Bolu'da bulunan bazı Crocus L. ( çiğdem) taksonlarının rDNA ıts ve cpDNA trnL-f bölgelerinin dizi analizleriyle çeşitliliğinin tanımlanması

In this preliminary study, taxonomic boundaries of Crocus abantensis T. Baytop & B. Mathew, C. ancyrensis (Herb.) Maw, C. biflorus Mill. subsp. pulchricolor (Herb.) B. Mathew, C. olivieri J. Gay subsp. olivieri, C. speciosus M. Bieb. subsp. speciosus and C. x paulineae Pasche & Kerndorff sampled from Bolu province were determined by using the base sequences of rDNA ITS and cpDNA trnL-F regions. The genetic status of the hybrid taxon, C. x paulineae was examined and its parents were determined in terms of molecular data. Existence of some other possible hybrids taxa between C. abantensis and C. olivieri subsp. olivieri were also investigated based on these molecular data in the studied area. Additionally, this study was carried out to determine the experimental protocols being involved in a phylogeographical research on Crocus taxa in Turkey. In moleculer analyses, gDNA isolated from the samples, and then ITS and trnL-F regions were amplified using the ITS-4, ITS-5A, trnL-e, and trnL-f primers in a PCR apparatus. After editing of base sequences by using CodenCode Aligner program, the multiple alignments of the sequences were made with the CLUSTAL W packed program. Consequently, all the phylogenetic trees based on the sequences were created by using the PAUP* 4.0b10 program. The phylogenetic trees of ITS and trnL-F produced slightly different results containing more or less similar clades. This result was interpreted that it occurred because of having short, 230-300 bp sequences of trnL-F in regard to ITS base sequences (680-700 bp). Therefore, the trees based on the ITS sequences were mostly considered during the interpretations of phylogenies of taxa. The results of distance and character based methods showed a quite positive congruencies between the phylogenetic trees such as parsimony, bootstrap, UPGMA, and NJ analyses. It was found that there were no differences between the groups of taxa obtained from this study and the groups of taxa obtained from the phenetic analyses by some other researchers. Finally, genetic status of the C. x paulineae taxon supported to be a hybrid of C. abantensis x C. ancyrensis species in terms of ITS and trnL-F analyses. Presence of other hybrid taxa between C. abantensis and C. olivieri subsp. olivieri could not be phylogenetically found in the research area.