The rising temperature, altering precipitation, and increasing extreme events under climate warming affect the stability and sustainability of grassland ecosystems. The dynamics of above-ground biomass (AGB), below-ground biomass (BGB), and biomass partitioning (BGB:AGB ratio) of grasslands are of fundamental importance to understand their feedback to climate change. In this study, we used grassland productivity data extracted from the Oak Ridge National Laboratory Distributed Active Archive Center, Tennessee, USA, in which the AGB was collected within a 1.0 m × 0.25 m quadrat and the BGB was sampled within the center of the quadrat. Using multiple pairwise tests and Pearson’s correlation analysis, we assessed the variations of grassland productivity and examined the response of single-harvest and annual biomass partitioning of C3- and C4-dominated grasslands to the growing-season and annual climatic variability and climate extremes in seven sites belonging to four ecoregions (i.e. cold steppe, humid temperate, humid savanna, and savanna). The results show that the annual and single-harvest BGB:AGB ratio varied significantly across the plant types and ecoregions. Overall, the C3-dominated grasslands exhibited a higher BGB:AGB ratio than that of C4-dominated grasslands. Growing-season temperatures (GSTs) were found to be the key determinants in explaining the single-harvest BGB:AGB ratio rather than growing-season precipitation. For instance, the single-harvest BGB:AGB ratio of C4-dominated grasslands increased, while that of C3-dominated grasslands decreased with elevated GSTs. The growing-season extreme dry climates significantly increased the single-harvest BGB:AGB ratio of C4 plants by a large reduction of AGB, potentially affecting the ecosystem functioning and stability. The C3-dominated grasslands in the cold steppe ecoregion are at great threat of drought-induced stress, as we observed that growing-season extreme dry climates reduced, albeit insignificantly, both the single-harvest AGB and BGB. This study provides key insights into factors influencing the biomass partitioning of C3- and C4-dominated grasslands and has important implications for assessing the grassland functioning and stability under increasing climate extremes.