Tatiana E. Samatadze

In karyotypes of Calendula officinalis L. (Asteraceae) (2n = 32) from various ecological and geographical places of growth, intraspecific polymorphism was studied based on patterns of chromosomal localization of molecular cytogenetic markers: C/DAPI bands and also 45S and 5S rDNA clusters. In all calendula samples, variable C/DAPI bands were found in the proximal-median regions of chromosomes. FISH revealed similar localization on chromosomes of the main 45S and 5S rDNA clusters: 45S rDNA clusters were localized on chromosomes 1 and 9, and 5S rDNA cluster was detected on the long arm of chromosome 10. Additionally, minor sites of 45S rDNA were identified in the karyotypes of calendula samples from Crimea and Belarus. Intraspecific variability in number and location of the minor 45S rDNA sites was revealed on chromosomes of C. officinalis plants from different places of growth.

Amaranthus cruentus L. and Amaranthus hypochondriacus L. are valuable and promising food crops for multi-purpose use that are distributed worldwide in temperate, subtropical, and tropical zones. However, their karyotypes and genomic relationships still remain insufficiently studied. For the first time, a comparative repeatome analysis of A. cruentus and A. hypochondriacus was performed based on the available NGS data; bioinformatic analyses using RepeatExplorer/TAREAN pipelines; and chromosome FISH mapping of 45S rDNA, 5S rDNA, and the most abundant satellite DNAs. In the repeatomes of these species, interspecific variations in the amount of Ty3/Gypsy and Ty1/Copia retroelements, DNA transposons, ribosomal, and satellite DNA were detected. In the repeatomes of both species, shared satDNAs with high sequence similarity were identified. The chromosome distribution patterns of four effective molecular markers, 45S rDNA, 5S rDNA, AmC4, and AmC9, allowed us to identify all chromosome pairs in the species karyotypes, construct unique karyograms of A. cruentus and A. hypochondriacus, and confirm the close relationship between their genomes. These results are important for comparative karyotypic studies within the genus Amaranthus. Our findings demonstrated that cytogenomic analyses might provide important data on genomic relationships within Amaranthus and increase knowledge on genome organization in these valuable crops.

The cosmopolitan genus Hedysarum L. (Fabaceae) is divided into sections Hedysarum, Stracheya, and Multicaulia. This genus includes many valuable medicinal, melliferous, and forage species. The species taxonomy and genome relationships within the sections are still unclear. We examined intra- and interspecific diversity in the section (sect.) Hedysarum based on repeatome analyses using NGS data, bioinformatic technologies, and chromosome FISH mapping of 35S rDNA, 5S rDNA, and the identified satellite DNA families (satDNAs). A comparison of repeatomes of H. alpinum, H. theinum, and H. flavescens revealed differences in their composition. However, similarity in sequences of most satDNAs indicated a close relationship between genomes within sect. Hedysarum. New effective satDNA chromosomal markers were detected, which is important for karyotype analyses within Hedysarum. Intra- and interspecific variability in the chromosomal distribution patterns of the studied markers were revealed, and species karyograms were constructed. These results provided new insight into the karyotype structures and genomic diversity within sect. Hedysarum, clarified the systematic position of H. sachalinense and H. arcticum, and confirmed the distant genomic relationships between species from sections Hedysarum and Multicaulia. Our findings are important for further comparative genome studies within the genus Hedysarum.

Background. Methods of artificial polyploidization are not used enough in medicinal crop production. The use of colchicine causes obvious morphological changes in the obtained P. caeruleum polyploids and changes in the productivity of raw materials and seeds. The research was conducted at the All-Russian Scientific Research Institute of Medicinal Plants, Moscow region. Purpose. Based on comparative biological, morphological and economically valuable features of diploid and tetraploid forms of the medicinal plant Polemonium caeruleum L. to identify and evaluate the obtained polyploid forms as a initial material in breeding. Materials and methods. Polyploid plants of P. caeruleum were obtained for the first time in 2019 [6, 14]. In 2021-2022 (in the 2nd and 3rd year of vegetation) in generation C2, the biological, morphological and economically valuable signs of colchicine-induced tetraploid plants were studied. The control (initial form) is the cultivated variety "Lazur". Results. For the first time, an assessment of the economic, biological and morphological parameters of the initial material of the medicinal culture of P. caeruleum obtained on the basis of colchicine-induced tetraploid plants is given. The economic and biological indicators of the initial material are higher than in the control: in terms of the yield of raw materials (rhizomes with roots) - by 21-52% and in terms of seed yield - by 44-53%. In terms of the amount of triterpene saponins in terms of β-escin, tetraploid plants met the requirements of the pharmacopoeia article (at least 10 %). The highest content of biologically active substances exceeded the control by 14-15 %. Biometric and descriptive data were also obtained on the height, size and shape of inflorescences and leaves, stem thickness, color of plant parts, including the severity of anthocyanin staining. These distinctive morphological characteristics are easily identified and can be used for a preliminary assessment of the putative polyploid forms of P. caeruleum. Tetraploid plants have numerous shoots with anthocyanin-colored stems of greater thickness than those of control plants. The study of morphological features showed that colchicine-induced plants are stunted – up to 86% lower than control plants. The inflorescences are also smaller in size, but more dense and branched than in the control. The leaves of tetraploids are larger, have a dark green color of the leaf blade. The excess over the control in the length of the stem leaves was 12-32 %, in the width of the leaves – 7-25 %. Fewer seeds were formed, but they were larger. The weight of 1000 seeds increased by 5.5 % compared to the control. Conclusion. As a result of research on a complex of biomorphological and economic and biological signs, promising numbers of P. caeruleum have been identified for further breeding work. The results of the study are relevant for the creation of new highly productive varieties of the P. caeruleum medicinal culture.

