[1]孟科,赵薇,郭晨浩,等.不同品种绵羊肌内脂肪沉积相关miRNA的筛选与功能预测[J].江苏农业学报,2023,(07):1554-1566.[doi:doi:10.3969/j.issn.1000-4440.2023.07.012]
　MENG Ke,ZHAO Wei,GUO Chen-hao,et al.Screening and functional prediction of miRNA related to intramuscular fat deposition in different sheep breeds[J].,2023,(07):1554-1566.[doi:doi:10.3969/j.issn.1000-4440.2023.07.012]
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不同品种绵羊肌内脂肪沉积相关miRNA的筛选与功能预测()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2023年07期
页码:
1554-1566
栏目:
畜牧兽医·水产养殖
出版日期:
2023-10-31

文章信息/Info

Title:
Screening and functional prediction of miRNA related to intramuscular fat deposition in different sheep breeds
作者:
孟科1; 2; 赵薇1; 郭晨浩1; 聂伟1; 陶毛孩1; 袁晓春1; 孙昊然1; 冯登侦1
(1.宁夏大学农学院，宁夏银川750021;2.宁夏中卫市沙坡头区畜牧水产技术推广服务中心，宁夏中卫755000)
Author(s):
MENG Ke1; 2; ZHAO Wei1; GUO Chen-hao1; NIE Wei1; TAO Mao-hai1; YUAN Xiao-chun1; SUN Hao-ran1; FENG Deng-zhen1
(1.School of Agriculture, Ningxia University, Yinchuan 750021, China;2.Animal Husbandry and Fishery Technology Extension and Service Center, Shapotou District, Zhongwei City, Zhongwei 755000, China)
关键词:
绵羊; 肌内脂肪沉积; 转录组; miRNA
Keywords:
sheep; intramuscular fat deposition; transcriptome; miRNA
分类号:
S826
DOI:
doi:10.3969/j.issn.1000-4440.2023.07.012
文献标志码:
A
摘要:
为明确不同品种绵羊(Ovis aries)肌肉组织miRNA的表达情况，筛选影响绵羊肌内脂肪沉积的关键miRNA，以滩羊、杜泊羊和小尾寒羊为研究对象，测定背最长肌肌内脂肪含量并对其进行转录组测序(RNA-Seq)，筛选绵羊脂肪沉积相关的差异表达miRNA。结果表明，滩羊肌内脂肪含量显著高于杜泊羊,极显著高于小尾寒羊；杜泊羊肌内脂肪含量显著高于小尾寒羊。转录组测序质量良好。3个品种绵羊共鉴定到134个已知miRNA和153个预测miRNA，这些 miRNA的长度主要介于21～23 nt，且首位碱基对U碱基具有明显偏好性。以P ≤0.05和|log2FC|>1(FC为差异倍数)为筛选条件，3个比较组共获得42个差异表达miRNA；杜泊羊与滩羊比较组中筛选出22个差异表达miRNA；小尾寒羊与杜泊羊比较组中筛选出21个差异表达 miRNA；小尾寒羊与滩羊比较组中筛选出12个差异表达miRNA。筛选出的42个miRNA靶向到326个靶基因，显著富集到减数分裂I、肌肽-寡糖1，2-α-甘露糖苷酶活性和甘露寡糖甘露糖苷酶活性等303个基因本体(GO)条目，以及富集到α-亚麻酸代谢、亚油酸代谢、多巴胺能突触、胰岛素信号传导途径及N-聚糖生物合成等320个京都基因与基因组百科全书(KEGG)代谢通路。根据GO和KEGG的功能描述进一步筛选出23个miRNA及其靶向的119个靶基因可能参与脂肪沉积。进一步构建miRNA靶向调控网络，最终筛选到oar-miR-133、oar-miR-485-5p、novel_6、novel_85和novel_103等绵羊肌内脂肪沉积的关键miRNA。随机选择的8个差异表达miRNA的实时荧光定量PCR结果与转录组测序结果一致，表明转录组测序结果可靠。本研究为深入开展绵羊肌内脂肪沉积的分子调控机理、高品质绵羊品种的选育提供理论依据。
Abstract:
In order to clarify the expression of miRNAs in muscle tissues of different breeds of sheep (Ovis aries) and screen the key miRNAs affecting intramuscular fat deposition in sheep, Tan sheep, Dorper sheep and Small Tail Han sheep were used as experimental materials. The intramuscular fat content of longissimus dorsi muscle was measured and transcriptome sequencing (RNA-Seq) was performed to screen the differentially expressed miRNAs related to fat deposition in sheep. The results showed that the intramuscular fat content of Tan sheep was significantly higher than that of Dorper sheep, and extremely significantly higher than that of Small Tail Han sheep, and the intramuscular fat content of Dorper sheep was significantly higher than that of Small Tail Han sheep. The quality of transcriptome sequencing was good. A total of 134 known miRNAs and 153 predicted miRNAs were identified in three breeds of sheep. The length of these miRNAs mainly ranged from 21 nt to 23 nt, and the first base had obvious preference for U base. With P≤0.05 and |log2FC|>1 (FC was the fold change)as the screening conditions, a total of 42 differentially expressed miRNAs were obtained from the three comparison groups. Twenty-two differentially expressed miRNAs were screened in the comparison group of Dorper sheep and Tan sheep, 21 differentially expressed miRNAs were screened in the comparison group of Small Tail Han sheep and Dorper sheep, and 12 differentially expressed miRNAs were screened in the comparison group of Small Tail Han sheep and Tan sheep. The screened 42 miRNAs targeted to 326 target genes, which were significantly enriched into 303 gene ontology(GO) entries such as meiosis I, mannosyl-oligosaccharide 1,2-α-mannosidase activity and mannosyl-oligosaccharide mannosidase activity, and 320 Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathways such as α-linolenic acid metabolism, linoleic acid metabolism, dopaminergic synapses, insulin signaling pathway and N-glycan biosynthesis. According to the functional description of GO and KEGG, 23 miRNAs and 119 target genes might be involved in fat deposition. The miRNA targeted regulatory network was further constructed, and finally the key miRNAs for fat deposition in sheep such as oar-miR-133, oar-miR-485-5p, novel_6, novel_85 and novel_103 were screened. The qRT-PCR results of the eight randomly selected differentially expressed miRNAs were consistent with transcriptome sequencing results, indicating that the transcriptome sequencing results were reliable. This study provides a theoretical basis for further research on the molecular regulation mechanism of intramuscular fat deposition in sheep and the breeding of high-quality sheep breeds.

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备注/Memo

备注/Memo:
收稿日期：2022-07-01基金项目：宁夏回族自治区自然科学基金项目(2020AAC03082); 宁夏回族自治区农业育种专项 (NXNYYZ20150101)作者简介：孟科(1996-)，男，宁夏中卫人，硕士研究生，主要从事动物遗传育种与分子育种研究。(E-mail)1131988962@qq.com通讯作者：冯登侦，(E-mail)fdzh126@sohu.com
更新日期/Last Update: 2023-11-17