Abstract: Split plot design was used to examine the effects of applying calcium and inoculating acid-tolerant rhizobium on growth, yield and quality of alfalfa planted in acid soil on Jinyun Mountains. The results showed that inoculating acid-tolerant rhizobium significantly improved the plant’s nodule weight, fresh root weight, plant height, fresh aboveground biomass weight, total nitrogen and rhizobium number in acid soil. Applying calcium significantly increased fresh root weight, plant height and fresh aboveground biomass weight, and nodule weight. Results of correlation analysis indicated that nodule weight was significantly correlated with alfalfa fresh aboveground biomass weight, fresh root weight, plant height, plant’s total nitrogen. We conclude that good nodulating ability of rhizobium can improve yield and quality of alfalfa in acid soil.