Species of Gentiana section Cruciata are highly similar in their morphological and anatomical characteristics, which complicates their taxonomic delimitation and phylogenetic interpretation. Molecular markers increasingly complement traditional morphology-based approaches, and the intergenic spacer of the 5S rRNA gene cluster (5S rDNA IGS) represents a promising candidate for such analyses. In this study, genetic variation among 10 species within section Cruciata was investigated using sequences of the 5S rDNA IGS region, and its potential utility for DNA barcoding and phylogenetic analysis was evaluated. Analysis of 38 cloned PCR products revealed a structural organization of the IGS region typical of other Gentiana species, including conserved regulatory elements, and a high level of nucleotide sequence similarity (91.2-99.5%). Interspecific polymorphism was comparable to intragenomic variation. Sequence alignment and clustering distinguished two groups of species that differed in their IGS nucleotide sequences. The resulting taxonomic grouping is generally consistent with previously reported phylogenetic relationships inferred from other molecular markers. The high similarity of IGS sequences among species of section Cruciata suggests their relatively recent divergence. Notably, G. cruciata, despite its migration from Asia to Europe, retained IGS region characteristics typical of the section.

Aim. The 5S rDNA intergenic spacer (IGS) region is a convenient tool for studying genome evolution and systematics of higher plants. The aim was to investigate the nucleotide sequence of 5S rDNA in some species of the sections Cruciata and Chondrophyllae, as well as to study the peculiarities of this region organisation by comparative analysis with other Gentiana species. Methods. Polymerase chain reaction, cloning, sequencing, bioinformatic analysis. Results. The cloned IGS regions of G. laciniata, G. cruciata and G. dahurica were sequenced and analysed. Typical regulatory elements were found in the regions of terminator and external promoter of 5S rRNA gene. Based on the comparative analysis, two groups of species were distinguished that differ significantly in the organisation of this region. One includes G. asclepiadea and species of the sections Gentiana, Ciminalis and Calathianae, and the other includes species of the sections Cruciata and Pneumonanthe. G. laciniata (sect. Chondrophyllae) differs in the IGS organization from species of both groups. Conclusions. The results of taxon grouping are generally consistent with the genus systematics and indicate that 5S rDNA IGS region can be a convenient tool for studying evolution and systematics of the genus.

Aim.To study molecular genetic aspects of stress tolerance in the extremophile plant D. antar ctica by exploring ICE transcription factors involved in cold stress response, regulation of stomatal development and flowering time.Methods.The DaICE genes were assembled from SRA reads homologous to orthologous genes used as a reference.The predicted genes and proteins were analyzed with bioinformatics methods.Gene expression was analyzed using the RNA-seq data.Results.Two variants of the DaICE1 gene were found: a variant that encodes a full-length protein and a variant with a large deletion, which led to the loss of the N-terminus and part of the bHLH domain in the protein product.Two variants of CDS differed by four single-nucleotide polymorphisms, three of which were synonymous, with more dissimilar promoter sequences were found for the DaICE2.The organization of promoters and the intronexon structure of the genes were studied.For the predicted TF proteins, physico-chemical properties were determined, nuclear localization and the possibility of post-translational modifications were shown, phylogenetic analysis was carried out, conservative motifs were found, 3D structure was predicted.Gene expression was detected in most or all plant tissues and increased expression of DaICE1 variants was shown in the plants from natural conditions compared to laboratory ones.Conclusions.DaICE1 and DaICE2 are the candidate regulators of the CBF gene expression under cold stress in D. antarctica.

