Abstract: Low winter temperature is one of the main factors that affect the development of grapes and the wine industry in China. Understanding the signal transduction pathway during cold stress will help in the breeding of high cold-resistance cultivars. Based on our previous transcriptome analysis, a gene that showed increased expression pattern in Vitis vinifera L. ‘Muscat Hamburg’ during cold treatment was identified and named VvCOR27 according to homological analysis. The whole length of VvCOR27 cDNA was 1082 bp, which contained a 909 bp open reading frame (ORF) and encoded 302 amino acids. Homological analysis of COR27s from thirteen species showed that they contained three COR27-specific conservative domains. Real time RT-PCR indicated that the transcript abundance of VvCOR27 was highly increased at 24 h after cold treatment. Four motifs, including EE, EEL, G-box and ABREL, were found in the promoter regions (from published Vitis vinifera genome sequences) of VvCOR27 but at less quantity than that in the promoter regions of AtCOR27. This may be why AtCOR27 showed more timely responses to cold treatment than did VvCOR27. Phenotypic analysis of three overexpression lines under cold treatment indicated that VvCOR27 was involved in responses to cold stress and enhanced cold tolerance in plants.