The use of the 5S rDNA IGS region for DNA barcoding and molecular taxonomy of two Ukrainian species of the genus Tulipa L.

Abstract

Taxonomic identification of closely related representatives of the genus Tulipa L. is complicated by the convergent similarity of morphological characters, interspecific hybridization, polyploidy and asexual reproduction. Among the tulips in the flora of Ukraine, the most problematic is the status of taxa of the complex species T. sylvestris L. Aim. To examine the organization and evolution of the nuclear 5S rDNA IGS region for four closely related Tulipa taxa and to evaluate the possibility of its use in the molecular taxonomy of this plant group. Methods. PCR amplification, cloning and sequencing of 5S rDNA repeat units, bioinformatic and phylogenetic analysis. Results. In this work, 5S rDNA repeats were sequenced for the species T. graniticola and T. ophiophylla, which are considered to be synonyms of T. sylvestris L. by many authors. We also assembled the main ribotypes from full-genome libraries of short Illumina reads for T. patens, and T. saxatilis, the species related to T. sylvestris. A comparative analysis of the structure of 5S rDNA sequences and a phylogenetic analysis based on the IGS region were carried out. Conclusions. A high level of 5S rDNA IGS variability has been demonstrated for representatives of the subgenus Eriostemones of the genus Tulipa. The use of 5S rDNA IGS for phylogenetic analysis enabled the clear delimitation of the closely related species T. saxatilis, T. patens, and the complex species T. sylvestris, as well as the delineation of the taxa with the controversial status, T. graniticola and T. ophiophylla. Therefore, the use of this region of the nuclear genome is a promising approach to the molecular taxonomy of the genus Tulipa.