The South Brazilian grasslands (Campos Sulinos) form the dominant vegetation in southern Brazil. They are species-rich ecosystems that occur under distinct geomorphological and climatic conditions but spatial variation of plant species diversity remains understudied. Here, we present a detailed description of plant communities across the region. Our data were obtained in 1080 plots, representing well-preserved grasslands in different ecological systems. Apart from describing alpha and beta diversity, we investigated the relations of plant communities with environmental features. We identified 759 plant species and found clear differences in community composition across the region. Northern and Southern highland grasslands, humid and dry coastal grasslands and the mesic Pampa grassland were clearly distinct, related to climatic and edaphic features. While species abundance distribution was markedly uneven, local species richness was high, above 20 species/m2, especially in the highlands and in mesic Pampa sites, on shallow soils. The predominant component of beta diversity was species turnover, which suggests that a network of well-conserved grasslands distributed across the region would be the best strategy to protect plant diversity. Our results establish regionalized reference values for richness and diversity that can be useful for initiatives of restoration and conservation of these grasslands.