Search Results (47)

Traditional methods for grapevine (Vitis spp.) breeding are marked by lengthy breeding cycles with usually low efficiency, rendering them inadequate for the demands of the rapidly evolving grapevine industry. While grapevine genetic transformation holds significant potential for improvement, its application is hampered by bottlenecks in efficiency, speed, and genotype dependence. In this context, this review systematically examines the factors influencing and challenges associated with key steps in grapevine genetic transformation—specifically, gene delivery and plant regeneration. It posits that the development and application of marker genes, the exploration and utilization of developmental regulators, and the establishment of novel genetic transformation systems are effective strategies to overcome current limitations. In this paper, we present a foundation and methodological guidance for creating efficient and stable genetic transformation systems for grapevine, with significant theoretical and practical implications. Full article

The genus Corydalis, belonging to the Papaveraceae family, is widely distributed across the Northern Hemisphere, primarily in Asia. This study aimed to investigate the effect of plant growth regulators (PGRs) on callus induction, and of light quality and intensity on indirect shoot organogenesis, dl-Tetrahydropalmatine (dl-THP) accumulation, and activities of antioxidant enzymes in Corydalis turtschaninovii Besser. Calli were successfully induced from the leaf, tuber, and petiole explants with different PGR combinations. The best callus induction from leaf, tuber, and petiole explants were obtained in the medium supplemented with 3 mg·L⁻¹ kinetin (Kn) combined with 0.8 mg·L⁻¹ naphthalene acetic acid (NAA), 3 mg·L⁻¹ benzyl adenine (BA) combined with 0.8 mg·L⁻¹ NAA, and 2 mg·L⁻¹ BA combined with 0.5 mg·L⁻¹ NAA, respectively. For indirect shoot organogenesis, calli were cultured on the Murashige and Skoog (MS) medium under dark (D), white (W), red (R), blue (B), or 1:1 mixture of red and blue (RB) light-emitting diodes (LEDs) at an intensity of 25 or 50 µmol·m⁻²·s⁻¹ photosynthetic photon flux density (PPFD) for six weeks. The RB treatment increased biomass accumulation of the callus, and promoted the induction of the shoot from the callus, whereas the R treatment promoted the dl-THP accumulation, especially with the higher light intensity. Light quality and intensity significantly influenced the activities of antioxidant enzymes and the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging capacity in calli, with the most pronounced effects observed under B or RB light treatments. Taken together, the application of monochromatic LED or combinations of red and blue LEDs could be used for the callus culture for different purposes in vitro. Full article

Generation of state-of-the-art highly productive cabbage genotypes (Brassica oleracea convar. capitata (L.) Alef.) with improved agronomic traits is attainable using modern biotechnological approaches. However, capitata cabbage is relatively recalcitrant to de novo shoot organogenesis from callus tissue, especially with loss of somatic cell totipotency during genetic transformation. An effective and rapid protocol for in vitro indirect shoot organogenesis from hypocotyl and cotyledon explants derived from 6-day-old aseptic donor seedlings of Russian cabbage genotypes (the DH line as well as cvs. Podarok and Parus) has been developed. In order to obtain standardized donor explants, aseptic cabbage seeds were germinated under dim light conditions (30–40 µmol m⁻² s⁻¹) with a 16 h light/8 h dark photoperiod. Multiple indirect shoot organogenesis (1.47–4.93 shoots per explant) from both cotyledonary leaves and hypocotyl segments with a frequency of 55.2–89.1% was achieved through 45 days of culture on the 0.7% agar-solidified (w/v) Murashige and Skoog (MS) basal medium containing 2 mg/L 6-benzylaminopurine (6-BAP), 0.02 mg/L 1-naphthalene acetic acid (NAA), and 5 mg/L AgNO₃. The regenerants were successfully rooted on an MS basal medium (69.2%) without plant growth regulators (PGRs), as well as supplemented with 0.5 mg/L NAA (86.8%). Subsequently, in vitro rooted cabbage plantlets were adapted to soil conditions with an efficiency of 85%. This rapid protocol, allowing for the performance of a full cycle from in vitro seed germination to growing adapted plantlets under ex vitro conditions over 95 days, can be successfully applied to induce an indirect shoot formation in various cabbage genotypes, and it is recommended to produce transgenic plants with improved quality traits and productivity. Full article

