Long-term cultivation in reclaimed mudflats significantly impacts soil properties through changes in salinity and organic carbon (OC) inputs, which subsequently affect soil physical, chemical, and biological characteristics. In addition, long-term rice cultivation can help optimize soil health and fertility management in paddy fields, mitigate climate change, and enhance ecosystem services. However, the impact of long-term cultivation on OC mineralization across a salinity gradient remains unclear. During an 80-d incubation experiment, we assessed 13C-labeled glucose mineralization in soils cultivated for 5, 20, 200, and 700 years, each with varying salinity levels. Our results demonstrated that glucose mineralization declined linearly with increasing soil salinity, from 76 % in low-salinity soils to 31 % in high-salinity soils, reflecting a clear physical impact. This process was closely linked with chemical properties, showing a positive correlation with Olsen P and levels of microbial biomass carbon and nitrogen. Soil organic carbon (SOC) mineralization was primarily controlled by dissolved OC regardless of soil salinity. Further investigation revealed significant shifts in the biological properties, notably the bacterial community composition, which varied with the duration of soil cultivation. The bacterial Shannon index increased steadily from 50 to 700 years, forming distinct clusters, influenced primarily by electrical conductivity and pH in high-salinity soils and by soil-labile nutrients in low-salinity soils. The abundance of Acidobacteria, Proteobacteria, and Verrucomicrobia—dominant phyla in soil cultivated for 700 years—is negatively correlated with soil salinity and pH and positively with labile nutrients, glucose, and OC mineralization, whereas the opposite was true for Bacteroidetes and Firmicutes dominant phyla in soil cultivated for 5 years, was affected positively by soil salinity and pH and negatively by labile nutrients and glucose mineralization. In conclusion, soil salinity and nutrients synergistically regulate OC mineralization in a 700-year cultivated saline soil chronosequence. Addressing these factors is critical to enhancing microbial activity and OC accumulation in saline soils, underscoring the importance of integrating measures to reduce salt and increase the import and accumulation of exogenous organic matter.