Functional Analysis of the Trehalose-6-phosphate Synthase Gene VvTPS10 in Salt Tolerance and Growth and Development of Grapevine

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

Abstract

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

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.

Figures/Tables 8

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

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

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

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

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

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

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

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

References 35

[1]	Han Y, Li XJ. Current progress in research focused on salt tolerance in Vitis vinifera L. [J]. Front Plant Sci, 2024, 15: 1353436.
[2]	Elbein AD, Pan YT, Pastuszak I, et al. New insights on trehalose: a multifunctional molecule [J]. Glycobiology, 2003, 13(4): 17R-27R.
[3]	Nawaz M, Hassan MU, Chattha MU, et al. Trehalose: a promising osmo-protectant against salinity stress-physiological and molecular mechanisms and future prospective [J]. Mol Biol Rep, 2022, 49(12): 11255-11271.
[4]	Cabib E, Leloir LF. The biosynthesis of trehalose phosphate [J]. J Biol Chem, 1958, 231(1): 259-275.
[5]	Mu M, Lu XK, Wang JJ, et al. Genome-wide identification and analysis of the stress-resistance function of the TPS (Trehalose-6-Phosphate Synthase) gene family in cotton [J]. BMC Genet, 2016, 17: 54.
[6]	Hu X, Wu ZD, Luo ZY, et al. Genome-wide analysis of the trehalose-6-phosphate synthase (TPS) gene family and expression profiling of ScTPS genes in sugarcane [J]. Agronomy, 2020, 10(7): 969.
[7]	Song JB, Mao HY, Cheng J, et al. Identification of the trehalose-6-phosphate synthase gene family in Medicago truncatula and expression analysis under abiotic stresses [J]. Gene, 2021, 787: 145641.
[8]	Guo WL, Sun YQ, Chai JQ, et al. Genome-wide identification of the trehalose-6-phosphate synthase gene family in alfalfa (Medicago sativa L.) reveals involvement of MsTPS16 in tolerance to saline-alkali stress [J]. Gene, 2025, 963: 149609.
[9]	刘丹丹, 王雷刚, 孙明慧, 等. 茶树海藻糖-6-磷酸合成酶(TPS)基因家族鉴定与表达分析 [J]. 生物技术通报, 2024, 40(8): 152-163.
	Liu DD, Wang LG, Sun MH, et al. Genome-wide identification and expression pattern profiling of the trehalose-6-phosphate synthase (TPS) gene family in tea plant (Camellia sinensis) [J]. Biotechnol Bull, 2024, 40(8): 152-163.
[10]	陈天池, 吴月燕, 沈乐意, 等. 葡萄TPS基因家族的鉴定与表达分析 [J]. 分子植物育种, 2024, 22(11): 3533-3544.
	Chen TC, Wu YY, Shen LY, et al. Identification and expression analysis of TPS genes family in grape [J]. Mol Plant Breed, 2024, 22(11): 3533-3544.
[11]	Almeida AM, Villalobos E, Araújo SS, et al. Transformation of tobacco with an Arabidopsis thaliana gene involved in trehalose biosynthesis increases tolerance to several abiotic stresses [J]. Euphytica, 2005, 146(1/2): 165-176.
[12]	Jiang T, Zhai H, Wang FB, et al. Cloning and characterization of a salt tolerance-associated gene encoding trehalose-6-phosphate synthase in sweetpotato [J]. J Integr Agric, 2014, 13(8): 1651-1661.
[13]	Vishal B, Krishnamurthy P, Ramamoorthy R, et al. OsTPS8 controls yield-related traits and confers salt stress tolerance in rice by enhancing suberin deposition [J]. New Phytol, 2019, 221(3): 1369-1386.
[14]	Yuan GP, Liu JP, An GL, et al. Genome-wide identification and characterization of the trehalose-6-phosphate synthetase (TPS) gene family in watermelon (Citrullus lanatus) and their transcriptional responses to salt stress [J]. Int J Mol Sci, 2022, 23(1): 276.
[15]	Wang PL, Lei XJ, Lü JX, et al. Overexpression of the ThTPS gene enhanced salt and osmotic stress tolerance in Tamarix hispida [J]. J For Res, 2022, 33(1): 299-308.
[16]	郭蓓, 胡磊, 何欣, 等. 海藻糖-6-磷酸合成酶转基因烟草提高耐盐性的研究 [J]. 植物学通报, 2008(1): 41-49.
	Guo B, Hu L, He X, et al. Trehalose-6-phosphate synthase transgenic tobacco enhanced tolerance to salt stress [J]. Chin Bull Bot, 2008(1): 41-49.
