Endemic species are especially vulnerable to biodiversity loss caused by isolation or habitat specificity, small population size, and anthropogenic factors. Endemic species biodiversity analysis has a critically important global value for the development of conservation strategies. The rare onion Allium ledebourianum is a narrow-lined endemic species, with natural populations located in the extreme climatic conditions of the Kazakh Altai. A. ledebourianum populations are decreasing everywhere due to anthropogenic impact, and therefore, this species requires preservation and protection. Conservation of this rare species is associated with monitoring studies to investigate the genetic diversity of natural populations. Fundamental components of eukaryote genome include multiple classes of interspersed repeats. Various PCR-based DNA fingerprinting methods are used to detect chromosomal changes related to recombination processes of these interspersed elements. These methods are based on interspersed repeat sequences and are an effective approach for assessing the biological diversity of plants and their variability. We applied DNA profiling approaches based on conservative sequences of interspersed repeats to assess the genetic diversity of natural A. ledebourianum populations located in the territory of Kazakhstan Altai. The analysis of natural A. ledebourianum populations, carried out using the DNA profiling approach, allowed the effective differentiation of the populations and assessment of their genetic diversity. We used conservative sequences of tRNA primer binding sites (PBS) of the long-terminal repeat (LTR) retrotransposons as PCR primers. Amplification using the three most effective PBS primers generated 628 PCR amplicons, with an average of 209 amplicons. The average polymorphism level varied from 34% to 40% for all studied samples. Resolution analysis of the PBS primers showed all of them to have high or medium polymorphism levels, which varied from 0.763 to 0.965. Results of the molecular analysis of variance showed that the general biodiversity of A. ledebourianum populations is due to interpopulation (67%) and intrapopulation (33%) differences. The revealed genetic diversity was higher in the most distant population of A. ledebourianum LD64, located on the Sarymsakty ridge of Southern Altai. This is the first genetic diversity study of the endemic species A. ledebourianum using DNA profiling approaches. This work allowed us to collect new genetic data on the structure of A. ledebourianum populations in the Altai for subsequent development of preservation strategies to enhance the reproduction of this relict species. The results will be useful for the conservation and exploitation of this species, serving as the basis for further studies of its evolution and ecology.

Sea buckthorn (Hippophae rhamnoides L.) is an ancient crop that has traditionally been used as a medicinal plant, health promotion agent and dietary supplement. The fruits of sea buckthorn, which are a source of carotenoids, which had been collected since ancient times and was introduced into cultivation only at the beginning of the 20th century. Currently, sea buckthorn berries are harvested on an industrial scale, as due to the high content of biologically active substances in them, consumer interest in sea buckthorn products has increased significantly, and their use in the food industry and medicine is expanding. Due to its biologically active compounds, high content of vitamin C, flavonoids, carotenoids and tocopherols, sea buckthorn is an excellent raw material for functional foods. The purpose of this work is to select conditions for cultivating explants of H. rhamnoides L. plants and obtaining micro-shoots for subsequent microclonal reproduction. The material for initiating a sterile culture was the seeds H. rhamnoides. The samples were introduced from natural populations in the RSE “Altai Botanical Garden” of the SC of MoES of the Republic of Kazakhstan (Ridder). Solutions of 0.1% sulema, 34% hydrogen peroxide, and 10% Domestos solution were used for seed sterilization, the exposure period was 20 minutes. Next, the seeds were planted on the surface of the WPM and ½ MS nutrient medium with the addition of phytohormones 6-benzylaminopurine (BA) and gibberellic acid (GA3) in various concentrations. Activated carbon in an amount of 2 g/l was used to reduce the accumulation of polyphenols in the medium. The beginning of germination was noted on days 7-8, depending on the genotype of sea buckthorn. The minimum number of infected seeds was observed in the variant using 0.1% sulema. When using hydrogen peroxide, the contamination of seeds with fungal and bacterial infections was at the level of 40.8-45.5%. The largest number of germinated seeds were observed when using WPM medium. The maximum values of seedling length were noted on WPM media with the addition of 1 mg/l BA + 1 mg/l GA3, where the average shoot length was 8.6±0.9 cm. In this variant, sterile seedlings were characterized by an intense green color and well-developed cotyledon leaves. In some genotypes, intensive growth and the appearance of a second internode were noted. In other variants, height indicators varied from 3.2 cm. up to 4.2 cm. Thus, we have found that WPM medium supplemented with 1 mg/l 6-benzylaminopurine and 1 mg/l gibberellic acid is better suited for obtaining sterile shoots from H. rhamnoides seeds. The obtained sterile seedlings of H. rhamnoides will be used in vitro culture for subsequent microclonal reproduction.

