Abstract: A simplified AFLP method, based on methylation-sensitive Alw44I restriction endonuclease, has been developed and evaluated for fingerprinting 15 wheat cultivars. The selected germplasms represented groups of spring and winter wheats with and without the 1BL.1RS translocation. Ten selective primers yielded 57 markers, including 19 polymorphic bands. Three markers (15.8%) were specific to wheat carrying the 1BL.1RS translocation, thus conflicting with the frequency expected by random marker distribution (2.4%), and suggesting qualitative differences in DNA methylation among winter wheat cultivars with the 1BL.1RS translocation. Mean Dice's similarities ranged from 0.85 to 0.99, thus all cultivars could be identified by the banding profile. Winter wheat cultivars, with and without the 1BL.1RS chromosome, were slightly more similar to one another (0.959) than spring wheat cultivars (0.952). Five (9%) specific markers were obtained from cultivars Sicco, Cheyenne, Fenman, Disponent and Chinese Spring.