An evaluation was performed of the potential use of AFLP markers to reveal polymorphisms among Lolium perenne plants with different degrees of kinship. Radioactive and fluorescent detection techniques were applied. The use of a fluorescent detection approach contributed greatly to the speed and ease of conducting and interpreting the AFLP patterns. The great discriminative power of AFLP markers and their capacity to represent genetic relationships among ryegrass plants was shown. Despite the high polymorphic value of the AFLP markers, standard statistical tests could not differentiate between two gene pools derived from different breeding programmes. It proved also impossible to correlate fodder and turf phenotypes with AFLP distance data. A very important point revealed by our data is the high degree of genetic diversity within commercial ryegrass varieties. Our findings are relevant to any outcrossing crop with a breeding strategy based on the production of synthetic populations.