孙宝盛, 张颜, 肖贵榜, 杨玉能, 杨泓涛, 李世春, 邓位喜. 2022: 黔北麻羊毛色差异主要调控基因及其通路分析. 南方农业学报, 53(6): 1483-1492. DOI: 10.3969/j.issn.2095-1191.2022.06.001
引用本文: 孙宝盛, 张颜, 肖贵榜, 杨玉能, 杨泓涛, 李世春, 邓位喜. 2022: 黔北麻羊毛色差异主要调控基因及其通路分析. 南方农业学报, 53(6): 1483-1492. DOI: 10.3969/j.issn.2095-1191.2022.06.001
SUN Bao-sheng, ZHANG Yan, XIAO Gui-bang, YANG Yu-neng, YANG Hong-tao, LI Shi-chun, DENG Wei-xi. 2022: Main wool color regulatory genes of Capra hircus and analysis of their pathways. Journal of Southern Agriculture, 53(6): 1483-1492. DOI: 10.3969/j.issn.2095-1191.2022.06.001
Citation: SUN Bao-sheng, ZHANG Yan, XIAO Gui-bang, YANG Yu-neng, YANG Hong-tao, LI Shi-chun, DENG Wei-xi. 2022: Main wool color regulatory genes of Capra hircus and analysis of their pathways. Journal of Southern Agriculture, 53(6): 1483-1492. DOI: 10.3969/j.issn.2095-1191.2022.06.001

黔北麻羊毛色差异主要调控基因及其通路分析

Main wool color regulatory genes of Capra hircus and analysis of their pathways

  • 摘要: 【目的】明确调控黔北麻羊毛色差异的候选基因及其通路,为揭示黔北麻羊特征毛色的形成机制提供理论依据。【方法】以1岁黔北麻羊公羊为研究对象,分别采集其肩部黑色羊毛下的皮肤组织块和腹部白色羊毛下的皮肤组织块,以RNA为模板合成双链cDNA,经末端修复等处理后进行PCR富集构建cDNA文库,采用Illumina HiSeqTM 4000测序平台完成转录组测序,经过滤及质控后以DESeq2筛选出差异表达基因(DEGs),然后分别进行GO功能注释分析和KEGG信号通路富集分析。【结果】与白色羊毛对照组(White)相比,黑色羊毛试验组(Black)存在509个显著上调表达基因、195个显著下调表达基因;在黑色羊毛试验组中的509个显著上调基因中筛选获得5个与黑色素生成相关且表达差异显著(基因表达差异倍数log2 Fold Change>1、校正后的P-adjust<0.05)的基因,分别是酪氨酸酶基因(TYR)、酪氨酸相关蛋白1基因(TYRP1)、黑素皮质素受体1基因(MC1R)、角蛋白19基因(KRT19)和角蛋白79基因(KRT79)。显著差异表达基因GO功能注释分析结果表明,除了色素代谢过程外,黑色羊毛试验组还显著富集于黑素小体、色素沉着、色素生物合成过程、黑色素生物合成过程和角质形成细胞分化等与黑色素形成相关的GO功能条目上; KEGG通路富集分析结果表明,在黑色羊毛试验组显著差异表达基因富集的通路中,与黑色素生成密切相关的是酪氨酸代谢通路,而不是Wnt/β-catenin、MAPK、PI3K/Akt和MC1R/α-MSH等经典的黑色素生成信号通路。【结论】 TYRTYRP1MC1RKTR19等是调控黔北麻羊毛色的主要基因,酪氨酸代谢通路是调控黔北麻羊毛色的主要通路。TYRTYRP1MC1RKTR19等基因的局部上调表达,激活酪氨酸代谢等通路而促使黑色素生成增加,是黔北麻羊肩部黑色羊毛和腹部白色羊毛存在差异的主要原因。

     

    Abstract: 【Objective】To explore dominant genes that regulated the wool color difference in Capra hircus and their pathways,so as to provide a theoretical basis for revealing the formation mechanism of characteristic wool color of the goat.【Method】 One-year-old male C.hircus goats were taken as research objects,and the skin tissue under the black wool on the shoulder and the skin tissue under white wool on the abdomen were collected respectively.The double strand cDNA was synthesized and purified with RNA as template,and then the cDNA library was constructed through PCR enrichment after processes such as end repair.Transcriptome sequencing was performed using Illumina HiSeqTM 4000 sequencing platform.After filtering and quality control,differentially expressed genes(DEGs)were screened by DESeq2,and then GO function annotation analysis and KEGG pathway enrichment analysis were performed respectively.【Result】Compared with the white wool control group,there were 509 significantly up-regulated genes and 195 significantly down-regulated genes in the black wool test group,and 5 significantly(log2 Fold Change>1,P-adjust<0.05)up-regulated genes related to melanin production were filtered from 509 significantly up-regulated genes,including genes of tyrosinase(TYR),tyrosinerelated protein 1(TYRP1),melanocortin receptor 1(MC1R),keratin 19(KRT19)and keratin 79(KRT79).The GO function annotation of significantly differential genes showed that the black wool test group was significantly enriched in melanosome,pigmentation,pigment biosynthetic process,melanin biosynthetic process,keratinocyte differentiation and other functions related to melanin formation.KEGG pathway analysis showed that,in black wool test group,the pathways closely related to melanin production were the tyrosinase metabolic pathway but not classical melanin production signal pathway such as Wnt/β-catenin,MAPK,PI3K/Akt or MC1R/α-MSH.【Conclusion】 TYRTYRP1MC1R and KTR19 genes are dominant genes regulating the wool color difference of C.hircus.Tyrosine metabolic pathway is the dominant pathway regulating the wool color difference of C.hircus.TYRTYRP1MC1R and KTR19 genes are up-regulated and activate the tyrosine metabolic pathway,resulting in the increase of melanin production,which is the major reason of color characteristics of black wool on the shoulder and abdominal white wool of C.hircus.

     

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