Introduction: Stevia rebaudiana Bertoni has recently gained significant attention due to the presence of intensely sweet yet low-calorie steviol glycosides (SGs) in its leaves, making it a promising natural sugar alternative with applications in the food, pharmaceutical, and cosmetics industries. The primary goal of this study was to determine whether generating somaclonal variation from plant material obtained by indirect regeneration results in further genetic changes identifiable using the SCoT marker (Start Codon Targeted). Methods: In the first stage, callus tissue was initiated from first-generation somaclones on MS medium supplemented with 4.0 mg/L 6-benzylaminopurine (BAP), 2.0 mg/L 1-naphthaleneacetic acid (NAA), and 2.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D). Their morphogenetic potential was analyzed on four media with different BAP and Kinetin concentrations. Donor plants, first and second generation somaclones, were also analyzed for genetic diversity using SCoT markers. Results: All first-generation somaclones demonstrated a very high callus initiation capacity, ranging from 95 to 100%. It was found that for most of the studied somaclones, the greatest number of shoots were developed by explants grown in a medium supplemented with 0.5 mg/L BAP and 0.25 mg/L Kin. The studied group of somaclones exhibits a high degree of polymorphism (55.2%). The analysis of genetic similarity of somaclones presented in the form of individual dendrograms indicates that in most cases, greater genetic diversity was revealed as a result of indirect regeneration in the first generation of somaclones compared to the second. Indirect organogenesis allows for the production of subsequent generations of genetically unstable somaclones, creating the potential for obtaining new phenotypic variants useful in plant breeding. Full article

This study evaluates the impact of biotic elicitors and hormone regimes on the in vitro establishment, shoot multiplication, and organogenesis of Solanum tuberosum L. cv. Fianna under controlled laboratory conditions. Explants derived from pre-treated tubers were cultured on Murashige and Skoog (MS) medium supplemented with vitamins and varying concentrations of growth regulators or elicitors. Aseptic establishment achieved a high success rate (~95%) using a 6% sodium hypochlorite disinfection protocol. Multiplication was significantly enhanced with a combination of 0.2 mg L⁻¹ naphthaleneacetic acid (NAA) and 0.5–1.0 mg L⁻¹ benzylaminopurine (BAP), producing the greatest number and length of shoots and roots. Direct organogenesis was stimulated by bio-elicitors Activane^®, Micobiol^®, and Stemicol^® in (MS) basal medium at mid-level concentrations (0.5 g or mL L⁻¹), improving shoot number, elongation, and root development. Activane^®, Micobiol^®, and Stemicol^® are commercial elicitors that stimulate plant defense pathways and morphogenesis through salicylic acid, microbial, and jasmonic acid signaling mechanisms, respectively. Indirect organogenesis showed significantly higher callus proliferation in Stemicol^® and Micobiol^® treatments compared to the control medium, resulting in the highest fresh weight, diameter, and friability of callus. The results demonstrate the potential of biotic elicitors as alternatives or enhancers to traditional plant growth regulators in potato tissue culture, supporting more efficient and cost-effective micropropagation strategies. Full article

