Identification of the Grape HCT Gene Family and Their Responses to Low-temperature Stress

doi:10.13560/j.cnki.biotech.bull.1985.2024-0597

Abstract

Abstract:

Objective Hydroxycinnamoyl-CoA shikimate hydroxycinnamoyl transferase (HCT) is a key enzyme for catalyzing chlorogenic acid synthesis in dicotyledonous plants and is widely involved in plant abiotic stress response. We identified members of the VvHCT gene family in the whole grape genome and analyzed the expression pattern of the related genes under low-temperature stress, which may provide a theoretical basis for subsequent studies. Method Based on the whole genome information of grapevine, bioinformatics methods was sued to identify the VvHCT family, and analyze the physical and chemical properties, chromosome distribution, gene structure, protein conserved structure, systematic evolution, promoter cis-acting elements and tissue expression characteristics of the family members. Fluorescence quantitative PCR was adapted to detect the expression pattern of VvHCT gene under low-temperature stress, and transient transformation method to verify the function of VvHCT8 gene. Result Eleven family members of VvHCT were identified in the whole genome of grapevine, encoding 398–457 amino acids, distributed on four chromosomes. They were classified into three subfamilies according to their phylogenetic characteristics. The analysis of the promoter cis-acting elements showed that the promoters of the VvHCT family members contained light-responsive elements, hormone-responsive elements, stress-responsive elements, tissue-specific elements and circadian response elements. The expression patterns of different VvHCT family members differed greatly. The expression of VvHCT5 was down-regulated after low-temperature stress treatment, while the expressions of the other 10 family members were up-regulated; the transient overexpression results of grape leaves showed that VvHCT8 alleviated the damages to chlorophyll and cell membranes of grape leaves by reducing the content of MDA and ROS. Conclusion VvHCT8 is involved in the process of low-temperature resistance in grapes and can be used as a candidate gene for cold resistance in grapes.

Key words: grape, VvHCT gene family, bioinformatics, low temperature

YAN Wei, CHEN Hui-ting, YE Qing, LIU Guang-chao, LIU Xin, HOU Li-xia. Identification of the Grape HCT Gene Family and Their Responses to Low-temperature Stress[J]. Biotechnology Bulletin, 2025, 41(2): 175-186.

Figures/Tables 12

References 32

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引物名称 Primer name	引物序列 Primer sequence （5′-3′）
VvActin-F	ATAGAAGCAGCAAGGGA
VvActin-R	TGAGGCTCTTACTAATG
VvHCT1-F	CCACCTTCGCCATTTCCAAC
VvHCT1-R	TAGGGAACACCCTGGTCAGT
VvHCT2-F	CCTCTCCAACCCCACATTCC
VvHCT2-R	CCCCTTATCATTGCATGGGTT
VvHCT3-F	TGAGTGCAACGATGAGGGAG
VvHCT3-R	TGCCGTGGAGATCAGAAACC
VvHCT4-F	GGAGGTTTGTGTTTGATGGGG
VvHCT4-R	TGGGCGCAAGTTTATGCTGT
VvHCT5-F	ACTTCCCTTGCGGTCCATTC
VvHCT5-R	TTATCGCAGTCGGGAACACA
VvHCT6-F	GGTTGTAAGCCGAGCGAAAC
VvHCT6-R	TTGCCACTGTCCGCCATAAA
VvHCT7-F	CTGCCAACTTCTTCGACCCT
VvHCT7-R	GTAGGGGCGAAATCACCGAA
VvHCT8-F	CGACAGGCAATAGAGAAGGCA
VvHCT8-R	AGATGGGAAGTCTAGCCCAG
VvHCT9-F	CTAGCGGGATGGTTGCACT
VvHCT9-R	CATCAGTCATGATATGCAATGCTC
VvHCT10-F	AGCAATTGGCTACGATGGGA
VvHCT10-R	AGCAATTGGCTACGATGGGA
VvHCT11-F	ATCGGAGCCCACTTTCGATG
VvHCT11-R	AGCGGGTAGAAATGCACCAA

引物名称 Primer name	引物序列 Primer sequence （5′-3′）
VvActin-F	ATAGAAGCAGCAAGGGA
VvActin-R	TGAGGCTCTTACTAATG
VvHCT1-F	CCACCTTCGCCATTTCCAAC
VvHCT1-R	TAGGGAACACCCTGGTCAGT
VvHCT2-F	CCTCTCCAACCCCACATTCC
VvHCT2-R	CCCCTTATCATTGCATGGGTT
VvHCT3-F	TGAGTGCAACGATGAGGGAG
VvHCT3-R	TGCCGTGGAGATCAGAAACC
VvHCT4-F	GGAGGTTTGTGTTTGATGGGG
VvHCT4-R	TGGGCGCAAGTTTATGCTGT
VvHCT5-F	ACTTCCCTTGCGGTCCATTC
VvHCT5-R	TTATCGCAGTCGGGAACACA
VvHCT6-F	GGTTGTAAGCCGAGCGAAAC
VvHCT6-R	TTGCCACTGTCCGCCATAAA
VvHCT7-F	CTGCCAACTTCTTCGACCCT
VvHCT7-R	GTAGGGGCGAAATCACCGAA
VvHCT8-F	CGACAGGCAATAGAGAAGGCA
VvHCT8-R	AGATGGGAAGTCTAGCCCAG
VvHCT9-F	CTAGCGGGATGGTTGCACT
VvHCT9-R	CATCAGTCATGATATGCAATGCTC
VvHCT10-F	AGCAATTGGCTACGATGGGA
VvHCT10-R	AGCAATTGGCTACGATGGGA
VvHCT11-F	ATCGGAGCCCACTTTCGATG
VvHCT11-R	AGCGGGTAGAAATGCACCAA

