The grassland of the Tibetan Plateau forms a globally significant biome, which represents 6% of the world’s grasslands and 44% of China’s grasslands. However, large uncertainties remain concerning the vegetation carbon storage and turnover time in this biome. In this study, we quantified the pool size of both the aboveground and belowground biomass and turnover time of belowground biomass across the Tibetan Plateau by combining systematic measurements taken from a substantial number of surveys (i.e. 1689 sites for aboveground biomass, 174 sites for belowground biomass) with a machine learning technique (i.e. random forest, RF). Our study demonstrated that the RF model is effective tool for upscaling local biomass observations to the regional scale, and for producing continuous biomass estimates of the Tibetan Plateau. On average, the models estimated 46.57 Tg (1 Tg = 10 ^12 g) C of aboveground biomass and 363.71 Tg C of belowground biomass in the Tibetan grasslands covering an area of 1.32 × 10 ^6 km ^2 . The turnover time of belowground biomass demonstrated large spatial heterogeneity, with a median turnover time of 4.25 years. Our results also demonstrated large differences in the biomass simulations among the major ecosystem models used for the Tibetan Plateau, largely because of inadequate model parameterization and validation. This study provides a spatially continuous measure of vegetation carbon storage and turnover time, and provides useful information for advancing ecosystem models and improving their performance.