The varietal differences of protein content in barley grain and its variation under different cultural conditions were studied. The varietal variation of protein content was significant with the range of 8～18%. The protein content of barley grain were increased by heavier nitrogen application and decreased by shading treatment and drill seeding. There was negative correlation between protein content and grain weight in hulled barley, while positive correlation between nitrogen content of plant and leaf at heading stage and protein content of barley grain was significant in Haganemugi and Buheung.