动物营养学报 >

2022 , Vol. 34 >Issue 2: 1247 - 1259

DOI: https://doi.org/10.3969/j.issn.1006-267x.2022.02.056

研究论文 RESEARCH PAPER

基于转录组测序技术研究饲粮赖氨酸水平调控贵州黄鸡采食量的差异表达基因和信号通路

  • 王小亚 ,
  • 饶永超 ,
  • 陈大海 ,
  • 唐宏 ,
  • 林家栋 ,
  • 张福平 ,
  • 施晓丽
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  • 1. 贵州大学高原山地动物遗传育种与繁殖教育部重点实验室, 贵阳 550025;
    2. 贵州省动物遗传育种与 繁殖重点实验室, 贵阳 550025;
    3. 贵州大学动物科学学院, 贵阳 550025;
    4. 贵州大学科研鸡场, 贵阳 550025
王小亚(1995-),女,贵州毕节人,硕士研究生,从事家禽育种研究。E-mail:1872745351@qq.com

收稿日期: 2021-08-22

  网络出版日期: 2022-02-15

基金资助

贵州省科技计划项目（黔科合支撑[2019]2288号）

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Differentially Expressed Genes and Signaling Pathways of Regulation of Dietary Lysine Level on Feed Intake of Guizhou Yellow Chickens Based on Transcriptome Sequencing Technology

  • WANG Xiaoya ,
  • RAO Yongchao ,
  • CHEN Dahai ,
  • TANG Hong ,
  • LIN Jiadong ,
  • ZHANG Fuping ,
  • SHI Xiaoli
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  • 1. Key Laboratory of Animal Genetics, Breeding and Reproduction in Plateau Mountainous Region, Ministry of Education, Guizhou University, Guiyang 550025, China;
    2. Guizhou Provincial Key Laboratory of Animal Genetics, Breeding and Reproduction, Guiyang 550025, China;
    3. College of Animal Science, Guizhou University, Guiyang 550025, China;
    4. Research Chicken Farm of Guizhou University, Guiyang 550025, China

Received date: 2021-08-22

  Online published: 2022-02-15

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摘要

本试验旨在利用转录组测序（RNA-Seq）技术研究饲粮赖氨酸（Lys）水平调控贵州黄鸡采食量的分子机制，挖掘与采食量相关的候选基因和信号通路，进一步提高家禽采食量以及促进养殖效益。选取健康的1日龄贵州黄鸡雏鸡400羽，随机分为4组，每组5个重复，每个重复20羽。各组饲粮Lys水平分别为0.7%、0.9%、1.1%和1.3%。试验期6周。在6周龄时，选取平均日采食量差异极显著（P<0.01）的0.9% Lys水平组（b组，n=3）和1.3% Lys水平组（d组，n=3）中的贵州黄鸡各3只，采集下丘脑组织进行RNA-Seq分析、差异表达基因GO功能富集和KEGG信号通路分析，利用实时荧光定量PCR （qRT-PCR）验证RNA-Seq测序数据。结果表明：1）0.7%、0.9%、1.1% Lys水平组1~6周龄平均日采食量极显著高于1.3% Lys水平组（P<0.01），其中，0.9% Lys水平组最高。2）每个样品所产的测序读数比对到鸡参考基因组上比对率均在89%以上，Q20、Q30含量分别在97%、93%以上；b组和d组共筛选到2 006个差异表达基因，其中，1 275个基因表达上调，731个基因表达下调。GO功能主要在调节神经过程、投射神经元形态发生、负调控神经发生等生物学过程极显著富集；KEGG通路显著富集到19条通路，富集到磷脂酰肌醇信号系统、叉头框转录因子O信号传导途径、哺乳动物雷帕霉素靶蛋白信号通路、丝裂原活化蛋白激酶信号通路、神经活性配体-受体相互作用信号通路和脂肪细胞因子信号通路6条通路可能影响贵州黄鸡的采食量。筛选到神经肽Y （NPY）、前阿黑皮素原（POMC）、刺鼠相关蛋白（AgRP）、可卡因和安非他明调控转录前肽（CARTPT）富集到神经活性配体-受体相互作用信号通路和脂肪细胞因子信号通路上。qRT-PCR结果显示6个随机选择的差异表达基因的表达趋势与RNA-Seq结果一致。综上所述，0.7%、0.9%、1.1%赖氨酸水平组1~6周龄平均日采食量极显著高于1.3%赖氨酸水平组。不同水平赖氨酸可能通过调控下丘脑组织NPY、AgRP、CARTPT和POMC等相关基因的表达，作用于神经活性配体-受体相互作用通路和脂肪细胞因子信号通路，以此来调控贵州黄鸡的采食量。

