葡萄线粒体转录终止因子（mTERF）家族基因鉴定与VamTERF5抗寒性研究

doi:10.16420/j.issn.0513-353x.2025-0302

摘要/Abstract

摘要：

线粒体转录终止因子（mTERF）是植物中高度保守的DNA结合蛋白，能够调控植物的生长发育和逆境胁迫。对山葡萄mTERF基因家族（VamTERF）进行生物信息学分析、qRT-PCR和基因功能验证，结果显示山葡萄基因组中存在7个mTERF成员，主要分布在6条染色体上，编码蛋白的理论等电点在4.92 ~ 9.07，不规则卷曲数值在27.12 ~ 44.42，α-螺旋、β-转角分别在41.26% ~ 57.55%和2.18% ~ 7.91%；系统进化结果表明该基因家族被聚类至3个亚族，其中第3亚族成员最多，且与杨树进化关系较为接近；VamTERF基因家族密码子使用偏好性较弱；与草莓等5个物种之间存在共线性关系；qRT-PCR分析结果显示VamTERF家族成员对非生物胁迫都有响应，其中VamTERF2、VamTERF3、VamTERF4、VamTERF5和VamTERF7在低温胁迫后显著上调。亚细胞定位显示这5个成员主要定位在细胞核中。过表达VamTERF5的葡萄愈伤组织在低温胁迫后，POD、SOD和CAT活性明显增强。表明VamTERF成员在低温胁迫响应中起重要作用。

关键词: 山葡萄, mTERF, 基因家族, 生物信息, 抗寒

Abstract:

Mitochondrial transcription termination factors（mTERF）are highly conserved DNA- binding proteins in plants that regulate plant growth，development，and responses to abiotic stress. In this study，comprehensive bioinformatics analysis，quantitative real-time PCR（qRT-PCR），and gene function validation were performed on the Vitis amurensis mTERF gene family（VamTERF）. A total of seven VamTERF members were identified in the Vitis amurensis genome，distributed across six chromosomes. The theoretical isoelectric points of the encoded proteins ranged from 4.92 to 9.07. Structural analysis revealed that the proportion of random coils ranged from 27.12% to 44.42%，while α-helices and β-turns accounted for 41.26%-57.55% and 2.18%-7.91%，respectively. Phylogenetic analysis clustered the VamTERF gene family into three subfamilies，with subfamily 3 containing the greatest number of members；Vitis amurensis mTERF genes showed a particularly close evolutionary relationship with those of poplar. Codon usage bias analysis indicated a relatively weak codon usage preference within the VamTERF gene family，and synteny analysis demonstrated co-linear relationships between Vitis amurensis and five other plant species，including strawberry. qRT-PCR analysis showed that all VamTERF members responded to abiotic stresses，with VamTERF2，VamTERF3，VamTERF4，VamTERF5，and VamTERF7 being significantly upregulated under low-temperature stress. Subcellular localization experiments revealed that these five members were predominantly localized in the nucleus. Overexpression of VamTERF5 in grapevine calli significantly enhanced the activities of peroxidase（POD），superoxide dismutase（SOD），and catalase（CAT）following low-temperature treatment. These findings indicate that VamTERF members play important regulatory roles in the response to low-temperature stress.

Key words: Vitis amurensis, mTERF, gene family, bioinformatics, cold tolerance

范基鸿, 陈继贤, 李阳阳, 曾宝珍, 郭艳兰, 毛娟, 陈佰鸿, 梁国平. 葡萄线粒体转录终止因子（mTERF）家族基因鉴定与VamTERF5抗寒性研究[J]. 园艺学报, 2026, 53(8): 2256-2274.

FAN Jihong, CHEN Jixian, LI Yangyang, ZENG Baozhen, GUO Yanlan, MAO Juan, CHEN Baihong, LIANG Guoping. Identification of Mitochondrial Transcriptional Termination Factor （mTERF）Gene Family and Cold Resistance of VamTERF5 in Vitis amurensis[J]. Acta Horticulturae Sinica, 2026, 53(8): 2256-2274.

