基于转录组数据的蒙古牛皮肤组织抗寒相关信号通路及候选基因的筛选

摘要

The aim of this study was to find out signaling pathways and candidate genes that related to cold tolerance of Mongolia cattle by RNA-sequencing and bioinformatics analysis of Mongolia cattle skin tissues in winter and summer.Transcriptome sequencing skin tissues of adult Mongolia cattle in winter and summer were carried out by Ion ProtonTM Sequencer,respectively.And then data processing,reference genome alignment,gene differential expression analysis,GO and KEGG enrichment analysis and candidate gene screening were applied on the transcriptome data;Additionally,the reliability of the transcriptome data was detected by Real-time PCR.The results showed that 86 954 060 and 69 837 009 reads were obtained from winter and summer groups,respectively;The percentage of the uniquely mapped reads was 73.21％ in winter group,and 75.55％ in summer group.There were 182 DEGs between winter and summer groups,compared to summer group,117 were up-regulated and 65 were down-regulated in winter group;Real-time PCR analysis showed that the transcriptome sequencing results were reliable;The results of GO analysis showed there were 21,39 and 224 terms enriched in the cellular components,molecular functions and biological processes,respectively;KEGG analysis showed that the DEGs were significantly enriched in lipid metabolism,blood coagulation,tyrosine metabolism,steroid hormone biosynthesis and retinol metabolism related pathways;8 candidate genes were screened out.In summary,lipid metabolism,blood coagulation and melanin synthesis related pathways and genes as well as hair synthesis related genes may play an important role in cold resistance of Mongolian cattle.%本研究旨在通过对冬夏两季蒙古牛皮肤组织进行转录组测序和生物信息学分析,找到与蒙古牛抗寒性状相关的信号通路及候选基因.用Ion ProtonTM Sequencer分别对冬季和夏季成年蒙古牛的皮肤组织进行转录组测序,随后对转录组数据进行处理、与参考基因组比对、差异表达分析、GO和KEGG富集分析以及候选基因筛选,并用Real-time PCR方法对转录组数据进行可靠性检测.结果显示,冬季组和夏季组分别得到86 954 060和69 837 009条reads;冬季组与参考基因组的唯一比对率为73.21％,夏季组为75.55％;冬夏两组共有182个差异表达基因(DEGs),与夏季组相比,其中117个在冬季组中上调,65个在冬季组中下调;Real-time PCR分析显示,转录组测序结果可靠;GO分析结果显示,细胞组分、分子功能、生物学过程分别富集到21、39和224个term;KEGG分析结果显示,差异表达基因(DEGs)显著地富集在脂类代谢、凝血、酪氨酸代谢、类固醇合成和视黄醇代谢相关通路上;共筛选出8个候选基因.综上表明,脂类代谢、凝血、黑色素合成相关通路和基因,以及与毛发发生相关的基因可能在蒙古牛抗寒过程中发挥重要作用.