The objective of this study was to determine the leaf rust resistance and leaf rust resistance genes of twelve main wheat cultivars(lines) in Shandong Province to provide theoretical basis for the promotion and reasonable layout, leaf rust control and resistance breeding. The resistance evaluation of twelve main wheat cultivars(lines) was carried out by using five mixed races of Puccinia triticina at the seedling stage. Meanwhile, fifteen Puccinia triticina races with different virulence patterns were used for seedling genes postulation. In addition, thirty molecular markers closely linked or co-segregated with 24 wheat leaf rust resistance genes were used to detect these wheat cultivars(lines). The results showed that these cultivars(lines) were highly susceptible to the mixed races. Gene postulation and molecular detection indicated that Ji'nan 17 carried Lr16. Aikang 58 and Shannong 20 carried Lr26. However, leaf rust resistance genes were not detected in the remaining nine cultivars(lines). In addition, this study also carried out the detection of three non main cultivars in Shandong. And the results showed that Zhongmai 175 carried Lr1 and the adult-plant resistance gene Lr37. Wanmai 38 carried Lr26. And leaf rust resistance genes were not detected in Jimai 20. Comprehensive above results, the abundance of leaf rust resistance genes in these main wheat cultivars(lines) was low, and no effective resistance genes were detected in them to against the main races of P. triticina in China, which need to be paid more attention. New leaf rust resistance genes should be introduced into new wheat cultivars to improve the leaf rust resistance.