Marina V. Protopopova

Anemone altaica Fisch. ex C. A. Mey., a component of the tertiary boreo-nemoral vegetation complex, exhibits a disjunct distribution from European Russia to Central China. The Khamar-Daban Ridge, extending along Lake Baikal’s southern coast, has served as a refugium preserving mesophilic forest remnants in South Siberia since the Pleistocene. This study aimed to elucidate the phylogenetic relationships and historical biogeography of A. altaica within the Khamar-Daban refugium using plastid DNA markers (trnL + trnL-trnF). Phylogenetic and mismatch distribution analysis revealed polyphyly (more specifically diphyly) among A. altaica lineages, suggesting past hybridization events with related species followed by backcrossing. Estimation of isolation by distance effect, spatial autocorrelation analysis, PCoA, and AMOVA indicated a clear spatial genetic structure for A. altaica on the Khamar-Daban Ridge. The most reliable geographical model suggests that during periods of Pleistocene cooling, A. altaica persisted in at least six microrefugia within the ridge. Populations associated with these microrefugia formed western, central, and eastern genetic supergroups with limited gene flow among them. Gene flow likely occurred more easily during glaciations or early interglacials when the subalpine zone shifted closer to Lake Baikal due to the depression of the snow boundary, allowing adjacent populations to intermingle along the glacial edges and terminal moraines in mountain forest belt.

Получение и первичная оценка фенотипа тополя берлинского, трансформированного геном AtGA20ox1 Получение и первичная оценка фенотипа тополя берлинского, трансформированного геном AtGA20ox1

Increasing the rate of biomass gain is one of the most important areas in the breeding of woody plants. However, the application of classical breeding approaches to woody plants is significantly limited due to the long breeding cycles of many species. The development of genetic engineering and genome editing technologies has made it possible to improve tree traits in a relatively short time. Genes for the biosynthesis of phytohormones are often the target of genetic manipulations aimed at accelerating plant growth. Gibberellin 20-oxidase is a key enzyme responsible for the active production of gibberellins in plants and is, therefore, a preferred target for genetic manipulation to increase growth rate. This paper presents an original study on the production of Populus × berolinensis K. Koch, transformed by gene encoding gibberellin 20-oxidase from Arabidopsis thaliana (L.) Heynh. (AtGA20ox1), and the initial evaluation of the phenotypic effects of the transformation. The main phenotypic manifestations of the transformation were a pronounced elongation of the stem due to an increase in the size of the internodes, a slight thinning of the stem, as well as lengthening and narrowing of the leaves. Our results showed that AtGA20ox1 gene overexpression in Berlin poplar leads to an acceleration of its growth by at least three times under in vitro conditions compared to control values. The negative effects of transformation, expressed in weak rooting or a high frequency of apical necrosis and observed in some transgenic strains at the initial stages of selection, do not appear in the three final selected strains.

Первая достоверная находка Galium odoratum (L.) Scop. и новые местонахождения Galium paradoxum Maxim. в Республике Бурятия

The rapid advancement of genetic technologies has made it possible to modify various plants through both genetic transformation and gene editing techniques. Poplar, with its rapid in vitro growth and regeneration enabling high rates of micropropagation, has emerged as a model system for the genetic transformation of woody plants. In this study, Populus × berolinensis K. Koch. (Berlin poplar) was chosen as the model organism due to its narrow leaves and spindle-shaped crown, which make it highly suitable for in vitro manipulations. Various protocols for the Agrobacterium-mediated transformation of poplar species have been developed to date. However, the genetic transformation procedures are often constrained by the complexity of the nutrient media used for plant regeneration and growth, which could potentially be simplified. Our study presents a cheaper, simplified, and relatively fast protocol for the Agrobacterium-mediated transformation of Berlin poplar. The protocol involved using internode sections without axillary buds as explants, which were co-cultivated in 10 µL droplets of bacterial suspension directly on the surface of a solid agar-based medium without rinsing and sterile paper drying after inoculation. We used only one regeneration Murashige and Skoogbased medium supplemented with BA (0.2 mg·L⁻¹), TDZ (0.02 mg·L⁻¹), and NAA (0.01 mg·L⁻¹). Acetosyringone was not used as an induction agent for vir genes during the genetic transformation. Applying our protocol and using the binary plasmid pBI121 carrying the nptII selective and uidA reporter genes, we obtained the six transgenic lines of poplar. Transgenesis was confirmed through a PCR-based screening of kanamycin-selected regenerants for the presence of both mentioned genes, Sanger sequencing, and tests for detecting the maintained activity of both genes. The transformation efficiency, considering the 100 explants taken originally, was 6%.

