Alien tree species have been introduced to Europe and often used as roadside trees. Currently, preference is given to species that are adapted to urban climate and drier conditions. Native epiphytic lichens are adapted to local tree species; however, little is known whether a shift toward more alien roadside trees would affect the diversity of epiphytic lichens within cities. We considered three genera of roadside trees that are common in Hamburg (Germany) Acer, Tilia, and Quercus, each with a native and an alien species per genus. Species numbers, frequency and diversity of epiphytic lichens were assessed and compared. Tree locations were grouped according to three classes of increasing traffic volume as a proxy for air pollution. Mean bark pH values have been recorded for each tree individual. The species pairs Quercus robur/rubra and Acer platanoides/saccharinum showed significant differences in the diversity and abundance of lichens, with the alien species showing lower values. The species pair Tilia cordata/tomentosa did not show any significant differences. The number of nitrogen-tolerant lichen species differed little among all tree species. Lichen species richness decreased with increasing traffic volume for all tree species pairs, thus demonstrating the inhibiting effect of traffic on the occurrence of lichens. Bark pH differed little between the species pairs yet high traffic volume resulted in an increased bark pH. In conclusion, two out of three alien species had negative effects on lichen diversity. Further tree species should be assessed to better estimate the effect of alien roadside trees on lichen diversity.