An investigation was carried out to estimate root respiration contribution to total soil respiration in a mixed C3 and C4 grassland in Japan. Root biomass showed linear relationships with total soil respiration. Further quantification of the relationship between the two in a regression equation yields a y-intercept value which estimates microbial respiration in the absence of root. We estimated that the contribution of root respiration to total soil respiration in the growing season ranged from 31% to 50%. Microbial biomass, soil organic carbon showed no significant correlation with soil respiration.