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Heterologous expression of the cysteine proteinase gene VaCP17 from the cold-adapted grapevine Vitis amurensis increases cold tolerance in Arabidopsis

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Abstract

Cysteine proteinases (thiol) carry out diverse and critical functions in plants through their ability to hydrolyze peptide bonds in target proteins. Here, we cloned a cysteine proteinase gene designated VaCP17 from a highly cold-resistant wild Vitis amurensis accession ‘Shuangyou’, and then its potential function in cold resistance was investigated. The results showed that the CDS of VaCP17 is 1404 bp, encoding 467 amino acids, the VaCP17 protein localized to the cell membrane. Expression of CP17 was highly distinctive among different structures of ‘Shuangyou’ and the cold-sensitive Vitis vinifera cultivar ‘Red Globe’, with the highest expression in the stem of ‘Shuangyou’ and the flower of ‘Red Globe’. Arabidopsis plants constitutively expressing a VaCP17-GREEN FLUORESCENT PROTEIN fusion (35S::VaCP17-GFP) showed increased survival after transient exposure to freezing (-6 °C), and showed lower electrolyte leakage and MDA content, higher soluble sugar content and SOD, POD and CAT activities, as compared with non-transgenic Arabidopsis controls. The expression of nine cold-resistance related genes (CBF1, CBF2, CBF3, RD29A, COR15A, KIN1, NCED3, AOC1 and JAZ10) in 35S::VaCP17-GFP plants was increased under cold treatment at 4 °C, relative to control plants. Using a yeast two-hybrid system, we identified VaNAC72, VaCAM7 and VaDi19 as potential interactors of VaCP17, and their interactions were demonstrated by a bimolecular fluorescence complementation assay. In conclusion, we revealed that VaCP17 can enhance cold resistance by influencing physiology and biochemistry and the expression of cold resistance related genes under cold stress.

Key message

VaCP17 gene was cloned from the high cold-resistant wild Vitis amurensis acc. ‘Shuangyou’. The heterologous expression of VaCP17 can enhance the cold tolerance of Arabidopsis thaliana.

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The data sets supporting the results of this article are included within the article and its additional files.

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Acknowledgements

This study was supported by Shaanxi Province Key Project-Agriculture of the People’s Republic of China [Grant No. 2017ZDXM-NY-026], and The National Science-Technology Support Plan Projects of the Ministry of Science and Technology of the People’s Republic of China [Grant No. 2013BAD02B04-06].

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Authors and Affiliations

  1. College of Horticulture, Northwest A & F University, Yangling, Xianyang, 712100, Shaanxi, China

    Xin Shu, Bao Gu, Hongjuan Zhang, Yunyun Tang & Jianxia Zhang

  2. Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China, Ministry of Agriculture, Yangling, Xianyang, 712100, Shaanxi, China

    Xin Shu, Bao Gu, Hongjuan Zhang, Yunyun Tang & Jianxia Zhang

  3. State Key Laboratory of Crop Stress Biology in Arid Areas, Northwest A & F University, Yangling, Xianyang , 712100, Shaanxi, China

    Xin Shu, Bao Gu, Hongjuan Zhang, Yunyun Tang & Jianxia Zhang

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  1. Xin Shu
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  2. Bao Gu
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  3. Hongjuan Zhang
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  4. Yunyun Tang
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Contributions

ZJX designed the experiments and revised the manuscript. SX performed experiments and wrote the manuscript. GB cloned VaCP17. ZHJ participated in RNA extraction from the 35S::VaCP17-GFP lines. TYY participated in the determination of physiological and biochemical indicators.

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Correspondence to Jianxia Zhang.

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The authors declare that the research was conducted in the absence of any commercial or fnancial relationships that could be construed as a potential confict of interest.

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Communicated by Henryk Flachowsky.

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Supplementary file1 (DOCX 15 kb) Oligonucleotide primers used in this study

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Shu, X., Gu, B., Zhang, H. et al. Heterologous expression of the cysteine proteinase gene VaCP17 from the cold-adapted grapevine Vitis amurensis increases cold tolerance in Arabidopsis. Plant Cell Tiss Organ Cult 150, 153–164 (2022). https://doi.org/10.1007/s11240-022-02256-x

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