Powdery mildew is a common disease in wheat production worldwide. The genetic improvement of powdery mildew resistance is limited by effective resistance genes. Although Chinese wheat landraces are known to be valuable sources of powdery mildew resistance genes, only a few of them are well-characterized for their genetic controls. We identified more than a hundred resistant landraces from about 1,500 accessions using a popular Blumeria graminis f. sp. tritici isolate E09. Tests of several segregating populations revealed that most of the powdery mildew resistance in the landraces examined were controlled by single genes. Molecular markers generated by the BSR-Seq and/or SNP arrays allowed the construction of genetic linkage maps and localization of the powdery mildew resistance genes, such as Pm24 (Hulutou), Pm47 (Hongyanglazi), Xuxusanyuehuang (Pm61), PmQ (Qingxinmai), and PmJ (Jiantouhong). Map-based cloning was used to isolate Pm5e from Fuzhuang 30 and Pm24 from Chiyacao, Baihulu and Hulutou. Pm5e encodes an NLR protein, and several other wheat landraces, including Xiaobaidongmai, Hongquanmang, Mazhamai, Bensanyuehuang, Youmangxiabadou, Ziganchangmang, Baiyouyantiao, and Youmailan, also carry Pm5e. A rare nonsynonymous SNP variant in the C-terminal LRR domain of Pm5e is associated with the gain of function for the powdery mildew resistance. Pm24 encodes a WHEAT TANDEM KINASE 3 (WTK3), which differs from the typical NLR resistance proteins. A rare 6-bp natural deletion of lysine-glycine codons was responsible for the powdery mildew resistance. Both Pm5e and Pm24 represent unique mechanisms and origin of the powdery mildew resistance genes from wheat landraces. The projects of cloning other powdery mildew resistance genes from wheat landraces, such as Pm61 and PmQ, are in progress. Isolation of more resistance genes will facilitate the exploration of disease resistance genes from wheat landraces and developing disease resistant wheat cultivars via marker-assisted selection and genome editing approaches.