Key Laboratory of Tropical & Subtropical Fisheries Resource Application & Cultivation, China Ministry of Agriculture, Pearl River Fisheries Institute, Chinese Academy of Fishery Sciences, Guangzhou, 510380, China.
Mol Biol Rep. 2013 Oct;40(10):5657-68. doi: 10.1007/s11033-013-2667-3. Epub 2013 Sep 26.
Tilapia is an important freshwater aquaculture species worldwide. In recent years, streptococcal diseases have severely threatened development of tilapia aquaculture, while effective prevention and control methods have not yet been established. In order to understand the immunological response of tilapia to infection by Streptococcus agalactiae (S. agalactiae), this study employed Solexa/Illumina RNA-seq and digital gene expression (DGE) technology to investigate changes in the tilapia transcriptome before and after S. agalactiae infection. We obtained 82,799 unigenes (mean size: 618 bp) using de novo assembly. Unigenes were annotated by comparing against databases including Nr, Swissprot, cluster of orthologous groups of proteins, Kyoto encyclopedia of genes and genomes, and gene ontology. Combined with DGE technology, transcriptomic changes in tilapia before and after bacteria challenging were examined. A total of 774 significantly up-regulated and 625 significantly down-regulated unigenes were identified, among which 293 were mapped to 181 signaling pathways including 17 immune-related pathways involving 65 differentially expressed genes. We observed a change in the expression of six genes in the Toll-like receptor signaling pathway, and this was subsequently confirmed via quantitative real-time PCR. This comparative study of the tilapia transcriptome before and after S. agalactiae infection identified important differentially-expressed immune-related genes and signaling pathways that will provide useful insights for further analysis of the mechanisms of tilapia defense against S. agalactiae infection.
罗非鱼是世界范围内重要的淡水养殖品种。近年来,链球菌病严重威胁着罗非鱼养殖业的发展,但尚未建立有效的预防和控制方法。为了了解罗非鱼对无乳链球菌(Streptococcus agalactiae,S. agalactiae)感染的免疫反应,本研究采用高通量测序和数字基因表达(digital gene expression,DGE)技术,研究了罗非鱼在无乳链球菌感染前后转录组的变化。我们通过从头组装获得了 82799 条 unigenes(平均大小:618bp)。通过与 Nr、Swissprot、蛋白质直系同源群簇、京都基因与基因组百科全书和基因本体等数据库的比对,对 unigenes 进行了注释。结合 DGE 技术,检测了细菌攻毒前后罗非鱼的转录组变化。共鉴定出 774 个显著上调和 625 个显著下调的 unigenes,其中 293 个 unigenes映射到 181 条信号通路,包括涉及 65 个差异表达基因的 17 条免疫相关通路。我们观察到 Toll 样受体信号通路中 6 个基因的表达发生了变化,随后通过定量实时 PCR 进行了验证。本研究对罗非鱼无乳链球菌感染前后的转录组进行了比较分析,鉴定出了重要的差异表达免疫相关基因和信号通路,为进一步分析罗非鱼抵抗无乳链球菌感染的机制提供了有用的信息。