Summary: | <i>Aegilops columnaris</i> Zhuk. is tetraploid grass species (2n = 4x = 28, U<sup>c</sup>U<sup>c</sup>X<sup>c</sup>X<sup>c</sup>) closely related to <i>Ae. neglecta</i> and growing in Western Asia and a western part of the Fertile Crescent. Genetic diversity of <i>Ae. columnaris</i> was assessed using C-banding, FISH, nuclear and chloroplast (cp) DNA analyses, and gliadin electrophoresis. Cytogenetically <i>Ae. columnaris</i> was subdivided into two groups, C-I and C-II, showing different karyotype structure, C-banding, and FISH patterns. C-I group was more similar to <i>Ae. neglecta</i>. All types of markers revealed significant heterogeneity in C-II group, although group C-I was also polymorphic. Two chromosomal groups were consistent with plastogroups identified in a current study based on sequencing of three chloroplast intergenic spacer regions. The similarity of group C-I of <i>Ae. columnaris</i> with <i>Ae. neglecta</i> and their distinctness from C-II indicate that divergence of the C-I group was associated with minor genome modifications. Group C-II could emerge from C-I relatively recently, probably due to introgression from another <i>Aegilops</i> species followed by a reorganization of the parental genomes. Most C-II accessions were collected from a very narrow geographic region, and they might originate from a common ancestor. We suggest that the C-II group is at the initial stage of species divergence and undergoing an extensive speciation process.
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