Abstract: Two varieties (DNDM 1 and JM22) with different cold resistance were used for experiment and sampled at cold-acclimation period and frozen period. In order to understand the antioxidant enzymes or active oxygen scavengers in low temperature, the contents of antioxidants (GSH, ASA) and the activities of antioxidant enzymes (SOD, POD) were examined, viewed to providing a theoretical basis for the identification of cold resistance on winter wheat cultivars in low temperatures. The results showed that, the relative conductivity of cold hardy variety was less than cold sensitive variety after frozen period. The content of MDA of cold resistance variety was significantly lower than that of cold sensitive variety only at depth frozen period (on January 14th). The SOD activity of cold sensitive variety decreased after freezing period for 15 days. The content of ASA of cold resistance variety was higher than that of cold sensitive variety significantly after freezing period for 30 days, the SOD activity and the ASA could be indicators to evaluate the cold hardiness of winter wheat varieties.