草莓FaJAZ5基因的克隆及其对盐胁迫的响应

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

摘要/Abstract

摘要：

目的鉴定‘红颜’草莓（Fragaria × ananassa Duch. ‘Benihoppe’）TIFY超家族成员FaJAZ5基因及其表达特性，揭示该基因在植物盐胁迫响应过程中的功能，为培育耐盐草莓新品种提供理论依据与基因资源。方法采用同源克隆法获得FaJAZ5基因序列，通过生物信息学分析其序列特征；利用RT-qPCR技术检测该基因在草莓不同组织中和盐胁迫下的表达模式；通过农杆菌介导的遗传转化法在烟草中异源表达FaJAZ5；对盐胁迫下的转基因烟草进行表型观测与相关生理指标测定。结果序列分析显示，FaJAZ5基因编码区全长543 bp，编码180个氨基酸，具有TIFY家族标志性基序；RT-qPCR分析表明，FaJAZ5基因表达具有组织特异性，在茎中表达量最高，盐胁迫能显著诱导FaJAZ5在根和叶中的表达；正常生长条件下，FaJAZ5过表达烟草的冠幅和叶绿素含量显著高于野生型，而株高和叶型与野生型无显著性差异。盐胁迫处理后，FaJAZ5过表达烟草出现迅速萎蔫现象，其丙二醛（MDA）和活性氧（ROS）含量显著高于野生型，同时渗透调节物质含量及抗氧化酶活性也显著高于野生型。结论盐胁迫下，FaJAZ5过表达烟草渗透调节物质含量和抗氧化酶活性提高，但盐胁迫造成的损伤程度已超过其抵御能力阈值，整体表现为对盐敏感性增加，表明FaJAZ5基因对植株的盐胁迫响应具有负调控作用。

关键词: 草莓, FaJAZ5, 序列特征, 表达模式, 异源表达, 盐胁迫, 生理响应

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. Result 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

黄姗, 陈言, 李桐, 林邦语, 梁玉梦, 苟倩, 林源秀, 汤浩茹, 张云婷. 草莓FaJAZ5基因的克隆及其对盐胁迫的响应[J]. 生物技术通报, 2026, 42(10): 160-168.

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, 2026, 42(10): 160-168.

图/表 7

图1 FaJAZ5蛋白序列多重比对（A）、系统发育树（B）和FaJAZ5基因在草莓不同组织和不同发育时期果实中的表达量（C）At：拟南芥；Mc：苦瓜；Cm：甜瓜；Bh：冬瓜；Rr：玫瑰；Fa：草莓；Aa：蕨麻；Pp：桃；Ms：欧洲野苹果；F：花；SG：小绿果；BG：大绿果；W：白果；HR：半红果；FR：全红果。柱状图上标的不同小写字母表示各实验处理间存在统计学显著差异（P<0.05）。下同

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

图2 FaJAZ5基因在草莓盐胁迫下的表达模式

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

图3 FaJAZ5过表达烟草的GUS（A）和PCR（B）检测WT：野生型；OE-1、OE-22、OE-27：3个FaJAZ5过表达株系。下同

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

图4 正常生长条件下过表达FaJAZ5对植株形态指标的影响

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

图5 过表达FaJAZ5对盐胁迫下植株表型（A）及膜脂过氧化指标（B‒E）的影响

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

图6 过表达FaJAZ5对植株渗透调节物质的影响

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

图7 过表达FaJAZ5对植株抗氧化酶活性的影响

Fig. 7 Effects of FaJAZ5 overexpression on plant antioxidant enzyme activities

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