Mining Key Genes of Muscle Growth and Development in Guizhou White Goats with Different Genders and Analysis of PI3K/ATK Signal Pathway Based on Transcriptome Sequencing

doi:10.7668/hbnxb.20192965

Abstract

Abstract:

Carcass muscle growth and development is an important evaluation index that affects the breeding efficiency of goats.The aim of this study was to analyze key candidate genes for muscle growth and development of Guizhou white goats with different genders by RNA-Seq,and provide new reference information for the research on the muscle growth and development of Guizhou white goat.We collected the longissimus dorsi muscle to extract RNA and determined the slaughter performance of 6 Guizhou white goats with different genders,which were two years old and were feeded in same level.Then screened differentially expressed genes,analyzed the signal pathway of related genes.Meanwhile,RT-qPCR was used to verify the screened differentially expressed genes.The results showed that the raw reads which obtained by sequencing were filtered,a total of 78.99 Gb Clean Data were obtained in six samples.Each single sample was obtained Clean reads between 83 030 104 and 95 739 024,and the comparison efficiency with the reference genome was between 93.75%-94.79%,a total of 25 089 transcripts were obtained,of which 1 077 were significant differentially expressed genes,and 194 were new transcripts.Among them,563 were up-regulated and 514 were down-regulated in longissimus doris tissue of male sheep.GO functional enrichment analysis was performed on 1 077 differentially expressed mRNAs,which 587 were significantly enriched(Q-value≥0.05) and concentrated in 35 groups of three major categories.KEGG signaling pathway analysis revealed that annotated differential genes participated in 243 signaling pathways,among which the PI3K/AKT signaling pathway were the most enriched,including 32 genes,among them,17 genes were significantly up-regulated and 1 gene was down-regulated,meanwhile,the coexpression score of COL4A1 and COL4A2 genes were the highest in this pathway,the ITGAV protein interaction with other proteins was the most abundant.6 genes that may be closely related to muscle growth and development in white goat were screened out,among which FHL3,WFIKKN2 and SOX6 genes were up-regulated in male longissimus doris tissue,QSOX2,MYH2 and LAP genes were down-regulated.RT-qPCR showed that the expression trend of the 6 candidate genes were consistent with high-throughput sequencing results,and the expression levels differences of these genes were significant,which showed the sequencing results were reliable.Totally,1 077 differentially expressed mRNAs,which 194 were new transcripts were obtained of longissimus doris tissue in Guizhou white goat with different genders by RNA-Seq technology,the screened differentially expressed genes and PI3K/AKT signaling pathway were related to the growth and development of longissimus dorsi muscle of Guizhou white goats.

Key words: Guizhou white goat, RNA-Seq technology, Different genders, Growth and development, Differentially expressed genes

AN Qingming, WANG Xing, WU Zhenyang, MENG Jinzhu, ZHAO Yuanyuan, SONG Xingchao. Mining Key Genes of Muscle Growth and Development in Guizhou White Goats with Different Genders and Analysis of PI3K/ATK Signal Pathway Based on Transcriptome Sequencing[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(3): 213-222. doi: 10.7668/hbnxb.20192965.

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Figures/Tables 11

References 41

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基因名称 Gene name	引物(5'-3') Primer sequences (5'-3')	产物大小/bp Size
FHL3	F:CCTAGCCGATGAGCCCTTC	85
	R:AGAGCACTGCGAGGAGAAG
WFIKKN2	F:TGTCATCTTGGGCCGAATCT	145
	R:CATGTGGAGCAGAGTGACCT
SOX6	F:CAAGCCACCTCTCCATTTGC	100
	R:CCACATGTTGATCACTGCCC
QSOX2	F:AGGACTTCGTGTACCACCTG	94
	R:CGGCTGTCCTCCATGAGATA
MYH2	F:TCTCCCTGATCCACTATGCG	97
	R:TACAGCCCAACCACGGTATC
LAP	F:AAGTCGTCGAAGCTGCCATA	81
	R:CAGGTGCCAATCTGTCTCATG
GAPDH	F:GGCCTCCAAGGAGTAAGGTC	117
	R:CGGGAGATTCTCAGTGTGGT

基因名称 Gene name	引物(5'-3') Primer sequences (5'-3')	产物大小/bp Size
FHL3	F:CCTAGCCGATGAGCCCTTC	85
	R:AGAGCACTGCGAGGAGAAG
WFIKKN2	F:TGTCATCTTGGGCCGAATCT	145
	R:CATGTGGAGCAGAGTGACCT
SOX6	F:CAAGCCACCTCTCCATTTGC	100
	R:CCACATGTTGATCACTGCCC
QSOX2	F:AGGACTTCGTGTACCACCTG	94
	R:CGGCTGTCCTCCATGAGATA
MYH2	F:TCTCCCTGATCCACTATGCG	97
	R:TACAGCCCAACCACGGTATC
LAP	F:AAGTCGTCGAAGCTGCCATA	81
	R:CAGGTGCCAATCTGTCTCATG
GAPDH	F:GGCCTCCAAGGAGTAAGGTC	117
	R:CGGGAGATTCTCAGTGTGGT

指标 Indexes	公羊 Ram	母羊 Ewe	P值 P value
宰前活质量/kg Live weight before slaughter	41.05±3.64a	26.12±3.64b	0.027
胴体质量/kg Carcass weight	19.50±1.86a	12.17±1.86b	0.032
屠宰率/% Yield of carcass	47.35±0.33a	46.55±0.32a	0.135
净肉质量/kg Meat weightg	6.81±0.70a	4.12±0.70b	0.036
净肉率/% Yield of meat	34.78±0.29a	33.85±0.29a	0.068

指标 Indexes	公羊 Ram	母羊 Ewe	P值 P value
宰前活质量/kg Live weight before slaughter	41.05±3.64a	26.12±3.64b	0.027
胴体质量/kg Carcass weight	19.50±1.86a	12.17±1.86b	0.032
屠宰率/% Yield of carcass	47.35±0.33a	46.55±0.32a	0.135
净肉质量/kg Meat weightg	6.81±0.70a	4.12±0.70b	0.036
净肉率/% Yield of meat	34.78±0.29a	33.85±0.29a	0.068

样品 Sample	基础数据/Gb Clean bases data	总 reads Total reads	可定位的 reads Mapped reads	唯一定位 reads Unique mapped reads	多点定位 reads Multiple mapped reads	Q30/%
R1	13.47	83 030 104	78 703 723 (94.79%)	69 863 817 (84.14%)	8 839 906 (10.65%)	89.86
R2	12.90	95 739 024	90 167 003 (94.18%)	82 561 093 (86.24%)	7 605 910 (7.94%)	90.42
R3	13.25	83 704 190	78 475 828 (93.75%)	72 994 411 (87.21%)	5 481 417 (6.55%)	89.97
E1	12.45	89 806 996	84 783 459 (94.41%)	77 295 726 (86.07%)	7 487 733 (8.34%)	90.81
E2	14.36	86 000 412	81 287 190 (94.52%)	74 457 221 (86.58%)	6 829 969 (7.94%)	92.86
E3	12.56	88 311 882	82 976 580 (93.96%)	74 701 217 (84.59%)	8 275 363 (9.37%)	92.77

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