Safflower is an important oilseed crop for the dry regions of the world. In the present study, we aimed to find out the seed oil content and composition of the wild relatives of safflower. For this purpose, 71 populations from 5 naturally growing Carthamus species were collected from Turkey: Carthamus dentatus (30 accessions), Carthamus lanatus (30 accessions), Carthamus persicus (2 accessions), Carthamus glaucus (6 accessions), and Carthamus tenuis (3 accessions). Our research covered all naturally growing safflower species in the flora of Turkey. The seeds belonging to 71 accessions were germinated in cabinets and transferred to vials in order to obtain seedlings and then sown in the field manually. After harvesting, the seed oil content (%) and fatty acid compositions were determined. The seed oil content ranged between 10.50% and 20.40% in all accessions. Thirteen fatty acids, mainly linoleic (58.8% to 82.6%), oleic (7.3% to 22.8%), and palmitic acids (4.8% to 8.8%) were determined. The fatty acid compositions were more or less similar in all investigated species; however, the seed oil contents were different. For the evaluation of fatty acid properties as a taxonomic character, cluster analysis was performed. The dendrogram based on the fatty acid ratios discriminated section Carthamus and section Atractylis of the genus Carthamus; however, the resulting tree was not compatible with the previously constructed phylogeny trees in detail.