Steamed bread is a fermented wheat flour product that is cooked by being steamed in a steamer. The properties of dough and the quality of steamed bread incorporated with barley flour at 10, 20 and 30% substitution levels are investigated. Increasing levels of barley flour correspondingly decreases the gluten content and increases the beta-glucan content. Upon incorporation of barley flour, the dough development time, departure time and stability time are decreased, but the mixing tolerance index is increased. The incorporation of barley flour into wheat flour also significantly decreases the extensibility, increases the value of R/E ratio and decreases the value of work input. In addition, increased levels of barley flour cause significant decreases in the specific volume, brightness and whiteness index of steamed bread, as well as increases in hardness and chewiness. This study also shows that six-row barley pearl flour contains more dietary fiber and has a more favorable influence than barley endosperm flour.