The expansion of land use for agriculture is among the main reasons for the reduction of natural grasslands around the world, and little is known about the recovery potential of original native grassland vegetation from seed banks. The aim of this study was to assess the seed bank potential of grassland areas with or without history of cultivation in southern Brazil. Specifically, we aimed at evaluating if agricultural use allowed for the persistence of species typical of natural grasslands in soil seed banks. We analyzed seed banks in two areas with a history of cultivation and in two natural grasslands. Samples were collected in spring and autumn in order to assess transient and persistent components, and were characterized by density, richness, presence of exotic species and relationship between seed bank and established vegetation. Both richness and density were high; we found a total of 114 species and a mean density of 61,796 seedlings/m2 for all four areas in the spring sampling. In the natural grasslands, the dominant species of established vegetation were mostly absent from the soil seed bank. Our study indicates that the seed bank is of little relevance for the recovery of typical grassland vegetation after disturbance.