甜瓜CmDREB基因的克隆及其表达分析

doi:10.16420/j.issn.0513-353x.2025-0358

摘要/Abstract

摘要：

DREB（dehydration responsive element binding protein）是植物中特有的一类转录因子，参与植物生长发育、代谢和非生物胁迫应答等过程，然而甜瓜中的CmDREB基因尚未被系统研究。以‘新密3号’甜瓜为试材，通过全基因组序列分析，鉴定出10个甜瓜CmDREB转录因子并进行了克隆与表达分析。结果表明，10个甜瓜CmDREB蛋白结构稳定，其主要二级结构为α-螺旋和无规则卷曲，氨基酸数量为179 ~ 428，分子量为44.1 ~ 104.2 kD；除CmDREB2C定位于叶绿体外，其余成员均定位于细胞核。启动子分析显示，10个CmDREB基因的启动子区域均含有响应低温、水杨酸（SA）、茉莉酸甲酯（MeJA）和脱落酸（ABA）等的顺式作用元件，但各基因响应特点不同。其中CmDREB2C和CmDREB4C在低温下表达量较高，而CmDREB4A和CmDREB4D受SA、MeJA和ABA诱导处理表达强烈。

关键词: 甜瓜, CmDREB, 非生物胁迫, 基因克隆, 表达分析

Abstract:

DREB（dehydration responsive element binding protein）is a plant-specific family of transcription factors involved in various processes such as plant growth and development，metabolism，and abiotic stress responses. However，the CmDREB genes in melon have not been systematically studied. In this study，using‘Xinmi 3’melon as the experimental material， 10 CmDREB transcription factors through genome-wide sequence analysis were identified，followed by cloning and expression analysis. The results showed that the 10 CmDREB proteins exhibited stable structures，with α-helix and random coil as the predominant secondary structures. Their amino acid lengths ranged from 179 to 428，with molecular weights ranging from 44.1 to 104.2 kD. Except for CmDREB2C，which was localized in the chloroplast，all other members were localized in the nucleus. Promoter analysis revealed that the promoter regions of all 10 CmDREB genes contained cis-acting elements responsive to low temperature，salicylic acid（SA），methyl jasmonate（MeJA），and abscisic acid（ABA）. Expression analysis further demonstrated distinct response patterns among these genes：CmDREB2C and CmDREB4C exhibited higher expression levels under low temperature，whereas CmDREB4A and CmDREB4D showed strong induction upon treatment with SA，MeJA，and ABA.

Key words: Cucumis melo, CmDREB, abiotic stress, gene cloning, expression analysis

克拉热木·克里木江, 张健, 赵志霞, 付慧鑫, 张婷. 甜瓜CmDREB基因的克隆及其表达分析[J]. 园艺学报, 2026, 53(8): 2319-2332.

KELAREMU · Kelimujiang, ZHANG Jian, ZHAO Zhixia, FU Huixin, ZHANG Ting. Cloning and Expression Analysis of CmDREB Genes in Cucumis melo[J]. Acta Horticulturae Sinica, 2026, 53(8): 2319-2332.

图/表 11

参考文献 38

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引物名称 Primer name	序列（5′-3′） Sequence	引物名称 Primer name	序列（5′-3′） Sequence
4268F	ATGGCAAAACCAAAGAATTCAAATCC	83P2F	ATGGTCGAGAAAATGGGTCACAGTG
4268R	TTAATCCCATAGTACGCCGTAA	83P2R	TCAAAATCCTAAATCACCAATTTCGTTCAG
9763F	ATGCCTAAATTAGGAAAACTAC	0977F	ATGTTGAAATCTTGTTCATCCAAC
9763R	CTAGACATCTTCAACTGCAC	0977R	TTAATACCATGGGATTGAAGAATCCC
4579F	ATGGCCAACGCCACCGT	4714F	ATGGAGAACGATTCTACAATTTTAAACT
4579R	TTACCACGAGAAAGATAAAGG	4714R	TTAATTTTGCCATAAGAAAGTATCGG
7769F	ATGGCAGCAGCTATAGATG	6024F	ATGACGTCTGAGAATTCTAGTGGA
7769R	TTAACACTCTGATTCGGCCA	6024R	TCATCCATGGAAGCATAAATCAAGC
83P1F	ATGATATCAGAGACGATTGAGA	1143F	ATGAGTAGTAGTAAAGAACAGA
83P1R	TCAAAATCCTAAATCACCAATTTCG	1143R	TCAATAATCAAACAGAAACCC

引物名称 Primer name	序列（5′-3′） Sequence	引物名称 Primer name	序列（5′-3′） Sequence
4268F	ATGGCAAAACCAAAGAATTCAAATCC	83P2F	ATGGTCGAGAAAATGGGTCACAGTG
4268R	TTAATCCCATAGTACGCCGTAA	83P2R	TCAAAATCCTAAATCACCAATTTCGTTCAG
9763F	ATGCCTAAATTAGGAAAACTAC	0977F	ATGTTGAAATCTTGTTCATCCAAC
9763R	CTAGACATCTTCAACTGCAC	0977R	TTAATACCATGGGATTGAAGAATCCC
4579F	ATGGCCAACGCCACCGT	4714F	ATGGAGAACGATTCTACAATTTTAAACT
4579R	TTACCACGAGAAAGATAAAGG	4714R	TTAATTTTGCCATAAGAAAGTATCGG
7769F	ATGGCAGCAGCTATAGATG	6024F	ATGACGTCTGAGAATTCTAGTGGA
7769R	TTAACACTCTGATTCGGCCA	6024R	TCATCCATGGAAGCATAAATCAAGC
83P1F	ATGATATCAGAGACGATTGAGA	1143F	ATGAGTAGTAGTAAAGAACAGA
83P1R	TCAAAATCCTAAATCACCAATTTCG	1143R	TCAATAATCAAACAGAAACCC

