Ungernia sewertzowii (US) and U. victoris (UV) are medicinal plants and sources of biologically active compounds for pharmaceutical needs. The leaves of US contain 0.29–0.81% sum of alkaloids with a predominance of lycorine, which is 0.04–0.46% in leaves and 0.15–0.38% in bulbs. Lycorine is used to treat acute and chronic bronchitis. The leaves of UV contain 0.27–0.71% sum of alkaloids with a predominance of galanthamine—0.13–1.15%. Galanthamine is used to treat mild-to-moderate dementia (Alzheimer’s disease). The natural populations of US and UV are in danger as sources of income for local people. To resolve this problem, two protocols for microclonal propagation were developed to replace natural raw materials with in vitro regenerated plants. Callusogenesis of US and UV was induced on Murashige and Skoog (MS) nutrient media with 2.4D (0.5 mg/L) in combination with BAP (0.5 mg/L), Kin (0.5 mg/L), or Zea (0.5 mg/L). Direct (for US) and indirect (for US and UN) organogenesis were observed on MS with BAP (0.5 mg/L) or Kin (0.5 mg/L) in combination with IAA (0.5 mg/L) or NAA (0.5 mg/L). Direct organogenesis resulted in 3–5 bulbs of US on one explant; indirect organogenesis resulted in up to 100–150 bulbs of US and UV on one explant within 6 months, or five to six subcultures after transferring the callus to the nutrient medium. The tissue cultures of US and UV were characterized by very low data on antioxidant activity based on IC50 values for DPPH and ABTS radical scavenging activities, whereas in vitro regenerated plants (leaves and bulbs) had higher data. We concluded that in vitro regenerated plants are valuable sources of lycorine and galanthamine, which allow the protection of the natural populations of these two species from extinction.

This study investigated the antioxidant potential of callus cultures derived from the endangered medicinal plant Ferula tadshikorum Pimenov (Apiaceae Lindl.), an endemic species in Central Asia. The plant’s long life cycle and overharvesting threaten its survival, necessitating the development of in vitro microclonal propagation protocols as a sustainable solution for bioactive compound production. Two propagation methods, indirect organogenesis and indirect somatic embryogenesis, were optimized using zygotic embryos as explants. Callus cultures at early stages, cultivated on Murashige and Skoog medium supplemented with 0.5 mg/l 2,4-dichlorophenoxyacetic acid, demonstrated antioxidant activity, with 2,2-diphenyl-1-picrylhydrazyl (DPPH) half-maximal inhibitory concentration (IC₅₀) values as low as 15.41 µg/mL, comparable to ascorbic acid (15.17 µg/mL), while the radical-scavenging activity in the 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) assay was even higher. Optimal bioactivity and extract yields were achieved on a Murashige and Skoog medium supplemented with 2.4-dichlorophenoxyacetic acid (0.5 mg/l) and Kinetin or Thidiazuron. Statistical analysis confirmed the influence of hormonal treatments on antioxidant properties. These findings highlight the dual potential of in vitro propagation for conserving F. tadshikorum and providing a sustainable source of bioactive compounds with significant antioxidant properties, supporting pharmaceutical applications. However, challenges in translating these findings into large-scale production or commercial applications must be addressed, including the optimization of protocols for industrial scalability and cost-effectiveness.

The genus Gagea exhibits high morphological diversity and complex taxonomy. In this study, we analyzed the phylogenetic relationships and morphological differentiation within Gagea using ITS sequences from 32 species across 9 sections. Bayesian analysis revealed 3 main clades, partially congruent with morphological traits. Our results highlight both the value of molecular data for species delimitation and the need for taxonomic revision due to cases of morphological convergence and incongruence. This study contributes to a deeper understanding of evolutionary patterns in Gagea.

