大型迪庆藏猪不同生长阶段肌内脂肪沉积差异表达基因及其调控通路分析

doi:10.16431/j.cnki.1671-7236.2022.08.002

摘要/Abstract

摘要： 【目的】筛选大型迪庆藏猪不同生长阶段肌内脂肪含量差异的关键基因并分析其调控途径。【方法】选择胎次相同、出生日期及体重相近的大型迪庆藏猪36头,随机分为3组,在相同条件下进行育肥试验,分别在体重达40、80及120 kg左右时屠宰,每组采集3头猪的背最长肌,采用RNA-Seq技术进行转录组测序,测序数据进行拼接、比对和注释,筛选与脂肪沉积相关的差异显著基因进行功能富集、STEM分析,构建基因互作网络,并进行实时荧光定量PCR验证。【结果】40 kg vs 80 kg、80 kg vs 120 kg与40 kg vs 120 kg阶段分别检测到730、981及735个基因差异显著表达;STEM分析共有4个差异显著模块,模块11、14的基因集先显著上调后轻微下调;模块9、10的基因集先显著上调后显著下调。差异基因互作网络显示,40 kg vs 80 kg阶段,EGR1、EGR2、PRKAG2、NOR-1和ATF3基因位于网络核心,EGR1和EGR2基因表达下调,PRKAG2、NOR-1和ATF3基因表达上调;80 kg vs 120 kg阶段,FOXO1、PDK4、PPARD、PPARGC-1、LIPE、ATF3和STAT1基因位于网络核心,FOXO1、PPARD、PPARGC-1基因表达下调,PPARG基因通过级联调控引起STAT1基因表达下调,LIPE和ATF3基因表达上调;40 kg vs 120 kg阶段,ATF3、NOR-1、EGR1、EGR2和STAT1基因位于网络核心,EGR1、EGR2和STAT1基因表达下调,ATF3和NOR-1基因表达上调。ATF3、FOXO1等10个基因的实时荧光定量PCR验证结果与转录组测序结果一致。【结论】EGR1、FOXO1等基因作为核心基因参与大型迪庆藏猪肌内脂肪调控,不同生长阶段参与调控的核心基因并不完全相同,结果可丰富中国地方猪肌内脂肪调控基础数据,为大型迪庆藏猪肌内脂肪含量的遗传改良提供参考。

关键词: 大型迪庆藏猪; RNA-Seq; 肌内脂肪沉积; 差异表达基因; 调控途径

Abstract: 【Objective】 This study was aimed to screen the key genes of the difference of intramuscular fat (IMF) content of large Diqing Tibetan pigs (TPs) at different growth stages and analyze its regulation pathway.【Method】 There were thirty-six TPs with the same parity,date of birth and weight were randomly divided into three groups.The fattening test was carried out under the same conditions.They were slaughtered when the weight was about 40,80 and 120 kg,respectively.The longissimus dorsi muscle (LD) of three pigs in each group was collected,and the transcriptome was sequenced by RNA-Seq.The sequencing data was spliced,compared and annotated,the significantly differentially expressed genes (DEGs) related to IMF deposition were screened,and the gene interaction network was constructed by functional enrichment and STEM analysis.The results were verified by Real-time quantitative PCR.【Result】 There were 730,981 and 735 genes were significantly differentially expressed in 40 kg vs 80 kg,80 kg vs 120 kg and 40 kg vs 120 kg stages,respectively.There were four modules with significant differences in STEM analysis.The gene sets of modules 11 and 14 were significantly up-regulated and then slightly down-regulated.The gene sets of modules 9 and 10 were significantly up-regulated and then down-regulated.The differential gene interaction network showed that at 40 kg vs 80 kg stage,EGR1,EGR2,PRKAG2,NOR-1 and ATF3 genes were located in the core of the network,EGR1 and EGR2 genes were down-regulated,PRKAG2,NOR-1 and ATF3 genes were up-regulated.At 80 kg vs 120 kg stage,FOXO1,PDK4,PPARD,PPARGC-1,LIPE,ATF3 and STAT1 genes were located in the core of the network,FOXO1,PPARD and PPARGC-1 genes were down-regulated,and PPARG gene caused STAT1 gene down-regulation through cascade regulation,LIPE and ATF3 genes were up-regulated.At 40 kg vs 120 kg stage,ATF3,NOR-1,EGR1,EGR2 and STAT1 genes were located in the core of the network,EGR1,EGR2 and STAT1 genes were down-regulated,ATF3 and NOR-1 genes were up-regulated.Real-time quantitative PCR results of ten genes such as ATF3 and FOXO1 were consistent with RNA-Seq results.【Conclusion】 This study screened some genes like EGR1 and FOXO1 participate in the regulation of IMF in TPs as core genes,which involved in regulation at different stages were not exactly the same.The results could enrich the basic data of IMF regulation in local pigs in China,and provide a reference for the genetic improvement of IMF content in TPs.

Key words: large Diqing Tibetan pigs; RNA-Seq; intramuscular fat deposition; differentially expressed gene; regulation pathway

中图分类号:

S828

聂靖茹, 马黎, 严达伟, 邓俊, 张浩, 张博, 刘金桥, 董新星. 大型迪庆藏猪不同生长阶段肌内脂肪沉积差异表达基因及其调控通路分析[J]. 中国畜牧兽医, 2022, 49(8): 2855-2868.

NIE Jingru, MA Li, YAN Dawei, DENG Jun, ZHANG Hao, ZHANG Bo, LIU Jinqiao, DONG Xinxing. Analysis of Differentially Expressed Genes and Regulation Pathways of Intramuscular Fat Deposition in Large Diqing Tibetan Pigs at Different Growth Stages[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 2855-2868.

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