The genus Hedysarum L. (Fabaceae) includes about 200 species of annual and perennial herbs distributed in Asia, Europe, North Africa, and North America. Many species of this genus are valuable medicinal, melliferous, and forage resources. In this review, we consider the taxonomic history of the genus Hedysarum, the chromosomal organization of the species from the sections Hedysarum and Multicaulia, as well as phylogenetic relationships between these sections. According to morphological, genetic, and phylogenetic data, the genus Hedysarum is divided into three main sections: Hedysarum (= syn. Gamotion), Multicaulia, and Stracheya. In species of this genus, two basic chromosome numbers, x = 7 (section Hedysarum) and x = 8 (sections Multicaulia and Stracheya), were determined. The systematic positions of some species within the sections are still uncertain due to their morphological similarities. The patterns of distribution of molecular chromosomal markers (45S rDNA, 5S rDNA, and different satellite DNAs) in karyotypes of various Hedysarum species made it possible to determine their ploidy status and also specify genomic relationships within the sections Hedysarum and Multicaulia. Recent molecular phylogenetic studies clarified significantly the taxonomy and evolutionary development of the genus Hedysarum.

Artificial hybrids between cultivated Avena species and wild Avena macrostachya that possess genes for resistance to biotic and abiotic stresses can be important for oat breeding. For the first time, a comprehensive study of genomes of artificial fertile hybrids Avena sativa × Avena macrostachya and their parental species was carried out based on the chromosome FISH mapping of satellite DNA sequences (satDNAs) and also analysis of intragenomic polymorphism in the 18S–ITS1–5.8S rDNA region, using NGS data. Chromosome distribution patterns of marker satDNAs allowed us to identify all chromosomes in the studied karyotypes, determine their subgenomic affiliation, and detect several chromosome rearrangements. Based on the obtained cytogenomic data, we revealed differences between two A. macrostachya subgenomes and demonstrated that only one of them was inherited in the studied octoploid hybrids. Ribotype analyses showed that the second major ribotype of A. macrostachya was species-specific and was not represented in rDNA pools of the octoploids, which could be related to the allopolyploid origin of this species. Our results indicate that the use of marker satDNAs in cytogenomic studies can provide important data on genomic relationships within Avena allopolyploid species and hybrids, and also expand the potential for interspecific crosses for breeding.

Впервые с использованием флуоресцентной гибридизации in situ проведено сравнительное исследование хромосомной организации кариотипов четырех видов календулы - Calendula L. (Asteraceae): С. officinalis, C. stellata, C. tripterocarpa и C. arvensis. Определены числа хромосом в кариотипах: C. stellatа (2n =2х = 14); C. officinalis (2n = 4х = 32); C. tripterocarpa (2n = 2х = 30); C. arvensis (2n = 4х = 44), и выявлена характерная специфическая локализация кластеров генов 45S и 5S рДНК для каждого вида. В кариотипе C. officinalis выявлен дополнительный полиморфный минорный сигнал гибридизации 45S рДНК, который располагался медианно в коротком плече одной хромосомы. Установлено, что рисунок распределения основных сайтов рибосомных генов на хромосомах в кариотипе C. stellatа (2n = 2х = 14) и C. tripterocarpa (2n = 2х = 30) свидетельствует в пользу гибридного происхождения C. arvensis (2n = 4х = 44) в результате возможной интрогрессивной гибридизации в процессе эволюции.

