It is substantially important to know genetic structure of fish populations for long term fishery and with regards to conservation strategies. The present study was aimed to reveal the fundamental population genetic data for continuity and conservation of the Yellow barbell (Carasobarbus luteus) stocks, being considered as an economically important endemic fish species in Southeastern Anatolia Region. Therefore, a total number of 108 fish specimens were sampled from 9 localities of Kueik, Euphrates and Tigris river systems and subsequently, genetically analyzed based on mtDNA D-loop region sequencing. In total, 15 polymorphic nucloetide sites and 7 haplotypes were established during the analysis carried out. The mean haplotype (Hd) and nucleotide variation (π) were calculated as 0.373 and 0.00536. The highest values of haplotype and nucleotide variation was observed in Birecik (Hd=0,65152) and Adıyaman population (π =0,01094); respectively. In turns, none haplotype and nucleotide variation was observed in Kilis and Diyarbakır populations. Obtained FST values for examined populations ranged between-0.12389 and 0.15630. All FST values were found to be statistically insignificant for the populations within the same river and significant between some populations originated from different rivers. According to analysis of neutrality tests, Tajima's D values were statistically significant in 4 populations (Samsat, Bozova, Hilvan and Bismil). Obtained, under the present study information on genetic variation and population structure of C. luteus will be useful to plan an effective strategies for conservation and fishery supplementation of the species in the region.