Fusarium Head Blight (FHB) is a devastating disease of wheat directly reducing yield and deteriorating quality due to mycotoxin contamination. Deploying genetic resistance is one of the most critical FHB management strategy. The majority of stable FHB resistance quantitative trait loci (QTL) have been derived from wild and non-adapted germplasm, which are not readily utilizable in the breeding programs due to the associated linkage drag. Therefore, it is crucial to identify and characterize native resistance derived from elite germplasm that could be available for immediate deployment in breeding programs. This study was conducted to fine map a native FHB resistance QTL on 1B chromosome of a high yielding moderately resistant soft red winter wheat cultivar Jamestown. Initial mapping of the QTL in a RIL population of 186 individuals identified two FHB QTL at long arm of 1B chromosome. In this study, genome-specific DNA markers were developed at 5 Mb interval spanning the flanking region of 1B FHB QTL. The original RIL population was genotyped, and a critical set of recombinant RILs were identified. The critical RILs were robustly phenotyped for FHB severity and DON under greenhouse conditions for two years. Genotypic and phenotypic analysis narrowed the region to 25 Mb between 330-355 Mb region on 1B chromosome. High-resolution mapping population developed by crossing resistant and susceptible RIL and genotyped using genome-specific Kompetitive allele specific polymerase chain reaction (KASP) assays marker revealed two recombinants. Subsequent phenotyping of F2 recombinants suggested a potential 15 Mb target region between 340-355 Mb. Genotyping and phenotyping of F3 families of recombinant and heterozygous F2 plants is underway. The fine mapping and KAPS markers developed in this study will enable precise selection for 1B FHB QTL and eventually facilitate cloning of the underlying candidate gene.