Using some of the chromosome substitution lines developed from the crosses of the donor Cheyenne to Chinese Spring we showed that the accumulation of water soluble carbohydrates during different stages of hardening was time dependent. Moreover there was a significant correlation between the rate of carbohydrate accumulation and the frost tolerance. The expression and regulation of a wheat gene homologous to the barley cold regulated cor14b gene was compared in frost sensitive and frost tolerant wheat genotypes at different temperatures. Studies made with chromosome substitution lines showed that the threshold induction temperature polymorphism of the cor14b wheat homologous gene was controlled by loci located on chromosome 5A of wheat, while cor14b gene was mapped, in Triticum monococcum, onto the long arm of chromosome 2Am. Our study on the effect of cold hardening on glutathione (GSH) metabolism showed that chromosome 5A of wheat has an influence on the GSH accumulation and on the ratio of reduced and oxidised glutathione as part of a complex regulatory function during cold hardening. In addition, the level of increase in GSH content during hardening may indicate the degree of the frost tolerance of wheat.