中国农业科学 ›› 2025, Vol. 58 ›› Issue (6): 1239-1258.doi: 10.3864/j.issn.0578-1752.2025.06.014

• 畜牧·兽医 • 上一篇    

脂质组与转录组联合揭示南阳牛不同肌肉组织脂质特征的差异表达模式

高岩浩1(), 王婷婷1, 白卫卫1, 杜兴杰1, 刘贤2, 秦本源2, 付彤1, 孙宇1, 高腾云1(), 张天留1()   

  1. 1 河南农业大学动物科技学院,郑州 450046
    2 河南省畜牧技术推广总站,郑州 450002
  • 收稿日期:2024-11-05 接受日期:2025-02-20 出版日期:2025-03-25 发布日期:2025-03-25
  • 通信作者:
    高腾云,Tel:13007530689;E-mail:
    张天留,Tel:19339965998;E-mail:
  • 联系方式: 高岩浩,Tel:15565032572;E-mail:gaoyanhao_eureka@163.com。
  • 基金资助:
    科技创新2030-重大专项(2023ZW0404802); 中国博士后科学基金面上资助项目(2023M741059); 河南省科技攻关项目(242102111002)

The Combination of Lipidome and Transcriptome Revealed the Differential Expression Patterns of Lipid Characteristics in Different Muscle Tissues for Nanyang Cattle

GAO YanHao1(), WANG TingTing1, BAI WeiWei1, DU XingJie1, LIU Xian2, QIN BenYuan2, FU Tong1, SUN Yu1, GAO TengYun1(), ZHANG TianLiu1()   

  1. 1 College of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046
    2 Animal Husbandry Technology Extension Station of Henan Province, Zhengzhou 450002
  • Received:2024-11-05 Accepted:2025-02-20 Published:2025-03-25 Online:2025-03-25

摘要:

【目的】 肌内脂质的组成与含量是影响风味和嫩度的重要因素,与牛肉品质紧密相关。通过比较南阳牛背最长肌与里脊组织脂质特征和基因表达模式,鉴定调控南阳牛不同肌肉组织脂质特征的潜在候选基因,为优质牛肉分子育种提供理论依据。【方法】 选用生长环境和遗传背景相同的成年南阳牛背最长肌和里脊组织为试验材料,采用气相色谱质谱联用(gas chromatography-mass spectrometry,GC-MS)、液相色谱质谱联用(liquid chromatography mass spectrometry,LC-MS)与转录组测序技术(RNA sequencing,RNA-seq),构建肌肉组织的脂质图谱与基因表达谱,鉴定差异脂质分子(differential lipid molecules, DLMs)和差异表达基因(differentially expressed genes, DEGs),进行功能富集分析及蛋白质互作网络分析(protein-protein interaction network, PPI)筛选潜在候选基因,并采用实时荧光定量PCR技术(real-time fluorescence quantitative PCR,RT-qPCR)进行表达水平验证。【结果】 在肌肉组织间共检测到19种脂肪酸,其中C18:0、C14:1n5和C16:1n7的含量在组织间存在显著差异。共检测到2 106种脂质分子,其中磷脂酰胆碱(phosphatidylcholine,PC)、甘油三酯(triacylglycerols,TG)和磷脂酰乙醇胺(phosphatidylethanolamine,PE)为主要成分。通过差异分析在肌肉组织间共筛选到39种DLMs和3 424个DEGs,经受试者工作特征曲线(receiver operating characteristic curve,ROC)分析鉴定到13种DLMs(如:DG(16:0_18:1)、DG(18:0_18:1)、DG(18:0_18:2))可作为组织间的潜在脂质生物标志物。PPI和MCODE分析获得3个与脂质代谢密切相关的核心基因模块,通路富集分析显示,DEGs与DLMs共同参与肌醇磷酸代谢、甘油酯代谢和甘油磷脂代谢通路。整合分析鉴定到19个调控脂质特征差异的潜在候选基因,其中7个基因(PLCG2SYNJ1LPIN1DGKZDGAT1LPLSELENOI)居于通路中的关键位置,对DLMs具有直接调控作用。RT-qPCR结果显示6个脂质候选基因的表达趋势与RNA-seq结果一致。【结论】 在南阳牛背最长肌与里脊组织间鉴定到13种潜在脂质生物标志物,筛选到19个潜在候选基因参与脂质特征调控的关键代谢通路,为今后进一步探究南阳牛脂质差异形成的调控机制及优质牛肉分子育种提供理论基础。

关键词: 南阳牛, 脂质生物标志物, 脂质组, 转录组, 候选基因

Abstract:

【Objective】 The composition and content of intramuscular lipids are important factors affecting flavor and tenderness of beef, and are closely related to beef quality. In this study, by comparing the lipid characteristics and gene expression patterns of longissimus dorsi muscle and tenderloin tissue of Nanyang cattle, the potential candidate genes regulating lipid characteristics of different muscle tissues in Nanyang cattle were identified. 【Method】 The longissimus dorsi muscle and tenderloin tissues of adult Nanyang cattle with the same growth environment and genetic background were selected as experimental materials, and then the lipid profile and gene expression profile of muscle tissue were constructed by gas chromatography-mass spectrometry (GC-MS), liquid chromatography-mass spectrometry (LC-MS) and transcriptome sequencing (RNA-seq) to identify differential lipid molecules (DLMs) and differentially expressed genes (DEGs) between different tissues. Functional enrichment analysis and protein-protein interaction network (PPI) were performed to screen potential candidate genes, and real-time fluorescent quantitative PCR (RT-qPCR) was used to verify their expression levels. 【Result】 In this study, 19 kinds of fatty acids were detected in muscle tissue, among which the content of C18:0, C14:1n5 and C16:1n7 were significantly different between tissues. A total of 2 106 lipid molecules were detected, of which Phosphatidylcholine (PC), Triacylglycerols (TG) and Phosphatidylethanolamine (PE) were the main components. A total of 39 DLMs and 3,424 DEGs were screened between two muscle tissues by difference analysis. According to receiver operating characteristic curve (ROC) analysis, 13 DLMs (e.g. DG(16:0_18:1), DG(18:0_18:1), DG(18:0_18:2)) could be used as potential lipid biomarkers between tissues. PPI and MCODE analysis obtained three core gene modules closely related to lipid metabolism. Pathway enrichment analysis showed that DEGs and DLMs were involved in Inositol phosphate metabolism, Glycerolipid metabolism and Glycerophospholipid metabolism. Integration analysis identified 19 potential candidate genes with different lipid characteristics, among which 7 genes (PLCG2, SYNJ1, LPIN1, DGKZ, DGAT1, LPL and SELENOI) were located in key positions in the pathway, and had direct regulatory effects on DLMs. RT-qPCR showed that the expression trend of six candidate genes was consistent with that of RNA-seq. 【Conclusion】 In this study, 13 potential lipid biomarkers were identified and 19 potential candidate genes were screened for the key metabolic pathways involved in the regulation of lipid characteristics between longissimus dorsi muscle and tenderloin tissue for Nanyang cattle, which provided a theoretical basis for further exploration of the regulatory mechanism for lipid difference formation in Nanyang cattle and molecular breeding for high-quality beef.

Key words: Nanyang cattle, lipid biomarkers, lipidome, transcriptome, candidate gene