引物名称 Primer name	序列（5′-3′） Sequence	引物名称 Primer name	序列（5′-3′） Sequence
Q4268F	TGAGTGAGATCGTCGAGTTG	Q83P2R	GATCCTTTAGCTGGAGCTTTACGG
Q4268R	GTACGCCGTAAAAACTGCTC	Q0977F	GGAGGTCCTGAAAATGCTTC
Q9763F	CTCGTAGAGTTCCAGCCAAG	Q0977R	TCAGAGCCATAAAGACGACG
Q9763R	GAAATGTACCCAACCAGAGC	Q4714F	ATTTTAAACTCATCTCCGCCG
Q4579F	GTAGTATTGCCGAGTTTGGG	Q4714R	GAAACCCATTTTCCCCATGC
Q4579R	AGTGGGAATCACACACTCTC	Q6024F	GGAGGAGTAGAGGAAACTCG
Q7769F	GGTTTTGAGCAATCAGGTCC	Q6024R	GTTACTCGCAATTGGCTCTC
Q7769R	GTAGGTTTAGGTGGAGAGCC	Q1143F	GTGAGATTGTTGAGTTGCCG
Q83P1F	CTCTACGTGTTCCGAGTCCACTACTG	Q1143R	TCCCATTCCCAACTTACCAC
Q83P1R	CAATCCTCATGTTTCACTTGATTCTC	EF1aF	AAATACTCCAAGGCAAGGTAC
Q83P2F	GGTCACAGTGATCAATATTCTG	EF1aR	TCATGTTGTCACCCTCGAAACCAG

引物名称 Primer name	序列（5′-3′） Sequence	引物名称 Primer name	序列（5′-3′） Sequence
Q4268F	TGAGTGAGATCGTCGAGTTG	Q83P2R	GATCCTTTAGCTGGAGCTTTACGG
Q4268R	GTACGCCGTAAAAACTGCTC	Q0977F	GGAGGTCCTGAAAATGCTTC
Q9763F	CTCGTAGAGTTCCAGCCAAG	Q0977R	TCAGAGCCATAAAGACGACG
Q9763R	GAAATGTACCCAACCAGAGC	Q4714F	ATTTTAAACTCATCTCCGCCG
Q4579F	GTAGTATTGCCGAGTTTGGG	Q4714R	GAAACCCATTTTCCCCATGC
Q4579R	AGTGGGAATCACACACTCTC	Q6024F	GGAGGAGTAGAGGAAACTCG
Q7769F	GGTTTTGAGCAATCAGGTCC	Q6024R	GTTACTCGCAATTGGCTCTC
Q7769R	GTAGGTTTAGGTGGAGAGCC	Q1143F	GTGAGATTGTTGAGTTGCCG
Q83P1F	CTCTACGTGTTCCGAGTCCACTACTG	Q1143R	TCCCATTCCCAACTTACCAC
Q83P1R	CAATCCTCATGTTTCACTTGATTCTC	EF1aF	AAATACTCCAAGGCAAGGTAC
Q83P2F	GGTCACAGTGATCAATATTCTG	EF1aR	TCATGTTGTCACCCTCGAAACCAG

名称 Name	基因ID Gene ID	氨基酸数量 No. of amino acid	不稳定系数 Coefficient of instability	等电点 pI	平均亲水指数 Grand average of hydropathicity	分子量/kD Molecular weight	亚细胞定位 Prediction of subcellular localization
CmDREB4A	MELO3C011143P1	229	34.13	5.16	0.662	57.6	细胞核Nuclear
CmDREB4B	MELO3C024268P1	201	39.68	5.14	0.800	49.7	细胞核Nuclear
CmDREB4C	MELO3C004579P1	252	42.04	5.08	0.878	63.3	细胞核Nuclear
CmDREB4D	MELO3C024714P1	250	58.21	5.08	0.910	64.3	细胞核Nuclear
CmDREB6A	MELO3C007769P1	349	56.57	5.02	0.860	90.4	细胞核Nuclear
CmDREB2A	MELO3C010977P1	211	51.30	5.19	0.641	53.8	细胞核Nuclear
CmDREB2B	MELO3C026024P1	373	32.06	5.11	0.651	90.5	细胞核Nuclear
CmDREB2C	MELO3C019763P1	179	41.57	5.22	0.754	44.1	叶绿体Chloroplast
CmDREB2D	MELO3C008318P1	413	39.59	5.06	0.710	100.2	细胞核Nuclear
CmDREB2E	MELO3C008318P2	428	40.02	5.06	0.713	104.2	细胞核Nuclear