Waldsteinia is a small plant genus inhabiting the temperate regions of the Northern Hemisphere. According to the latest revisions, Waldsteinia is included in Geum. We have obtained a phylogenetic reconstruction based on the nuclear (ITS) and plastid (trnL-trnF) DNA to understand the phylogenetic structure of Waldsteinia and its relationships with other taxa of Geum s.l. Phylogenetic analysis based on the joint ITS + trnL-trnF dataset demonstrated Waldsteinia monophyly. The phylogenetic relationships of Waldsteinia species were better explained by their geographical distribution than their morphology. Hence, Euro-Siberian, Northeast Asian, and North American phylogeographic groups were distinguished, with East Asia having been suggested as the place of Waldsteinia origin. Considering the incongruence in W. geoides (a type species) position on the plastid and nuclear DNA trees, together with the discrepancy between the species morphology and its location on the plastid DNA tree, a hybrid origin was suggested for this species. Despite the fact that the position of W. maximowicziana is still not fully resolved, we support the point of view that claims it should be separated from the W. ternata aggregate (traditionally including W. trifolia, W. ternata s.str., and W. maximowicziana) and considered a separate species. The American W. doniana, W. fragarioides, and W. lobata belong to a single maternal lineage, but the observed genetic differences are too small to serve as a convincing argument for species segregation, so their relationships still remain unresolved.

Fritillaria Tourn. ex L. is a genus of Liliaceae including a little more than 150 species occurring in the temperate Holarctic. Fritillaria sonnikovae Shaulo & Erst is the most recently described Siberian species in the genus. In the affinity section of the F. sonnikovae diagnosis, only F. dagana Turcz. and F. roylei Hook. are mentioned. Our study is an original attempt to shed light on the F. sonnikovae origin and its evolutionary relationships with other Fritillaria using nuclear (ITS) and plastid (matK + rps16 + trnH-psbA) DNA markers. Our results showed that F. sonnikovae together with F. dagana and F. maximowiczii Freyn belongs to the North Asian lineage of the Liliorhiza subgenus and produced no evidence supporting relationship between F. sonnikovae and F. roylei. Monophyly of Fritillaria sonnikovae was not reliably confirmed in our study since its close affinity with F. maximowiczii was demonstrated by phylogenetic analysis and morphology. Fritillaria dagana was shown to be a sister to the F. maximowiczii + F. sonnikovae group. Most authors of the present study suggest considering F. sonnikovae as a synonym for F. maximowiczii. In this view, F. sonnikovae may be considered a narrow endemic and one of the light-perianth morphs of F. maximowiczii, which has emerged in the Western Sayan and remained there as a tertiary relict.

Eranthis Salisb. (Ranunculaceae) is a herbaceous plant genus, including few species disjunctively distributed throughout the temperate zone from Southeastern Europe to Eastern Asia. Until recently, only Eranthis sibirica DC. was known in South Siberia, being considered endemic and tertiary relict. Not long ago, Eranthis tanhoensis Erst was also described in Siberia. We report here a reconstruction of the phylogenetic relationships between the Siberian Eranthis species based on nuclear (ITS) and plastid (trnL + trnL-trnF + trnH-psbA) DNA. The phylogeographic structure of Siberian Eranthis is distinguished by the presence of the two “eastern” and “western” supergroups, which most likely formed as a result of disjunction caused by active mountain uplifts during the late Neogene–early Quaternary and subsequent progressive Pleistocene cooling. The eastern supergroup combines lineage I, containing populations from the eastern Khamar-Daban Ridge, the Eastern Sayan Mountains, and the Tannu-Ola Ridge, and lineage II containing western Khamar-Daban populations. The western supergroup includes only lineage III, containing Western Sayan populations. Our data clearly show that E. tanhoensis is nested in the E. sibirica clade, thereby indicating that its description as a separate species is unjustified, as it compromises the monophyletic status of E. sibirica. Therefore, we suggest here to consider E. tanhoensis as a synonym of E. sibirica.

The methods of biological species identification using nucleotide sequences of short genome regions (DNA barcoding) are actively developed. The universal DNA barcode for plants remains to be discovered, and one of the leading candidates is the plastid gene of the large subunit of ribulose-bisphosphate carboxylase gene (rbcL). In our study, we estimated the part of rbcL gene as a possible marker for molecular identification of Rhaponticum carthamoides (Willd.) Iljin. Due to its officinal properties, the species is susceptible to uncontrolled and illegal harvesting from natural populations. Today, the species needs to be protected and therefore is included into the Red Data Books of the Russian Federation and certain regions. The study was carried out using plants from the natural populations sampled from the Khamar-Daban Ridge (South Siberia) and considering now as Rh. carthamoides var. chamarense (Peschkova) O S Zhirova. It was shown that rbcL gene can be used to identify Rh. carthamoides at least from the populations of the Khamar-Daban Ridge using a fragment of the maximum length or its 3’ region. Apparently, the 5’ region of the gene (rbcLa) most often used as DNA barcode for plants may be of lesser importance for Rh. carthamoides. The rbcL gene sequences can be also used for the development of approaches for Rh. carthamoides identification in the medicinal preparations and products containing dried tissues to prevent their falsification and illegal harvesting of this species. The combination of rbcL gene with additional markers seems to be highly desirable to create effective DNA barcodes for Rhaponticum species.