In central Europe, barnyardgrass [Echinochloa crus-galli (L.) P. Beauv.], has commonly been found in humid lowland areas. As a result of the introduction of new crops and farming practices, in the northwest Carpathians, E. crus-galli has spread from lowland (<200 m altitude) to highland (>400 m altitude) areas. We collected seed samples from local populations lying at a distance of approximately 5 km from each other and lined up along transects following the flows of two rivers. The rivers first flow through the valleys separated by mountain ridges and eventually flow into a common lowland. After ripening, the seeds of all populations were germinated at 25 C under long-day conditions. Only the seeds of some lowland populations germinated up to 75%. The frequency of germinated seeds decreased as the altitude where the population was collected increased, and above 200 m above sea level, germination was mostly zero. We then studied the phenological and morphological differentiation of plants from the original (lowland) and recently occupied (highland) areas. Seeds of the lowest and the highest localities lying on the transect of each river were sown in a common garden experiment. In plants from the highland localities, heading and seed dispersal were earlier, while tiller height and tiller mass were lower than in plants from the lowland localities. Seed mass produced per tiller in the lowland and highland plants was similar, and as a result, highland plants allocated a larger proportion of body mass to seed production than did lowland plants. Echinochloa crus-galli populations from highland localities thus produce their progeny earlier and at a lower energy cost than populations from lowland localities. The plasticity of phenological characters likely facilitated adaptation during E. crus-galli spread from lowlands to highlands. Similar adaptations in plant phenology may contribute to the spread of E. crus-galli in other geographic areas.