We successfully generated clear amplified fragment length polymorphism (AFLP) profiles of Anemone coronaria using only a single restriction enzyme (EcoR I). Four primer combinations generated 571 bands in 37 anemone accessions with an average 142.75 bands per primer combination. Polymorphism among the 37 samples was very high (98.15%). This high polymorphism showed that the one-enzyme AFLP method is effective for the assessment of genetic diversity as well as cultivar differentiation of anemone. We observed 26 bands which differentiated between fingerprints derived from leaf and corm DNA of eight plant individuals. However, the low percentage of such bands showed that anemone corms can also be used to evaluate anemone germplasm using AFLP markers.