DNA barcoding is a molecular approach for species identification and germplasm conservation in the Origanum genus. This study proposes a DNA- based method to accurately identify the numerous species within this diverse genus. Four chloroplast genes (matK, rbcL, rpoB, trnL) and a non-coding sequence (psbA) were analyzed. Among the applied barcoding loci, matK exhibited the longest aligned sequence length, followed by rpoB, rbcL, psbA-trnH, and trnL. The rbcL gene showed the highest number of informative sites, while psbA-trnH displayed the highest proportion of variable sites within the Origa- num genus. Phylogenetic analysis indicated that rbcL, rpoB, trnL, and psbA sequences were more effective than matK for identifying the Origanum genus, spe- cies, and subspecies. Combining multiple sequences improved accuracy. This study provides a molecular reference for herbal product identification and valu- able tools for species identification and germplasm conservation within the Lamiaceae family.