Functional Analysis of Chrysanthemum CmJAZ1 Gene in Negative Regulation of Drought Stress

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

Abstract

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

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.

Figures/Tables 7

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

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

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

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

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

Fig. 5 Phenotype of tobacco under drought stress

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

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