The process of urbanization has resulted in an expansion of alien plant species and declines of native species, in particular already rare species. These processes may cause a greater similarity between different urban regions, i.e. biotic homogenization. We explored the relationship between urban regions and homogenization for plant species in Germany using (i) all plant species, (ii) only native species, (iii) all alien species and only those introduced (iv) before or (v) after 1500, respectively. We used the database FLORKART for species distribution on a 6 min latitude × 10 min longitude (ca. 130 km²) scale. We calculated mean similarities for the 60 “most urbanized” cells. We then resampled 60 randomly drawn “less urbanized” cells and 60 “rural” cells and compared these results to the “most urbanized” cells taking distance effects into account. Urbanization does not have an overall effect on homogenization of all species, but native species as well as pre-1500 alien plant assemblages show effects of homogenization while post-1500 alien plant assemblages show the opposite effect. On a regional scale, urbanization is not unequivocally related to homogenization. This might be different when extending the analysed range across several bioclimatic regions. Specific urban habitats, or what remains of them, require special protection and management if the trend towards homogenization is to be avoided.