Twelve rice (Oryza sativa L.) cultivars and breeding lines developed by the Rice Breeding Program at the National Institute of Agriculture Research (INIA),Quilamapu Regional ResearchCenter, were genetically characterized by amplified fragment length polymorphism (AFLP). Sixteen of 21 primer combinations evaluated were informative, generating a total of 667 amplicons, 94 of them polymorphic (14.4%), with a range between 9 to 33% of polymorphism per primer combination. This indicates that the material handled by the breeding program at INIA has low genetic diversity in comparison to wild germplasm of the species, or even compared to the material managed by other rice breeding programs. The detected polymorphic amplicons were used to prepare a dendrogram of the genetic distances, detecting four differentiable groups that only partially coincide with the information available in their pedigrees. In spite of the low genetic diversity detected, it was determined that with the use of three AFLP+3 combinations (PE1G/PM1I, PE1H/PM1A and PE1G/PM1H), it is possible to differentiate among the cultivars studied.