The intergenic spacer (IGS) of 5S ribosomal RNA genes (5S rDNA) (that are present in the genome of all living organisms) is characterized by a high variability, due to which it is a convenient and widely used object for clarifying the issues of genome evolution, population genetics, systematics, etc. The aim of this work was to study the IGS of 5S rDNA of the Antarctic pearlwort Colobanthus quitensis. The nucleotide sequence was determined by the methods of molecular genetic analysis, and molecular organization of the C. quitensis 5S rDNA IGS was studied. It was demonstrated that it contains the regulatory elements typical for other vascular plants. The existence of at least two classes of 5S rDNA repeats in the individual genome (that differ significantly in the length and nucleotide sequence of IGS) was established. In addition, based on differences in the length and nucleotide sequence, two subclasses of repeats with a long IGS and three subclasses with a short IGS were identified. A comparison of 5S rDNA IGS sequences in C. quitensis and Silene latifolia Poir. (a member of another section of the Caryophyllaceae family) demonstrated significant differences in the structure of a spacer region of 5S rRNA genes (except for its parts containing regulatory elements). In general, data obtained indicate a significant level of intragenomic polymorphism of C. quitensis 5S rDNA IGS.

This review presents the results of multi-year efforts for the development and integral investigation of the collection of Deschampsia antarctica e. Desv. plants originated from the maritime Antarctic. To create the collection, we have optimized the procedures and conditions for germination of seeds and generation of aseptic plants in vitro together with their subsequent clonal propagation, long-term in vitro culture, and acclimatization of plants to pot culture, and finally produced fertile plants able to set viable seeds. The collection of plants cultivated in vitro and in pots is consisted of clones of 22 different genotypes originated from eight island populations from the Argentine Islands - Kyiv Peninsula region. Some of these clones have been successfully cultured for more than ten years and were found to be genetically stable over this period. The collection was shown to be heterogeneous in a number of variables. The morphological and anatomical characteristics of the plants, as well as the data of karyological, molecular cytogenetic, molecular genetics, and biochemical analysis are presented. The created collection of D. antarctica plants can provide a valuable plant material with well-studied characteristics grown under controlled conditions to perform experimental research on the mechanisms for plant adaptation and resistance to various abiotic factors of Antarctic environment. Furthermore, development of the collection and its comprehensive characterization form the basis for further biotechnological developments focused on the use of this Antarctic plant as a source of biologically active substances, which can potentially be used for therapeutic and prophylactic purposes.

Aim.To characterize a new hormone-independent strain К-27М of R. serpentina tissue culture.Methods.Plant tissue culture, biochemical and cytological analysis, statistical methods.Results.The strain is maintained on the specially designed simple-composition hormone-free 10C medium.The maximum biomass yield was at day 69 of subculture (693.5 g/l of live biomass, 43.0 g/l of dry biomass), the weight of cell biomass increased 15-18 times over a subculture period.The maximum content of indole alkaloids was observed from 88 to 108 days of subculture.The dry biomass contained 4.0% of total indole alkaloids, 1.64% of ajmaline-like alkaloids, 0.789% of ajmaline, 0.337% of vomilenine, and 0.006% of each yohimbine and reserpine.The strain is a heterogeneous mixoploid cell population with a modal class of 22 to 33 chromosomes (2n = 22), which characterized by significant cell and nuclear polymorphism.Conclusions.The K-27M strain of R. serpentina cultured tissue is a prospective producer of indole alkaloids.

The organization of promoters of DREB2B TF ortholog genes involved in the response to abiotic stresses was studied in D. antarctica E. Desv. extremophile plant and 12 more grass species with different cold and drought resistance. The evolutionary distances between the sequences of promoters were 0.621 on average, those between coding sequences (including introns) were 0.442, and the values of nucleotide diversity (π) for these regions were 0.410 and 0.274, respectively. Clustering of sequences corresponded in general to the accepted systematic division of the Poaceae family into subfamilies. Fifty-four cis -elements involved in the response to abiotic and biotic stresses, light, hormones (particularly, abscisic acid, auxin, methyl jasmonate, ethylene, gibberellin, and salicylic acid), and tissue-specific cis -elements were identified. Most of the cis -elements were associated with the response to abiotic stress that is consistent with data on DREB2B TF functions. Except for single peculiarities, representatives of different grass subfamilies and D. antarctica were similar in the total set of cis -elements in the DREB2B promoter that indicates the similarity of the regulation of expression of this gene and its potential functions in the response to stress in the studied species.

