Polymorphism of 5S rDNA intergenic spacer in some Gentiana species
Aim. The aim was to determine the nucleotide sequence of non-transcribed spacer (NTS) of 5S rRNA genes in some Gentiana species of the Ukrainian flora and to study the organization of this region. Methods. DNA was isolated from leaf tissue. DNA isolation, PCR-analysis, and agarose gel electrophoresis were performed according to standard procedures. Isolated PCR-fragments were cloned and sequenced. DNA sequences were processed and aligned in Unipro UGENE software. Results. The size and nucleotide sequence of 5S rDNA NTS were determined for G. acaulis, G. lutea, and G. asclepiadea. On the basis of alignment and comparative analysis of 5S rDNA NTS of these and additional 11 gentians found in GenBank, two groups of species were identified, which differed significantly in the organization of this region. The first group included three East-Asian species of section Pneumonanthe, and the second included 7 species of section Calathianae, mainly from Europe, and three species studied here. The revealed peculiarities of the structure indicate the independent evolution of the NTS region in identified groups of the species. Conclusions. Nucleotide sequence of 5S rDNA NTS region was determined for three Gentiana species of Ukrainian flora. The features of the organization of the 5S rDNA NTS region are identified in European and Asian species of gentians.
Keywords: Gentiana, non-transcribed spacer, 5S rRNA genes, sequencing.
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