Allium microdictyon Prokh. is a rare, endemic species possessing good taste qualities and listed in the Red Book of Kazakhstan; therefore, it is subject to anthropogenic impact (food gathering, grazing, logging, fires, etc.), which leads to a substantial reduction of its area. The aim of the study was to develop a protocol for microclonal propagation of A. microdictyon. Mature seeds of A. microdictyon collected from natural habitats in the Kazakhstani Altai were used as explants. Optimization of seed sterilization methods, selection of growth regulators for inducing adventitious shoot formation and microclonal propagation, and optimization of conditions for adaptation of regenerants to ex vitro conditions were carried out. Surface sterilization of seeds with 70% EtOH and 0.01% HgCl₂ is optimal for obtaining sterile and viable A. microdictyon seedlings. Sterile seedlings obtained in vitro on ½ Murashige and Skoog medium supplemented with 10 mg L⁻¹ gibberellic acid and 0.1 mg L⁻¹ indole-3-acetic acid (IAA) were used as a source for obtaining micropropagation cultures. Induction of adventitious organogenesis of A. microdictyon was effective on media containing 0.5 mg L⁻¹ 6-benzylaminopurine (BAP) and 1.5–2 mg L⁻¹ zeatin. On these variants, leaf conglomerates consisting of abundantly overgrown thin leaves were formed. The effect of 0.2 mg L⁻¹ indole-3-butyric acid (IBA) on further development of organogenesis and formation of microbulbs in A. microdictyon was shown in comparison with IAA, NAA, and PAC. Regenerated A. microdictyon plants were adapted to ex vitro conditions and resumed growth after 16–20 weeks of relative dormancy. The developed micropropagation protocol can be used to preserve germplasm and propagate for subsequent restoration of A. microdictyon populations in natural habitats.

The analysis of genetic diversity in natural populations of valuable medicinal plant species experiencing overexploitation is a key aspect of their natural conservation strategy. Mobile genetic elements and other interspersed repeats, which are major components of eukaryotic genomes, serve as effective tools for studying plant biodiversity and variability. The genetic diversity of four valuable medicinal plant Rhodiola rosea L. populations was investigated using the inter-repeat amplified PCR method with inter-priming binding sites (iPBSs) for genome profiling. At the interpopulation level, unique amplicons characteristic of specific R. rosea populations were identified. Molecular variance analysis revealed that the biodiversity of R. rosea populations in the Kazakh Altai region is 56% attributed to interpopulation differences and 44% to intrapopulation differences. It was shown that populations located in favorable environmental conditions have greater genetic diversity compared to those in extreme habitats. This study identified a high degree of polymorphism among R. rosea populations using the inter-repeat amplified PCR method. The genetic diversity of the populations ranged from 0.105 to 0.156, with an average heterozygosity of 0.134. The findings provide new insights into the population structure of R. rosea in the Kazakh Altai, enabling the identification of different genotypes, which will significantly complement traditional methods for conserving this valuable medicinal plant.

Rhaponticum carthamoides (Willd.) Iljin. (Leuzea carthamoides, Maral root), a medicinally valuable species listed in the Red Book of Kazakhstan, is known for its rich phytochemical profile. However, limited data exist on its microclonal propagation. This study aimed to optimize in vitro and medium-term storage conditions using biotechnological methods. Mature seeds collected from natural populations in the Kazakhstani Altai were germinated, and tissues from the seedlings were used as explants. Sterile shoots were cultured on Murashige and Skoog (MS) medium supplemented with 3.0 mg L⁻¹ −6-benzylaminopurine and 3.0 mg L⁻¹ kinetin. For shoot induction, MS medium supplemented with 0.5 mg L⁻¹ meta-Topolin and using stem apices as explants yielded optimal results. Medium-term storage with chlorocholine chloride at 0.1–0.4 g/L effectively preserved regenerative capacity for further rooting. After 12 months of storage, plantlets were transferred to half-strength MS medium with 3.0 g/L activated carbon and at 2.0 or 5.0 mg L⁻¹ indole-3-butyric acid for rooting. Regenerated plants were successfully acclimatized ex vitro. The 20-hydroxyecdysone content in field-grown plants post-storage reached 9.24 mg/mL, 2.4-fold higher than in wild plants. Inter simple sequence repeat analysis confirmed genetic stability. Our optimized protocol ensures high-yield metabolite production and genetic fidelity, enabling in vitro conservation, nursery-scale cultivation, and the restoration of R. carthamoides natural populations.

