Introduction Cytotaxonomy is a branch of cytogenetics in which karyological features are systematically evaluated for evolutionary purposes (Siljak-Yakovlev and Peruzzi, 2012). Chromosomes, especially plant chromosomes, are useful materials in nearly any type of cytogenetic research (Guerra, 2012). Because the genetic information of an organism is transmitted through its chromosomes, any changes in numbers (e.g., polyploid or diploid) and structures (e.g., inversion, deletion, or translocation) contribute significantly to plant evolution and speciation; however, to interpret the evolutionary history of a group, the number of chromosomes alone is not enough (Weiss-Schneeweiss and Schneeweiss, 2003) and karyomorphology might supply additional information. In some cases, ecological and morphological data might not be sufficient for analyzing the line of descent among the species. In these situations, cytotaxonomic relationships along with molecular data might be more influential in the analyses (Venora et al., 2008). The karyotype reveals phenotypic appearance in terms of number, size, arm ratio, centromere position, and other basic characteristics of chromosomes (Levin, 2002). Centaurea, which belongs to the tribe Cardueae, is a large genus with approximately 250 species (Susanna and Garcia-Jacas, 2007) and the highest rate of endemism, with 112 endemics among the total 181 species in Turkey (Uysal, 2012). Cyanus, a subgenus, is represented by approximately 25 species worldwide (Hellwig, 2004). According to recent definitions of Centaurea (Susanna and Garcia-Jacas, 2007), the following 3 subgenera are included: Acrocentron, Centaurea, and Cyanus. Sister relationships of Cyanus and Centaurea are firmly established, but the connections between these subgenera and Acrocentron are unclear (Susanna and Garcia-Jacas, 2009). Based on current molecular studies, for the aims of our research, Cyanus is considered a subgenus. Taxonomically, Cyanus was first described as a genus by Miller (1754). The group was subsequently reassigned as a section of Centaurea by de Candolle (1838) and this was widely accepted by taxonomists (Bentham, 1873; Boissier, 1875; Wagenitz, 1975). It is now generally accepted that Cyanus is a subgenus (Hilpold et al., 2014) or, rarely, a group (Wagenitz and Hellwig, 1996; Garcia-Jacas et al., 2001) within Centaurea; however, some authors still assert that it is a genus (Greuter, 2003; Bancheva and Greilhuber, 2006). The Cyanus group is distributed across central and southern Europe, North Africa, Anatolia, and the Caucasus, and some species have spread to Iran and Afghanistan (Boršić et al., 2011). The floret colors are blue or purplish blue in this group, with a few exceptions of taxa that are cream or Research Article This work is licensed under a Creative Commons Attribution 4.0 International License.