菊花CmJAZ1基因负调控干旱胁迫的功能分析

doi:10.13560/j.cnki.biotech.bull.1985.2025-1228

摘要/Abstract

摘要：

目的探究菊花（Chrysanthemum morifolium）在干旱胁迫中CmJAZ基因家族特征并验证CmJAZ1的功能，为后续深入研究菊花耐旱分子育种提供理论依据。方法基于菊花基因组数据，利用生物信息学方法对CmJAZ基因家族成员进行筛选鉴定，对其编码蛋白质的特征、系统进化关系等进行分析，并利用实时荧光定量PCR（RT-qPCR）分析其在干旱胁迫和茉莉酸甲酯（MeJA）下的表达模式。将筛选出来的目的基因（JAZ1）进行遗传转化获得转基因烟草株系，对其干旱胁迫下生理指标进行测定。结果从菊花基因组中鉴定出9个CmJAZ基因，其编码蛋白分子量为15.30‒34.55 kD，等电点6.44‒9.61，均含保守的TIFY结构域和Jas基序，亚细胞定位预测多位于细胞核。序列比对显示，CmJAZ1与青蒿AaJAZ1氨基酸同源性最高（96.38%）。系统发育分析表明其与菊科植物JAZ蛋白亲缘关系较近。表达谱分析发现，20% PEG-6000模拟干旱胁迫和MeJA处理诱导9个CmJAZ基因表达，其中CmJAZ1表达量上调约9倍。通过农杆菌介导法获得CmJAZ1过表达（OE）、基因沉默（RNAi）转基因烟草株系。干旱胁迫下与野生型（WT）相比，RNAi株系相对电导率和丙二醛含量显著降低，超氧化物歧化酶（SOD）、过氧化物酶（POD）、过氧化氢酶（CAT）活性显著升高，脯氨酸和可溶性糖含量显著增加，而OE株系呈现相反趋势。结论 CmJAZ1基因通过负调控细胞膜稳定性、抗氧化酶活性和渗透调节物质积累影响植株抗旱性，抑制其表达可增强菊花干旱耐受性。

关键词: 菊花, CmJAZ1, 干旱胁迫, 抗氧化酶, 渗透调节

Abstract:

Objective The characteristics of the CmJAZ gene family in response to drought stress in chrysanthemum (Chrysanthemum morifolium) were studied, and the function of CmJAZ1 was verified, providing a theoretical basis for further research on drought-resistant molecular breeding of chrysanthemum. Method Based on the chrysanthemum genome data, the JAZ gene family members were screened and identified using bioinformatics methods. The characteristics of the encoded proteins and phylogenetic relationships were analyzed. The expression patterns of the JAZ gene family under drought stress and methyl jasmonate (MeJA) were analyzed using real-time quantitative PCR (RT-qPCR). The screened target gene (CmJAZ1) was subjected to genetic transformation to obtain transgenic tobacco lines, and the physiological indices of these lines under drought stress were determined. Result A total of 9 CmJAZ genes were identified from the chrysanthemum genome. The molecular weights of their encoded proteins ranged from 15.30 to 34.55 kD, with isoelectric points between 6.44 and 9.61. All these proteins contained conserved TIFY domains and Jas motifs, and subcellular localization prediction indicates that they are mostly in the nucleus. Sequence alignment showed that CmJAZ1 shares the highest amino acid homology (96.38%) with AaJAZ1 from Artemisia annua. Phylogenetic analysis indicated that it has a close genetic relationship with JAZ proteins from Asteraceae plants. Expression profile analysis revealed that both 20% PEG-6000-simulated drought stress and MeJA treatment significantly induced the expression of the CmJAZ genes, among which the expression level of CmJAZ1 was upregulated by approximately 9-fold. Transgenic tobacco lines with CmJAZ1 overexpression (OE) and RNAi-mediated gene silencing (RNAi) were obtained via the Agrobacterium-mediated method. Under drought stress, compared with the wild type (WT), the RNAi lines exhibited significantly reduced relative electrical conductivity and malondialdehyde (MDA) content, significantly increased activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), as well as significantly elevated proline and soluble sugar contents; whereas the OE lines showed the opposite trends. Conclusion The CmJAZ1 gene affects plant drought resistance by negatively regulating cell membrane stability, antioxidant enzyme activity, and the accumulation of osmotic adjustment substances, and inhibiting its expression can enhance drought tolerance in chrysanthemums.

Key words: chrysanthemum, CmJAZ1, drought stress, antioxidant enzymes, osmotic regulation

李家群, 张煜婷, 张新仪, 保嘉佳, 严瑞. 菊花CmJAZ1基因负调控干旱胁迫的功能分析[J]. 生物技术通报, 2026, 42(11): 145-153.

