Molecular Insights into the Centaurea Calocephala Complex (Compositae) from the Balkans - Does Phylogeny Match Systematics?

Groups of recent speciation are characterized by high levels of introgression and gene flow, which often confounds delimitation of species on a DNA basis. We analyzed nuclear DNA sequences (ETS spacer and the AGT1 gene) obtained from a large sample of the C. calocephala complex from the Balkan clade of Centaurea sect. Acrocentron (Compositae, Cardueae-Centaureinae) together with a wide representation of other species from the section. Our main goals were to verify the monophyly of the complex as currently defined and to examine the possible presence of introgression and gene flow. Within the complex, species are well-delimited from a morphological point of view and probably originated by allopatric speciation in the Balkan Peninsula. Our results confirm that the Balkan¿Eurasian complex is a natural group, but the Centaurea calocephala complex shows a very complicated pattern and its phylogeny is not resolved. Our hypothesis suggests that altitudinal shifts in the transits from glacial to interglacial periods caused successive hybridization events, which are very evident from the DNA networks, between taxa not currently sympatric. As a result, confirmation of interspecific boundaries using molecular markers is extremely complicated.This research was funded by Serbian Ministry of Education, Science and Technological Development, grant no. 451-03-68/2020-14/200178, and the Catalan Government (“Ajuts a grups consolidats” 2017-SGR1116)1. Introduction 2. Materials and Methods 2.1. Plant Material 2.2. DNA Extraction, Amplification, Cloning, and Sequencing 2.3. Phylogenetic Analyses 2.4. Chromosome Number 3. Results 3.1. Phylogenetic Analysis 3.2. Network Analyses 3.3. Chromosome Numbers 4. Discussion 4.1. Utility of Nuclear-Ribosomal ETS and Low-Copy AGT1 in Acrocentron 4.2. Phylogeny of the Balkan Clade: Introgression and Gene Flow 5. Conclusions Supplementary Materials Author Contributions Funding Reference