Catalpa bungei C.A.Mey is an economically significant deciduous tree valued for timber production and landscaping applications. An efficient regeneration system is crucial for clonal propagation and serves as a foundation for future molecular breeding in C. bungei. This study established two in vitro regeneration pathways—indirect somatic embryogenesis and shoot organogenesis utilizing mature zygotic embryos as explants. Primary callus was induced from cotyledon, hypocotyl, and plumule explants. A high frequency (45.73%) of yellow-green compact callus was achieved on De-Klerk and Walton (DKW) medium supplemented with 2.0 mg/L 6-BA, 1.0 mg/L zeatin (ZT), and 0.1 mg/L NAA. Subsequent transfer to 1.5× Murashige and Skoog (MS) medium containing 1.5 mg/L 6-BA, 0.2 mg/L ZT, and 0.1 mg/L NAA yielded the highest embryogenic callus induction rate (16.67%). Embryogenic callus demonstrated bipotent potential, generating both adventitious shoots and somatic embryos under specific hormonal conditions. Histological analyses confirmed the typical developmental stages of somatic embryos, from globular to cotyledonary forms, validating the embryogenic origin of regenerated structures. Furthermore, hormone or osmotic additives such as abscisic acid (ABA), Phytagel, and polyethylene glycol 4000 (PEG4000) significantly enhanced somatic embryo induction, with Phytagel at 5.0 g/L achieving the highest rate (76.31%). For shoot organogenesis, the optimal hormonal combination of the 0.6 mg/L 6-BA, 0.4 mg/L KT, and 0.15 mg/L NAA achieved the highest bud induction rate (88.89%) and produced an average of 4.07 adventitious buds per explant. This study presents an efficient regeneration system for C. bungei, providing a practical platform for large-scale propagation and basis for biotechnological applications in woody plants. Full article

Bombax ceiba is an important medicinal and ornamental tree widely distributed in tropical and subtropical areas. However, its seeds lose viability rapidly after harvest, which has created hurdles in large-scale propagation. Here, we describe the development of a rapid and efficient de novo organogenesis system for Bombax ceiba, incorporating both indirect and direct regeneration pathways. The optimal basal medium used throughout the protocol was ½ MS supplemented with 30 g/L glucose, with all cultures maintained at 26–28 °C. For the indirect pathway, callus was induced from both ends of each hypocotyl on basal medium supplemented with 0.2 mg·L⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg·L⁻¹ 6-Benzylaminopurine (6-BA) under dark conditions. The induced calluses were subsequently differentiated into adventitious shoots on basal media containing 0.5 mg·L⁻¹ Indole-3-butyric acid (IBA), 0.15 mg·L⁻¹ Kinetin (KIN), and 1 mg·L⁻¹ 6-BA under a 16 h photoperiod, resulting in a callus induction rate of 140% and a differentiation rate of 51%. For the direct regeneration pathway, shoot buds cultured on medium with 0.5 mg·L⁻¹ IBA and 1 mg·L⁻¹ 6-BA achieved a 100% sprouting rate with a regeneration coefficient of approximately 3.2. The regenerated adventitious shoots rooted successfully on medium supplemented with 0.5 mg·L⁻¹ Naphthylacetic acid (NAA) and were acclimatized under greenhouse conditions to produce viable plantlets. This regeneration system efficiently utilizes sterile seedling explants, is not limited by seasonal or environmental factors, and significantly improves the propagation efficiency of Bombax ceiba. These optimized micropropagation methods also provide a robust platform for future genetic transformation studies using hypocotyls and shoot buds as explants. Full article

The use of in vitro cultures in plant breeding allows for obtaining cultivars with improved properties. In the case of Stevia rebaudiana Bert., genotypes with an appropriate rebaudioside A/stevioside ratio are desirable. The use of indirect organogenesis allows for the induction of somaclonal variation, which, consequently, results in obtaining variability within the regenerants. The Murashige and Skoog medium containing 4.0 mg × dm⁻³ 6-benzylaminopurine (BAP), 2.0 mg × dm⁻³ 1-naphthaleneacetic acid (NAA), and 2.0 mg × dm⁻³ 2,4-dichlorophenoxyacetic acid (2,4-D) resulted in obtaining plants that were biochemically and genetically diverse. The obtained regenerants were characterized by an increased content of rebaudioside A and a better rebaudioside A/stevioside ratio. Genetic analysis using SCoT (start-codon-targeted) markers showed their diversity at the molecular level. Moreover, this study showed that genotype multiplication through six subsequent re-cultures does not cause variability at the genotype level and does not affect the steviol glycoside profile. This study is the first report on obtaining genotypes with higher rebaudioside A content and a more attractive rebaudioside A to stevioside ratio through the use of in vitro cultures. The improved regenerants can be used as parents in hybridization programs or directly as valuable new genotypes. Full article