基因名称 Gene name	基因 ID Gene ID	染色体位置 Chromosome location	编码区 CDS/bp	氨基酸 Amino acid/bp	分子量 Molecular weigh/kD	等电点 Theoretical isoelectric points	不稳定系数 Instability index	总平均亲水性 Grand average of hydropathicity	KEGG	亚细胞定位 Subcellular localization
VvHCT1	Vitvi01g01514	Chr1：20494476：20495771	1 296	431	48.45	6.04	38.66	-0.018	2.3.1.133	细胞质
VvHCT2	Vitvi01g01517	Chr1：20532349：20533650	1 197	398	44.78	5.76	41.1	-0.096	2.3.1.133	细胞质
VvHCT3	Vitvi01g02214	Chr1：20566690：20568324	1 290	429	47.97	6.87	40.5	-0.139	2.3.1.133	细胞质
VvHCT4	Vitvi03g00077	Chr3：1057145：1058434	1 290	429	47.75	6.84	42.89	-0.257	2.3.1.133	细胞质
VvHCT5	Vitvi03g01800	Chr3：16887244：16888554	1 311	436	47.96	6.39	35.16	-0.128	2.3.1.133	细胞质
VvHCT6	Vitvi03g01816	Chr3：17430346：17432051	1 284	427	47.51	7.6	44.02	-0.171	2.3.1.133	细胞质
VvHCT7	Vitvi09g01229	Chr9：19655320：19657669	1 290	429	47.86	6.13	47.54	-0.166	2.3.1.133	细胞质
VvHCT8	Vitvi11g00730	Chr11：8730745：8732578	1 338	445	49.41	6.33	44.21	-0.273	2.3.1.133	细胞质
VvHCT9	Vitvi11g00735	Chr11：8783989：8786081	1 374	457	50.73	6.51	49.25	-0.224	2.3.1.133	细胞质
VvHCT10	Vitvi11g00742	Chr11：8868521：8869858	1 338	445	49.63	7.67	46.99	-0.252	2.3.1.133	细胞质
VvHCT11	Vitvi11g01099	Chr11：15978906：15980276	1 371	456	50.25	5.33	44.68	-0.163	2.3.1.133	细胞质

基因名称 Gene name	基因 ID Gene ID	染色体位置 Chromosome location	编码区 CDS/bp	氨基酸 Amino acid/bp	分子量 Molecular weigh/kD	等电点 Theoretical isoelectric points	不稳定系数 Instability index	总平均亲水性 Grand average of hydropathicity	KEGG	亚细胞定位 Subcellular localization
VvHCT1	Vitvi01g01514	Chr1：20494476：20495771	1 296	431	48.45	6.04	38.66	-0.018	2.3.1.133	细胞质
VvHCT2	Vitvi01g01517	Chr1：20532349：20533650	1 197	398	44.78	5.76	41.1	-0.096	2.3.1.133	细胞质
VvHCT3	Vitvi01g02214	Chr1：20566690：20568324	1 290	429	47.97	6.87	40.5	-0.139	2.3.1.133	细胞质
VvHCT4	Vitvi03g00077	Chr3：1057145：1058434	1 290	429	47.75	6.84	42.89	-0.257	2.3.1.133	细胞质
VvHCT5	Vitvi03g01800	Chr3：16887244：16888554	1 311	436	47.96	6.39	35.16	-0.128	2.3.1.133	细胞质
VvHCT6	Vitvi03g01816	Chr3：17430346：17432051	1 284	427	47.51	7.6	44.02	-0.171	2.3.1.133	细胞质
VvHCT7	Vitvi09g01229	Chr9：19655320：19657669	1 290	429	47.86	6.13	47.54	-0.166	2.3.1.133	细胞质
VvHCT8	Vitvi11g00730	Chr11：8730745：8732578	1 338	445	49.41	6.33	44.21	-0.273	2.3.1.133	细胞质
VvHCT9	Vitvi11g00735	Chr11：8783989：8786081	1 374	457	50.73	6.51	49.25	-0.224	2.3.1.133	细胞质
VvHCT10	Vitvi11g00742	Chr11：8868521：8869858	1 338	445	49.63	7.67	46.99	-0.252	2.3.1.133	细胞质
VvHCT11	Vitvi11g01099	Chr11：15978906：15980276	1 371	456	50.25	5.33	44.68	-0.163	2.3.1.133	细胞质

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