The genus Rubus belongs to the Rosaceae family and is comprised of 600-800 species distributed worldwide. Understanding the genetic relationships and genetic structure in Rubus species is important for enabling efficient management, conservation, characterization and utilization of the species. However, as a minor crop, genetic research foundation was limited to explore genetic diversity and relationships in Rubus species. The present study shows the results of application SSR markers that were developed from SSR-enriched libraries of the one Rubus species (Rubus coreanus Mique.) in our previous study. We used 34 polymorphic microsatellite markers to analysis of genetic diversity within the Rubus species, including redraspberry, blackraspberry, blackberry and mountainberry. All the 34 SSR primers pairs produced 483 polymorphic and reproducible amplification fragments. The largest number of alleles per primer pair was confirmed at GB-RC-167, GB-RC-100, GB-RC-076 and GB-RC-245, which contained 26, 25, 23 and 21, respectively. An average value of polymorphic information contents (PIC) were 0.74 with a range of 0.36 to 0.92. Population structure and phylogenetic analyses showed that all Rubus species formed three largely distinct clusters, which were confirmed by principal coordinate analysis (PCoA). We obtained the results that the developed SSR markers showed a substantial degree of genetic diversity in the various Rubus species distributed in Korea.