图/表 15

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基因 Gene	引物序列（5′-3′） Primer sequence	扩增产物长度/bp Length of amplification product	用途 Application
VamTERF1-F	TCGGCAGAGAGCATAGGCAAGG	132	qRT-PCR
VamTERF1-R	ACGGTGGAGAAGAATGGCAACATC	132
VamTERF2-F	GGTTCGCAGGTCTGATCGCATC	186
VamTERF2-R	AGCACCCTCCTCCTCTTCTTCTTC	186
VamTERF3-F	ACTCCGTCCTCGCACTCCTTAG	118
VamTERF3-R	AGTTTGGGCAAAAGGGTCTTCTCAG	118
VamTERF4-F	ATGCCAGTGTTGTAGTAGACCTTGC	147
VamTERF4-R	AAGCCACAGATGTGCTCATAGTTCC	147
VamTERF5-F	CCATCTCCCACAAGCATCACCATC	147
VamTERF5-R	GCGATTTCCACCGATTCCTCTTCC	147
VamTERF6-F	ATCTACATTCAGCCGTCGCAATCG	86
VamTERF6-R	TCGTCATCGTCTTCATCGCTTTCTC	86
VamTERF7-F	AGAAGAAGCAGTTGCAGAACCAGTG	92
VamTERF7-R	CCGTAGCGTGACCTGCGATTG	92
VaGAPDH-F	TTCTCGTTGAGGGCTATTCCA	70
VaGAPDH-R	CCACAGACTTCATCGGTGACA	70
VamTERF2-F	gagctcggtacccggggatccATGGTTCGCAGGTCTGATCG	1 239	亚细胞定位 Subcellular localization
VamTERF2-R	catgtcgactctagaggatcccATGATACATTTCCAGTCACAATATTG	1 239
VamTERF3-F	gagctcggtacccggggatccATGAACTCATGTGGGTTGTCCC	1 065
VamTERF3-R	catgtcgactctagaggatccTCCCCTGACAGACAATGAACAA	1 065
VamTERF4-F	gagctcggtacccggggatccATGATACAGACCATTATGAACAT	1 593
VamTERF4-R	catgtcgactctagaggatccCTATGCATATTCATCATCACT	1 593
VamTERF5-F	gagctcggtacccggggatccATGTTCTCCAAACTCCTCCCAA	1 806
VamTERF5-R	catgtcgactctagaggatccATGAGGAGAAATGGGAGGAGAGG	1 806
VamTERF7-F	gagctcggtacccggggatccATGGCTGCTTTCTATCTCTACACG	1 545
VamTERF7-R	catgtcgactctagaggatccTCTTCGCTTCTCATACTTCTTTGC	1 545
Kan-F	CTAAAACAATTCATCCAGTAA	795	阳性愈伤组织检测 Identification of positive callus
Kan-R	ATGGCTAAAATGAGAATATCAC	795	阳性愈伤组织检测 Identification of positive callus

基因 Gene	引物序列（5′-3′） Primer sequence	扩增产物长度/bp Length of amplification product	用途 Application
VamTERF1-F	TCGGCAGAGAGCATAGGCAAGG	132	qRT-PCR
VamTERF1-R	ACGGTGGAGAAGAATGGCAACATC	132
VamTERF2-F	GGTTCGCAGGTCTGATCGCATC	186
VamTERF2-R	AGCACCCTCCTCCTCTTCTTCTTC	186
VamTERF3-F	ACTCCGTCCTCGCACTCCTTAG	118
VamTERF3-R	AGTTTGGGCAAAAGGGTCTTCTCAG	118
VamTERF4-F	ATGCCAGTGTTGTAGTAGACCTTGC	147
VamTERF4-R	AAGCCACAGATGTGCTCATAGTTCC	147
VamTERF5-F	CCATCTCCCACAAGCATCACCATC	147
VamTERF5-R	GCGATTTCCACCGATTCCTCTTCC	147
VamTERF6-F	ATCTACATTCAGCCGTCGCAATCG	86
VamTERF6-R	TCGTCATCGTCTTCATCGCTTTCTC	86
VamTERF7-F	AGAAGAAGCAGTTGCAGAACCAGTG	92
VamTERF7-R	CCGTAGCGTGACCTGCGATTG	92
VaGAPDH-F	TTCTCGTTGAGGGCTATTCCA	70
VaGAPDH-R	CCACAGACTTCATCGGTGACA	70
VamTERF2-F	gagctcggtacccggggatccATGGTTCGCAGGTCTGATCG	1 239	亚细胞定位 Subcellular localization
VamTERF2-R	catgtcgactctagaggatcccATGATACATTTCCAGTCACAATATTG	1 239
VamTERF3-F	gagctcggtacccggggatccATGAACTCATGTGGGTTGTCCC	1 065
VamTERF3-R	catgtcgactctagaggatccTCCCCTGACAGACAATGAACAA	1 065
VamTERF4-F	gagctcggtacccggggatccATGATACAGACCATTATGAACAT	1 593
VamTERF4-R	catgtcgactctagaggatccCTATGCATATTCATCATCACT	1 593
VamTERF5-F	gagctcggtacccggggatccATGTTCTCCAAACTCCTCCCAA	1 806
VamTERF5-R	catgtcgactctagaggatccATGAGGAGAAATGGGAGGAGAGG	1 806
VamTERF7-F	gagctcggtacccggggatccATGGCTGCTTTCTATCTCTACACG	1 545
VamTERF7-R	catgtcgactctagaggatccTCTTCGCTTCTCATACTTCTTTGC	1 545
Kan-F	CTAAAACAATTCATCCAGTAA	795	阳性愈伤组织检测 Identification of positive callus
Kan-R	ATGGCTAAAATGAGAATATCAC	795	阳性愈伤组织检测 Identification of positive callus