LI Jia-qun, ZHANG Yu-ting, ZHANG Xin-yi, BAO Jia-jia, YAN Rui. Functional Analysis of Chrysanthemum CmJAZ1 Gene in Negative Regulation of Drought Stress[J]. Biotechnology Bulletin, 2026, 42(11): 145-153.

图/表 7

表1 菊花CmJAZ家族成员基本理化性质分析

Table 1 Analysis of basic physicochemical properties of CmJAZfamily members in chrysanthemum

蛋白 Protein	氨基酸数目 Amino acid number	分子质量 Molecular weight （Da）	等电点 Isoelectric point	不稳定系数 Instability index	脂肪指数 Aliphatic index	亲水性 GRAVY	亚细胞定位 Subcellular localization	稳定性 Stability
CmJAZ1	219	24 231.17	6.44	39.94	67.76	-0.605	细胞核	稳定蛋白
CmJAZ2	185	20 556.44	9.39	45.24	81.19	-0.450	线粒体	不稳定蛋白
CmJAZ3	185	20 072.58	9.22	45.52	54.38	-0.659	细胞核	不稳定蛋白
CmJAZ4	187	20 653.67	9.39	55.37	73.48	-0.456	细胞核	不稳定蛋白
CmJAZ5	139	15 300.25	7.86	44.13	66.12	-0.428	细胞核	不稳定蛋白
CmJAZ6	237	26 121.55	8.85	46.79	77.05	-0.607	线粒体	不稳定蛋白
CmJAZ7	326	34 548.01	9.36	53.11	64.02	-0.331	叶绿体	不稳定蛋白
CmJAZ8	255	27 669.43	9.25	51.69	62.78	-0.343	细胞核	不稳定蛋白
CmJAZ9	188	20 346.19	9.61	63.34	78.88	-0.387	细胞核	不稳定蛋白

图1 干旱胁迫（A）和MeJA处理（B）下菊花JAZ基因表达量

Fig. 1 Expression of JAZ gene in chrysanthemum under drought stress (A) and MeJA treatment (B)

图2 菊花CmJAZ1蛋白序列比对（A）与系统进化树构建（B）Cs：茶树；Pa：银白杨；Mt：苜蓿；Gh：陆地棉；Bo：甘蓝；Nt：烟草；Pt：毛果杨；Hb：橡胶树；Cn：椰子；Ma：香蕉；Tu：小麦；Os：水稻；Ob：短药稻；Cl：油茶；Ha：向日葵；Cm：菊花；Aa：青蒿；At：拟南芥

Fig. 2 Protein sequence alignment (A) and phylogenetic tree construction (B) of chrysanthemum CmJAZ1Cs: Camellia sinensis. Pa: Populus alba. Mt: Medicago truncatula. Gh: Gossypium hirsutum. Bo: Brassica oleracea. Nt: Nicotiana tabacum. Pt: Populus trichocarpa. Hb: Hevea brasiliensis. Cn: Cocos nucifera. Ma: Musa acuminata. Tu: Triticum urartu. Os: Oryza sativa. Ob: Oryza brachyantha. Cl: Camellia lanceoleosa. Ha: Helianthus annuus. Cm: Chrysanthemum morifolium. Aa: Artemisia annua. At: Arabidopsis thaliana

图3 CmJAZ1亚细胞定位GFP：绿色荧光通道；Merged：混合场，叠加后的照片；Bright：明场

Fig. 3 CmJAZ1 subcellular localizationGFP: Green fluorescence channel. Merged: Mixed field, which means superimposed photos. Bright: Bright field

图4 转基因烟草株系表型A：CmJAZ1-2300转化植株的不同时期；B：CmJAZ1-RNAi转化植株的不同时期；a：外植体制备；b：外植体预培养；c：愈伤诱导及分化；d：生根培养

Fig. 4 Phenotypes of transgenic tobacco strainsA: Different stages of CmJAZ1-2300 transformed plants. B: Different stages of CmJAZ1-RNAi transformed plants. a: Explant preparation. b: Explant pre-culture. c: Callus induction and differentiation. d: Rooting culture

图5 干旱胁迫下烟草表型

Fig. 5 Phenotype of tobacco under drought stress

图6 干旱胁迫对转基因菊花生理指标的影响不同小写字母代表差异显著（P<0.05）

Fig. 6 Effects of drought stress on physiological indicators of transgenic chrysanthemumsDifferent lowercase letters indicate significant difference (P<0.05)

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