Enhancing resistance to Fusarium head blight (FHB) has become one of important breeding objectives in the major wheat-growing regions in China. A prominent locus Fhb1 conferring stable FHB resistance with the largest effect is the major source of resistance in wheat breeding. Understanding the distribution and putative donor of Fhb1 in Chinese wheat cultivars will facilitate the application of this gene and thus benefit FHB resistance breeding in China. Haplotype analysis of PFT (pore-forming toxin-like), HC (HCBT-like defense response protein) and His (histidine-rich calcium-binding protein) genes in the Fhb1 region of 229 wheat cultivars and advanced lines indicated that PFT-I/His-I was a resistant haplotype. Both pedigree and marker (or sequence) information revealed that Fhb1 in Chinese wheat cultivars was mainly derived from Sumai 3 and Ningmai 9, in which Ningmai 9 was the major donor. The Fhb1 diagnostic markers PFT-CAPS and His-InDel developed in this study can be used effectively in marker-assisted selection for improving FHB resistance.