Алғышарттар мен мақсат. Кәдімгі шырғанақ (Hippophae rhamnoides) қосүйлі жапырақты бұта. Оның жапырақтары мен жидектеріндегі биологиялық қосылыстардың көптігіне байланысты бұл өсімдік қазіргі уақытта адамдар мен жануарлардың денсаулығын сақтауда қолданылатын өсімдік ретінде танымал. Жұмыстың мақсаты шырғанақ эксплантарын in vitro жағдайына енгізу, зарарсыздандыру режимдері мен қоректік орта құрамын таңдау болып табылады. Зерттеу материалы ретінде шырғанақтың тұқымдары пайдаланылды. Материалдар мен әдістер. Зарарсыздандыру үшін әртүрлі антисептиктер қолданылды: этанол, сынап (ІІ) хлориді, сутегінің асқын тотығы және натрий гипохлориді. Зарарсыздандыру уақыты әр антисептик үшін эксперименталды түрде таңдалып алынды. Зарарсыз өскіндерді өсіру үшін әртүрлі өсу реттегіштері қолданылды. Фенолды қосылыстардың бөлінуін тежеу үшін антиоксидант ретінде белсендірілген көмір қолданылды. Нәтижелер. Зарарсыз және өміршең экспланттарының ең жоғарғы пайызы (83,3%) 0,1% сынап (ІІ) хлориді ерітіндісімен зарарсыздандыру нұсқасында алынды. Өсу деңгейі сорттардың түріне және қоректік орта құрамына байланысты болды. Ш-9-81(4-5) сортының максималды өскін ұзындығы (4,0 см) құрамында 1,0 мг/л BAР, 5,0 мг/л IAA және 2 г/л белсендірілген көмір қосылған ортада болатыны анықталды. Қорытынды. Hippophae rhamnoides өскіндердің пролиферациясы үшін ең тиімді орта ретінде құрамына 1,0 мг/л BAP, 5,0 IAA және 2,0 г/л белсендірілген көмір қосылған орта екені анықталды, ең жоғарғы нәтиже Ш-9 81(4-5) сортында байқалды, максималды өскін ұзындығы 4,0 см құрады.

Daphne altaica Pall. is a rare medicinal shrub of Kazakhstan with poor natural regeneration and no reported in vitro propagation protocol. We established in vitro cultures from nodal explants of wild plants and evaluated the effects of collection season, basal medium, and cytokinin type. Spring-collected explants outperformed autumn-collected explants at every stage. Disinfection with 0.1% HgCl₂ for 5 min gave 8.61% contamination and 61.65% explant viability in spring material (12.52% and 34.22% in autumn material). On Woody Plant Medium (WPM) with 1.0 mg L⁻¹ 6-benzylaminopurine and 0.1 mg L⁻¹ 1-naphthaleneacetic acid (NAA), axillary shoots emerged after 3.3 (spring) and 10.3 days (autumn), with induction rates of 100% and 77.78%. Shoot proliferation was highest on WPM with 1.0 mg L⁻¹ meta-topolin and 0.1 mg L⁻¹ NAA (5.97 and 3.11 shoots per explant in spring and autumn material); basal medium, cytokinin, and season all had significant effects (p < 0.001). Rooting remained the main bottleneck (maximum 20.63% on WPM with 2 mg L⁻¹ indole-3-butyric acid), and ex vitro acclimatization was not assessed. These results provide the first baseline conditions for the in vitro establishment and multiplication of D. altaica.

Analysis of the genetic diversity of natural populations of economically valuable plants is important for conservation and selection strategies. In this study, the genetic diversity of 11 natural populations of Hippophae rhamnoides L.—sea buckthorn from different regions of Kazakhstan—was studied using Inter-Primer Binding Site Polymorphism (iPBS) markers based on conserved sequences of tRNA primer-binding sites (PBSs) that initiate retrotransposon replication. Universal PBS primers yielded reproducible and informative amplicons, forming unique profiles for each sample. Analysis of molecular variance showed that 60% of the total genetic variation was due to intrapopulation differences and 40% was due to interpopulation differentiation. The highest genetic diversity was found in the Shetlasty and Tersayryk sea buckthorn populations, whereas the Karatal and Topkain populations were characterised by minimal values, although unique alleles were observed in the latter population, indicating possible adaptation to local environmental conditions or genetic isolation. Principal coordinate analysis, UPGMA clustering, and Bayesian structure analysis (K = 4) confirmed geographical structuring. This study provides insights into the genetic structure of sea buckthorn populations in Kazakhstan and demonstrates the effectiveness of iPBS markers for assessing intraspecific diversity. The obtained results provide a basis for the conversation of H. rhamnoides gene pool and underscore the need for both in situ conservation of genetically rich populations and ex situ protection of vulnerable groups.

This study presents a comprehensive identification and characterization of fungal pathogens that affect sea buckthorn (Hippophae rhamnoides). Utilizing microbiological methods, biochemical analyses, molecular genetic diagnostics – including PCR and DNA sequencing – and pathogenicity testing, three species of phytopathogenic fungi were identified: Aureobasidium pullulans, Didymella glomerata, and Epicoccum nigrum. The pathogenicity of these fungi towards sea buckthorn was experimentally confirmed. An evaluation of the isolates' sensitivity to six antifungal agents (terbinafine, nystatin, ketoconazole, clotrimazole, itraconazole, and fluconazole) indicated notable species-specific differences in susceptibility to fungicidal effects. The findings enhance our understanding of the etiology of fungal diseases affecting sea buckthorn and can inform the development of effective strategies for phytosanitary control and plant protection.