Gaillardia pulchella is an important plant species with pharmacological and ornamental applications. It contains a wide array of phytocompounds which play roles against diseases. In vitro propagation requires callogenesis and differentiation of plant organs, which offers a sustainable, alternative synthesis of compounds. The morphogenetic processes and the underlying mechanisms are, however, known to be under genetic regulation and are little understood. The present study investigated these events by generating transcriptome data, with de novo assembly of sequences to describe shoot morphogenesis molecularly in G. pulchella. The RNA was extracted from the callus of pre- and post-shoot organogenesis time. The callus induction was optimal using leaf segments cultured onto MS medium containing α-naphthalene acetic acid (NAA; 2.0 mg/L) and 6-benzylaminopurine (BAP; 0.5 mg/L) and further exhibited a high shoot regeneration/caulogenesis ability. A total of 68,366 coding sequences were obtained using Illumina150bpPE sequencing and transcriptome assembly. Differences in gene expression patterns were noted in the studied samples, showing opposite morphogenetic responses. Out of 10,108 genes, 5374 (53%) were downregulated, and there were 4734 upregulated genes, representing 47% of the total genes. Through the heatmap, the top 100 up- and downregulating genes’ names were identified and presented. The up- and downregulated genes were identified using the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. Important pathways, operative during G. pulchella shoot organogenesis, were signal transduction (13.55%), carbohydrate metabolism (8.68%), amino acid metabolism (5.11%), lipid metabolism (3.75%), and energy metabolism (3.39%). The synthesized proteins displayed phosphorylation, defense response, translation, regulation of DNA-templated transcription, carbohydrate metabolic processes, and methylation activities. The genes’ product also exhibited ATP binding, DNA binding, metal ion binding, protein serine/threonine kinase -, ATP hydrolysis activity, RNA binding, protein kinase, heme and GTP binding, and DNA binding transcription factor activity. The most abundant proteins were located in the membrane, nucleus, cytoplasm, ribosome, ribonucleoprotein complex, chloroplast, endoplasmic reticulum membrane, mitochondrion, nucleosome, Golgi membrane, and other organellar membranes. These findings provide information for the concept of molecular triggers, regulating programming, differentiation and reprogramming of cells, and their uses. Full article

Calendula officinalis L. is an important medicinal and ornamental plant possessing multiple bioactive compounds. The in vitro plant regeneration method has recently replaced traditional field cultivation practices of calendula due to its fascinating phytochemical profile. In this study, callus formation and indirect organogenesis were described to establish an effective in vitro propagation strategy in C. officinalis. Using a gas chromatography–mass spectrometry (GC–MS) approach, the phytochemical content of tissues developed in vitro and field-grown was studied, and the biochemical contents were quantified and compared in various tissues. The incidence of callus formation from leaf explants was highest (94.44%) on MS medium fortified with 1.0 mg/L BAP and 1.0 mg/L NAA, which later became organogenic. On MS, 1.0 mg/L BAP and 1.0 mg/L NAA showed the highest indirect shoot proliferation (88.88%) efficiency. After being sub-cultured, the regenerated shootlets were cultured onto rooting medium containing different IAA/IBA concentrations; the best rooting percentage (94.44%) was achieved with 1.0 mg/L IBA. The biochemical parameters, like total phenolic content, flavonoid content, and DPPH scavenging activity, were measured. When compared to callus and field-grown developed leaf (donor) samples, all the biochemical characteristics of in vitro-produced leaf were noted to be higher. The methanolic extracts of leaf-callus and field-grown and in vitro-developed leaf tissues were subject to GC–MS-based phytocompound investigation. More than 45 therapeutically significant bioactive chemicals, like n-hexadecanoic acid, vitamin E, stigmasterol, and squalene were found in these samples. These results showed that the callus that is formed from in vitro leaf is a reliable and powerful source of several bioactive compounds with a wide range of medicinal uses. The successful stimulation of callus development, indirect organogenesis, biochemical analysis, and GC–MS confirmation of the presence of significant phytocompounds are all described in this study. This work provides a different avenue for ongoing and sustained synthesis of chemicals without endangering the surrounding ecosystem or native vegetation. Full article