The taxonomically challenging genus Calendula L. (Asteraceae) includes lots of medicinal species characterized by their high morphological and karyological variability. For the first time, a repeatome analysis of a valuable medicinal plant Calendula officinalis L. was carried out using high-throughput genome DNA sequencing and RepeatExplorer/TAREAN pipelines. The FISH-based visualization of the 45S rDNA, 5S rDNA, and satellite DNAs of C. officinalis was performed on the chromosomes of C. officinalis, C. stellata Cav., C. tripterocarpa Rupr., and C. arvensis L. Three satellite DNAs were demonstrated to be new molecular chromosome markers to study the karyotype structure. Karyograms of the studied species were constructed, their ploidy status was specified, and their relationships were clarified. Our results showed that the C. officinalis karyotype differed from the karyotypes of the other three species, indicating its separate position in the Calendula phylogeny. However, the presence of common repeats revealed in the genomes of all the studied species could be related to their common origin. Our findings demonstrated that C. stellata contributed its genome to allotetraploid C. tripterocarpa, and C. arvensis is an allohexaploid hybrid between C. stellata and C. tripterocarpa. At the same time, further karyotype studies of various Calendula species are required to clarify the pathways of chromosomal reorganization that occurred during speciation.

Aim. Comparative study of the source material of four populations of Sedum roseum (L.) Scop. belonging to different ecological and geographical zones of origin and assessment of their suitability for cultivation in the conditions of the non‐chernozem zone of the Russian Federation.Materials and methods. The following populations were studied: the cultivated population of VILAR (Altai, 1980), the cultivated population of the Botanical Garden of Pitirim Sorokin Syktyvkar State University (Syktyvkar, Komi Republic), an introduced population from Kirovsk, N. A. Avrorin Polar‐Alpine Botanical Garden‐Institute and a cultivated population of Innsbruck Botanical Garden, Austria.Results. Morphological description of the samples was carried out and the productivity of raw materials and seeds and the main biological and karyological features of the populations studied were determined. The results of made it possible to identify Rhodiola populations characterized by high winter hardiness (A and D) and low winter hardiness (B and C). A high productivity of roots and rhizomes in the fourth year of life in populations C and A (30–31 g) was established, and in terms of the content of active substances (phenylpropanoids) in population B (Komi) – 5.4%. The numbers of chromosomes (2n=22) were determined and their morphometric analysis was carried out and formulas of karyotypes of the Rhodiola populations studied were compiled. Chromosomal analysis using C/DAPI‐differential staining showed that no chromosomal abnormalities were detected in the karyotypes, which indicates their stability.Conclusion. Morphological variability indicates the ecological plasticity of the populations studied, which indicates the prospects for their field cultivation.

Background. Currently, it is quite relevant in breeding to create varieties with high yields of raw materials and the content of biologically active compounds. This work aim is to identify promising forms of P. caeruleum L. is for further inclusion in the breeding process based on biomorphological, anatomical, cytogenetic studies and the bioproductivity of colchicine-induced and wild-growing forms in comparison with the Lazur variety. Materials and methods. To obtain polyploids in 2018, the seeds of the P. caeruleum L. the Lazur variety were germinated in Petri dishes on filter paper at room temperature. When the roots reached the length of the seed, they were placed in different concentrations of colchicine solution 0.05 %, 0.1 %, 0.2 % and 0.52 % for 12 and 24 hours, then washed in running water for three hours. Results. The article for the first time presents data on a comparative study of three forms of Polemonium caeruleum L. in terms of morphological characteristics, microscopy of the leaf epidermis and cytogenetic analysis. It was established that the raw materials productivity of the underground part of the plants tetraploid forms in the C1 generation significantly exceeded the diploid and wild forms of plants by 32 %, as well as the yield of seeds by 22 %. In the C2 generation, the yield of raw materials (rhizomes with roots) in tetraploid plants increases in relation to the control by 56 %, seeds by 34 %. Quantitative determination of the amount of triterpene saponins in terms of β-escin in the plant material of three forms of P. caeruleum L. showed an excess of this indicator in the colchicine form by 24 % in the grass and by 36 % in rhizomes with roots, in the wild-growing form - by 15 % in the aerial part and decrease in the underground part by 9 % in relation to the control. It was found that plants of colchicine and wild forms differ in the presence of larger stomata compared to the control. Based on the pattern of DAPI-bandin and the distribution of 45S rDNA and 5S rDNA genes, a generalized species idiogram of blue cyanosis was constructed, taking into account polymorphic variants of DAPI-bandin patterns and indicating the location of the sites of 45S rDNA and 5S rDNA genes. The cytogenetic stability of the resulting tetraploid form was revealed. Conclusion. The article presents comparative studies on morphological characteristics, bioproductivity, microscopy of the leaf epidermis and cytogenetic analysis of the tetraploid form (colchicine) of P. caeruleum L. in comparison with the variety Lazur (diploid form) and the wild form of P. caeruleum L. plants.