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

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