Roadsides are preferential migration corridors for invasive plant species and can act as starting points for plant invasions into adjacent habitats. Rapid spread and interrupted distribution patterns of introduced plant species indicate long-distance dispersal along roads. The extent to which this process is due to species' migration along linear habitats or, alternatively, to seed transport by vehicles has not yet been tested systematically. We tested this by sampling seeds inside long motorway tunnels to exclude nontraffic dispersal. Vehicles transported large amounts of seeds. The annual seed rain caused by vehicles on the roadsides of five different tunnel lanes within three tunnels along a single urban motorway in Berlin, Germany, ranged from 635 to 1579 seeds/m(2)/year. Seeds of non-native species accounted for 50.0% of the 204 species and 54.4% of the total 11,818 seeds trapped inside the tunnels. Among the samples were 39 (19.1%) highly invasive species that exhibit detrimental effects on native biodiversity in some parts of the world. By comparing the flora in the tunnel with that adjacent to the tunnel entrances we confirmed long-distance dispersal events (>250 m) for 32.3% of the sampled species. Seed sources in a radius of 100 m around the entrances of the tunnels had no significant effect on species richness and species composition of seed samples from inside the tunnels, indicating a strong effect of long-distance dispersal by vehicles. Consistently, the species composition of the tunnel seeds was more similar to the regional roadside flora of Berlin than to the local flora around the tunnel entrances. Long-distance dispersal occurred significantly more frequently in seeds of non-native (mean share 38.5%) than native species (mean share 4.1%). Our results showed that long-distance dispersal by vehicles was a routine rather than an occasional mechanism. Dispersal of plants by vehicles will thus accelerate plant invasions and induce rapid changes in biodiversity patterns.