Relatively unmanaged interstitial areas at the residential–wildland interface can support the development of novel woody plant communities. Community assembly processes in urban areas involve interactions between spontaneous and cultivated species pools that include native, introduced (exotic/non-native) and invasive species. The potential of these communities to spread under changing climate conditions has implications for the future trajectories of forests within and beyond urban areas. We quantified woody vegetation (including trees and shrubs) in relatively unmanaged “interstitial” areas at the residential–wildland interface and in exurban reference natural areas in six metropolitan regions across the continental USA. In addition, we analyzed soil N and C cycling processes to ensure that there were no major anthropogenic differences between reference and interstitial sites such as compaction, profile disturbance or fertilization, and to explore effects of novel plant communities on soil processes. We observed marked differences in woody plant community composition between interstitial and reference sites in most metropolitan regions. These differences appeared to be driven by the expanded species pool in urban areas. There were no obvious anthropogenic effects on soils, enabling us to determine that compositional differences between interstitial and reference areas were associated with variation in soil N availability. Our observations of the formation of novel communities in interstitial spaces in six cities across a very broad range of climates, suggest that our results have relevance for how forests within and beyond urban areas are assessed and managed to provide ecosystem services and resilience that rely on native biodiversity.