Although there has been research on managing Fusarium head blight (FHB) in spring barley, little has been published on cultivar resistance and optimal fungicide timing for FHB management in winter barley. A 3-year (2015 to 2017) field experiment was conducted to measure FHB resistance of winter barley varieties, gauge the potential benefit from a fungicide, and help determine the optimal timing for fungicide application. The split-plot experiment took place in a misted, inoculated nursery in Raleigh, North Carolina using main plots of four winter barley cultivars (Atlantic, Endeavor, Nomini, and Thoroughbred). Three fungicide treatments were applied to subplots: prothioconazole + tebuconazole at full spike emergence, the same fungicide 6 days later, or no fungicide. The late applications significantly reduced FHB index in each of 3 years and significantly reduced deoxynivalenol (DON) in harvested grain in 2 of the 3 years. Applications at full spike emergence also yielded significant benefit in 1 of the 3 years for each parameter. Neither disease symptoms nor DON gave reason to prefer one of the fungicide timings over the other. Across the 3 years, DON ranked the cultivars Endeavor < Nomini = Thoroughbred < Atlantic. Combining the moderate resistance of Endeavor with a fungicide application and averaging the two timings resulted in a 75% DON reduction compared with unsprayed Atlantic. Taken together, our results indicate that barley growers concerned about minimizing DON should both plant moderately resistant varieties and apply fungicide if there is scab risk. During the same period, 16 commercial winter barley cultivars were tested in from three to seven Virginia and North Carolina environments each, and the DON results were compared after standardization across environments. The winter two-row malting barley cultivars Endeavor and Calypso displayed superior and robust DON resistance across environments.