高原雨点鸽与詹森鸽胸肌转录组差异表达基因筛选与功能分析

doi:10.3969/j.issn.1004-1524.2022.03.11

浙江农业学报 ›› 2022, Vol. 34 ›› Issue (3): 507-516.DOI: 10.3969/j.issn.1004-1524.2022.03.11

高原雨点鸽与詹森鸽胸肌转录组差异表达基因筛选与功能分析

兰国湘¹(), 金思琪¹, 李星润², 刘喜雨², 李国美¹, 董新星¹^,^*

1.云南农业大学动物科学技术学院,云南昆明 650201
2.大理农林职业技术学院动物科学学院,云南大理 671003

收稿日期:2021-07-02 出版日期:2022-03-25 发布日期:2022-03-30
作者简介:*董新星,E-mail:dong-xinxing@qq.com
兰国湘(1978—),男,云南保山人,硕士,实验师,研究方向为动物遗传育种。E-mail: 2304205650@qq.com
通讯作者: 董新星
基金资助:
云南农业大学教育教学改革研究项目(2018YAUKG07)

Screening and functional analysis of differentially expressed genes in breast muscle transcriptome between Plateau raindrop pigeon and Janssen pigeon

LAN Guoxiang¹(), JIN Siqi¹, LI Xingrun², LIU Xiyu², LI Guomei¹, DONG Xinxing¹^,^*

1. College of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, China
2. Department of Animal Science, Dali Vocational and Technical College of Agriculture and Forestry, Dali 671003, Yunnan, China

Received:2021-07-02 Online:2022-03-25 Published:2022-03-30
Contact: DONG Xinxing

摘要/Abstract

摘要：

筛选高原雨点鸽与詹森鸽胸肌飞行能力差异的关键基因,为赛鸽选育奠定基础。选择性别相同、体况与日龄相近的高原雨点鸽、詹森鸽各3只,屠宰后取胸肌,进行转录组测序,筛选差异表达基因,对差异基因进行Gene Ontology(GO)分析、Kyoto Encyclopedia of Genes and Genomes(KEGG)分析与蛋白质互作网络分析。结果表明:高原雨点鸽与詹森鸽胸肌转录组比较共检测到75个显著差异基因,49个基因上调表达,26个基因下调表达;GO功能分析显示,差异基因主要富集在骨骼肌细胞分化、细胞代谢过程调节等条目;KEGG通路分析显示,差异基因显著富集在胰岛素信号通路、AMPK信号通路等。与詹森鸽相比,高原雨点鸽胸肌SMYD1、STAT1、VEGFA、PPM1K、PLCE1基因上调,MYOD1、SOCS3、MGLL基因下调。STAT1、MYOD1可能导致高原雨点鸽胸肌生长慢于詹森鸽;SOCS3可能导致高原雨点鸽胸肌肌纤维直径变小,爆发力下降;PLCE1可能导致高原雨点鸽胸肌肌纤维增多;SMYD1可能导致肌纤维分化形成更多的红肌纤维;VEGFA可能导致高原雨点鸽体内白色脂肪转化为棕色脂肪;PPM1K、MGLL可能催化支链氨基酸分解,为高原雨点鸽长距离负重飞行提供足够能量,更适宜远距离飞行。

关键词: 高原雨点鸽, 詹森鸽, 转录组, 差异表达基因, 胸肌

Abstract:

