Very little is known about the impact of atmospheric carbon dioxide pressure (pCO2) on the shaping of biomes. The development of pCO2 throughout the Brunhes Chron may be considered a natural experiment to elucidate relationships between vegetation and pCO2. While the glacial periods show low to very low values (~ 230 to ~ 190 ppmv, respectively), the pCO2 levels of the interglacial periods vary from intermediate to relatively high (~250 to ~ 270, respectively). To study the influence of pCO2 on the Pleistocene development of SE African vegetation, we used the pollen record of a marine core (MD96-2048) retrieved from Maputo Bay south of the Limpopo River mouth in combination with stable isotope and geochemical proxies. Applying endmember analysis, four pollen assemblages could be distinguished representing different biomes: heathland, mountain forest, shrubland and woodland. We find that the vegetation of the Limpopo River catchment and the coastal region of southern Mozambique is not only influenced by hydroclimate but by also temperature and atmospheric pCO2. Our results suggest that the extension of either open ericaceous vegetation including C4 sedges or mountain forest depended on glacial pCO2 levels, and that the main development of woodlands in the area took place after the Mid-Brunhes Event when interglacial pCO2 levels rose over 270 ppmv.