Exploring ADSL gene regulation of inosine monophosphate content in Bashang long tail chicken based on transcriptome and next-generation sequencing technology

doi:10.3969/j.issn.1004-1524.20240222

Acta Agriculturae Zhejiangensis ›› 2024, Vol. 36 ›› Issue (12): 2676-2686.DOI: 10.3969/j.issn.1004-1524.20240222

• Animal Science • Previous Articles Next Articles

Exploring ADSL gene regulation of inosine monophosphate content in Bashang long tail chicken based on transcriptome and next-generation sequencing technology

HE Lingyu¹(), QIAO Xian¹^,², WANG Xinyue¹, LI Xianglong¹^,²^,^*()

1. College of Animal Science and Technology, Hebei Normal University of Science and Technology, Qinhuangdao 066000, Hebei, China
2. Hebei Provincial Key Laboratory of Characteristic Animal Germplasm Resources Mining and Innovation, Qinhuangdao 066000, Hebei, China

Received:2024-03-08 Online:2024-12-25 Published:2024-12-27

Abstract

Abstract:

In order to search and improve the genes involved in the muscle inosine monophosphate (IMP) content of Bashang long tail chickens, a high-quality meat and egg dual-use breed of Bashang long tail chickens in Hebei Province was selected as the experimental subjects. The inosine content of 60 chickens was measured. The high and low inosine monophosphate content of Bashang long tail chickens was used as the indicator for grouping, and transcriptome sequencing was performed for differential expression gene analysis and KEGG pathway enrichment. PCR amplification and sequencing technology were used to screen for polymorphic sites. Use one-way ANOVA in SPSS to conduct correlation analysis with inosine monophosphate content. The results showed that:among the significantly differentially expressed genes in transcriptome sequencing analysis, there were 251 up-regulated genes and 241 down-regulated genes, while 18 300 genes were not significantly different. KEGG pathway enrichment analysis was conducted on the differentially expressed genes, and it was found that the nucleotide metabolism pathway has an important impact on the content of inosine monophosphate. The ADSL gene is involved in this pathway and ultimately affects the synthesis of inosine monophosphate. the 2nd, 7th, 9th, and 11th exons of the ADSL gene were successfully amplified, and g. 3713G>A, g. 3797C>T, g. 8282C>G, g. 10191C>T, and g. 12827C>A were screened. Among them, the IMP content in exon 2 (g. 3797C>T) CT individuals was significantly (P<0.05) higher than that in CC and TT individuals, while in exon 7 (g. 8282C>G), the IMP content in CG individuals was significantly (P<0.05) higher than that in GG individuals and CC individuals. The dominant genotypes of the ADSL gene in Bashang long tail chickens are located at exon 2, g. 3797C>T site CT type, and exon 7, g. 828282C>G site CG type, which can be used as molecular markers for IMP content in breeding practice.

Key words: Bashang long tail chicken, transcriptome sequencing, ADSL, polymorphic locus, inosine monophosphate

CLC Number:

S831

HE Lingyu, QIAO Xian, WANG Xinyue, LI Xianglong. Exploring ADSL gene regulation of inosine monophosphate content in Bashang long tail chicken based on transcriptome and next-generation sequencing technology[J]. Acta Agriculturae Zhejiangensis, 2024, 36(12): 2676-2686.

Figures/Tables 14

References 31

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样品编号(A组) Sample name (Group A)	肌苷酸含量 Inosine monophosphate content/(mg·kg^-1)	样品编号(B组) Sample name (Group B)	肌苷酸含量 Inosine monophosphate content/(mg·kg^-1)
1	1 183.73	6	11.28
2	1 601.84	7	17.71
3	1 173.14	8	12.43
4	1 246.67	9	13.21
5	1 153.34	10	14.01

样品编号(A组) Sample name (Group A)	肌苷酸含量 Inosine monophosphate content/(mg·kg^-1)	样品编号(B组) Sample name (Group B)	肌苷酸含量 Inosine monophosphate content/(mg·kg^-1)
1	1 183.73	6	11.28
2	1 601.84	7	17.71
3	1 173.14	8	12.43
4	1 246.67	9	13.21
5	1 153.34	10	14.01

