The increasing demand for food has driven the conversion of native areas into agricultural land, a transformation with significant implications for soil carbon (C) balance due to current cultivation systems. Developing and implementing cropping systems that promote C accumulation is crucial for maintaining agricultural sustainability. In this context, this study aimed to understand land management in the state of Piauí, located within the Brazilian Cerrado, investigating biological indicators of soil quality in a cultivation chrono sequence on an Oxisol, in order to support the development and implementation of cropping systems that promote carbon accumulation. The chrono sequence comprised four areas: one native area and three cultivated areas for 12, 18, and 24 years. The biological quality indicators included soil organic carbon (SOC), microbial biomass carbon (MBC), basal soil respiration (C-CO₂), microbial quotient (qMic), and metabolic quotient (qCO₂), as well as C stock and CO₂ sequestration across the 0.0-10, 10-20, 20-30, 30-40, and 40-50 cm soil layers. The results showed no significant differences among areas for MBC, qMic, and qCO₂, but the SOC stock, C-CO₂, and C stock were increased with longer cultivation periods. CO₂ sequestration rates were 0.96, -0.06, and 1.69 Mg ha-¹ year-¹ in the areas cultivated for 12, 18, and 24 years, respectively. Overall, the duration of cultivation enhanced microbial activity and C stock, contributing to CO₂ sequestration compared to the native area. Our findings evidenced that research focusing on region-specific management systems are essential to ensure the long-term agriculture sustainability, since each region has distinct environmental and climatic conditions that directly influence the outcomes of implemented agricultural practices.