We used inter-simple sequence repeats and inter-retrotransposon amplified polymorphism markers to assess genetic variation in Deschampsia antarctica populations in the context of its uneven distribution in the northern and central maritime Antarctic. Genetic diversity and population structure and differentiation were assessed in nine populations from geographically isolated population groups of D. antarctica, including the South Shetland Islands, Anvers Island and the Argentine Islands regions. In total, 265 amplified DNA fragments were scored, of which 220 (83.0%) were polymorphic. The total sample showed low genetic diversity (unbiased expected heterozygosity = 0.081 and Shannon diversity index = 0.115) and high population differentiation (molecular variance among populations = 0.659). We also found a trend toward a decrease in genetic diversity and an increase in population differentiation toward the southern edge of the species range. Principal coordinates analysis of polymerase chain reaction data and Bayesian population structure analysis showed three main clusters, which included plants originating from three spatially isolated population groups. The unweighted pair group method with arithmetic mean clustering of populations based on Nei’s genetic distances was mainly in agreement with this pattern. Testing of isolation by distance using the Mantel test demonstrated a significant correlation between genetic and ln-transformed geographical distance (r = 0.703). The data obtained indicate that the geographically isolated D. antarctica populations in the maritime Antarctic might form genetic clusters within the total range.

Aim. The aim of the study was to determine the quantitative and qualitative content of phenolic compounds and flavonoids in Deschampsia antarctica E. Desv. tissue cultures obtained from plants originating from different islands of the maritime Antarctic. Methods. In vitro tissue culture, Folin-Ciocalteu method, spectrophotometry, HPLC analysis. Results. The quantitative content of phenolic compounds and flavonoids in D. antarctica tissue cultures obtained from plants of six genotypes (DAR12, DAR13, G/D12-2a, Y66, R30 and L57) was determined. The highest content of phenolic compounds (4.46 and 3.75 mg/g) was found in tissue cultures obtained from root and leaf explants of plant genotype L57. The highest amount of flavonoids (7.17 mg/g) was accumulated in G/D12-2a tissue culture of root origin. The content of the studied biologically active compounds (BACs) did not change with increasing number of subculture generations (from passage 10 to 19). HPLC analysis showed that in D. antarctica tissue cultures, a shift in the biosynthesis of BACs occurred towards the synthesis of more polar metabolites compared to explant donor plants. Conclusions. It was found that the transition of cells to undifferentiated growth affected the content of BACs, the amount of which decreased 2–5 times simultaneously with a significant change in their profile. This provided a basis for further biochemical studies, as well as for careful selection of tissue culture of D. antarctica to use it as a potential source of BACs.

Aim. The study was aimed at cloning and analysis of molecular organization of 5S rDNA intergenic spacer (IGS) in two Gentiana species of Ukrainian flora, G. pneumonanthe L. and G. punctata L. Methods. 5S rDNA IGS sequence was amplified using polymerase chain reaction (PCR) with a pair of primers specific for the gene coding region. The produced PCR products were fractionated by gel-electrophoresis, isolated, ligated into plasmid pUC18, cloned into E. coli, and then sequenced. Nucleotide sequences were aligned using the Muscle algorithm and analyzed in the Unipro UGENE software. Results. The intergenic spacer region of the 5S rRNA genes was cloned and sequenced for two Gentiana species of Ukrainian flora, G. pneumonanthe and G. punctata. Based on the analysis of the alignment of the IGS sequences of five Gentiana species from three sections, some features of molecular organization of IGS of 5S rRNA genes in the studied species were established. In particular, motifs typical for other angiosperm families were identified, such as conservative oligo-dT motif at the IGS 3'-end that served as a transcription termination site and AT-rich region preceding the coding region of 5S rRNA gene. However, in the region of transcription initiation, conservative GC-element in position -13 is changed to AC. Conclusions. The interspecific variation of molecular organization of 5S rDNA IGS was identified among Gentiana species that can be used to clarify the phylogenetic relationships between members of this genus.Keywords: Gentiana species, 5S rDNA intergenic spacer, molecular organization, phylogeny.

