Background : Ixeris dentata is the perennial herbaceous plant in the Ixeris genera within the Compositae family. The whole plant has been used traditionally as herbal medicine. It is widely distributed in South Korea and the genetic difference among the plants harvested from different regions may differ due to the disparity in cultivation climate. Therefore, this research was performed to discriminate the I. dentata that are collected from four locations in South Korea based on sequence analysis of nrDNA-ITS region. Methods and Results : Genomic DNA was extracted from I. dentata obtained from Goesan, Dangjin, Yangpyeong, and Chuncheon. I. stonolifera was included for a comparison of genetic distance with I. dentata. PCR amplification was performed by using an universal barcode nrDNA-ITS primer for DNA barcoding. After sequencing, the data was aligned using ClustalW multiple alignment tool in BioEdit version 7.2.5 software. SNP and phylogenetic analysis were conducted with the MEGA7 program. Phylogenetic analysis was presented with Neighbor-joining method using the K2P model. Statistical significance are evaluated using bootstrap (1,000 replicates). PCR products were amplified with about 800bp length for the ITS1-4 sequences of all samples. Nineteen SNPs were detected within a 578bp fragment of the aligned sequences. The mean GC content was 51.88% for ITS1-4 sequences of them. The interspecific genetic distance between I. dentata and I. stonolifera was 0.029%. The highest region-specific distance was confirmed to 0.010% between the plants from Dangjin with Goesan and Chuncheon group. Meanwhile, the mean intraspecific distance among the plants from Yangpyeong was 0.002%. Conclusion : In the phylogenetic analysis, the plants from Goesan and Chuncheon were placed within the same clade, while the plants from Dangjin formed independent clade. On the other hand, the plants from Yangpyeong were not grouped into one clade followed by the intraspecies variation. In conclusion, the data from the research will be useful for the regional identification of Dangjin, even if it is required to perform some additional researches.