不同发育阶段斯氏副柔线虫比较转录组学分析  

The Comparative Transcriptome Analysis of Parabronema skrjabini at Different Developmental Stages

作　　者：王文龙 冯陈晨 红梅 岳建伟 呼和巴特尔 刘春霞[2] 

机构地区：[1]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,呼和浩特010018 [2]内蒙古农业大学生命科学学院,呼和浩特010018

出　　处：《中国农业科学》2017年第23期4644-4655,共12页Scientia Agricultura Sinica

基　　金：国家自然科学基金(31260603)

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【Objective】 The objective of this study was to identify the differentially expressed genes（DEG） and describe biological characteristics involved in functional classifications and metabolic pathways at different developmental stages of Parabronema skrjabini infecting camel, which is necessary to better understand functional genes involved in growth and development and enrich the transcriptome data of parasitical nematodes.【Method】Eggs, the third-stage larvae（L3 s） and females of Parabronema skrjabini were sequenced by Illumina Hi Seq2000 TM sequencing platform and constructed their c DNA libraries after quality filtering. De novo assembling and assembly efficiency assessment were carried out using Trinity, a short-read assembly program. Then all the effective sequential data obtained were assigned to the relevant databases to perform functional annotation and bioinformatic analysis.【Result】The results showed that 47 717, 76 342 and 54 624 unigenes were obtained respectively in eggs,the third-stage larvae and female stages. 33 579 differentially expressed genes（DEGs） were identified by comparing the unigenes obtained from eggs and L3 s stages, of which 20 477 were up-regulated and 13 102 were down-regulated. There were 32 199 differentially expressed genes between L3 s and female stages, of these genes, 9 293 were up-regulated genes and 22 906 were down-regulated genes. The differentially expressed genes of two pairwise comparisons were respectively enriched in Gene Ontology. 6 617, 3 891 and 8 755 differentially expressed genes comparing eggs and L3 s stages were annotated into database of biological process, cellular component and molecular function respectively, while the number by comparing L3 s and female stages were 7 043, 3 686 and 10 177 respectively. In KEGG pathways identification, 6 521 differentially expressed genes comparing eggs and L3 s stages were assigned to 251 KEGG pathways, and clustered significantly in MARK, Wnt signaling pathways and oxidative phosphorylation. In compa

关 键 词：斯氏副柔线虫 转录组 差异表达基因 发育相关基因 

分 类 号：S852.731[农业科学—基础兽医学]