The distribution of root biomass and its relationship with soil environmental factors are important for the study of grassland conservation and the degraded grassland restoration. This study was based on the alpine meadows in Damxung County, northern Tibet. The root biomass, root carbon and nitrogen content, soil carbon content (SOC, DOC, MBC), nitrogen content (DTN, MBN, TN), carbon to nitrogen ratio (MBC/MBN, SOC/TN), pH, conductivity of 0-50 cm soil depth were measured at three altitudes (4300 m, 4500 m, 4700 m) to explore the distribution pattern of root biomass and carbon and nitrogen in the soil at different altitudes in the alpine meadow, as well as the relationship between them. The results showed that:(1) the various forms of carbon and nitrogen measured in soil presented a downward trend with the increase of soil depth. The total DOC and SOC (0-50 cm depth) showed an increasing trend with the increase of altitude. (2) As the soil depth increased, the root biomass decreased exponentially. With the increase of altitude, the more concentrated the root biomass was in the upper layer of soil, the less the distribution of root biomass in the lower layer, but the change tends to be gentle. (3) The root biomass was positively correlated with the measured carbon and nitrogen indexes and conductivity, while it showed a significant negative correlation with pH. Root nitrogen pool was a main factor affecting the distribution of root biomass in soil. pH, conductivity and soil carbon and nitrogen indicators were important factors affecting the distribution of root biomass in soil.