Screening key genes for the difference in pectoral flight ability between Plateau raindrop pigeon and Janssen pigeon can lay foundation for breeding pigeons. Each 3 Plateau raindrop pigeons (JG) and Janssen pigeons (JS) with the same sex, similar body condition and age were selected. After slaughter, breast muscle tissues were taken for transcriptome sequencing and then differentially expressed genes (DEGs) were screened, Gene Ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis and protein interaction network analysis were performed on the DEGs. The results showed that there were 75 significantly DEGs in the breast muscle transcriptome between JG and JS, including 49 up-regulated genes and 26 down-regulated genes. GO enrichment analysis results showed that DEGs were enriched in items such as skeletal muscle cell differentiation, regulation of cellular metabolic process. KEGG pathway analysis showed that DEGs were significant enriched in insulin signaling pathway, AMPK signaling pathway, etc. Compared with the JS,SMYD1, STAT1, VEGFA, PPM1K and PLCE1 genes in breast muscle of the JG were up-regulated, while MYOD1, SOCS3 and MGLL genes were down-regulated. The results showed that STAT1 and MYOD1 might cause breast muscle growth of JG to be slower than that of JS. SOCS3 might cause muscle fiber diameter of the breast muscle of the JG to become smaller and explosive power to decrease at the same time. PLCE1 might cause increase of muscle fibers in the breast muscle of JG. SMYD1 might cause muscle fiber differentiation to form more red muscle fibers. VEGFA might cause white adipose in JGs to be converted into brown adipose. PPM1K and MGLL might catalyze the decomposition of branched chain amino acids, providing enough energy for the long-distance load-bearing flight of the JG, which made they more suitable for long-distance flight.

Key words: Plateau raindrop pigeons, Janssen pigeons, transcriptome, differentially expressed genes, breast muscle

中图分类号:

S836

兰国湘, 金思琪, 李星润, 刘喜雨, 李国美, 董新星. 高原雨点鸽与詹森鸽胸肌转录组差异表达基因筛选与功能分析[J]. 浙江农业学报, 2022, 34(3): 507-516.

LAN Guoxiang, JIN Siqi, LI Xingrun, LIU Xiyu, LI Guomei, DONG Xinxing. Screening and functional analysis of differentially expressed genes in breast muscle transcriptome between Plateau raindrop pigeon and Janssen pigeon[J]. Acta Agriculturae Zhejiangensis, 2022, 34(3): 507-516.

图/表 8

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基因名 Gene name	上游引物序列(5'→3') Forward primer sequence(5'→3')	下游引物序列(5'→3') Reverse primer sequence(5'→3')	产物长度 Length of product/bp
MGLL	GAGCTGCCCGTTCTCATTCT	CGACAGGTTTGGGAGGACAA	189
SOCS3	GCCACCAGAGAACGGAAAGA	TTAGTCCCCCGGAAAATGGC	201
PPM1K	AGATGGCTGCTGATGCAACT	AGCCACCGCACTTCCTAATC	201
PLCE1	ATGCTTCCTTCACCTGGGC	TGGTAGGTGTGGTGGGGTTG	163
SMYD1	ATCTGTCATACCTGCTTCAAACG	CCAGCCTGATGTTTTCGGT	169
β-actin	TTACCCACACTGTGCCCATC	AGGGCAACATAGCACAGCTT	187

基因名 Gene name	上游引物序列(5'→3') Forward primer sequence(5'→3')	下游引物序列(5'→3') Reverse primer sequence(5'→3')	产物长度 Length of product/bp
MGLL	GAGCTGCCCGTTCTCATTCT	CGACAGGTTTGGGAGGACAA	189
SOCS3	GCCACCAGAGAACGGAAAGA	TTAGTCCCCCGGAAAATGGC	201
PPM1K	AGATGGCTGCTGATGCAACT	AGCCACCGCACTTCCTAATC	201
PLCE1	ATGCTTCCTTCACCTGGGC	TGGTAGGTGTGGTGGGGTTG	163
SMYD1	ATCTGTCATACCTGCTTCAAACG	CCAGCCTGATGTTTTCGGT	169
β-actin	TTACCCACACTGTGCCCATC	AGGGCAACATAGCACAGCTT	187

样品 Sample	原始读段 Raw reads	高质量序列所占比例 Clean reads rate/%	Q30/%	比对基因组所占比例 Mapping rate/%
JG1	50 784 230	94.47	93.57	83.09
JG2	49 764 078	95.51	93.26	84.60
JG3	48 035 956	93.37	94.08	83.03
JS1	51 604 076	91.46	93.81	85.33
JS2	49 022 332	91.97	93.85	85.90
JS3	47 948 950	94.94	93.56	84.03

样品 Sample	原始读段 Raw reads	高质量序列所占比例 Clean reads rate/%	Q30/%	比对基因组所占比例 Mapping rate/%
JG1	50 784 230	94.47	93.57	83.09
JG2	49 764 078	95.51	93.26	84.60
JG3	48 035 956	93.37	94.08	83.03
JS1	51 604 076	91.46	93.81	85.33
JS2	49 022 332	91.97	93.85	85.90
JS3	47 948 950	94.94	93.56	84.03