Tulipa auliekolica and Tulipa turgaica have been recently described as endangered species endemic to Kazakhstan, which require urgent conservation amid rising human impact and climate change. Biotechnology offers effective tools for conserving such rare species; however, species-specific in vitro protocols tailored to their biological traits remain largely unreported. This study aimed to develop an in vitro propagation protocol for these rare Tulipa species by investigating the effects of different temperature regimes and phytohormone treatments. We conducted a study on the in vitro propagation of two recently described species, T. auliekolica and T. turgaica. Species-specific temperature regimes for seed stratification were established. Maximum germination of T. auliekolica was achieved at alternating temperatures of 4/10 °C, and of T. turgaica at 10/20 °C. No seed germination from either species occurred at a constant temperature of 20 °C. Bulbs cultured on Murashige & Skoog (MS) medium supplemented with 90 g/L sucrose and the growth regulators mT (meta-topolin) and BAP (6-benzylaminopurine) were effective in stimulating the formation of up to 4–7 microbulbs. Cultivation on a medium supplemented with 0.5 mg L⁻¹ IBA (indole-3-butyric acid) resulted in the formation of mature bulbs covered with scales. These results can be successfully used in biodiversity conservation programs for the endemic Tulipa species. In addition, they provide a valuable basis for future biotechnological research, including microclonal propagation, the establishment of gene banks, and the development of reintroduction methods for Kazakh endemic Tulipa species.

Background: Rhodiola quadrifida (Pall.) Fisch. & C.A. Mey. (Crassulaceae) is a rare medicinal species in the Kazakh Altai, yet information on its population structure and genetic diversity remains limited. This study presents findings from an investigation of natural R. quadrifida populations. Methods: The morphometric characteristics, population age structure, and genetic diversity of the plants were analysed using PCR-based genome profiling. Genetic diversity within R. quadrifida populations was assessed using PCR primers for binding sites complementary to a specific region at the 3′ terminus of a particular tRNA. Results: The populations exhibited variations in morphological traits, age structures, and reproductive strategies. The IVA-1, IVA-2 and KOK populations represent the most mature stages, characterized by a dominance of vegetative reproduction and a disturbed age structure, with a predominance of senile and virgin individuals. In contrast, the LIN-1 and LIN-2 population is characterized by a balanced age structure, encompassing all ontogenetic groups, and a mixed reproductive system that includes both sexual and vegetative propagation. Genetic diversity, as measured by Shannon’s information index, ranged from 0.194 to 0.247, indicating low genetic diversity in R. quadrifida. Analysis of molecular variance (AMOVA) revealed significantly greater variation within populations (62%) than among populations (38%). Genetic diversity was higher in the LIN-1 and LIN-2 populations, which employs a mixed reproductive system (clonal and seeds), than in populations dominated by vegetative reproduction. Both LIN populations, characterized by a mixed reproductive system, exhibited higher genetic diversity than the KOK, IVA-1 and IVA-2 populations, where vegetative reproduction predominated. Conclusions: These results underscore the necessity for priority conservation measures, including monitoring population size and age structure in populations with low levels of seed reproduction and disturbed age structure. Additional measures include supporting in situ and ex situ conservation, such as clonal collection, seed banks, and tissue cultures, as well as restricting the harvesting of medicinal raw materials.

Optimization of the method of medium-term storage of the rare and endangered plant species Allium ivasczenkoae under in vitro conditions using modern approaches of biotechnology is an urgent task. Aseptic microplants Allium ivasczenkoae were used as starting material. The cultivation of microplants was carried out on nutrient media supplemented in various combinations with an increased concentration of sucrose, mannitol, chlorocholine chloride, ABA and BAP at various temperature and light conditions of deposition. The survival dynamics of A. ivasczenkoae varied depending on cultivation regimes. After 9 months of deposition, the survival of test-tube plants in mode 1 (control) was 20%, mode 2 - 60-80%, mode 3 - 30-50%. Under experimental conditions, the best results were obtained by depositing aseptic cultures for 9 months at a low positive temperature and low light intensity (mode 2) on nutrient media with the addition of CCC 120 mg/l and a sucrose concentration of 60 g/l and (B-1) and MS medium containing 0.5 mg/l BAP and 3 g/l mannitol (MSВМ). This mode of deposition made it possible to lengthen the period between transplants, as well as to maintain the viability of A. ivasczenkoae microplants from 60 to 80%. The data obtained make it possible to optimize the mode of medium-term storage of Allium ivasczenkoae under in vitro conditions for the mass production of rare species of bulbous plants in order to restore, reproduce and preserve the number of valuable gene pool and natural populations, create gene banks in vitro, reintroduce and green building.