Aidana Nurtaza

Amygdalus ledebouriana (Schltdl.) Y.Y. Yao is a rare, endangered wild almond species native to Eastern Kazakhstan, listed in both Kazakhstan’s Red Book and the IUCN Red List, and valued as a source of drought- and frost-tolerance genes for stone fruit breeding. This study developed an efficient droplet-vitrification cryopreservation protocol for A. ledebouriana shoot tips and assessed regenerant genetic stability using simple sequence repeat (SSR) markers. The protocol comprised five stages: preculture, osmotic conditioning (loading), vitrification with plant vitrification solutions 2 and 3 (PVS2/PVS3), liquid nitrogen storage, and unloading/regeneration. Each stage was optimized by stepwise variation of a single factor while holding the remaining conditions constant, using generalized linear models (quasi-binomial distribution, Holm-corrected post hoc comparisons). Optimal conditions included preculture on 0.75 M sucrose for 3 days (31.1% survival), 30 min loading (38.9%), and 120 min dehydration with PVS3, which significantly outperformed PVS2 in terms of shoot tip survival after cryopreservation (50.0% vs. 30.0%). Unloading with 0.8 M sucrose under mild conditions (22 °C, 20 min) and recovery medium supplemented with ascorbic acid achieved the highest survival (61.1%) and regrowth (60.0%). SSR analysis at five loci revealed no genetic changes in regenerants following cryopreservation. The developed protocol is the first reported for A. ledebouriana and can be recommended for the long-term ex situ conservation of its genetic resources.

Biotechnology-based conservation of the endemic and endangered Amygdalus ledebouriana , listed in the Red Book of Kazakhstan and IUCN Red List, can support biodiversity conservation and sustainable ecosystem management. This poorly studied species grows in eastern Kazakhstan in the foothills of the southwestern Altai, Tarbagatai and Dzungarian Alatau mountains, which are negatively affected by overgrazing, spring and autumn burning, flower harvesting and moisture deficits. As a rootstock, it is used for plum and peach tree cultivation (conferring drought resistance), for almond breeding; further, the species is used for soil protection. An effective micropropagation protocol was developed using the plant growth regulators meta-topolin (1.0 mg L ^− 1 ), gibberellic acid (GA ₃ ) (1.0 mg L ^− 1 ) and indole-3-butyric acid (IBA) (0.01 mg L ^− 1 ), generating 4.96 microshoots per explant. Effective in vitro rooting was achieved via shoot cultivation for 10 days in the dark on half-strength Quoirin and Lepoivre (QL) medium supplemented with 1.0 mg L ^− 1 IBA, followed by transfer to hormone-free half-strength QL medium, generating 4.2 roots per explant (68% rooting rate). Slow-growth storage of shoots was achieved at a low temperature (4 °C) in the dark on QL medium with either 2.0 g L ^− 1 ABA or 1.5% sucrose; this suppressed microshoot growth while maintaining up to 100% plant viability for four months without sub-cultivation. Molecular genetic analysis using eight SSR markers confirmed the complete identity of the regenerants and the maternal plant, demonstrating their genetic stability. The micropropagation and slow-growth storage protocol developed in this study may serve as a useful tool for the ex situ conservation of this endemic and endangered species.

The conservation of rare and endangered plant species requires the development of effective approaches for obtaining viable planting material suitable for ex situ preservation and subsequent reintroduction. Berberis iliensis M. Pop. is a rare endemic species with limited natural regeneration, which necessitates the application of biotechnological methods. The aim of this study was to obtain viable planting material of Berberis iliensis using in vitro culture followed by adaptation and ex vitro acclimatization. Microshoots were established from shoot explants and cultured on Murashige and Skoog (MS) medium supplemented with growth regulators. Rooting was induced on MS medium containing indole-3-butyric acid (IBA). The effect of substrate composition on plant adaptation was evaluated. The highest growth and development parameters were obtained on a soil:sand substrate, where shoot height reached 6.53±0.14 cm, the number of leaves was 6.30±0.23, root number was 4.93±0.14, and root length was 5.11±0.20 cm, with all differences being statistically significant compared to the control. During acclimatization, the influence of mineral nutrition was assessed. The most pronounced positive effect was observed with calcium nitrate treatment, resulting in maximum plant height (16.84±0.42 cm), number of leaves (20.45±0.61), and crown diameter (15.83±0.62 cm). The results demonstrate that the successful transition of in vitro-derived plants to ex vitro conditions depends on substrate properties and mineral nutrition, which affect plant growth and survival. The proposed approach enables the production of viable planting material and can be applied for conservation and restoration of Berberis iliensis populations.

Malus niedzwetzkyana is a rare and valuable wild species characterised by a high anthocyanin content. This species is of considerable interest as a promising source of bioactive compounds. In this study, we investigated the effect of explant type on callus induction and its morphological characteristics. Stems and leaves from 40-day-old plants grown in vitro on Murashige-Skoog (MS) medium were used as explants. The results showed that stem explants demonstrated a higher ability to form callus compared to leaf explants - 100% and 86.7%, respectively. MS medium with various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) (2.5; 3.0; 3.5 mg/l) and 6-benzylaminopurine (BAP) (0.5; 1.0 mg/l) had a significant effect on diameter, fresh and dry weight, as well as dry matter (p < 0.001). The combination of MS medium with 2,4-D at a concentration of 3.5 mg/L and BAP at 0.5 mg/L proved optimal, as it resulted in the formation of a uniform, loose callus of a pinkish burgundy colour. Thus, our results provide a basis to produce suspension cultures and the biotechnological production of biologically active substances from Malus niedzwetzkyana.

