Abstract: Taking two winter wheat cultivars Ji’nan 17 and Shannong 8355 as test materials, this paper measured the superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) activities and the malondialdehyde (MDA) and soluble protein contents in the functional leaves and sheaths of the tillers at different positions at stem elongation stage under low temperature stress, and then, the freezing resistance of the tillers was comprehensively evaluated by the methods of principal component analysis and cluster analysis. The results showed that under low temperature stress, the SOD, POD, and CAT activities in the functional leaves and sheaths of each tiller at stem elongation stage increased, but the MDA and soluble protein contents increased or decreased to some extent. By using principal component analysis and cluster analysis, the tillers of each cultivar were grouped into three kinds of freezing resistance type. For Ji’nan 17, the main stem, tiller Ⅰ, and tiller Ⅱ belonged to high freezing resistance type, the tiller Ⅲ, tiller Ⅳ, and tiller Ⅰp belonged to medium freezing resistance type, and the tiller Ⅱ p belonged to low freezing resistance type. For Shannong 8355, the main stem, tiller Ⅰ, tiller Ⅱ, and tiller Ⅲ belonged to high freezing resistance type, the tiller Ⅳ and tiller Ⅰp belonged to medium freezing resistance type, and the tiller Ⅱp belonged to low freezing resistance type. It was concluded that the freezing resistance of the winter wheat tillers at different positions at stem elongation stage differed, with the lower position tillers being more resistant than the higher position tillers.