Sesame is queen of oil seed crops and widely cultivated in Asia and Africa. The aim of this study was to develop a mini sub core set representing the diverse germplasm of sesame and to assess the genetic diversity, population structure and phylogenetic relationship of the resulted sub core set to be used in whole genome resequencing platform. One hundred twelve accessions out of 277 accessions were selected by the PowerCore program. A total of 155 alleles were captured from the 158 alleles detected in the primary core population, and rare alleles and specific alleles were also maintained in the sub core set accessions representing almost 100% of the primary core population. Among the sub core set accessions, four sub populations were observed with some admixture accessions. Although the genetic diversity of Pop-1 which includes most accessions from Korea is relatively lower than that of other three sub populations, it can maintain maximum number of accessions in the sub core set with the same percentage as in the primary core set probably because of the specific features of these accessions. Based on this framework of genetically defined populations, the effective use and conservation management of Sesamum indicum for crop improvement might be possible.