Amplified fragment length polymorphism (AFLP) analysis was used to assess the genetic relationships among 46 accessions of sorghum (Sorghum bicolor (L.) Moench) collected from the north-western, western and central parts of Ethiopia as well as South Africa. Six AFLP primer combinations were used for the analysis of DNA fragment amplification. Dice similarity coefficients were calculated and a dendrogram was constructed following the UPGMA method of cluster analysis. A total of 186 fragments were amplified of which 78 (43.10%) were polymorphic. The number of polymorphic fragments amplified per primer combination varied from 9 to 21. Genetic polymorphism present among sorghum accessions was low, as evidenced by the high level of similarity in the AFLP marker profiles of different sorghum accessions. Pair-wise genetic similarity coefficients ranged from 0.87 to 0.99, with an average of 0.92. This indicates low levels of genetic diversity among tested sorghum accessions. The landraces were genetically very similar, while the differences between landraces, the Ethiopian cultivars and the South African cultivars were relatively higher. Genetic similarity within the South African and the Ethiopian cultivars was very high. Almost all accessions clustered according to the geographical origin. Results of this study indicate that the landraces were related, and this most likely results from the exchange of seeds between farmers in the collection regions, although no duplications were found in the material.Key words: Amplified fragment length polymorphism (AFLP), accessions, genetic diversity, sorghum.