森林草莓FveTCP5基因在盐胁迫中的功能分析

doi:10.13560/j.cnki.biotech.bull.1985.2026-0311

摘要/Abstract

摘要：

目的 TCP转录因子在植物抵御逆境胁迫过程中发挥重要作用，解析森林草莓FveTCP5基因在响应盐胁迫方面的功能，为培育草莓耐盐新品种提供基因资源。方法以森林草莓‘Ruegen’为材料，克隆FveTCP5基因，结合生物信息学手段分析其理化性质、结构特征和系统进化关系；利用RT-qPCR技术测定FveTCP5基因在森林草莓不同组织中的表达模式；构建FveTCP5基因的过表达载体，利用花序浸染法获得转基因拟南芥过表达株系；用150 mmol/L NaCl处理转基因拟南芥株系，测定相关生理指标和盐胁迫响应相关基因的表达水平，开展耐盐功能验证。结果 FveTCP5基因CDS全长1 131 bp，编码376个氨基酸；系统进化分析表明，FveTCP5与蔷薇科其他物种的TCP5蛋白间亲缘关系较近；RT-qPCR结果显示，FveTCP5基因在森林草莓各组织中均普遍表达，在叶片中的表达量最高。在NaCl处理下，过表达FveTCP5拟南芥株系的子叶绿化率和根长均显著高于野生型；测定相关生理指标显示，转基因植株的丙二醛含量较野生型显著降低，而叶绿素、脯氨酸含量和超氧化物歧化酶（SOD）、过氧化物酶（POD）和过氧化氢酶（CAT）活性较野生型均显著升高。此外，盐胁迫响应相关基因AtSOS1、AtSOS2、AtNHX1、AtHKT1、AtCOR15A和AtRD29A的表达水平较野生型均有所提高。结论过表达FveTCP5可通过促进盐胁迫响应相关基因的表达来提高拟南芥的耐盐性，推测FveTCP5在草莓盐胁迫响应中发挥重要功能。

关键词: 草莓, TCP5, 盐胁迫, 生物信息学分析, 转基因拟南芥, 基因表达, 生理生化指标, 功能分析

Abstract:

Objective TCP transcription factors play important roles in plant stress responses. This study aims to characterize the function of the FveTCP5 gene in Fragaria vesca under salt stress, thereby providing genetic resources for breeding new salt-tolerant strawberry varieties. Method Using woodland strawberry ‘Ruegen’ as the experimental material, we cloned FveTCP5 and analyzed its physicochemical properties, structural characteristics, and phylogenetic relationships via bioinformatics approaches. The expression pattern of FveTCP5 in different tissues of woodland strawberry was determined by RT-qPCR. An overexpression vector of FveTCP5 was constructed, and transgenic Arabidopsis lines were obtained through floral dip transformation. These transgenic lines were treated with 150 mmol/L NaCl, and relevant physiological indices as well as the expression levels of salt stress-responsive genes were measured to validate salt tolerance function. Result The full-length CDS of FveTCP5 is 1 131 bp, encoding 376 amino acids. Phylogenetic analysis showed that FveTCP5 is closely related to TCP5 proteins from other Rosaceae species. RT-qPCR revealed that FveTCP5 was ubiquitously expressed in various tissues of woodland strawberry, with the highest transcript level in leaves. Under NaCl treatment, Arabidopsis lines overexpressing FveTCP5 exhibited significantly higher green cotyledon rates and root lengths than wild-type plants. Physiological measurements indicated that, compared with wild-type, transgenic plants had significantly reduced malondialdehyde content and significantly increased levels of chlorophyll, proline, superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT). Moreover, the expression of salt stress-responsive genes, including AtSOS1, AtSOS2, AtNHX1, AtHKT1, AtCOR15A, and AtRD29A, were upregulated in the transgenic lines. Conclusion Overexpression of FveTCP5 enhances salt tolerance in A. thaliana by promoting the expression of salt stress-responsive genes, suggesting that FveTCP5 plays an important role in the salt stress response of strawberry.

Key words: strawberry, TCP5, salt stress, bioinformatics analysis, transgenic Arabidopsis, gene expression, physiological and biochemical indices, functional analysis

董向向, 缪百灵, 陈娟娟, 张坤鹏, 李亮杰, 周厚成, 朱庆松. 森林草莓FveTCP5基因在盐胁迫中的功能分析[J]. 生物技术通报, doi: 10.13560/j.cnki.biotech.bull.1985.2026-0311.

DONG Xiang-xiang, MIAO Bai-ling, CHEN Juan-juan, ZHANG Kun-peng, LI Liang-jie, ZHOU Hou-cheng, ZHU Qing-song. Functional Analysis of FveTCP5 Gene in Fragaria vesca under Salt Stress[J]. Biotechnology Bulletin, doi: 10.13560/j.cnki.biotech.bull.1985.2026-0311.

图/表 6

图1 不同物种TCP5蛋白的系统进化分析

Fig. 1 Phylogenetic analysis of TCP5 proteins in different species

图2 FveTCP5的表达模式分析不同小写字母代表差异显著（P<0.05）。下同

Fig. 2 Expression pattern analysis of FveTCP5Different lowercase letters indicate significant differences at P<0.05. The same below

图3 过表达FveTCP5拟南芥的PCR鉴定（A）和基因表达量分析（B）1：野生型拟南芥；2-4：3个转基因株系；WT：野生型拟南芥；OE1-3：3个转基因株系；下同

Fig. 3 PCR identification (A) and gene expression analysis (B) of FveTCP5-overexpressing A. thaliana1: Wild type A. thaliana; 2-4: three transgenic lines. WT: Wild type A. thaliana; OE1-3: three transgenic lines. The same below

图4 转基因拟南芥的耐盐表型分析

Fig. 4 Analysis of salt tolerance phenotype of transgenic A. thaliana

图5 转基因拟南芥在盐胁迫下的生理指标测定

Fig. 5 Determination of physiological indices of transgenic A. thaliana under salt stress

图6 转基因拟南芥中盐胁迫响应基因的表达分析

Fig. 6 Expression analysis of salt stress-response genes in transgenic A. thaliana

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