大型迪庆藏猪不同生长阶段肌肉生长差异基因及其调控通路分析  

Screening of differential genes for muscle growth and regulatory pathways analysis in large Diqing Tibetan pig at different growth stages

作　　者：聂靖茹 张博[2] 马黎[3] 张浩[2] 李国美 严达伟[1] 董新星[1] NIE Jingru;ZHANG Bo;MA Li;ZHANG Hao;LI Guomei;YAN Dawei;DONG Xinxing(College of Animal Science and Technology,Yunnan Agricultural University,Kunming 650201,China;College of Animal Science and Technology,China Agricultural University,Beijing 100193,China;Department of Animal Husbandary and Veterinary Medicine,Yunnan Vocational and Technical College of Agriculture,Kunming 650212,China)

机构地区：[1]云南农业大学动物科学技术学院,昆明650201 [2]中国农业大学动物科学技术学院,北京100193 [3]云南农业职业技术学院畜牧兽医学院,昆明650212

出　　处：《中国农业大学学报》2022年第6期132-144,共13页Journal of China Agricultural University

基　　金：云南省乡村振兴科技专项(202104BI090021);云南省重大科技专项(2018BB003)。

摘　　要：为筛选大型迪庆藏猪不同生长阶段肌肉生长差异的关键基因并分析其调控途径,以40、80和120kg的大型迪庆藏猪为对象,基于RNA-seq技术对背最长肌组织进行转录组测序,对获得的测序数据进行拼接、比对、注释和差异分析,筛选与肌肉生长相关的差异基因进行STEM趋势分析、功能分析并构建差异基因互作网络。结果表明:1)10~40kg阶段与40~80kg阶段、40~80kg阶段与80~120kg阶段比较共检测到730个、981个基因显著差异表达;2)差异表达基因STEM趋势分析显示,共有4个差异显著模块,模块11和14的差异基因先上调表达后轻微下调;模块9和10的差异基因先上调后下调表达;3)差异基因互作网络显示,10~40kg vs 40~80kg,FOS基因位于调控网络核心,与EGRs、CCL2和NOR-1等基因有互作关系,NOR-1基因上调导致肌肉生长速度加快,FOS基因下调抑制肌源性分化,EGRs基因下调导致胶原纤维束减少,CCL2基因下调导致肌肉再生受损;40~80kg vs 80~120kg,FOXO1、PDK4和PPARD等基因位于网络中心,SFRPs基因上调阻止成肌细胞终末分化,FZD7基因下调减缓肌纤维肥大速度,FOXO1下调导致肌生长抑制素mRNA下降减缓肌肉萎缩进程,PPARD与FOXO1均降低PDK4含量从而使肌肉丙酮酸脱氢酶复合物活性增强,对碳水化合物氧化和葡萄糖摄取增加,肌纤维变粗,与大型迪庆藏猪40至80kg生长速度显著快于10至40kg阶段,而80kg之后缓慢下降的规律吻合;4)挑选了12个差异基因进行qPCR验证,EGR1、SFRP1和FOXO1等12个基因定量验证结果与转录组测序结果一致。综上,本研究利用RNA-seq技术筛选出12个影响大型迪庆藏猪不同生长阶段肌肉生长的差异基因,可为解析地方猪肌肉生长的遗传调控机制、应用分子标记辅助选择提高迪庆藏猪瘦肉产量提供参考。In order to screen the key genes of muscle growth in different growth stages of large Diqing Tibetan pig(TP)and analyze their regulatory pathways,the transcriptome of longissimus dorsi(LD)of TPs of 40,80and 120kg was sequenced based on RNA-seq,and the sequencing data were spliced,compared,annotated and analyzed.The differential genes related to muscle growth were screened for STEM analysis and function analysis,and the differential gene interaction network was constructed.The results showed that:1)There were respectively 730and 981genes significantly differentially expressed in 10-40kg stage and 40-80kg stage and 40-80kg stage and 80-120kg stage.2)STEM analysis of differential expression genes(DEGs)showed that there were four modules with significant differences.The DEGs in modules 11and 14were up-regulated first and then slightly down-regulated.The DEGs of modules 9and 10 were up-regulated first and down-regulated later.3)The differential gene interaction network analysis showed that at 10-40kg vs 40-80kg,FOS gene was located in the center of regulatory network and interacted with EGRs,CCL2 and NOR-1.The up-regulation of NOR-1leads to the acceleration of muscle growth,the down-regulation of FOSinhibits myogenic differentiation,the down-regulation of EGRslead to the decrease of collagen fiber bundles,and the down-regulation of CCL2 leads to the damage of muscle regeneration.At 40-80kg vs 80-120kg,FOXO1,PPARD,PDK4 and other genes were located in the center of the network.The up-regulation of SFRPs genes prevent myoblasts from entering the terminal differentiation.The down-regulation of FZD7 slows down muscle fiber hypertrophy,down-regulation of FOXO1 leads to the decrease of myostatin mRNA and slows down the process of muscle atrophy.PPARD and FOXO1 both reduce the content of PDK4,so as to enhance the activity of muscle pyruvate dehydrogenase complex,increase carbohydrate oxidation and glucose uptake,and thicken muscle fibers,which is consistent with the law that the growth rate of TPs is faster at 40-80kg,signifi

关 键 词：大型迪庆藏猪 RNA-SEQ 肌肉生长 差异表达基因 调控途径 

分 类 号：S828[农业科学—畜牧学]