In order to understand the physiological response of wheat to post-anthesis high temperature, 26 wheat cultivars including Luohan2, Zhengmai366, etc were screened. And 11 physiological indexes including the content of leaf relative water content, soluble protein, malondialdehyde(MDA), etc were analyzed under post-anthesis high temperature stress(plastic-covered tunnel) and natural growth conditions(CK). The results showed that under high temperature stress, the change trend of the photosynthetic rate, transpiration rate, stomatal conductance, chlorophyll content, and leaf relative water content decreased as a whole, while the change trend of soluble protein content and malondialdehyde(MDA) content increased as a whole, but the change trend of superoxide dismutase(SOD), peroxidase(POD), and catalase(CAT) were not obvious in 26 wheat cultivars. And we took each high temperature resistance coefficient as the index of high temperature resistance, and evaluated different cultivars wheat comprehensively through principal components analysis. The results showed that 11 indices were transformed to 5 independent comprehensive indices, which total cumulative rate reached to 80.561% through analysis of principal component. Based on the relevant indexes of yield, 26 wheat cultivars were divided into 4 groups. The first category included 11 cultivars, such as Zhoumai22, Xinmai11 belonged to the medium-level high temperature resistant wheat cultivars. The second category included 10 cultivars, such as Zhengmai366, Lankaoaizao8 belonged to high-level high temperature resistant wheat cultivars. The last category included 4 cultivars, such as Xinong979, Luohan6, belonged to low-level high temperature resistant wheat cultivars. Among them, Luohan2 was high temperature sensitive cultivar and it’s heat sensitivity index of 1000-grain weight and yield were higher than 2, the index and coefficient of resistance of 1000-grain weight were less than 80%. Comprehensive judgment based on D-value clustering classification results, Zhengmai366, Lankaoaizao8 and Jimai47 belonged to the high temperature resistant wheat varieties; Xinmai11, Zhoumai19, Zhengmai7698 and Fengdecunmai1 belonged to the medium-high temperature resistant wheat varieties; Xinong979, Luohan2 and Beijing8 belonged to low and high temperature resistant wheat cultivars.