The paper presents data on the total phenolic and flavonoid content in Deschampsia antarctica È. Desv.plants collected from natural habitats, plants cultured in vitro, regenerated plants, and plants cultivated in a growth chamber.It was found that the shoots (aerial parts) had higher phenolic and flavonoid contents compared to the roots.The largest amount of these substances was found in wild plants from Great Yalour Island.The content of phenolics and flavonoids in plants cultured in vitro was generally comparable to that in plants collected from natural habitats except for some clones.HPLC analysis revealed five main metabolites present in similar ratios in the studied samples of wild and in vitro plants.Minor substances varied slightly in different extracts, however their presence did not depend on plant growth conditions.No significant qualitative differences in HPLC profiles were found between the wild and in vitro plants.The studied samples did not contain quercetin, kaempferol and luteolin, whereas orientin was found in all studied samples of D. antarctica.

The use of microalgae for the economic needs and the commercial goals determines the areas of the scientific researches that will make it possible to increase their productivity. It is also important to direct the metabolism of the algae to the activating of certain synthetic processes in order to obtain the desired compounds. The metals and non-metals, entering into the cell, have a high biochemical activity. These elements modify the metabolic reactions in general and the metabolic reactions related to the functioning of the genome of microalgae cells. Aim. The aim was to study the genetic polymorphism of Chlorella vulgaris under the action of such trace elements as selenium, zinc and chromium in order to optimize the methods of algae cultivation and the obtaining of the beneficial compounds. Methods. The hydrobiological methods of algae cultivation, DNA isolation method by Rogers S. and Bendich A. (1985), PCR-analysis with ISSR (inter simple sequence repeats)- and IRAP-markers (inter-retransposon amplified polymorphism) have been used. Results. For all samples of C. vulgaris 109 DNA-fragments were obtained and 42 of them were polymorphic (38.5%). Jacquard distances (DJ) between the samples of C. vulgaris culture (cultures are grown on the media with different elements compositions and control (standard conditions) were 0.232 (only selenite), 0.206 (selenite and zinc) and 0.300 (selenite and chromium). Conclusions. Probably the genetic modifications of C. vulgaris cells are caused by the additional introduction of the microelements into the culture medium. The genetic polymorphism of the algae grown on media with various trace elements and their combinations was like the genetic polymorphism of the unicellular green algae grown in the natural conditions. It indicates the absence of significant genotoxic effects of the trace elements and high metabolic and genetic plasticity of algal culture.

Retrotransposons make up a significant part of plant genome and are probably the most dynamic part of it, so they play a significant role in the generation of genetic variation. In particular, their activation can lead to structural reorganization of genome and changes in genome size, the emergence of novel genetic and phenotypic variants, as well as changes in gene expression, thus providing the raw material for adaptation and evolution. This review summarizes literature data on the activation of LTR-retrotransposons of the superfamilies Ty1/Copia and Ty3/Gypsy during in vitro culture and under various abiotic and biotic stress conditions. Their structure, classification, and significance for the organization and functioning of plant genome are reviewed. The main mechanisms of activation of LTR-retrotransposons under stress conditions are explored, including changes in DNA methylation and interaction of stress-induced transcription factors with retrotransposon promoters due to the presence of specific binding sites and other regulatory elements. The review also discusses consequences of activation of retrotransposons and control of their activity by self-inactivation mechanisms and the epigenetic regulation of genome.Keywords: retrotransposons, Ty1/Copia, Ty3/Gypsy, in vitro culture, abiotic and biotic stress.

Cytogenetic analysis of D. antarctica plants from the Argentine Islands of the Maritime Antarctic region was performed. Chromosome number 2n = 26 was determined for most of the samples. New forms of chromosome polymorphism for the species were demonstrated for the first time. In particular, the plants from Darboux Island were found, to have some cells that contained one supernumerary B-chromosome along with 26 chromosomes of a regular set. The plants from Great Yalour Island were determined to be mixoploid – the number of chromosomes ranged from 13 to 39. The occurrence of new and unknown earlier karyotypic forms of D. antarctica might have resulted from increased genome instability due to the extreme environmental conditions in the Argentine Islands region.

