高温胁迫条件下大菱鲆肾脏转录组研究

Transcriptome Study of Kidney of Turbot under High-Temperature Stress

为探索大菱鲆(Scophthalmus maximus)耐高温的分子机制,筛选耐高温相关基因,采用高通量测序平台(IlluminaHiSeq-2500)分别对5个不同高温处理组的大菱鲆肾脏组织开展转录组测序,进行生物信息学分析,包括GO(基因功能注释)、SSR(简单重复序列)分析等。结果显示,通过组装得到Unigenes总数目为68525,长度范围为201~23456 bp,平均长度为1124 bp,N50长度为2316 bp。将Unigenes分别在Nr、Swissprot、KEGG、KOG、GO数据库进行序列比对及功能注释,共注释25498条,其中,Nr数据库注释到的Unigenes最多;按GO功能分类,共分为细胞组分、分子功能及生物学过程3类,包括56个功能组,其中,大量Unigenes与细胞进程、代谢过程、催化活性、生物调节、应激反应相关。将Unigenes进行pathway注释,归属于218条代谢通路,分为5类KEGG途径:代谢途径、遗传信息处理、细胞过程、环境信息和生物系统。进行转录因子分析,共检测到65类转录因子,其中,C2H2锌指蛋白家族的基因数目最多。通过对不同温度胁迫下基因表达谱结果进行分析,不同温度组之间存在着显著差异,在不同温度胁迫组中,20℃组与28℃组存在差异最大,差异基因达到4734个,其中,上调基因3386个,下调基因1348个。本研究建立了大菱鲆热应激肾脏转录组数据库,为大菱鲆高温胁迫分子机理研究提供了参考数据。

In order to explore the molecular mechanism of high-temperature tolerance in turbot,Scophthalmus maximus L.,and to screen heat tolerance related genes,the IlluminaHiSeq-2500 platform was used to sequence the transcriptome of five turbot kidneys from five different heat treatments,through bioinformatics analysis,including GO(gene function annotation),SSR(simple repeat sequence)analysis,etc.The main results obtained were as follows:the total number of Unigenes obtained by assembly was 68525,the length range was 201~23456 bp,the average length was 1124 bp,and the length of N50 was 2316 bp.Sequence alignment and function annotation of Unigenes was conducted in Nr,Swissprot,KEGG,KOG,and GO databases.A total of 25,498 clusters were obtained,of which the Nr database noted the most Unigenes;and according to GO function classification,had three kinds of cell components,molecular functions,and biological processes,including 56 functional groups.A large number of Unigenes were related to cell processes,metabolic processes,catalytic activities,biological regulation,and stress response.Further,Unigenes were classified into 218 metabolic pathways and five KEGG pathways:metabolic pathway,genetic information processing,cellular processes,environmental information,and biological systems.A total of 65 transcription factors were detected by transcription factor analysis,with the C2H2 Zn finger protein family having the highest number of genes.Through the analysis of gene expression profiles under different temperature stress,there were significant differences among different temperature groups,In the 20℃ and 28℃ groups,the difference was the largest;4734 genes,of which 3386 were up-regulated and 1348 were down-regulated.In this study,we established a database of renal transcriptional groups in turbot under heat stress,which provides abundant data for the study of molecular mechanisms of heat stress in turbot.