80 accessions of sugarcane germplasm (32 parental species, 48 cultivars or clones) were analyzed by random amplify microsatellite polymorphisms(RAMP) markers.4 pairs of primers were selected from 30 pairs, and used to establish the RAMP fingerprints of the germplasm, which amplified 84 polymorphic bands, and were accounting for 91.7%.80 accessions were clustered with RAMP data using UPGMA method. The genetic similarity coefficients among 80 germplasm ranged from 0.433 to 0.988 with a mean of 0.710. Based on the difference of the similarity coefficients,the sugarcane germplasm could be distinguished among intergenus(Saccharum and E.arundinaceus),wild species(S. spontaneum, S. barberi ,S. sinense and S. robustum),cultivated species( S. officinarum).cultivated species and cultivars derived from interspecific hybridization. The similarity between cultivars and parental species S. officinarum were the highest, and followed by S. barberi,S. sinense,S. robustum, S. spontaneum and E. arundinaceus in the order from high to low. Clustering based on RAMP showed that the groupings of most cultivars were corresponded to their pedigrees in which a given cultivar and one of its parents was always clustered into the same group. In the meantime, we found some specific DNA fragments of S. officinarum, wild species of Saccharum spp. and E.arundinaceus ,which could be recovered in their derived cultivars with different probabilities.