海藻糖-6-磷酸合成酶基因VvTPS10在葡萄耐盐和生长发育中的功能分析

doi:10.13560/j.cnki.biotech.bull.1985.2025-1248

摘要/Abstract

摘要：

目的探究VvTPS10基因在葡萄中对盐胁迫的响应，为葡萄耐盐机制研究提供理论依据。方法葡萄盆栽苗经盐胁迫处理后，利用实时荧光定量PCR（RT-qPCR）对VvTPS10的表达模式进行检测。盐胁迫处理后观察VvTPS10过表达拟南芥和葡萄愈伤组织表型，并进行生理生化指标测定和抗性基因表达分析。结果葡萄VvTPS10基因表达量在盐胁迫处理下显著增加。在150和200 mmol/L NaCl处理下，VvTPS10过表达拟南芥发芽率比野生型分别高18.04%和50.57%。盐胁迫下，VvTPS10过表达拟南芥的电导率、MDA和超氧阴离子含量均显著低于野生型，而叶绿素含量、SOD、POD和CAT活性均显著高于野生型。RT-qPCR检测发现，VvTPS10过表达植株中，COR15A、COR47、RD29B、SOS1、NHX1和DREB2A抗性基因在盐胁迫下被诱导表达，表达量显著高于野生型。同时，过表达VvTPS10不仅导致拟南芥提早抽薹开花，同时植株响应外源蔗糖处理，表现为花色苷积累显著增加。盐胁迫下，VvTPS10过表达葡萄愈伤组织的生长状态更好，鲜重更高，相对电导率更低。结论 VvTPS10基因被盐胁迫诱导表达，并通过维持细胞膜稳定性、增强抗氧化酶活性和抗性基因表达等提高拟南芥耐盐性。VvTPS10基因促使拟南芥提前开花，并在蔗糖处理下增加花色苷的积累。在葡萄愈伤组织中，过表达VvTPS10同样能够增强其耐盐性。

关键词: 葡萄, VvTPS10, 海藻糖, 基因表达, 转基因, 耐盐性, 生长发育, 生理生化指标

Abstract:

Objective This study aimed to investigate the response of the VvTPS10 gene to salt stress in grapevine (Vitis vinifera), providing theoretical insights for the study of salt tolerance mechanism in grapevine. Method After subjecting potted grape seedlings to salt stress treatment, the expression pattern of VvTPS10 was detected by RT-qPCR. Phenotypic observations of VvTPS10-overexpressing Arabidopsis thaliana and grape calli were performed after salt stress treatment, followed by the determination of physiological and biochemical indices and the analysis of stress-resistant gene expression. Result The expression of the VvTPS10 gene in grapevines was significantly up-regulated under salt stress. Under the treatments of 150 mmol/L NaCl and 200 mmol/L NaCl, the germination rates of VvTPS10-overexpressing A. thaliana were 18.04% and 50.57% higher than those of the wild-type, respectively. Under salt stress, the relative electrical conductivity, MDA content and superoxide anion content of VvTPS10-overexpressing A. thaliana were significantly lower than those of the wild-type, whereas the chlorophyll content, and the activities of SOD, POD and CAT were significantly higher than those of the wild-type. RT-qPCR analysis showed that under salt stress, the expression levels of stress-resistant genes including COR15A、COR47、RD29B、SOS1、NHX1 and DREB2A in VvTPS10-overexpressing plants were significantly higher than those in the wild-type. Meanwhile, we found that overexpression of VvTPS10 not only resulted in an early bolting and flowering in A. thaliana but also responded to exogenous sucrose treatment, as evidenced by a significant increase in anthocyanin accumulation. Finally, under salt stress, VvTPS10-overexpressing grape calli exhibited better growth status, higher fresh weight and lower relative electrical conductivity. Conclusion The VvTPS10 gene is induced by salt stress, and it enhances the salt tolerance of A. thaliana by maintaining cell membrane stability, improving antioxidant enzyme activities and promoting the expression of stress-resistant genes. In addition, this gene promotes early flowering of A. thaliana and increases anthocyanin accumulation under sucrose treatment. Similarly, overexpression of VvTPS10 enhanced salt tolerance in grape callus.

Key words: grapevine, VvTPS10, trehalose, gene expression, genetic transformation, salt tolerance, growth and development, physiological and biochemical indexes

王建波, 李诗雅, 邢鹤炫, 周瑞金, 李桂荣, 王玲. 海藻糖-6-磷酸合成酶基因VvTPS10在葡萄耐盐和生长发育中的功能分析[J]. 生物技术通报, 2026, 42(8): 188-196.

WANG Jian-bo, LI Shi-ya, XING He-xuan, ZHOU Rui-jin, LI Gui-rong, WANG Ling. Functional Analysis of the Trehalose-6-phosphate Synthase Gene VvTPS10 in Salt Tolerance and Growth and Development of Grapevine[J]. Biotechnology Bulletin, 2026, 42(8): 188-196.

图/表 8

图1 葡萄VvTPS10在NaCl处理后的表达量分析数值来自3个生物学重复的平均值±SD，*和**表示差异显著（* P<0.05，** P<0.01）。下同

Fig. 1 Analysis of VvTPS10 gene expression after NaCl treatment in grapevineThe values represent the mean ±SD of three biological replicates. The asterisk (*) and double asterisk (**) indicate significant differences (*P<0.05, **P<0.01). The same below

图2 野生型（WT）和VvTPS10过表达拟南芥（OE）在盐胁迫下的表型（A）及发芽率（B）

Fig. 2 Phenotypic characterization (A) and germination rate analysis (B) of wild-type (WT) and VvTPS10-overexpressing Arabidopsis thaliana (OE) under salt stress

图3 野生型和VvTPS10过表达拟南芥株系在盐胁迫下的表型（A）及抗逆生理指标（B）

Fig. 3 Phenotypic changes (A) and stress physiological indexes (B) of wild-type and VvTPS10-overexpressing A. thaliana lines under salt stress

图4 野生型和VvTPS10过表达拟南芥在盐胁迫下的NBT染色（A）及O2·-含量和SOD、POD、CAT活性（B）分析

Fig. 4 Analysis of NBT staining (A) and O2·- content, SOD, POD, and CAT activities (B) of wild-type and VvTPS10-overexpressing A. thaliana lines under salt stress

图5 野生型和VvTPS10过表达拟南芥株系在盐胁迫下的抗性基因表达分析

Fig. 5 Expression analysis of stress-responsive genes in wild-type and VvTPS10-overexpressing A. thaliana lines under salt stress

图6 野生型和VvTPS10过表达拟南芥的表型（A）及抽薹率（B）

Fig. 6 Phenotypes (A) and bolting rate (B) of wild-type and VvTPS10-overexpressing A. thaliana

图7 野生型和VvTPS10过表达拟南芥对蔗糖响应的表型（A）及花色苷含量（B）

Fig. 7 Phenotypic (A) and anthocyanin content (B) of wild-type and VvTPS10-overexpressing A. thaliana under sucrose treatment

图8 野生型和VvTPS10过表达葡萄愈伤组织在盐胁迫下的表型（A）、鲜重（B）和相对电导率（C）

Fig. 8 Phenotypic (A), fresh weight (B), and relative electrical conductivity (C) analyses of wild-type and VvTPS10-overexpressing grape calli under salt stress

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