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In order to search out chromosomes involved in cold resistance, reciprocal sets of chromosome substitution lines in duplicate between two winter wheat cultivars, Cheyenne and Wichita, were used. Two experiments were carried out in a complete block design with two replications for each duplicate. Crown and leaf water content, crown and leaf wet weigth were measured in the filed. Crown survival, electrolyte leakage and L TSO were measured in laboratory. The results showed that Cheyenne is more resistant than Wichita. Crown survival had significant correlations with crown water content, crown wet weight and electrolyte leakage. Chromosomes 6A, 3B, SD substituted from Wichita into Cheyenne decreased the crown survival, membrane stability and increased crown water content as well as crown wet weight of Cheyenne. Therefore, these crhomosomes decreased cold hardiness in Cheyenne. Reciprocally, chromosomes SA, SD, 3B, 4A and 4D from Cheyenne into Wichita increased crown survival, decreased crown water conetnt and crown weight in Wichita. It is concluded that these chromosomes causing cold hardiness in Wichita and carring cold resistance genes.