Objective: Low-temperature fermentation (10-15 ℃) has been recognized as a valuable tool to improve the varietal aroma and fermentation aroma of wine， which has a positive impact on improving wine quality. In order to improve the low-temperature tolerance of Saccharomyces cerevisiae and identification analysis genes related to low-temperature tolerance. Methods: A low-temperature-tolerant native yeast (ZX11) was screened in our laboratory in previous study. In this study， ZX11△ard1 and ARD1-ZX11△ard1 mutant strains were constructed through gene knockout and overexpression techniques. By using techniques such as gene knockout and gene replenishment， mutant strains of ZX11△ard1 and ARD1-ZX11△ard1 were constructed to analyze their growth status at low and normal temperatures. To screen and validate the target protein of Ard1p， GST-Pull down， LC-MS/MS and yeast two hybrid was used. Result: The research results indicate that the ARD1 gene plays a crucial role in the low temperature tolerance of ZX11. The Cbk1p protein encoded by CBK1 is an interaction protein of Ard1p， and the interaction between Ard1p and Cbk1p was further verified in vivo and in vitro. Conclusion: This study laid a foundation of further research on the biological function of ARD1 gene and the molecular regulation mechanism of low temperature tolerance of S. cerevisiae.