不同腹脂率番鸭回肠转录组差异分析

Analysis of Ileum Transcriptome Differences in Muscovy Ducks with Different Abdominal Fat Rates

【目的】解析番鸭回肠组织中与腹脂沉积相关的主效基因。【方法】从2000只番鸭中随机选取200只70日龄的番鸭进行屠宰,采集腹脂并称重,计算腹脂率。按腹脂率大小排序,选取腹脂率最高的5只和最低的5只番鸭的回肠组织进行转录组测序分析。利用Illumina NovaSeq 6000高通量RNA-Seq测序技术对2种腹脂率番鸭的回肠进行转录组测序,使用RSEM软件将测序得到的reads序列与Trinity拼接的参考基因组进行序列比对,筛选出差异表达基因,利用GO和KEGG数据库进行功能注释、富集分析和聚类分析,并利用皮尔森相关性分析方法分析了腹脂重和腹脂率与差异表达基因的相关性。【结果】高腹脂率和低腹脂率番鸭的回肠差异表达基因共有602个;与低腹脂率番鸭相比,高腹脂率番鸭回肠中有285个基因上调,317个基因下调。GO和KEGG富集分析显示,有5个与脂肪代谢相关的GO条目(脂质应答、类固醇激素介导的信号通路、类固醇激素应答、类固醇激素刺激的细胞应答和脂质的细胞应答)和4个KEGG通路(PPAR信号通路、初级胆汁酸的生物合成、花生四烯酸代谢和胆汁分泌),并筛选出与脂肪代谢显著相关的6个基因(P<0.05),包括NR5A2、ACBP、ACOX2、FABP2、FABP4和FABP5,其中,ACOX2和FABP5基因与腹脂重和腹脂率呈显著正相关(P<0.05),ACBP基因与腹脂重和腹脂率呈极显著正相关(P<0.01)。【结论】不同腹脂率番鸭回肠转录组存在差异,发现5个GO条目和4个KEGG通路,NR5A2、ACBP、ACOX2、FABP2、FABP4和FABP5等基因与番鸭腹部脂肪沉积相关。

【Objective】This study was conducted to identify the major genes related to abdominal fat deposition in ileum of muscovy ducks.【Method】20070-day-old muscovy ducks were randomly selected from 2000 muscovy ducks for slaughter.The abdominal fat was collected and weighed to calculate abdominal fat percentage.Sorted by abdominal fat rate,the ileum tissues of 5 muscovy ducks with the highest abdominal fat percentage and 5 muscovy ducks with the lowest abdominal fat percentage were selected for transcriptome sequencing analysis.Illumina NovaSeq 6000 high-throughput RNA-Seq sequencing technology was used to perform transcriptome sequencing on ileum of 2 kinds of muscovy ducks with abdominal fat rate.RSEM software was used to compare the sequenced reads sequence with the reference genome of Trinity to screen out differentially expressed genes.GO and KEGG databases were used for functional annotation,enrichment analysis and cluster analysis,and Pearson correlation analysis was used to analyze the correlation between abdominal fat weight and abdominal fat percentage and differentially expressed genes.【Result】There were 602 differentially expressed genes in ileum of muscovy ducks with high and low abdominal fat rates.Compared with the muscovy ducks with low abdominal fat rate,285 genes were up-regulated and 317 genes were down-regulated in ileum of muscovy ducks with high abdominal fat rate.GO and KEGG enrichment analysis showed that there were 5 GO terms(lipid response,steroid hormone-mediated signaling,steroid hormone response,steroid hormone-stimulated cellular response,and lipid cellular response)and 4 KEGG pathways(PPAR signaling,biosynthesis of primary bile acids,arachidonic acid metabolism,and bile secretion)related to lipid metabolism.In addition,6 genes were found to be significantly correlated with fat metabolism(P<0.05),including NR5A2,ACBP,ACOX2,FABP2,FABP4 and FABP5,among which ACOX2 and FABP5 genes were significantly positively correlated with abdominal fat weight and abdominal fat percentage(P<0.05),ACBP gene was significantly positively correlated with abdominal fat weight and abdominal fat percentage(P<0.01).【Conclusion】There were differences in the ileum transcriptome of muscovy ducks with different abdominal fat rates.It was found that 5 GO terms and 4 KEGG pathways,NR5A2,ACBP,ACOX2,FABP2,FABP4 and FABP5 genes were related to abdominal fat deposition in muscovy ducks.