Passive steady-state chlorophyll fluorescence (Fs) provides a direct diagnosis of the functional status of vegetation photosynthesis. With the prospect of mapping Fs using remote sensing techniques, field measurements are mandatory to understand to which extent Fs allows detecting plant stress in different environments. Trees of four common species in Valencia were classified in either a low or a high local traffic exposure class based on their leaf magnetic value. Upward and downward hyperspectral fluorescence yield (FY) and indices based on the two Fs peaks (at 687 and 741 nm) were calculated. FY indices of P. canariensis and P. x acerifolia were significantly different between the two traffic exposure classes defined, but not for C. australis nor M. alba. While chlorophyll content could not indicate the difference between low and high traffic exposure, the FY(687)/FY(741) peak ratio increased significantly (p < 0.05) for both leaf sides for the higher traffic exposure class.