[17]	Wang L, Zhang X, Qu ZY, et al. Grapevine VviERF105 promotes tolerance to abiotic stress and is degraded by the E3 ubiquitin ligase VviPUB19 [J]. Environ Exp Bot, 2022, 201: 105001.
[18]	Zhao MY, Wang L, Feng MJ, et al. Trehalose-6-phosphate synthase VvTPS10 regulates cold tolerance by participating in the CBF pathway and VvPUB19-mediated ubiquitination in grapevine [J]. Plant Cell Environ, 2025, 48(11): 8086-8102.
[19]	高俊凤. 植物生理学实验指导 [M]. 北京: 高等教育出版社, 2006.
	Gao JF. Experimental guidance for plant physiology [M]. Beijing: Higher Education Press, 2006.
[20]	靳容, 刘明, 赵鹏, 等. 甘薯丝裂原活化蛋白激酶MPK6对低温胁迫的响应 [J]. 中国农业科学, 2021, 54(20): 4265-4273.
	Jin R, Liu M, Zhao P, et al. IbMKP6, a mitogen-activated protein kinase, confers low temperature tolerance in sweetpotato [J]. Sci Agric Sin, 2021, 54(20): 4265-4273.
[21]	杜姣林, 蔺新兰, 马豫皖, 等. 植物海藻糖-6-磷酸合成酶基因研究进展 [J]. 植物科学学报, 2023, 41(3): 411-420.
	Du JL, Lin XL, Ma YW, et al. Research progress in plant trehalose-6-phosphate synthase genes [J]. Plant Sci J, 2023, 41(3): 411-420.
[22]	张莹莹, 吕燕, 宣雯燕, 等. 龙须菜(Gracilariopsis lemaneiformis)海藻糖-6-磷酸合成酶(TPS)对逆境胁迫的响应 [J]. 海洋与湖沼, 2021, 52(3): 777-785.
	Zhang YY, Lyu Y, Xuan WY, et al. Response of trehalose-6-phosphate synthase (TPS) to abiotic stress in Gracilariopsis lemaneiformis [J]. Oceanol Limnol Sin, 2021, 52(3): 777-785.
[23]	Zhou WY, Sheng C, Koffi Dossou SS, et al. Genome-wide identification of TPS genes in sesame and analysis of their expression in response to abiotic stresses [J]. Oil Crop Sci, 2023, 8(2): 81-88.
[24]	曹婕, 卢秋连, 翟健平, 等. 大豆TPS基因家族在盐胁迫下的表达变化及单倍型选择规律分析 [J]. 植物学报, 2025, 60(2): 172-185.
	Cao J, Lu QL, Zhai JP, et al. Changes in the expression of the soybean TPS gene family under salt stress and haplotype selection analysis [J]. Bull Bot, 2025, 60(2): 172-185.
[25]	Li HW, Zang BS, Deng XW, et al. Overexpression of the trehalose-6-phosphate synthase gene OsTPS1 enhances abiotic stress tolerance in rice [J]. Planta, 2011, 234(5): 1007-1018.
[26]	张桂华, 岳春雷, 李贺鹏, 等. 木麻黄海藻糖-6-磷酸合酶家族鉴定及表达分析 [J]. 农业生物技术学报, 2025, 33(3): 581-593.
	Zhang GH, Yue CL, Li HP, et al. Identification and expression analysis of trehalose-6-phosphate synthase family in Casuarina equisetifolia [J]. J Agric Biotechnol, 2025, 33(3): 581-593.
[27]	杨永超, 张海斐, 杨小振, 等. 甜瓜海藻糖-6-磷酸合成酶基因鉴定及表达 [J]. 西北植物学报, 2017, 37(6): 1066-1072.
	Yang YC, Zhang HF, Yang XZ, et al. Expression analysis and genome-wide identification of trehalose-6-phosphate synthase gene family in melon (Cucumis melo L.) [J]. Acta Bot Boreali Occidentalia Sin, 2017, 37(6): 1066-1072.
[28]	Cortina C, Culiáñez-Macià FA. Tomato abiotic stress enhanced tolerance by trehalose biosynthesis [J]. Plant Sci, 2005, 169(1): 75-82.
[29]	Zhang XT, Zhang YX, Yan ML, et al. Trehalose metabolism contributed to abiotic stress tolerance of Larix gmelinii seedlings [J]. Plant Physiol Biochem, 2025, 229: 110654.
[30]	Wahl V, Ponnu J, Schlereth A, et al. Regulation of flowering by trehalose-6-phosphate signaling in Arabidopsis thaliana [J]. Science, 2013, 339(6120): 704-707.
[31]	Lin MF, Ma SY, Quan KH, et al. Comparative transcriptome analysis provides insight into the molecular mechanisms of long-day photoperiod in Moringa oleifera [J]. Physiol Mol Biol Plants, 2022, 28(5): 935-946.
[32]	赵美丽, 倪军, 陈茂盛, 等. 超量表达小桐子JcTPS1促进拟南芥开花并增加叶片花青素含量 [J]. 分子植物育种, 2018, 16(1): 255-261.
	Zhao ML, Ni J, Chen MS, et al. Over-expression of JcTPS1 from Jatropha curcas induces early flowering of Arabidopsis and improves anthocyanin accumulation in leaves [J]. Mol Plant Breed, 2018, 16(1): 255-261.
[33]	吴新亮, 吴慧玲, 张绥林, 等. 蓝莓VcTPS/VcTPP基因家族鉴定及VcTPS24对花诱导的功能分析 [J]. 中国农业科学, 2025, 58(13): 2663-2681.
	Wu XL, Wu HL, Zhang SL, et al. Identification of the VcTPS/VcTPP gene family in blueberries and functional analysis of VcTPS24 in floral induction [J]. Sci Agric Sin, 2025, 58(13): 2663-2681.
[34]	Van Leene J, Eeckhout D, Gadeyne A, et al. Mapping of the plant SnRK1 kinase signalling network reveals a key regulatory role for the class II T6P synthase-like proteins [J]. Nat Plants, 2022, 8(11): 1245-1261.
[35]	Wingler A, Delatte TL, O’Hara LE, et al. Trehalose 6-phosphate is required for the onset of leaf senescence associated with high carbon availability [J]. Plant Physiol, 2012, 158(3): 1241-1251.