A high level of endemic flowering plant species is highlighted for the first time for the southern Brazilian flora. We present a comprehensive list of 1020 endemic taxa and analyse their distribution in different biomes, focusing on the Subtropical Highland Grasslands (Campos de Cima da Serra). Considering all biomes represented in southern Brazil, c. 13% of the flowering plant species are endemic, which accounts for more than 5% of the total endemic taxa of Brazil and c. 12% of the endemic taxa from the Atlantic Forest hotspot. Like the High Altitude Tropical Grasslands that are found further north in the south-eastern Brazil region, the Subtropical Highland Grasslands are home to an assemblage of tropical and temperate plant lineages. The Subtropical Highland Grasslands are characterized by their transitional nature and by having 296 endemic plant taxa (25% of its flora). © 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 167, 378–393.