Environmental pollution is an important issue in metropolitan areas, and roadside trees are directly affected by various sources of pollution to which they exhibit numerous responses. The aim of the present study was to identify morpho-physio-biochemical attributes of maidenhair tree (Ginkgo biloba L.) and American sycamore (Platanus occidentalis L.) growing under two different air quality conditions (roadside with high air pollution, RH and roadside with low air pollution, RL) and to assess the possibility of using their physiological and biochemical parameters as biomonitoring tools in urban areas. The results showed that the photosynthetic rate, photosynthetic nitrogen-use efficiencies, and photochromic contents were generally low in RH in both G. biloba and P. occidentalis. However, water-use efficiency and leaf temperature showed high values in RH trees. Among biochemical parameters, in G. biloba, the lipid peroxide content was higher in RH than in RL trees, but in P. occidentalis, this content was lower in RH than in RL trees. In both species, physiological activities were low in trees planted in areas with high levels of air pollution, whereas their biochemical and morphological variables showed different responses to air pollution. Thus, we concluded that it is possible to determine species-specific physiological variables affected by regional differences of air pollution in urban areas, and these findings may be helpful for monitoring air quality and environmental health using trees.