石淳元, 赵彦玲. 2024: 基于转录组测序的藏猪睾丸组织性成熟相关基因鉴定. 南方农业学报, 55(3): 680-688. DOI: 10.3969/j.issn.2095-1191.2024.03.009
引用本文: 石淳元, 赵彦玲. 2024: 基于转录组测序的藏猪睾丸组织性成熟相关基因鉴定. 南方农业学报, 55(3): 680-688. DOI: 10.3969/j.issn.2095-1191.2024.03.009
SHI Chun-yuan, ZHAO Yan-ling. 2024: Identification of genes related to sexual maturation in Tibetan pig testis tissue based on transcriptome sequencing. Journal of Southern Agriculture, 55(3): 680-688. DOI: 10.3969/j.issn.2095-1191.2024.03.009
Citation: SHI Chun-yuan, ZHAO Yan-ling. 2024: Identification of genes related to sexual maturation in Tibetan pig testis tissue based on transcriptome sequencing. Journal of Southern Agriculture, 55(3): 680-688. DOI: 10.3969/j.issn.2095-1191.2024.03.009

基于转录组测序的藏猪睾丸组织性成熟相关基因鉴定

Identification of genes related to sexual maturation in Tibetan pig testis tissue based on transcriptome sequencing

  • 摘要: 【目的】基于转录组测序分析筛选出藏猪睾丸组织中的性成熟相关基因,为提高藏猪的繁殖育种性能提供参考依据。【方法】以性成熟前(2月龄)和性成熟后(24月龄)的藏公猪为研究对象,通过人工阉割法采集藏猪睾丸组织进行转录组测序,依据错误发现率(FDR)<0.05且|log2 Fold Change|>1的标准筛选差异表达基因(DEGs),综合GO功能注释分析和KEGG信号通路富集分析筛选出藏公猪性成熟相关候选基因,并通过实时荧光定量PCR验证转录组测序结果的准确性。【结果】经转录组测序,从性成熟前后藏猪睾丸组织样品中筛选出5929个DEGs,其中有3822个DEGs呈上调表达、2107个DEGs呈下调表达。筛选获得的5929个DEGs主要注释到生精过程、纤毛运动、鞭毛精子运动、运动纤毛、精子主块、精子顶囊、动力蛋白轻链结合、动态蛋白中间链结合等与精子发生和精子运动等有关的GO功能条目;KEGG信号通路富集分析发现,DEGs主要富集于受体相互作用通路、轴突引导通路、神经活性配体—受体相互作用通路、钙信号通路等;综合DEGs的GO功能注释分析和KEGG信号通路富集分析结果,共筛选出9个与藏公猪性成熟相关的基因,分别是BAX、DMRT1、FSHR、IGF1R、IZUMO1、LYZL4、LYZL6、SYCP3TNP2。实时荧光定量PCR检测结果显示,BAX、DMRT1、FSHR、IGF1R等4个基因在性成熟后藏猪睾丸组织中的相对表达量较性成熟前呈明显的下降趋势,且下降幅度均在50%以上;IZUMO1、LYZL4、LYZL6、SYCP3、TNP2等5个基因的相对表达量较性成熟前上升了1.07~30.25倍,其中IZUMO1、LYZL4LYZL6基因的上升倍数达极显著水平(P<0.01),与转录组测序结果基本一致。【结论】基于转录组测序分析从藏猪睾丸组织中筛选获得9个与性成熟相关的DEGs,即BAX、DMRT1、FSHR、IGF1R、IZUMO1、LYZL4、LYZL6、SYCP3TNP2,可作为研究藏猪繁殖育种能力的主要候选基因。

     

    Abstract: 【Objective】Based on transcriptome sequencing analysis, the genes related to sexual maturation were screened out in testicular tissue of Tibetan pigs so as to provide reference for improving the breeding performance of Tibetan pigs. 【Method】Tibetan boars before sexual maturity (2 months old) and after sexual maturity (24 months old) were selected as the study objects, testicular tissues of the Tibetan boars were collected by artificial castration method for transcriptomic sequencing, and differentially expressed genes (DEGs) were screened according to the criteria of false discovery rate (FDR)<0.05 and |log2 Fold Change|>1. GO functional annotation analysis and KEGG signaling pathway enrichment analysis were combined to select the candidate genes related to sexual maturity in Tibetan boars. And the accuracy of transcriptome sequencing results were verified by real-time fluorescence quantitative PCR. 【Result】According to transcriptome sequencing, 5929 DEGs were screened from testicular tissue samples of Tibetan pigs before and after sexual maturity, among which 3822 DEGs were up-regulated and 2107 DEGs were down-regulated. The 5929 DEGs were selected to include spermatogenesis, ciliary motility, flagellate sperm motility, motile cilia, sperm mass, sperm apical sac, kinetic protein light chain binding, dynamic protein intermediate chain binding and other GO functional entries related to spermatogenesis and sperm motility. KEGG signaling pathway enrichment analysis showed that the DEGs was mainly enriched in receptor interaction pathway, axon-guided pathway, neural active ligand-receptor interaction pathway, calcium signaling pathway, etc. Based on the GO functional annotation analysis of DEGs and the enrichment analysis of KEGG signaling pathway, a total of nine genes related to sexual maturation in Tibetan boars were selected, which were BAX, DMRT1, FSHR, IGF1R, IZUMO1, LYZL4, LYZL6, SYCP3 and TNP2. Real-time fluorescence quantitative PCR detection results showed that the relative expression levels of the four genes, including BAX, DMRT1, FSHRand IGF1R, were decreased after sexual maturity compared with that before sexual maturity, all decreased by more than 50%. The relative expression levels of the five genes, including IZUMO1, LYZL4, LYZL6, SYCP3 and TNP2, were increased by 1.07-30.25 times compared with that before sexual maturity, and the increasing times of IZUMO1,LYZL4and LYZL6 genes reached extremely significant level(P<0.01), which was basically consistent with the transcriptome sequencing results. 【Conclusion】Nine DEGs related to sexual maturity are detected in testicular tissue of Tibetan pigs before and after sexual maturity, namely BAX, DMRT1, FSHR, IGF1R, IZUMO1, LYZL4, LYZL6, SYCP3 and TNP2, which can be used as the main candidate genes to study the breeding ability of Tibetan pigs.

     

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