. [Conclusion] 3 with RNA-seq technology is used. By sequencing and bioinformatics analysis, the biological characteristics of differential expression genes of different developmental stages of insect body in GO function classification, KEGG metabolic pathway and gene function clustering were studied. The research on insect functional genomics provides a theoretical basis for the interaction between host and host, pathogenesis and immune evasion.
【作者单位】： 内蒙古农业大学兽医学院/农业部动物疾病临床诊疗技术重点实验室;内蒙古农业大学生命科学学院;
【基金】：国家自然科学基金(31260603)
【分类号】：S852.731
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本文编号：1942813