类别 Category	登记号 Accession	条目 Term	P值 P value	差异表达基因数量 Number of DEGs
肌肉生长	GO:0007259	JAK-STAT级联JAK-STAT cascade	0.003 5	2
Muscle growth	GO:0035914	骨骼肌细胞分化Skeletal muscle cell differentiation	0.005 7	2
	GO:0010830	肌管分化调控Regulation of myotube differentiation	0.008 4	2
	GO:0045661	成肌细胞分化调控Regulation of myoblast differentiation	0.011 0	2
	GO:0051153	横纹肌细胞分化调控Regulation of striated muscle cell differentiation	0.024 0	2
	GO:0006937	肌肉收缩调节Regulation of muscle contraction	0.044 0	1
能量利用	GO:0009083	支链氨基酸分解过程Branched-chain amino acid catabolic process	0.000 9	2
Energy utilization	GO:0016042	脂质分解过程Lipid catabolic process	0.001 0	5
	GO:0009081	支链氨基酸代谢过程Branched-chain amino acid metabolic process	0.001 3	2
	GO:0006651	甘油二酯生物合成过程Diacylglycerol biosynthetic process	0.007 1	1
	GO:0036155	酰基甘油链重构Acylglycerol acyl-chain remodeling	0.007 1	1
	GO:0006638	中性脂质代谢过程Neutral lipid metabolic process	0.017 0	2
	GO:0006639	酰基甘油代谢过程Acylglycerol metabolic process	0.017 0	2
	GO:0006633	脂肪酸生物合成过程Fatty acid biosynthetic process	0.030 0	2
	GO:0046339	二酰基甘油代谢过程Diacylglycerol metabolic process	0.031 0	2
	GO:0019433	甘油三酯分解过程Triglyceride catabolic process	0.038 0	2
	GO:0006629	脂质代谢过程Lipid metabolic process	0.041 0	7
	GO:0006631	脂肪酸代谢过程Fatty acid metabolic process	0.047 0	3
肌肉生长与能量利用	GO:0031323	细胞代谢过程调节Regulation of cellular metabolic process	8.5×10^-6	27
Muscle growth and	GO:0080090	初级代谢过程调节Regulation of primary metabolic process	1.8×10^-5	26
energy utilization	GO:0019222	代谢过程调节Regulation of metabolic process	3.5×10^-5	27
	GO:0060255	大分子代谢过程调控Regulation of macromolecule metabolic process	0.000 2	26
	GO:0051171	氮化合物代谢过程的调节	0.000 8	24
		Regulation of nitrogen compound metabolic process
	GO:0044238	初级代谢过程Primary metabolic process	0.003 5	29
	GO:0071704	有机物代谢过程Organic substance metabolic process	0.008 2	28
	GO:0008152	代谢过程Metabolic process	0.018 0	28

高原雨点鸽与詹森鸽胸肌转录组差异表达基因筛选与功能分析

Screening and functional analysis of differentially expressed genes in breast muscle transcriptome between Plateau raindrop pigeon and Janssen pigeon

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功能ID Function ID	功能描述 Description of function	P值 P value	差异表达基因 DEGs
map04910	胰岛素信号通路Insulin signaling pathway	0.000 4	SOCS3、ACACB、PPP1R3C
map04931	胰岛素抵抗Insulin resistance	0.003 3	SOCS3、ACACB、PPP1R3C
map04919	甲状腺激素信号通路Thyroid hormone signaling pathway	0.003 8	STAT1、HIF1A、PLCE1
map04920	脂肪细胞因子信号通路Adipocytokine signaling pathway	0.014 0	SOCS3、ACACB
map00061	脂肪酸生物合成Fatty acid biosynthesis	0.039 0	ACACB
map04152	AMPK信号通路AMPK signal pathway	0.042 0	ACACB
map04630	Jak-STAT信号通路Jak-STAT signaling pathway	0.050 0	SOCS3、STAT1

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