It is widely accepted that old-aged forest can accumulate soil organic carbon (SOC). How microbial physiological traits respond to forest age and whether they drive SOC sequestration in old-aged forest remain elusive. Therefore, we compared the microbial C use efficiency (CUE), biomass turnover rate (rB), microbial biomass C (MBC) and necromass C (MNC) across soil profiles from middle and old-aged forest and evaluated how these microbial traits are related to SOC storage. The results revealed that both forests could accumulate SOC and old-aged forest supported higher SOC storage than middle-aged forest from 2005 to 2020. Moreover, SOC was concentrated on the surface soils of middle-aged forest, whereas it was more distributed across the deeper soil profile in old-aged forest. Compared with middle-aged forest, the O, A and B soil layers of old-aged forest presented increases in microbial CUE (17.8%, 36.9% and 25.0%, respectively), rB (43.7%, 39.7% and 10.8%, respectively), MBC (114.8%, 81.1% and 122.9%, respectively), and MNC content (47.0%, 22.2% and 21.6%, respectively). Random forest analysis suggested that SOC accumulation is controlled mainly by microbial physiological traits rather than other factors including environmental variables. Specifically, microbial CUE and turnover rates increased in old-aged forest, resulting in higher MBC and MNC contents, which in turn led to SOC accumulation. Moreover, the effects of plant and soil properties on SOC storage are regulated mainly by microbial-physiological parameters and the size of microbial C pools. Our findings provide valuable insights into the microbial mechanisms underlying SOC storage in old-aged forest.