Fusarium head blight (FHB) becomes a major disease in the wheat production of the Yellow-Huai Rivers Valley Winter Wheat Zone of China. Improving FHB resistance is one of the major breeding targets in this region. The donors of Fhb1 gene, including Ningmai 9, Shengxuan 6, Jianyang 798, Jianyang 84, Sumai 3, and Ningmai 13, were moderately or highly resistant to FHB. Six backcrossing populations were developed by crossing these donors with a dwarfing and male-sterile wheat near isogenic line Zhoumai 16, and backcrossing with Zhoumai 16. The progenies were subjected to phenotypic evaluation of FHB by both the floret-inoculation method and natural infection in a field disease nursery. Fhb1 was detected by a functional Kompetitive Allele Specific PCR (KASP) marker. The backcrossing progenies with Fhb1 gene were moderately susceptible to FHB. Compared with the backcrossing progenies without Fhb1 gene, mean number of diseased spikelets and disease index reduced by 4.2 (P < 0.01) and 4.0 in the progenies with Fhb1 gene, respectively. The mean number of diseased spikelets and disease index for the backcrossing progenies with Fhb1 gene were 8.1 (P < 0.01) and 28.4 (P < 0.01) lower than those of the recurrent parent Zhoumai 16. There were significant differences in resistance among the backcrossing progenies from different donors. The progenies from Shengxuan 6 had better performance in number of diseased spikelets and disease index compared with those from any other donors. The results from this study indicate that Fhb1 gene can efficiently improve the FHB resistance of wheat cultivars grown in the Yellow-Huai Rivers Valley Winter Wheat Zone.