基因名称 Gene name	基因全长/bp Full length	CDS长度/bp Length of CDS	氨基酸数量No. of amino acids	分子量/kD Molecular weight	等电点 pI	α-螺旋/% Alpha helix	β-转角/% Beta turn	不规则卷曲/% Random coil	亚细胞定位预测Subcellular location prediction
VamTERF1	1 911	1 488	495	160 228.81	4.96	55.76	6.87	29.49	叶绿体Chloroplast
VamTERF2	2 989	1 239	412	249 214.26	4.92	41.26	2.18	44.42	细胞核Nucleus
VamTERF3	2 774	1 065	344	232 135.57	4.92	51.98	7.91	27.12	叶绿体Chloroplast，细胞核Nucleus
VamTERF4	1 593	1 593	530	59 929.75	6.17	57.55	5.28	28.68	细胞核Nucleus
VamTERF5	5 432	1 806	601	69 035.95	7.28	55.57	5.49	29.28	细胞核Nucleus
VamTERF6	4 343	1 713	570	65 407.95	8.09	47.02	5.61	38.07	叶绿体Chloroplast
VamTERF7	9 385	1 545	514	59 217.08	9.07	55.25	6.03	31.52	细胞核Nucleus

基因名称 Gene name	基因全长/bp Full length	CDS长度/bp Length of CDS	氨基酸数量No. of amino acids	分子量/kD Molecular weight	等电点 pI	α-螺旋/% Alpha helix	β-转角/% Beta turn	不规则卷曲/% Random coil	亚细胞定位预测Subcellular location prediction
VamTERF1	1 911	1 488	495	160 228.81	4.96	55.76	6.87	29.49	叶绿体Chloroplast
VamTERF2	2 989	1 239	412	249 214.26	4.92	41.26	2.18	44.42	细胞核Nucleus
VamTERF3	2 774	1 065	344	232 135.57	4.92	51.98	7.91	27.12	叶绿体Chloroplast，细胞核Nucleus
VamTERF4	1 593	1 593	530	59 929.75	6.17	57.55	5.28	28.68	细胞核Nucleus
VamTERF5	5 432	1 806	601	69 035.95	7.28	55.57	5.49	29.28	细胞核Nucleus
VamTERF6	4 343	1 713	570	65 407.95	8.09	47.02	5.61	38.07	叶绿体Chloroplast
VamTERF7	9 385	1 545	514	59 217.08	9.07	55.25	6.03	31.52	细胞核Nucleus

物种 Species	编号 No.	名称 Gene name	基因登录号 Gene accession number	物种 Species	编号 No.	名称 Gene name	基因登录号 Gene accession number
山葡萄 Vitis amurensis	1	VamTERF1	VIT_214s0171g00230.1.v2.1	拟南芥 Arabidopsis thaliana	1	AtmTERF1	XP_028953475.1
	2	VamTERF2	VIT_210s0116g01530.1.v2.1		2	AtmTERF2	XP_050136630.1
	3	VamTERF3	VIT_207s0104g01550.1.v2.1		3	AtmTERF3	XP_017178176.1
	4	VamTERF4	VIT_207s0104g00720.1.v2.1		4	AtmTERF4	XP_008368884.2
	5	VamTERF5	VIT_209s0054g01030.1.v2.1		5	AtmTERF5	XP_028953495.1
	6	VamTERF6	VIT_203s0063g02590.4.v2.1		6	AtmTERF6	XP_021686388.2
	7	VamTERF7	VIT_219s0090g01770.2.v2.1		7	AtmTERF7	XP_008385796.2
苹果 Malus × domestica	1	MdmTERF1	XP_028953475.1	杨树 Populus tremula	1	PtmTERF1	KAJ6860499.1
	2	MdmTERF2	XP_050136630.1	杨树 Populus tremula	2	PtmTERF2	XP_034903054.1
	3	MdmTERF3	XP_017178176.1		3	PtmTERF3	KAJ6970000.1
	4	MdmTERF4	XP_008368884.2		4	PtmTERF4	KAJ6973063.1
	5	MdmTERF5	XP_028953495.1		5	PtmTERF5	KAJ6915659.1
	6	MdmTERF6	XP_021686388.2		6	PtmTERF6	KAJ6985531.1
	7	MdmTERF7	XP_008385796.2		7	PtmTERF7	XP_034925224.1
草莓 Fragaria × ananassa	1	FamTERF1	XP_004305919.1	柑橘 Citrus sinensis	1	CitmTERF1	KPL70_018202
	2	FamTERF2	XP_011463401.1		2	CitmTERF2	KDO54693.1
	3	FamTERF3	PRQ51409.1		3	CitmTERF3	XM_006444194.2
	4	FamTERF4	XP_004290690.1		4	CitmTERF4	KAH9730041.1
	5	FamTERF5	XP_004305797.1		5	CitmTERF5	XP_006450033.1
	6	FamTERF6	XP_011461117.1		6	CitmTERF6	KDO49079.1
	7	FamTERF7	XP_059647673.1		7	CitmTERF7	XP_006452528.1
	8	FamTERF8	XP_011461117.1