2种不同生长规格墨瑞鳕肌肉组织转录组测序分析

Analysis of muscle transcriptome of two different growth specifications of Maccullochella peelii

  • 摘要: 【目的】掌握不同生长规格墨瑞鳕个体间差异表达基因的表达特点,为其功能相关基因深度挖掘及分子遗传育种提供科学依据。【方法】挑选同一养殖条件下极大个体和极小个体的墨瑞鳕,构建肌肉组织cDNA文库后,采用Illumina HiSeqTM 4000测序平台对存在生长差异的墨瑞鳕肌肉组织进行转录组测序分析,获得的Unigenes在Nr、Nt、Pfam、KOG/COG、Swiss-Prot、KEGG和GO等数据库中进行比对;通过FPKM及DEGseq筛选出差异表达基因,以GOseq和KOBAS对差异表达基因分别进行GO功能注释及KEGG信号通路富集分析,并采用MISA进行SSR鉴定分析。【结果】从墨瑞鳕肌肉组织中共测序获得39749条Unigenes,其长度范围在301~55230 bp,平均长度为1705 bp。注释到Nt、Nr、Swiss-Prot、Pfam数据库的Unigenes分别有27046、20824、18268和17772条,在7个数据库中均得到注释的Unigenes共计6742条,占Unigenes总数的16.96%。根据差异表达基因筛选条件P<0.05且|log2 Fold Change|>1,共筛选出722个差异表达基因,其中上调基因308个、下调基因414个。差异表达基因GO功能注释分析结果表明,注释基因数目较多的GO功能条目包括细胞过程、代谢过程、膜、细胞器及结合等;KEGG信号通路富集分析发现,差异表达基因被成功富集到234条信号通路上,主要涉及磷脂酰肌醇3激酶/蛋白激酶信号通路、MAPK信号通路、胰岛素信号通路及FoxO信号通路等。在39749条Unigenes中鉴定筛选出22120个SSRs,占Unigene总数的55.65%,SSR的平均间距为3063 bp。【结论】基于转录组测序分析获得的墨瑞鳕肌肉组织差异表达基因以发挥结合、细胞过程及代谢过程等功能为主,且主要富集在PI3K-Akt信号通路、核糖体信号通路、FoxO信号通路及细胞凋亡等能量代谢相关通路上,通过共同协调而对墨瑞鳕的生长发育起调控作用。

     

    Abstract: 【Objective】To identify differentially expressed genes(DEGs)between small and large individuals of Maccullochella peelii,to provide a scientific basis for the deep study of genes related to growth and development,molecular genetics and breeding.【Method】Significantly large and small M. peelii individuals obtained under the same breeding conditions were selected and their muscle tissue sampled to construct cDNA libraries. Transcriptome sequencing of M. peelii muscle tissues was conducted using the Illumina HiSeqTM 4000 sequencing platform. The obtained unigenes were compared in Nr,Nt,Pfam,KOG/COG,Swiss-Prot,KEGG and GO databases. The differentially expressed genes were screened using FPKM and DEGseq. GOseq and KOBAS were used to perform GO function annotation and KEGG signaling pathway enrichment,then identified by SSR by MISA.ResultsA total of 39749 unigenes were generated from muscle tissue of M. peelii,with lengths ranging from 301 to 55230 bp and an average length of 1705 bp. There were 27046, 20824,18268 and 17772 unigenes annotated to Nt,Nr,Swiss-Prot and Pfam databases,respectively. In total,6742 unigenes were annotated by seven databases,accounted for 16.96% of the total unigenes. Differentially expressed genes were defined as those displaying an adjusted P<0.05 and|log2 Fold Change|>1. A total of 722 DEGs were selected,consisting of 308 up-regulated genes and 414 down-regulated genes. The results of GO function annotation showed that the GO function items with a large number of annotation genes including cellular process,metabolic process,membrane, organelle and binding. KEGG signaling pathway enrichment revealed that DEGs were enriched in 234 signal pathways, mainly involving phosphatidylinositol 3 kinase/protein kinase signal pathway,MAPK signal pathway,insulin signal pathway and the FoxO signal pathway. A total of 22120 SSRs were identified from 39749 unigenes,accounted for 55.65% of the total number of unigenes. The average spacing of SSRs was 3063 bp.【Conclusion】The obtained DEGs in M. peelii muscle tissue based on transcriptome sequencing mainly play the functions of binding,cellular process and metabolic process,and mainly enriched in PI3K Akt signal pathway,ribosomal signal pathway,FoxO signal pathway,apoptosis and other energy metabolism related pathways,which together regulate the growth and development of M. peelii.

     

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