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Cytogenetic and Phylogenetic studies of Diploid and Polyploid Members of Tribe Anemoninae (Ranunculaceae)

The ancestry, phylogenetic differentiation and systematic classification of worldwide distributed Anemone genus have been debated for years. In this paper eleven Anemone, three Pulsatilla species and Hepatica nobilis were subjected to detailed karyotype analysis with the aim to obtain new cytogenetic data that will contribute to the karyotype evolutionary studies of the tribe Anemoninae. Obtained results are interpreted in a phylogenetic context established from intergenic nontranscribed spacer (NTS) of the 5S rDNA and internal transcribed spacer (ITS) of the 35S rDNA. One to three 35S and one to three 5S rDNA loci are present in diploid and polyploid taxa. The 35S rDNA loci are located terminally on the short arm of acrocentric chromosomes, while for 5S rDNA there is no preferential chromosomal position as it exhibits terminal, subterminal, interstitial or pericentromeric position, and is located either on acrocentric or metacentric chromosomes. The karyotype of hexaploid A. baldensis (2n = 6x = 48) is presented for the first time and A. sylvestris is proposed as one of its putative parental species. The chromosome fusion/translocation is proposed as key mechanism involved in the reduction of the basic chromosome number from 8 in the Anemone subgenus to 7 in the Anemonidium subgenus. The cytogenetic data obtained in this study are mainly supported by ITS and NTS phylogeny. Diversification of anemones was accompanied with the large reduction of heterochromatin, from the Mediterranean anemones that have large amount of heterochromatin to the New World anemones without any detectable heterochromatic blocks.

35S and 5S rDNA; 5S rDNA spacer phylogeny; Anemone; fluorescence in situ hybridization; karyotype evolution; polyploidy; Pulsatilla.

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