The objective of this study is to determine the potential resistance genes in wheat landraces and provide reference information for the selection and utilization of leaf rust resistance resources. In this study, 15 wheat landraces, susceptible control Zhengzhou 5389 and 36 resistance donor lines were tested for leaf rust resistance. The genotypes were tested in the greenhouse to 19 Puccinia triticina pathotypes for a seedling resistance, and in the field to a mixed pathotypes for slow rusting resistance in two cropping seasons (2016-2017 and 2017-2018) in Baoding, Hebei Province. Meanwhile, twelve molecular markers targeting the resistance loci were used for genotyping in wheat cultivars. Resulted from the resistance spectra and marker analysis, seven leaf rust resistance genes were detected in 15 landraces. For instance, the landrace Honggoudou may contain both Lr1 and Lr46; Huanghuamai may have both Lr13 and Lr34; Dabaimai may carry Lr14b and Lr26; Yangmai may carry Lr37 and Lr46; Chengduguangtou may carry Lr34 and Lr46. Two resistance genes Lr26 and Lr46 were found to be present in genotypes Motuomai and Xishanbiansui. Furthermore, the slowing rust resistance gene Lr34 was detected in Tongjiabaxiaomai, Wudubaijianer, Bianbachunmai-6 and Baihuamai, while another partial resistance gene Lr46 was detected in Hongqiangmai, Baibiansui and Baihuomai. These genotypes effective against leaf rust might provide resistance genes for disease-resistant breeding in hexaploid wheat.