Obesity is a widespread problem within modern society, serving to increase the risk of cardiovascular, metabolic, and neurodegenerative disorders. Peroxisome proliferator-activated receptor gamma (PPARγ) and PPARγ coactivator 1 α (PGC1α) play a key role in the regulation of cellular energy metabolism and is implicated in the pathology of these diseases. This study examined the association between polymorphisms of the PPARG and PPARGC1A genes and individual variability in weight loss in response to physical activity intervention. 39 obese Ukrainian women (44.4 ± 7.5 years, BMI > 30.0 kg/m2) undertook a 3-month fitness program whilst following a hypocaloric diet (~ 1500 cal). Anthropometric and biochemical measurements took place before and after the program. Single nucleotide polymorphisms within or near PPARG (n = 94) and PPARGC1A (n = 138) were identified and expression of PPARG mRNA was measured via reverse transcription and amplification. The association between DNA polymorphisms and exercise-induced weight loss, initial body mass, biochemistry and PPARG expression was determined using one-way analysis of variance (ANOVA). The present intervention induced significant fat loss in all participants (total fat: 40.3 ± 5.3 vs 36.4 ± 5.7%; P < 0.00001). Only one polymorphism (rs17650401 C/T) within the PPARGC1A gene was found to be associated with fat loss efficiency after correction for multiple testing, with T allele carriers showing the greatest reduction in body fat percentage (2.5-fold; P = 0.00013) compared to non-carriers. PPARGC1A (rs17650401) is associated with fat loss efficiency of the fitness program in obese women. Further studies are warranted to test whether this variation is associated with fat oxidation.

Habitat fragmentation is one of serious threats to biodiversity of nature in today's world. The present study of a typical steppe species Iris pumila L. (Iridaceae) has analyzed the impacts of geographical isolation and population size on genetic diversity and population structure in conditions of habitat fragmentation. The key indices of population genetic variability calculated from the ISSR markers data were on average as follows: Shannon diversity index (S) - 0.188; unbiased Nei's gene diversity (H-e) - 0.123; and the average measure of Jaccard's genetic distances between individuals within populations - 58.4%. Although the largest population had significantly higher values of S and He, the small and marginal populations also showed a comparable level of variation. Most of the genetic variation of I. pumila was distributed within the populations. A strong correlation was found between Nei's genetic distances and geographic distances between the populations. According to the Bayesian analysis, genetic structure of the populations was highly homogeneous; however, the presence of admixed genotypes indicated the possibility of gene flow between the populations at present.

Habitat fragmentation is one of serious threats to biodiversity of nature in today's world. The present study of a typical steppe species Iris pumila L. (Iridaceae) has analyzed the impacts of geographical isolation and population size on genetic diversity and population structure in conditions of habitat fragmentation. The key indices of population genetic variability calculated from the ISSR markers data were on average as follows: Shannon diversity index (S) – 0.188; unbiased Nei ’ s gene diversity (H e ) – 0.123; and the average measure of Jaccard ’ s genetic distances between individuals within populations – 58.4%. Although the largest population had significantly higher values of S and He, the small and marginal populations also showed a comparable level of variation. Most of the genetic variation of I. pumila was distributed within the populations. A strong correlation was found between Nei ’ s genetic distances and geographic distances between the populations. According to the Bayesian analysis, genetic structure of the populations was highly homogeneous; however, the presence of admixed genotypes indicated the possibility of gene flow between the populations at present.