Sesuvium portulacastrum L. is a dicotyledonous halophyte belonging to the family Aizoaceae. Its young leaves are highly nutritious, and many ecotypes are used as leafy vegetable and medicinal crops. Additionally, due to their tolerance to soil salinity, flooding, and high temperatures, some ecotypes are used for the remediation of saline soils. As a result, there is an increasing need for a large number of disease-free S. portulacastrum propagules. This study developed an efficient protocol for the regeneration of S. portulacastrum through indirect shoot organogenesis. Leaf explants were cultured on Murashige and Skoog basal medium supplemented with different concentrations of zeatin (ZT) and indole-3-acetic acid (IAA). Callus was induced in all explants cultured with 1.5 mg/L ZT only or 1.5 mg/L ZT with 0.5 mg/L IAA. The callus was cut into small pieces and cultured on the same medium on which it was initially induced. ZT at 1.5 mg/L induced 73.7% of callus pieces to produce adventitious shoots, and the shoot numbers per callus piece were up to 20. To improve the in vitro rooting of adventitious shoots, commonly known as microshoots or microcuttings, an endophytic fungus, Cladosporium ‘BF-F’, was inoculated onto the rooting medium. ‘BF-F’ substantially enhanced rooting and plantlet growth, as the root numbers were three times more and plantlet heights were 70% greater than those without ‘BF-F’ inoculation. To detect the genes involved in the enhanced rooting and plantlet growth, qRT-PCR analysis was performed. Results showed that genes related to auxin responses and nitrogen uptake and metabolism were highly upregulated in ‘BF-F’-inoculated plantlets. Plants inoculated with ‘BF-F’ grew vigorously after being transplanted into a sand–soil substrate. Thus, this study not only established an efficient protocol for the regeneration of S. portulacastrum but also developed a novel method for improving the rooting of microshoots and plantlet growth. The established propagation system could be used for producing a large number of S. portulacastrum plantlets for commercial use and also for genetic transformation. Full article

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. Full article

Gaillardia pulchella Foug. is a widely studied plant because of its high pharmacological and ornamental value. The leaves of G. pulchella were used for inducing callus and subsequent plant regeneration as it is the primary source of phytocompounds. The purpose of the present investigation was to formulate an in vitro propagation method for Gaillardia by using leaf explants in MS (Murashige and Skoog) medium. The best callus induction was observed on high (2.0 mg/L) α-naphthalene acetic acid (NAA) and a low (0.5 mg/L) 6-benzylaminopurine (BAP) with callus induction frequency of 91.66%. The leaf callus also demonstrated high caulogenesis ability (95.83%), with an average 5.2 shoots/callus mass at 0.5 mg/L BAP and 2.0 mg/L NAA. Indole Acetic acid (IAA) at 1.0 mg/L had the maximum rooting percentage (79.17%) with 12.4 roots per shoot. Rooted plantlets were later transferred to greenhouse conditions, showing a survivability rate of 75–80%. The physiological parameters, i.e., phenolic compounds and the flavonoids’ level, in the DPPH assay were higher in leaves obtained in vitro compared to callus formed from leaves and field-obtained (mother) leaves. Gas chromatography–mass spectrometry (GC–MS) analysis of methanol extracts of leaves (in vivo and in vitro) and leaf callus presented a wide array of compounds. In callus extract, some 34 phytocompounds were identified. Some of them were 3-hydroxy-2,3-dihydromaltol (25.39%), isoamyl acetate (11.63%), palmitic acid (11.55%), 4-methyloxazole (7.54%), and 5-methoxypyrrolidin-2-one (7.49%). Leaves derived in vivo and in vitro had 45 and 28 phytocompounds, respectively, belonging to different classes like lignans, phenols, terpenoids, alkaloids and fatty acids, etc. Those findings demonstrated that the leaf derived callus and the leaves are the potential stable source of several compounds with medicinal importance. The developed protocol may provide an alternative source of compounds without affecting wild flora. Full article

