This study was carried out to investigated optimum additive ratio of barley flour for substitution of rice bran atcultivation of winter mushrooms. Mycerial growth was faster according to increase of barley flour ratio than those of controls, butonly some slow at the addition of 10% barley flour. Yields of fruiting body show the highest to 165.4g/850ml of medium whichare the addition 30% of barley flour and not significant difference of yields up to the addition of 70% barley flour. Diameter ofpileus was the highest at the addition of 30% barley flour. Hardness of pileus and stipes were the highest at the addition of 10%barley flour. The L value of stipes were the hight at the addition of barley flour, but the L value of pileus were decreased at theaddition of barley flour, but there was no significant difference in the a-value and the b-value.