Aim. The work was aimed at the development of conditions for introduction into in vitro culture of two species of irises, Iris attica and I. pseudopumila to obtain aseptic seedlings with subsequent reintroduction into natural environment, as well as at cytogenetic analysis of the obtained plants. Methods. In vitro seed germination and seedling cultivation. Cytogenetic analysis of cells of root meristem, determination of chromosome number and morphology in mitotic metaphase plates, anaphase analysis. Results. The plants of I. attica and I. pseudopumila were introduced in vitro. Aseptic seedlings were obtained, which were actively growing on MS/2 medium without phytohormones. The experiments on the adaptation of the plants to greenhouse conditions revealed the high survival rate for both species. The chromosome number 2n = 16 was established for the obtained plants of both I. attica and I. pseudopumila. Mixoploidy was detected in root meristem of some of the plants, the incidence of which was 10.9 % for I. pseudopumila and 30.4 % for I. attica. The frequency of cells with chromosomal rearrangements revealed by anaphase analysis in root meristem of I. pseudopumila seedlings was 2.6 %; in I. attica plants, chromosome aberrations were not detected. Conclusions. The plants of I. attica and I. pseudopumila were introduced into in vitro culture, aseptic seedlings were obtained, which showed a high survival level when adapted to greenhouse conditions. Chromosome number 2n = 16 was established for the obtained plants of both species. The root apical meristems of the seedlings were found to be mixoploid, with the incidence of mixoploidy in I. attica identified as three times higher than in I. pseudopumila plants.Keywords: I. attica Boiss. & Heldr., I. pseudopumila Tineo, aseptic seedlings, mixoploidy, anaphase aberration.

Aim. Identification of Ty1/Copia-like transposable elements in Deschampsia antarctica E Desv. in silico. Methods. Bioinformatic analysis of sequence read archives of D. antarctica genome and transcriptome contained in the GenBank database was conducted. The search was carried out using the rice Ty1/Copia TE Tos 17 as a reference sequence. Results. The search revealed the sequences of Ty1/Copia retrotransposons in the D. antarctica genome to show a high level of identity (up to 75% in the homologous regions) to Tos 17. The uneven distribution of the found reads along the reference sequence indicates the existence of a group of the sequences in the genome to contain elements typical of the family and have varying degrees of identity to Tos 17, with more conservative ones being represented in correspondingly greater numbers among the found reads. The presence of the reads identical to Tos 17 in transcriptome indicates that the TE sequences identified in genome have a certain background level of transcriptional activity. Conclusions. Transcriptionally active Ty1/Copia-like transposable elements were identified in silico in D. antarctica genome using methods of bioinformatics. The sequences found can be used to construct primers for PCR and to analyze the activity of the TE of this family in D. antarctica in further studies. Keywords: bioinformatic analysis, Deschampsia antarctica E Desv., transposable elements, Ty1/Copia.

Aim. The study was aimed at cytogenetic analysis of Iris pumila, I. attica, and I. pseudopumila, comparative study of the karyotypes of these species, as well as identification of putative origin of I. pumila karyotype. Methods. Cytogenetic analysis of root apical meristem, determination of chromosome number in mitotic metaphase plates, anaphase analysis. Results. The chromosome numbers observed were 2n=32 for I. pumila plants from different localities in Ukraine and 2n=16 for I. attica and I. pseudopumila plants from Greece and Italy, respectively. Some of the plants were mixoploids, the smallest proportion of mixoploids was in I. pseudopumila (10.9%) and the largest in I.pumila from all studied populations (60-80%). Anaphase analysis showed the presence of chromosomal aberrations in 2.6% of cells in roots of I. pseudopumila seedlings. The highest level of structural chromosomal aberrations (9.2%) was found in root apical meristem cells of I. pumila seedlings. Conclusions. The chromosome number was established as 2n=32 for I.pumila plants and 2n=16 for I. attica and I. pseudopumila plants. The high level of mixoploidy (60–80% of mixoploid plants) and anaphase chromosomal aberrations (up to 9.2%) was found in apical meristem of I. pumila seedlings. The amphidipiloid nature of I. pumila was established; the karyotype of the species could be formed as a result of a combination of chromosome sets from hypothetical ancestral species I. attica and I. pseudopumila. Keywords: Iris pumila L., Iris attica Boiss. & Heldr., Iris pseudopumila Tineo, chromosome number, amphidiploid, mixoploidy.