Hyalomma scupense, H. excavatum and H. marginatum are globally important tick species, as well as in Turkey, both in terms of human and animal health. The genome sizes of these tick species were determined in this study for the first time. From flow cytometric measurements, diploid genome sizes of female and male H. scupensewere found to be 2.13 pg and 1.75 pg, respectively, while H. excavatum were 2.21 pg and 1.94 pg, and H. marginatum were 2.48 pg and 1.98 pg, respectively. Differences in diploid genome size indicate X chromosome size of females and males in these ticks because they have an XX:XO sex determination system. Thus, it was estimated that the X chromosome of H. scupense, H. excavatum, and H. marginatum may be composed of as much as 0.38 pg, 0.27 pg, and 0.50 pg of DNA, respectively. These findings indicate suitability of these three species for genome sequencing due to the relatively small size of their genomes compared with other tick species.