Background and Aims:Atmospheric nitrogen deposition and natural fire regime suppression are key drivers of vegetation change in urbanizing grasslands. Some species thrive under these conditions, while others face local extinction. In the natural grasslands that surround Melbourne, Australia, biotic homogenization has occurred with intensifying urbanization. Some native species have become rarer (decreaser species) across the landscape, while others have become more widespread (increaser species). This study experimentally examined the response of increaser and decreaser plant species to nitrogen addition/depletion, and examined the presence/absence of annual disturbance to the vegetation. Methods:Decreaser and increaser species were planted into 60 field plots established in an urban Melbourne grassland and examined over 2 years. Annual removal of above-ground biomass occurred in half the plots to simulate biomass removal via fire, with the remaining plots undisturbed. Soil nitrogen was depleted in one-third of plots, one-third received no nitrogen treatment and one-third were fertilized with nitrogen. Increaser plant species were predicted to persist in the absence of disturbance, and thrive when fertilized. In contrast, high mortality was predicted for decreaser species in the absence of disturbance, with fertilization providing no advantage. Key Results:Seedling mortality for increaser and decreaser species was unrelated to the treatments. The mortality of decreaser species was high (69 %), and the mortality of increaser species low (20 %). However, seedling growth was related to the treatments. The total biomass of decreaser species was highest in annually disturbed plots, with growth suppressed in undisturbed plots. In contrast, the total biomass of increaser species was unrelated to the disturbance regime, but responded positively to nitrogen enrichment. Conclusions:The results provide evidence that by affecting plant growth, declines in biomass removal and atmospheric nitrogen deposition could be key drivers of biotic homogenization in urban grasslands.