Fusarium head blight (FHB) is a devastating disease of wheat worldwide. Novel sources of resistance are critical for improving FHB resistance levels in wheat. From a large-scale evaluation of germplasm for reactions to FHB, we identified one wheat accession (PI 277012) that consistently showed a high level of resistance in both greenhouse and field experiments. To characterize the FHB resistance in this accession, we developed a doubled haploid (DH) mapping population consisting of 130 lines from the cross between PI 277012 and the hard red spring wheat cultivar 'Grandin'. The DH population was then evaluated for reactions to FHB in three greenhouse seasons and five field environments. Based on a linkage map that consisted of 340 SSR markers spanning 2,703 cM of genetic distance, two major quantitative trait loci (QTLs) for FHB resistance were identified on chromosome arms 5AS and 5AL, with each explaining up to 20 and 32% of the variation in FHB severity, respectively. The two QTLs also showed major effects on reducing the percentage of Fusarium damaged kernels (FDK) and deoxynivalenol (DON) accumulation in seeds. FHB resistance has not previously been reported to be associated with this particular genomic region of chromosome arm 5AL, thus indicating the novelty of FHB resistance in PI 277012. Plant maturity was not associated with FHB resistance and the effects of plant height on FHB resistance were minor. Therefore, these results suggest that PI 277012 is an excellent source for improving FHB resistance in wheat. The markers identified in this research are being used for marker-assisted introgression of the QTLs into adapted durum and hard red spring wheat cultivars.