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The development of Fusarium head blight (FHB)-resistant winter wheat cultivars using the gene Fhb1 has been conducted in northern China. Sumai 3, a Chinese FHB-resistant spring wheat cultivar, carries three FHB resistance genes: Fhb1, Fhb2 and Fhb5. To better use these genes for increasing FHB resistance in northern China, it is necessary to elucidate the pyramiding effects of Fhb1, Fhb2 and Fhb5 in winter wheat backgrounds. Eight gene combinations involving Fhb1, Fhb2 and Fhb5 were identified in a double haploid (DH) population, and the effects on FHB resistance were evaluated in six tests. At the single gene level, Fhb1 was more efficient than the other two genes in single-floret inoculation tests, whereas Fhb5 showed better resistance than Fhb1 and Fhb2 under a natural infection test. Pyramiding Fhb1, Fhb2 and Fhb5 showed better FHB resistance than the other gene combinations. Forty-nine DH lines showing consistently better resistance than the moderately susceptible control Huaimai 20 in multiple tests were evaluated for main agronomic traits, and no difference in grain yield was found between the mean values of DH lines and the recipient parents Lunxuan 136 and Lunxuan 6, which are higher than those of recipient parent Zhoumai 16 and the donor parent Sumai 3 (p < 0.05). Based on the phenotypic and genomic composition analyses, five promising DH lines fully combined the FHB resistance of donor Sumai 3 and the elite agronomic traits from the recipient parents. This study elucidates the pyramiding effects of three FHB resistance genes and that the promising DH lines with resistance to FHB can be directly applied in wheat production or as parents in winter wheat breeding programs. [ABSTRACT FROM AUTHOR]