乌鳢感染鰤诺卡氏菌的转录组反应(英文)

Northern snakehead Channa argus de novo transcriptomic response to Nocardia seriolae infection

利用转录组技术对鰤诺卡氏菌(Nocardia seriolae)感染的乌鳢(Channa argus)进行分析与比较,以揭示可能的免疫机制,尤其在Toll样受体和Nod样受体的免疫反应.从乌鳢的头肾提取总RNA,采用Illumina Hi Seq TM2500进行测序和de novo转录组分析.乌鳢感染鰤诺卡氏菌后,对照组和试验组的clean reads分别是33 556 284(93.79%)和34 202 766(93.52%).利用Trinity软件对unigenes进行注释,从133 999条unigenes里,一共106319条unigenes得到注释,占79.34%;NR(NCBI non-redundant protein database)注释54 886(40.97%)条unigenes;39 795(29.69%)的Swiss-Prot注释;5 885(4.39%)条unigenes被KEGG(Kyoto encyclopedia of genes and genomes)注释到335条通路;5 753(4.29%)被注释到GO(Gene ontology)里.3 912条unigenes表达差异,其中1552条表达上调(39.67%)和2 360条表达下调(60.33%).Toll样受体和Nod样受体的免疫信号通路分别有28和13条差别表达基因发现有调制,12和3个上调,11和6个下调,5和4个unigenes没有显著的变化.经过转录组测序分析显示,乌鳢感染鰤诺卡氏菌后的转录复杂程度高,导致感染个体重诱导表达的基因产品的验证发现,尤其在非特异性免疫系统方面.

The head kidney transcriptional response of Northern snakehead fish （Channa argus ） with Nocardia seriolae infection, expecially Toll and Nod like receptors, were analyzed and compared in order to reveal mechanisms that might confer infection resistance. Total RNA was extracted from the head kidney with or without N. seriolae, subjected to transcriptome analysis using Illumina HiSeqTM 2500 sequencing followed by de novo Transcriptome analysis. Clean reads of the control group and the experimental group of snakehead fish infected by N. seriolae were 33 556 284 （93. 79% ） and 34 202 766 （93.52%）, respec- tively. Annotation was performed by using Trinity software, 106 319 unigenes were annotated from the to- tal of 133 999 unigenes, account for 79.34% ; the NCBI non-redundant protein database （NR） annota- tions yielded 54 886 unigenes （40. 97% ） ; Swiss-Prot annotations had 39 795 unigenes （29.69%） ; the annotation by Kyoto encyclopedia of genes and genomes （KEGG） has 5 885 （4.39%） unigenes were an-notated into 335 channels, and the Gene ontology （GO） annotations has 5 753 unigenes （4.29%）. Ap- proximately, 3 912 genes were differentially expressed, among them, 1 552 unigenes were up-regulated （39.67%）, and 2 360 unigenes were down-regulated （60. 33% ）. Subsequently, there were 28 or 13 differentially expressed genes found to be modulated, from that 12 or 3 unigenes were up-regulated, 11 or 6 unigenes were down-regulated, 5 and 4 unigenes did not showed significant changes from the Toll and Nod like receptor immune signaling pathways, respectively. In conclusion, our deep sequencing analysis revealed in depth the high degree of transcriptional complexity of the C. argus immune response to N. se-riolae infection and resulted in the discovery and validation of new gene products with induced expression in infected individuals, especially in the non-specific immune system. Key words： Northern snakehead fish （ Channa argus） ; Nocardia seriolae ; transcriptome ; Toll like recep- tors （TLR） ; Nod like receptors （NLR） histopathology