Cloning of Strawberry FaJAZ5 Gene and Its Response to Salt Stress

doi:10.13560/j.cnki.biotech.bull.1985.2025-1288

Abstract

Abstract:

Objective To identify the TIFY superfamily member FaJAZ5 gene and its expression characteristics in strawberry (Fragaria × ananassa Duch. ‘Benihoppe’), and to elucidate its role in salt stress response, providing a theoretical basis and genetic resources for breeding salt-tolerant strawberry varieties. Method The FaJAZ5 gene sequence was obtained via homologous cloning, and its sequence characteristics were analyzed using bioinformatics. The expression patterns of this gene in different strawberry tissues and under salt stress were detected by RT-qPCR. FaJAZ5 was heterologously expressed in tobacco via an Agrobacterium-mediated genetic transformation method. Phenotypic observation and determination of related physiological indices were performed on the transgenic tobacco plants under salt stress. Results Sequence analysis revealed that the FaJAZ5 gene had a complete coding region of 543 bp, encoding 180 amino acids and harboring the signature motif of the TIFY family. RT-qPCR analysis showed that the expression of FaJAZ5 was tissue-specific, with the highest level detected in stems. Salt stress significantly induced the expression of FaJAZ5 in roots and leaves. Under normal growth conditions, the plant canopy width and chlorophyll content of FaJAZ5-overexpressing tobacco plants were significantly higher than those of the wild type. However, no significant differences were observed in plant height and leaf morphology between the overexpressing lines and the wild type. After salt stress treatment, the overexpressing tobacco plants showed a rapid wilting phenotype. The levels of malondialdehyde (MDA) and reactive oxygen species (ROS) in these plants were significantly higher than in the wild type. The contents of osmotic adjustment substances and the activities of antioxidant enzymes were also significantly elevated in the overexpressing lines compared with the wild type. Conclusion Although FaJAZ5 overexpression in tobacco enhanced its stress resistance by increasing the contents of osmotic adjustment substances and the activities of antioxidant enzymes under salt stress, the extent of damage caused exceeded its defensive capacity. Overall, this resulted in an increased sensitivity to salt stress, indicating that the FaJAZ5 gene plays a negative regulatory role in the plant response to salt stress.

Key words: strawberry, FaJAZ5, sequence characteristics, expression pattern, heterologous expression, salt stress, physiological response

HUANG Shan, CHEN Yan, LI Tong, LIN Bang-yu, LIANG Yu-meng, GOU Qian, LIN Yuan-xiu, TANG Hao-ru, ZHANG Yun-ting. Cloning of Strawberry FaJAZ5 Gene and Its Response to Salt Stress[J]. Biotechnology Bulletin, doi: 10.13560/j.cnki.biotech.bull.1985.2025-1288.

Figures/Tables 7

Fig. 1 Multiple alignment of FaJAZ5 protein sequences (A), phylogenetic tree (B), and expression analysis of the FaJAZ5 gene in different tissues of strawberry and fruits at different developmental stages (C)At: Arabidopsis thaliana; Mc: Momordica charantia; Cm: Cucumis melo; Bh: Benincasa hispida; Rr: Rosa rugosa; Fa: Fragaria × ananassa; Aa: Argentina anserina; Pp: Prunus persica; Ms: Malus sylvestris. F: Flower; SG: small green fruit; BG: big green fruit; W: white fruit; HR: half-red fruit; FR: full-red fruit. Bars with different lowercase letters indicate significant differences at P<0.05. The same below

Fig. 2 Expression pattern of the FaJAZ5 gene in strawberry under salt stress

Fig. 3 GUS (A) detection and PCR (B) of FaJAZ5-overexpressing tobaccoWT: Wild type; OE-1, OE-22, OE-27: three FaJAZ5 overexpression lines. The same below

Fig. 4 Effects of FaJAZ5 overexpression on plant morphological indicators under normal growth conditions

Fig. 5 Effects of FaJAZ5 overexpression on plant phenotype (A) and membrane lipid peroxidation indices (B‒E) under salt stress

Fig. 6 Effects of FaJAZ5 overexpression on osmotic adjustment substances in plants

Fig. 6 Effects of FaJAZ5 overexpression on plant antioxidant enzyme activities

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