Wheat Leaf rust seriously distablizes the wheat production in China. Breeding wheat cultivars with durable resistance is considered as one of the most economical and effective methods in disease control. In this study，the Leaf rust resistance in 63 advanced wheat lines were analyzed by combining marker-assisted identification of known resistance genes，pedigree analysis and the field test upon Leaf rust infection. These genotypes together with 36 donor lines（carrying known Leaf rust resistance genes）were inoculated with 17 Puccinia triticina（Pt）races with specific virulence at the seedling stage，and also genotyed using 10 molecular markers associating with known resistance genes. Moreover，these 63 wheat advanced lines were evaluated for Leaf rust severity in Zhoukou city of Henan province and Baoding of Hebei province in the crop season（2018- 2019），followed by analyzing variance and calculating the LSD value to identify wheat lines that showed adult plant resistance. Out of 63 lines，18 lines were detected with Lr26，14 harboring Lr45，11 harboring Lr17，as well as 10 lines harboring Lr1. Any of three genes Lr10，Lr11，and Lr46 was found in 2 lines，while either of Lr2a， Lr2b，Lr15 and Lr37 was present in one line. Interestingly，a total of 17 lines showed slow rusting resistance at adult plant stage. Altogether，this study identified many wheat advanced breeding lines containing Leaf rust resistance genes，thus valuable in genetic improvement of wheat cultivars showing Leaf rust resistance.