In this study, soil organic carbon content, organic carbon density, and carbon sequestration potential in tobacco- growing soils in the main tobacco growing areas of Guizhou Province (Zunyi City, Bijie City, Tongren City, and Southwest Guizhou Autonomous Prefecture) were analyzed. Results showed that there were significant differences in organic carbon contents and organic carbon densities of tobacco-growing soils in the main tobacco-growing areas of Guizhou Province. Organic carbon content of tobacco-growing soils in Southwest Guizhou (19.39 g/kg) was significantly higher than that in Bijie (15.35 g/kg), Zunyi (15.53 g/kg), and Tongren (13.65 g/kg) (P<0.05). Organic carbon density of tobacco-growing soils in Zunyi was significantly higher than that in Bijie, Southwest Guizhou, and Tongren(P<0.05). Organic carbon storage in tobacco-growing soils of different tobacco-growing areas showed the order of Zunyi (12.30 Tg) > Bijie (11.10 Tg) > Southwest Guizhou (5.83 Tg) > Tongren (2.48 Tg). Carbon sequestration potentials of tobacco-growing areas in Zunyi, Tongren, Southwest Guizhou and Bijie were measured at 2.23, 2.36, 1.25, and 1.51 kg/m², respectively. Random forest model analysis showed that soil alkali-hydrolyzable nitrogen, total nitrogen, total phosphorus, altitude, and pH were the important factors affecting soil organic carbon density. In conclusion, the tobacco-growing soils in Guizhou Province demonstrated substantial potential for carbon sequestration. This research can serve as a reference for formulating strategies aimed at enhancing carbon sequestration and increasing carbon sinks in the main tobacco-growing areas of Guizhou.