基因 Gene	SNP位点 SNP site	引物序列 Primer sequences(5'→3')	产物大小 Product size/bp	退火温度 Annealing temperature/℃
ADSL(AY665559.1)	37133797	F:CTTTTGACTGCCCCATTGCC R:AAAGGGAGAAGCCACAGGAG	278	57.2
	8282	F:CTTTCATTCCCACTTGCGGC R:GCTTGGACCAATTCAGCACA	333	56.0
	10191	F:TCCCCACCGCATTTAGATGT R:GTCTGCTTTTCATCAAGACGGT	192	56.4
	12827	F:TCCACAGTAACGTTCCAGACA R:TTCACAGAAGGAAAGCGCCT	341	56.9

基因 Gene	SNP位点 SNP site	引物序列 Primer sequences(5'→3')	产物大小 Product size/bp	退火温度 Annealing temperature/℃
ADSL(AY665559.1)	37133797	F:CTTTTGACTGCCCCATTGCC R:AAAGGGAGAAGCCACAGGAG	278	57.2
	8282	F:CTTTCATTCCCACTTGCGGC R:GCTTGGACCAATTCAGCACA	333	56.0
	10191	F:TCCCCACCGCATTTAGATGT R:GTCTGCTTTTCATCAAGACGGT	192	56.4
	12827	F:TCCACAGTAACGTTCCAGACA R:TTCACAGAAGGAAAGCGCCT	341	56.9

样品名称 Sample name	浓度 Concentration/ (ng·μL^-1)	D₂₆₀/D₂₈₀	RNA完整值 RNA integrity number(RIN)
A1	139	2.006	6.8
A2	156	2.012	7.5
A3	128	2.000	8.2
A4	132	1.982	7.9
A5	152	2.003	8.3
B1	161	2.036	7.7
B2	137	1.908	8.5
B3	166	2.016	8.3
B4	159	2.027	9.1
B5	118	1.968	8.6

Exploring ADSL gene regulation of inosine monophosphate content in Bashang long tail chicken based on transcriptome and next-generation sequencing technology

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样本 Sample	原始reads Raw reads	过滤后reads Clean reads	过滤后碱基 Clean bases/Gb	错误发现率 False discovery rate/%	Q20%	Q30%	GC含量 GC content/%
A1	50 728 000	48 889 916	7.33	0.03	97.39	93.09	51.85
A2	42 785 508	40 886 724	6.13	0.03	97.28	92.85	51.58
A3	47 588 060	45 744 878	6.86	0.03	97.19	92.63	52.19
A4	52 503 284	50 632 380	7.59	0.03	96.72	91.73	50.37
A5	43 447 046	41 692 442	6.25	0.03	97.44	93.39	52.31
B1	46 702 784	44 980 912	6.75	0.03	97.29	92.8	52.23
B2	41 813 830	40 168 072	6.03	0.03	97.28	92.84	52.61
B3	43 513 932	42 487 038	6.37	0.03	96.88	91.9	52.94
B4	45 702 690	44 466 824	6.67	0.03	97.03	92.29	50.88
B5	46 914 640	45 820 392	6.87	0.03	97.37	93.1	52.39

SNP位点 SNP site	基因型频率(个体数) Genotype frequency							基因频率 Allele frequency				PIC	χ²	P值 P value
	AA	AG	GG	CG	CC	CT	TT	A	G	C	T
g.3713G>A	0.61(32)	0.30(16)	0.09(5)					0.75	0.25			0.30	15.07	0.00
g.3797C>T					0.54(28)	0.15(8)	0.31(16)			0.61	0.39	0.36	23.82	—
g.8282C>G			0.46(25)	0.39(21)	0.15(8)				0.66	0.34		0.35	1.48	0.48
g.10191C>T					0.92(49)	0.08(4)				0.96	0.04	0.07	1.00	—
g.12827C>A	0.06(3)				0.94(50)			0.06		0.94		0.11	5.33	0.07

SNP位点 SNP site	基因型 Genotype	样本 Sample size	肌苷酸含量 IMP content/(g·kg^-1)
g.3713G>A	AA	32	2.2±3.5
	AG	16	4.2±4.8
	GG	5	1.1±1.2
g.3797C>T	CC	28	2.1±3.2 a
	CT	8	6.3±6.0 b
	TT	16	2.1±3.1 a
g.8282C>G	CC	8	2.1±2.2 a
	CG	21	4.2±5.1 b
	GG	25	1.5±2.4 a
g.10191C>T	CC	49	2.8±4.0
	CT	4	1.4±1.1
g.12827C>A	AA	3	4.3±3.6
	CC	50	2.6±3.9

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