Forest ecosystems play an important role in regulating the global carbon, a substantial portion of terrestrial carbon pool which is stored in biomass stocks. However, how multiple biotic (i.e. topography) and abiotic (biodiversity, stand structure, and functional traits) influence forest biomass in natural forests, the relative important of these factors determine biomass is still controversial for subtropical natural forests. We used forest inventory data from nine 1-ha plots at different altitude gradients in China’s subtropical forests. We used multiple analyse to quantify the relative importance of multiple facets of diversity, key functional traits, stand structural attributes, and topography variables in determining forest biomass. We found that multiple facets of diversity and stand structure variables enhances biomass. Specifically, large-diameter trees had a strong positive effect on biomass and were the most important factor in determining biomass. Plant functional traits were closely related to biomass. Community-weighted mean value (CWM) of maximum height positively correlated with biomass, but CWM of wood density negatively correlated biomass. Topographic factors including elevation and slope, had a positive effect on biomass. Moreover, among the aforementioned four types of variables, stand structure had the greatest impact on biomass and is linked to diversity-biomass relationship. Topography mainly indirectly affected biomass by altering multiple diversity and stand structure. Functional traits also directly and indirectly affected biomass. Overall, these results support niche complementarity effect and mass-ratio hypothesis. Our results indicate that biodiversity is essential for maintaining ecosystem functions of species-rich subtropical natural forests. Further, adjusting stand structure may be an effective forest management approach to increase forest carbon storage.