Abstract: In order to identify the resistance and distribution of leaf rust resistance genes of wheat cultivars in Henan Province to provide theoretical basis for the promotion and reasonable layout, leaf rust control and resistance breeding, the resistance evaluation of sixteen main wheat cultivars of Henan was carried out using five mixed races of Puccinia triticina at the seedling stage. In addition, the resistance genes of the 16 wheat cultivars at seedling stage were postulated by inoculating 12 races of P. triticina. Meanwhile, thirty molecular markers closely linked or co-segregated with 24 wheat leaf rust resistance genes were used to detect these cultivars. The results showed that these cultivars were highly susceptible to the mixed races examined. The data of gene postulation and molecular detection indicated that four leaf rust resistance genes, Lr1, Lr16, Lr26 and Lr30, were found in these cultivars. Among them, Xianmai 8 carried Lr1 and Lr26, Zhengmai 366 and Zhengmai 9023 carried Lr1. Xinong 979 and Huaichuan 916 carried Lr16. Zhongmai 895, Yanzhan 4110, Zhengmai 7698, Ping'an 8, Zhongmai 1, Zhoumai 16, Hengguan 35 and Aikang 58 carried Lr26. The gene Lr26 was detected in Zhoumai 22, and Lr1 and Lr30 were also postulated in Zhoumai 22. Besides, Yumai 49-198 and Luomai 23 might carry unknown or untested resistance genes. Therefore, the abundance of leaf rust resistance genes in the main wheat cultivars of Henan was low, and the new leaf rust resistance genes should be introduced into new wheat cultivars to improve the leaf rust resistance and control wheat leaf rust effectively.