Powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is one of the most serious wheat diseases worldwide, leading to severe reduction in grain yield and quality. Breeding and deploying disease-resistant cultivars is the most effective, economical and environmentally friendly strategy. However, because of the rapid evolution of Bgt isolates, the resistance of applied resistant genes would be conquered by a new virulence isolate very shortly. Thus, continued efforts should be made to mine new disease resistance genes from wheat germplasms and closely related species. Wheat landraces are known as important gene pool for durable and broad-spectrum disease resistance genes after thousand years natural and artificial selections. Here we reported the map-based cloning of powdery mildew resistance genes Pm5e and Pm24 from Chinese wheat landraces. A CC-NBS-LRR (NLR) protein confers the powdery mildew resistance locus Pm5 and a wheat tandem kinase (WTK) provides broad spectrum powdery mildew resistance of Pm24. Haplotype analyses of diversified worldwide wheat germplasm collection were performed to study the genetic diversity of the two loci. The results revealed single A/G SNP in the NLR was responsible for the resistant difference between Pm5e and Pm5a, and a 6 bp deletion in the WTK was responsible for Pm24 resistance. The rare natural variations in Pm5e and Pm24 loci only identified in a few Chinese wheat landraces provide evidences of natural and human selections in preserving the wheat germplasm diversity and disease resistance genes evolution under pathogen stress. This finding also provides useful gene resources for future wheat improvement, especially target genes and sites for genome editing breeding for disease resistance in wheat.