Functional Analysis of VvHSP18.2 Overexpression in Regulating Salt-alkali Resistance in Grapevines

doi:10.13560/j.cnki.biotech.bull.1985.2025-0692

Abstract

Abstract:

Objective Saline-alkali stress severely affects grape growth, development, and quality formation. This study aims to evaluate the function of VvHSP18.2 in regulating grape saline-alkali resistance, identify key genes involved in grape salt-alkali tolerance, and establish a theoretical foundation for the genetic improvement of stress resistance in grapes. Method Phylogenetic analysis of VvHSP18.2 was conducted using bioinformatics tools, and its promoter was analyzed. RT-qPCR was used to assess the expression of VvHSP18.2 under various stress treatments, including NaHCO₃. VvHSP18.2 was overexpressed in Arabidopsis and grape, and its function was evaluated through phenotypic observation and physiological indicators. Result VvHSP18.2 belonged to the heat shock protein CI subfamily and is mainly expressed in grape callus, fruit, and roots. Its promoter region contained multiple cis-acting elements responsive to drought, salt, and high-temperature stresses. VvHSP18.2 rapidly responded to multiple abiotic stresses: its expression was significantly induced by NaCl, especially NaHCO₃, at 6 and 12 h post-treatment; it was significantly induced by PEG6000 at 1 hour post-treatment; and its expression significantly decreased at 3 and 6 h after AlCl₃ treatment. The overexpression of VvHSP18.2 in grape roots and Arabidopsis alleviated phenotypic damage caused by NaHCO₃ treatment, reduced electrolyte leakage and MDA content, and enhanced resistance to NaHCO₃ stress. Furthermore, VvHSP18.2 overexpression increased the activities of SOD, CAT, and POD enzymes in Arabidopsis, reduced the accumulation of O^2- and H₂O₂, indicating that it enhanced stress resistance by improving reactive oxygen species (ROS) scavenging capacity. In contrast, VvHSP18.2 overexpression had no significant effect on NaCl resistance in Arabidopsis. Conclusion VvHSP18.2 is involved in regulating the stress response in grapes. The overexpression of VvHSP18.2 enhances saline-alkali stress resistance in both grape and Arabidopsis by increasing antioxidant enzyme activities and improving ROS scavenging capacity, thereby regulating the balance of ROS metabolism.

Key words: grapevines, VvHSP18.2, alkaline salt stress, reactive oxygen species, Arabidopsis thaliana

XU Yu-jiao, SUN Yu-shuai, LIU Dao-qi, ZHANG Li, ZHANG Zhi-chang, YAO Yu-xin. Functional Analysis of VvHSP18.2 Overexpression in Regulating Salt-alkali Resistance in Grapevines[J]. Biotechnology Bulletin, 2026, 42(4): 161-169.

Figures/Tables 6

Table 1 Primer sequences for RT-qPCR

引物

Primer

正向引物

Forward primer （5′‒3′）

反向引物

Reverse primer （5′‒3′）

GAGATTCCGTTGTCCAGAAGTC

CAATGTTGCCATAGAGGTCCTT

Fig. 1 Phylogenetic evolution and expression analysis of VvHSP18.2A: Phylogenetic tree of HSP proteins in Arabidopsis thaliana (At) and Vitis vinifera (Vv). B: Analysis of the 2 000 bp promoter before VvHSP18.2

Fig. 2 VvHSP18.2 tissue-specific analysis and abiotic stress response analysisA: VvHSP18.2 tissue-specific analysis. B: NaHCO3 treatment. C: NaCl treatment. D: AlCl3 treatment. E: PEG6000treatment. Different lowercase letters indicate significant differences (P<0.05). The same below

Fig. 3 Overexpression of VvHSP18.2 alleviates alkaline salt stress injury in grapevine rootsA: Root system identificationof VvHSP18.2 transgenic grape (NC: Negative control). B: Phenotype of damage to the root system of genetically modified grapes under alkaline salt stress. C: The expressions of VvHSP18.2 in the root system of genetically modified grapes. D: MDA content. E: Root activity

Fig. 4 Identification and overexpression of VvHSP18.2 transgenic Arabidopsis thaliana to enhance Arabidopsis NaHCO3 toleranceA: Identification of VvHSP18.2 transgenic Arabidopsis. B: Expressions of VvHSP18.2 in transgenic Arabidopsis phenotype chart. C: Phenotype chart. D: Root activity content. E: MDA content. F: Relative electrical conductivity. G: SOD activity. H: CAT activity. I: POD activity. J: NBT staining (left) and DAB staining (right). WT: Wild-type Arabidopsisthaliana background. L1, L2, and L3: Three transgenic lines harboring the VvHSP18.2 overexpression construct

Fig. 5 Effect of overexpression of VvHSP18.2 on NaCl resistance in Arabidopsis thalianaA: Phenotypes. B: Fresh weight. C: Root length. D: Number of lateral roots

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