关键词： 转录组测序; 贵州黄鸡; 赖氨酸; 下丘脑; 采食量

本文引用格式

王小亚 , 饶永超 , 陈大海 , 唐宏 , 林家栋 , 张福平 , 施晓丽 . 基于转录组测序技术研究饲粮赖氨酸水平调控贵州黄鸡采食量的差异表达基因和信号通路[J]. 动物营养学报, 2022 , 34(2) : 1247 -1259 . DOI: 10.3969/j.issn.1006-267x.2022.02.056

Abstract

This experiment aims to study the molecular mechanism of dietary lysine (Lys) level regulated feed intake of Guizhou yellow chickens by transcriptome sequencing (RNA-Seq) technique, and to explore the candidate genes and signaling pathways related to feed intake, to further increase the intake of poultry and to promote the efficiency of farming. A total of 400 one-day-old Guizhou yellow chickens were randomly divided into 4 groups with 5 replicates per group and 20 chickens per replicate. The dietary Lys levels of each group were 0.7%, 0.9%, 1.1% and 1.3%, respectively. The experiment lasted for 6 weeks. At 6 weeks of age, three Guizhou yellow chickens in each of the 0.9% Lys level group (b group, n=3) and 1.3% Lys level group (d group, n=3) with significant differences (P<0.01) in average daily feed intake were selected, collected the hypothalamus tissue and used for RNA-Seq analyze, GO functional enrichment and KEGG pathways of differential expression genes, ultimately, the RNA-Seq results were verified by real-time quantitative PCR (qRT-PCR). The results showed as follows:1) the average daily feed intake during 1 to 6 weeks of age of 0.7%, 0.9% and 1.1% Lys level groups was significantly higher than that of 1.3% Lys level group (P<0.01), and the 0.9% Lys level group was the highest. 2) The comparison rate of the sequencing reads produced by each sample to the chicken reference genome was greater than 89%, Q20 and Q30 contents were 97% and 93% respectively; a total of 200 6 differentially expressed genes were screened in b group and d group, which 1 275 genes were up-regulated, while 731 genes were down-regulated. The GO function was highly enriched in biological processes such as neuron projection morphogenesis, positive regulation of synaptic transmission and negative regulation of neurogenes; the KEGG pathway was significantly enriched to 19 pathways, among these pathways, the enriched phosphatidylinositol signaling system, forkhead box O signaling pathway, mammalian target of rapamycin signaling pathway, mitogen-activated protein kinase signaling pathway, neuroactive ligand-receptor interaction signaling pathway and adipocytokine signaling pathway, and these pathways could affect the feed intake of Guizhou yellow chickens. The screening of neuropeptide Y (NPY), proopiomelanocortin (POMC), agouti-related protein (AgRP), cocaine and amphetamine-regulated prepropeptide (CARTPT) enriched in the neuroactive ligand-receptor interaction and adipocytokine signaling pathway. The qRT-PCR results showed that the expression trends of 6 differentially expressed genes randomly selected were consistent with the RNA-Seq results. In summary, the average daily feed intake of 1 to 6 weeks of age of 0.7%, 0.9% and 1.1% Lys level group was significantly higher than that of 1.3% Lys level group. Different Lys levels may regulate the expression of NPY, AgRP, CARTPT, POMC and other related genes in hypothalamic tissue, acting on the neural active ligand-receptor interaction pathway and adipocytokine signaling pathway, thereby regulate the feed intake of Guizhou yellow chickens.

Key words： RNA-Seq; Guizhou yellow chickens; lysine; hypothalamus; feed intake

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