Background: Flavonoids, which are bioactive compounds found in medicinal plant extracts, are recognized for their significant therapeutic properties. To efficiently isolate these compounds from fresh plant materials, it is essential to optimize extraction methods. The most widely used approach involves solvent extraction using solvents such as methanol, ethanol, acetone, and hexane. This study aimed to determine the optimal extraction methods for isolating phenolic compounds and flavonoids from callus and regenerating plants of two medicinal species of the genus Ferula cultivated in vitro with or without indole-3-butyric acid (IBA) and 6-benzylaminopurine (BAP). This study focuses on Ferula tadshikorum and Ferula sumbul, with extracts analyzed quantitatively using high performance liquid chromatography (HPLC). The results revealed the presence of flavonoids such as kaempferol, quercetin, p-coumaric acid, and cinnamic acid at varying concentrations across different solvents. Notably, extracts from F. sumbul plants grown with Murashige and Skoog (MS) medium containing IBA and BAP exhibited higher levels of p-coumaric acid (9.588 µg/mL) in methanol, and kaempferol (9.595 µg/mL) in hexane. For F. tadshikorum, ethanol extraction yielded a significant amount of p-coumaric acid (29.9 µg/mL). Quercetin and cinnamic acid were detected in minimal quantities in both extracts, whereas kaempferol was notably higher (2.61 µg/mL) than in the methanol extract (0.617 µg/mL). Objectives: The present study represents the first attempt to determine the phenolic compounds in regenerating plants obtained through in vitro methods. Methods: In vitro regenerated plants of F. tadshikorum and F. sumbul, cultured on MS basal medium supplemented with IBA (0.5 mg/mL) and BAP (0.5 mg/mL), were ground into a fine powder in liquid nitrogen and then dried in a drying oven at 50°C before chemical analyses. The bioactive compounds were extracted by soaking the powders in methanol, ethanol, and hexane overnight. The extracts were filtered and evaporated under a vacuum at 35 - 36°C. The resultant slurry was partitioned with equal volumes of ethyl acetate (EA) and water to remove excess polar compounds. The EA fraction was vacuum-dried, and the mass of the crude extract was recorded. The extract was subsequently dissolved in methanol, ethanol, and hexane to achieve a concentration of 1 mg/mL. Finally, the solution was filtered through a 0.45 µm PTFE filter (German Acrodisc 13 CR) before analysis by HPLC. Results: We then analyzed methanol, ethanol, and hexane extracts from in vitro F. tadshikorum and F. sumbul plants grown under various combinations of hormones and hormone-free conditions using HPLC. When comparing extracts prepared with 3 solvents from 2 different samples of the F. sumbul, plants obtained in a combination of IBA and BAP hormones showed a higher content of p-coumaric acid in methanol extraction (9.58 µg/mL) and a higher content of kaempferol in the hexane extraction (9.59 µg/mL). During the extraction of callus and plant regenerants from in vitro F. tadshikorum plants using methanol and ethanol, a higher content of p-coumaric acid (29.9 µg/mL) was observed in the ethanol extract than in methanol. Quercetin and cinnamic acid were detected in very low quantities in both the extracts. Notably, kaempferol content was higher (2.61 µg/mL) than in the methanol extract (0.617 µg/mL). Conclusions: This study focused on the declining wild medicinal herbs in natural populations, providing an opportunity for systematic in vitro propagation year-round. This approach aims to establish these herbs as alternative sources for extracting biologically active secondary metabolites such as flavonoids.

In vitro microclonal reproduction protocols have been developed for some high-value medicinal ferule species, for example, F. ferulaeoides (Steud.) Korov., F. assa-foetida L., F. gummosa Boiss., F. jaeschkeana Vatke, F. orientalis L. and Ferula sinkiangensis K. M. Shen. To date, there are no reports of reproduction of F. sumbul in vitro. These species are vulnerable or threatened with extinction due to low seed germination, the duration of the dormant period of seeds, poor regeneration in nature, overexploitation by humans, as well as the lack of organized cultivation, limited geographical range, etc. These factors lead to the threat of extinction of the listed species. The Institute of Botany of the Academy of Sciences of the Republic of Uzbekistan is conducting research on the development of a protocol for microclonal reproduction of two valuable medical species of the genus Ferula L. (Apiaceae Lindl.): F. tadshikorum Pimenov and F. sumbul (Kauffm.) Hook. f. within the framework of the A-FA-2021-146 project "Creation of technology for the organization and reproduction of medicinal plants by in vitro method" with a implementation period of 2021-2024. Within the framework of the project, a dissertation work is being prepared. F. sumbul - perennial, herbaceous. polycarpic. They could not germinate seeds in the laboratory. Therefore, mature seed germs were used as an explant. In all trea xplants (80-90%). only the consistency and color of the calluses differed. In order to proliferate somatic embryos, the embryogenic callus was passioned onto an MS nutrient medium that did not contain phytohormones. Some embryos were transferred to a nutrient medium of MS with IBA and BAP in low concentrations, as well as to nutrient media containing only phytohormone 2,4 D. In order to form regenerating plants by indirect somatic embryogenesis, the sixth and seventh passaging of somatic embryos was carried out on a hormone-free nutrient medium. It was at the stage 6-7 of passage that the maturation of somatic embryos and the development of regenerating plants in 42.1% of embryos were observed. The formation of regenerating plants for species of the genus ferula was observed on the 5th month after introduction into culture in vitro, while for nutmeg ferula, the formation of regenerating plants was noted on the 7th month after introduction into culture in vitro.

This study describes tissue culture protocols for plantlet regeneration through indirect somatic embryogenesis and indirect organogenesis of Dorema microcarpum Korov. and Dorema sabulosum Litv. (Apiaceae) for the first time. The highest percentage of callusogenesis (90%) was obtained by using 2,4 -D (0.2 and 0.5 mg/l), followed by 2,4 -D (0.5 mg/l) + TDZ (0.5 mg/l) and 2,4-D (0.5 m g/l) + Kin (0.5 mg/l). Additionally, IAA (0.2 mg/l) + BAP (0.5 mg/l) showed the high percentage of callusogenesis (60–70%) for both Dorema species. In D. microcarpum , the highest percentage of callusogenesis was obtained from 2,4-D (0.5 mg/l) + TDZ (0.5 mg/l), while the highest percentage of embryogenesis was achieved with 2,4-D (0.5 mg/l) + Kin (0.5 mg/l). Hypocotyl explants yielded the highest percentage of embryogenesis. For somatic embryo maturation, 2,4-D had to be removed from a nutrient medium. Embryo maturation occurs after the embryo was transferred to a hormone-free medium. For D. sabulosum , morphogenesis occured through indirectly organogenesis, and the most optimal combination for the process of hemogenesis was IAA (0.5 mg/l) + BAP (1.0 mg/l). In vitro propagation of Dorema species in the future will become the basis for continuous year-round propagation of rare, endemic and medicinal species of this genus by biotechnological methods, and will become an alternative to obtaining biologically active compounds of medicinal plants.