With the extreme changes of the global climate, winter freezing injury has become an important limiting factor for the development of the global grape industry. Therefore, there is a significant need for the screening of cold-resistant wine grape germplasms and cold regionalization for cold-resistant breeding and the development of grapevine cultivation in cold regions. In this study, the low-temperature half-lethal temperature (LT50) values were determined for the annual dormant branches of 124 wine grape germplasms (V. vinifera) to evaluate their cold resistance. The LT50 values of the 124 tested germplasms ranged from −22.01 °C to −13.18 °C, with six cold-resistant germplasms below −20 °C. Based on the LT50 values, the 124 germplasms were clustered into four types, with cold resistance from strong to weak in the order of type Ⅱ > type Ⅰ > type Ⅳ > type Ⅲ, corresponding to the four cold hardiness zones. Zones 1, 2, 3, and 4 included 6, 22, 68, and 28 germplasms, respectively, with decreasing cold resistance. The number of germplasms in different hardiness zones followed a normal distribution, with the most in zone 3. In Type Ⅱ, the fruit skin color of germplasms was positively correlated with cold hardiness, while the temperature of origin was negatively correlated with cold hardiness. The average LT50 of germplasms in different origin regions ranged from −17.44 °C to −16.26 °C, with differences among some regions. The cold regionalization analysis resulted in the distribution of 124 germplasms in four temperature regions in China with six germplasms in region A (−22 °C ≤ LT50 ≤ −20 °C), 30 germplasms in region B (−20°C ≤ LT50 ≤ −18°C), 71 germplasms in region C (−18 °C ≤ LT50 ≤ −15 °C), and 17 germplasms in region D (−15 °C ≤ LT50 ≤ −13 °C). Strong cold-resistant wine grape germplasms (V. vinifera) were identified, and these could be used as parental material for cold-resistant breeding. In some areas in China, soil-burial over-wintering strategies are used, but our results suggest that some wine grapes could be cultivated without requiring winter burial during overwintering. The results of this study should provide guidance for the selection of promising strains for cold-resistant breeding for expanded cultivation of improved varieties for wine grape production in China.