Land-cover change has significant influence on carbon storage and fluxes in terrestrial ecosystems. The southern United States is thought to be the largest carbon sink across the conterminous United States. However, the spatial and temporary variability of carbon storage and fluxes due to land-cover change in the southern United States remains unclear. In this study, we first reconstructed the annual data set of land-cover of the southern United States from 1860 to 2003 with a spatial resolution of 8 km. Then we used a spatially explicit process-based biogeochemical model (Terrestrial Ecosystem Model [TEM] 4.3) to simulate the effects of cropland expansion and forest regrowth on the carbon dynamics in this region. The pattern of land-cover change in the southern United States was primarily driven by the change of cropland, including cropland expansion and forest regrowth on abandoned cropland. The TEM simulation estimated that total carbon storage in the southern United States in 1860 was 36.8 Pg C, which likely was overestimated, including 10.8 Pg C in the southeast and 26 Pg C in the south-central. During 1860-2003, a total of 9.4 Pg C, including 6.5 Pg C of vegetation and 2.9 Pg C of soil C pool, was released to the atmosphere in the southern United States. The net carbon flux due to cropland expansion and forest regrowth on abandoned cropland was approximately zero in the entire southern region between 1980 and 2003. The temporal and spatial variability of regional net carbon exchange was influenced by land-cover pattern, especially the distribution of cropland. The land-use analysis in this study is incomplete and preliminary. Finally, the limitations, improvements, and future research needs of this study were discussed.