Results
The ITS sequences (ITS1, complete 5.8S rDNA gene, ITS2 and a small part of 26S rDNA gene) in the dataset of Tulipa species ranged from 644 to 657 bp. The alignment of the ingroup included 45 ambiguous positions, including the outgroup another 7 positions were ambiguous. 52 positions were potentially informative. Parsimony analysis of ITS sequences included 132 potentially informative indels, MP tree lengths were 213 bp, with RI of 0.945, CI of 0.855, composite index 0.897 and 60.1 % G + C content (Table 2). The sequence lengths of ITS1 were 229-233 bp, 5.8S rDNA were 162-166 bp, ITS2 were 225-231 bp and 26S rDNA (partial) were 26 bp, while the alignments were found to be 238 bp for ITS1, 166 bp for 5.8S rDNA, 233 bp for ITS2 and 26 bp for 26S rDNA. The average G + C content for ITS1 was 59.5%, while for ITS2 it was 60.8%, respectively. Tulip samples showed an average of 141 and 143 conserved sites for ITS1 and ITS2, respectively. ITS1 contained 24 and ITS2 contained 27 potentially parsimony informative sites, respectively. The partial sequences of the 26S rDNA gene included only 26 alignment characters without indels. The average G + C content in 5.8S rDNA was 57.0% and 76.9% in 26S rDNA. The number of potentially parsimony informative sites was 1 for the 5.8S rDNA gene and none for the 26S rDNA gene, respectively.
The trnL-trnF sequences of Tulipa species in the dataset ranged from 720 to 775 bp in length. The complete alignment (including outgroup) contained 86 ambiguous positions, within the in-group 51 positions were ambiguous. Analysed sequences showed ten potentially informative characters, 47 potentially informative indels, MP tree length was 58 bp, CI was 0.769, RI 0.928, composite index 0.897 and the G + C content was 31.2% (Table 2). The trnL-trnF (trnL631-692 bp, trnF 57-64 bp and IGS 25 bp for each sequence, respectively) alignment included 717 bp for trnL, 64 bp fortrnF and 25 bp for the IGS. G + C content was 30.8% fortrnL, 44.5% for trnF and 11.8% for the IGS, respectively. Ten potentially informative characters were counted for this region.
The combined ITS + trnL-trnF sequences ranged from 1377 to 1430 pb in length. The complete alignment including the outgroup showed 93 ambiguous positions, within the ingroup 91 positions were ambiguous. The alignment included 1229 conserved sites, 241 variable sites, 62 potentially informative characters and 179 potentially informative indels (Table 2). The average G+C content was 44.5%, the CI was 0.840, RI was 0.940, and the composite index was 0.890.
Table 2. Data set and parsimony-based tree characteristics for ITS andtrnL-trnF analyses.