Haploid and doubled haploid (DH) technologies are important tools for accelerated wheat (Triticum aestivum L.) breeding, enabling the rapid production of fully homozygous lines and increasing the efficiency of genetic analysis of complex traits. This review presents a comprehensive analysis of the main approaches to producing haploid and DH wheat plants, with particular emphasis on androgenesis-mediated and chromosome elimination methods, including wheat-maize hybridisation. The biological basis of androgenesis is discussed in relation to stress-induced microspore reprogramming; however, the primary focus is on the methodological factors determining the efficiency of DH production, including the donor plant genotype, microspore development stage, pretreatment conditions, composition of the induction and regeneration media, and chromosome doubling. However, its widespread application remains limited by pronounced genotypic dependence, low responsiveness of many commercial varieties, albinism, and a lack of universally effective protocols. In contrast, distant hybridisation systems, particularly wheat-maize hybridisation, are generally characterised by greater reproducibility and less genotypic dependence, although they remain labour-intensive and require precise embryo rescue and chromosome doubling procedures. Overall, further progress in producing DH in wheat will be associated with the optimisation of protocols for difficult-to-respond genotypes and the integration of classical haploidisation systems with omics approaches, genomic selection, and genome-editing.

Early blight, caused by Alternaria alternata , is a key factor that reduces tomato yield and tomato product quality. Owing to the quantitative nature of resistance and the limited number of characterized loci, identification of genomic regions associated with resistance remains an urgent task for breeding programs. Here, bulked segregant analysis with whole-genome resequencing (BSA-Seq) was used to identify candidate regions for resistance to early blight in the F₂ population obtained by crossing a resistant line (Gloria×BSS335) and a susceptible cultivar (Samaladay). A total of 14 overlapping candidate genomic regions were identified on chromosome 2 by intersecting SNP- and InDel-associated regions (totaling 0.26 Mb). Functional annotation of the 33 genes in this locus revealed the involvement of membrane proteins, components of signaling pathways, transcriptional regulators, and Rab GTPase family proteins, which are potentially involved in stress and immune responses. This is the first genome-wide study of early blight resistance in tomato in Kazakhstan. These results expand our understanding of the genetic architecture of early blight resistance and may serve as a basis for the functional validation of key candidate genes and their application in marker-assisted selection (MAS).

Androgenesis in anther culture is a promising method for the accelerated production of homozygous wheat (Triticum aestivum L.) lines; however, the widespread application of this technology is limited by strong genotype dependence and low regeneration efficiency. The aim of this study was to optimize the conditions for androgenesis induction and plant regeneration by selecting the appropriate culture medium and hormonal composition. Twelve medium variants based on N6, C17, and MS were tested with the addition of different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D). The highest embryogenic activity was observed on media containing 2.0 mg/L 2,4-D and 0.5 mg/L kinetin, particularly on N6 and C17 bases. Plant regeneration also varied depending on the conditions, with the highest efficiency observed on hormonal media containing auxin and cytokinin combinations. The results confirm the necessity of a comprehensive selection of medium components and demonstrate that the proper combination of nutrients and phytohormones can significantly increase the yield of embryoids and viable regenerants.

The article presents the results of a study on the in vitro conservation of the rare and endangered species Amygdalus ledebouriana. This almond species is noted for its high resistance to drought and low temperatures, as well as the nutritional value of its fruits. Prior to this study, no research had been conducted on the micropropagation of Amygdalus ledebouriana. In the present work, the authors developed an effective protocol for explant sterilization and establishment in vitro culture. A 9% hydrogen peroxide solution applied for 5 minutes was selected as the most suitable sterilizing agent, ensuring up to 75% explant viability. For shoot multiplication, the QL nutrient medium supplemented with 0.25 mg/L 6-benzylaminopurine (BAP) was optimized, resulting in an average of 7.24 shoots per explant. To date, 400 microshoots have been successfully propagated. An in vitro collection has been established from three natural populations of Amygdalus ledebouriana.

The ongoing genetic erosion of natural Prunus armeniaca populations in their native habitats underscores the urgent need for targeted conservation and restoration strategies. This study provides the first comprehensive characterization of P. armeniaca populations in the Almaty region of Kazakhstan, integrating morphological descriptors (46 parameters), molecular markers, geobotanical, and remote sensing analyses. Geobotanical and remote sensing analyses enhanced understanding of accession distribution, geological features, and ecosystem health across sites, while also revealing their vulnerability to various biotic and abiotic threats. Of 111 morphologically classified accessions, 54 were analyzed with 13 simple sequence repeat (SSR) markers and four DNA barcoding regions. Our findings demonstrate the necessity of integrated morphological and molecular analyses to differentiate closely related accessions. Genetic analysis identified 11 distinct populations with high heterozygosity and substantial genetic variability. Eight populations exhibited 100% polymorphism, indicating their potential as sources of adaptive genetic diversity. Cluster analysis grouped populations into three geographic clusters, suggesting limited gene flow across Gorges (features of a mountainous landscape) and greater connectivity within them. These findings underscore the need for site-specific conservation strategies, especially for genetically distinct, isolated populations with unique allelic profiles. This study provides a valuable foundation for prioritizing conservation targets, confirming genetic redundancies, and preserving genetic uniqueness to enhance the efficiency and effectiveness of the future conservation and use of P. armeniaca genetic resources in the region.