This study aimed to evaluate the rate of ecogenotoxicity caused by traffic emissions at sites in the vicinity of roads and at sites near planned highway construction using a pollen grain abortion assay with higher wild plant species. The relationship between pollen grain abortivity and distance from a road also was assessed. The highest values were found at a distance of 30 m, where the prevailing wind direction was parallel to the road. Wind blowing away from the road shifted this boundary to a distance of 350 m. The results showed the highest genotoxicity at the “Ring road” site, where the frequency of abortive pollen grains was 4.05 times higher than at the control site, and at the “Dubná Skala” site, with induction factor 3.48. Based on our results we can conclude that Chelidonium majus, Cichorium intybus, and Melilotus albus are suitable species for the detection of genotoxicity in the environment.