Aseptic seedlings of different ages derived from surface-sterilized mature seeds were applied as an explant source. Various explants such as 7- and 21-day-old hypocotyl fragments, 42-day-old nodal stem segments, and transverse nodal segments of stem, as well as leaf petioles, were cultured on the agar-solidified Murashige and Skoog (MS) basal medium supplemented with 0.1 mg/L IAA, 5 mg/L AgNO₃ and different types and concentrations of cytokinin (1 mg/L zeatin, 0.25 mg/L thidiazuron (TDZ), and 5 mg/L 6-benzylaminopurine (6-BAP)). Consequently, it was found that 7- and 21-day-old hypocotyl fragments, as well as nodal stem segments obtained from adult aseptic seedlings, are characterized by a high explant viability and callus formation capacity with a frequency of 79.7–100%. However, the success of in vitro somatic shoot organogenesis was significantly determined not only by the culture medium composition and explant type but also depending on its age, as well as on the size and explant preparation in cases of hypocotyl and age-matched nodal stem fragments, respectively. Multiple somatic shoot organogenesis (5.7 regenerants per explant) with a frequency of 67.5% was achieved during 3 subcultures of juvenile hypocotyl-derived callus tissue on MS culture medium containing 0.25 mg/L TDZ as cytokinin source. Castor bean regenerants were excised from the callus and successfully rooted on ½ MS basal medium without exogenous auxin (81%). In vitro plantlets with well-developed roots were adapted to ex vitro conditions with a frequency of 90%. Full article

The efficiency and method of regeneration in in vitro culture conditions depend primarily on the plant growth regulators (PGRs) used. Even growth regulators belonging to one group may have different effects, stimulating the process of direct or indirect organogenesis, thus possibly disturbing the genetic stability among regenerants. The main aim of this study was to identify the genetic stability of Scutellaria baicalensis regenerates obtained by in vitro culture method using start codon targeted (ScoT) markers. S. baicalensis nodal explants were regenerated on MS medium supplemented with kinetin (KIN) at concentrations of 0.25, 0.5, 1.0, and 2.0 mg × dm⁻³ or benzylaminopurine (BAP)—0.25, 0.5, 1.0, and 2.0 mg × dm⁻³. The effects of the number of propagated shoots, length, number of nodes, and fresh mass of regenerants were assessed. Moreover, the genetic stability of the regenerants was analyzed using start codon targeted (SCoT) markers. Direct shoot organogenesis was observed on an MS medium containing kinetin, while indirect shoot induction occurred on an MS medium supplemented with BAP. The highest average number of shoots (3.6) was achieved for the MS + KIN medium at a concentration of 0.25 and 5.8 for the MS + BAP 1.0 medium. The average length and average number of nodes were the highest on the MS + BAP 0.25 medium (50.0 and 6.0, respectively), while the lowest values of these features were observed on the MS + KIN 2.0 medium (40.3 and 4.9, respectively). A total of 111 amplified bands were exhibited by SCoT primers. Three of the analyzed primers revealed four unique genotype-specific markers. The average percentage of polymorphism obtained was 36.7%. The analysis of genetic similarity revealed a high level of genetic similarity between the donor plant and regenerants obtained on MS “0” (medium without the addition of phytohormones). A slightly lower value of genetic similarity was observed for regenerants obtained by direct organogenesis (MS + KIN medium at all concentrations). Indirect shoot organogenesis observed on the MS + BAP medium (all concentrations) resulted in the highest differentiation, both in relation to the donor plant and MS “0” regenerants. The results of our work indicate that, in the case of S. baicalensis, the maintenance of genetic stability depends primarily on the presence of the cytokinin type in the medium. Full article