Enhanced nitrogen (N) availability is one of the main drivers of biodiversity loss and degradation of ecosystem functions. However, in very nutrient-poor ecosystems, enhanced N input can, in the short-term, promote diversity. Mediterranean Basin ecosystems are nutrient-limited biodiversity hotspots, but no information is available on their medium- or long-term responses to enhanced N input. Since 2007, we have been manipulating the form and dose of available N in a Mediterranean Basin maquis in south-western Europe that has low ambient N deposition (<4 kg N ha(-1) yr(-1)) and low soil N content (0.1%). N availability was modified by the addition of 40 kg N ha(-1) yr(-1) as a 1∶1 NH4Cl to (NH4)2SO4 mixture, and 40 and 80 kg N ha(-1) yr(-1) as NH4NO3. Over the following 5 years, the impacts on plant composition and diversity (richness and evenness) and some ecosystem characteristics (soil extractable N and organic matter, aboveground biomass and % of bare soil) were assessed. Plant species richness increased with enhanced N input and was more related to ammonium than to nitrate. Exposure to 40 kg NH4+-N ha(-1) yr(-1) (alone and with nitrate) enhanced plant richness, but did not increase aboveground biomass; soil extractable N even increased under 80 kg NH4NO3-N ha(-1) yr(-1) and the % of bare soil increased under 40 kg NH4+-N ha(-1) yr(-1). The treatment containing less ammonium, 40 kg NH4NO3-N ha(-1) yr(-1), did not enhance plant diversity but promoted aboveground biomass and reduced the % of bare soil. Data suggest that enhanced NHy availability affects the structure of the maquis, which may promote soil erosion and N leakage, whereas enhanced NOx availability leads to biomass accumulation which may increase the fire risk. These observations are relevant for land use management in biodiverse and fragmented ecosystems such as the maquis, especially in conservation areas.