Search Results (86)

To overcome the seasonal constraints of explant availability and facilitate genetic improvement in Catalpa ovata, this study established a dual-pathway in vitro regeneration system (encompassing adventitious shoot organogenesis and somatic embryogenesis) using mature zygotic embryos. We systematically evaluated the synergistic effects of maternal genotypes, plant growth regulators (PGRs), basal media, and the histone deacetylase inhibitor Trichostatin A (TSA). Genotype screening revealed significant divergence in regenerative potential, with the half-sib family 32F17 exhibiting superior responsiveness (84.7% callus induction). A high cytokinin-to-auxin ratio (ZA3 medium) optimally drove direct shoot organogenesis. For adventitious shoot proliferation, the addition of TDZ significantly improved the multiplication coefficient (up to 2.99 on ZB4 medium), although a physiological trade-off with shoot elongation was observed. In parallel, the application of 10 µM TSA significantly enhanced somatic embryogenesis from embryogenic calli, effectively alleviating the inhibitory constraints of exogenous PGRs. For rhizogenesis, the DKW basal medium proved superior to half-strength MS, with the ZE3 treatment (0.1 mg·L⁻¹ NAA + 0.1 mg·L⁻¹ IBA) yielding the highest rooting frequency (69.6%) and robust root architecture. Notably, while somatic embryo conversion remained recalcitrant, plantlets derived exclusively from the adventitious shoot organogenesis pathway were successfully acclimatized ex vitro. These transplanted plantlets exhibited consistently high survival rates (83.1–84.4%) across all tested genotypes, effectively overcoming the initial genotype-dependent recalcitrance. Collectively, this optimized protocol provides a reliable technical platform for the large-scale clonal propagation and biotechnological breeding of C. ovata. Full article

The ornamental plant industry faces escalating threats from erratic climate patterns and post-harvest perishability. Phytomelatonin (N-acetyl-5-methoxytryptamine) has emerged as a potent biostimulant capable of addressing these critical bottlenecks. This review synthesizes current knowledge on melatonin’s multifaceted roles in ornamental horticulture, clarifying the molecular pathways where it acts as both a direct Reactive Oxygen Species (ROS) scavenger and a signaling molecule orchestrating crosstalk with auxins, abscisic acid, and ethylene. We highlight applications in propagation, where melatonin synergizes with auxins to enhance rhizogenesis and promotes seed germination via hormopriming. Furthermore, we examine melatonin-mediated tolerance to abiotic stresses including drought, salinity, and temperature extremes emphasizing its role in preserving photosynthetic machinery and ion homeostasis. Crucially, the review addresses the post-harvest sector, demonstrating how melatonin extends vase life by repressing senescence-associated genes (SAGs) and antagonizing ethylene biosynthesis. Finally, we discuss future perspectives on genetic bio-fortification and commercial formulations, positioning phytomelatonin as a sustainable tool for securing the resilience and quality of ornamental crops. Full article

Marijuana (Cannabis sativa L.) has a great economic potential due to its phytotherapeutic properties. Its propagation, however, faces numerous challenges due to the limited availability of standardized technical protocols for the crop. Vegetative propagation represents a, or even the, viable method for multiplying the genetically identical individuals while preserving their phytochemical profile, at lower costs and with shorter production times. This study investigated the morphological and colorimetric attributes associated with vegetative propagation success, aiming to develop sustainable cultivation strategies. Four cutting lengths (5, 10, 15 and 20 cm) were evaluated after 21 days of rooting, considering fresh mass, basal diameter, presence of apical meristem, number of root primordia, root length, and foliar and stem color parameters. Logistic regressions indicated that longer cuttings (p = 0.0101), greater fresh mass (p = 0.073) and the presence of apical meristem (p = 0.065), as well as greener leaves (p = 0.089), were positively associated with rooting probability (p < 0.10). Positive correlations between morphological and colorimetric variables were confirmed by Principal Component Analysis, with the first two principal components explaining 31.2% of the total variance in the dataset. The results provide support for the development of more efficient and low-cost vegetative propagation protocols, promoting uniformity and autonomy in local cutting production of marijuana. Full article

Sweet potato (Ipomoea batatas) is a globally important crop and one of a growing number of plants recognized as naturally transgenic, harboring Agrobacterium-derived T-DNA genes whose functions remain largely uncharacterized. In this proof-of-concept study, we applied CRISPR/Cas9 technology to generate targeted knockouts of the Ib-rolB/C and Ib-rolD-like genes located within the sweet potato cellular T-DNA2 (IbT-DNA2) region. Mutations were introduced into sweet potato callus cultures using an optimized genome editing protocol, with most edits consisting of single-nucleotide insertions. Knockout of Ib-rolB/C did not affect callus growth but significantly reduced levels of chlorogenic acid derivatives. Validation in planta using transient expression in I. batatas leaves confirmed the suppressive effect of Ib-rolB/C disruption on polyphenol content. In contrast, Ib-rolD-like knockout lines showed reduced biomass accumulation and downregulation of cell cycle–related genes, but did not display significant changes in metabolite content in either callus cultures or leaf tissues. These findings suggest that Ib-rolB/C and Ib-rolD-like may differentially contribute to growth and secondary metabolism in sweet potato. Full article

Background/Objectives: Licorice (Glycyrrhiza glabra L.) is a highly important medicinal plant that is widely used in China owing to its active ingredients. Its main active components are flavonoids, including liquiritigenin, liquiritin and licochalcone A. The hairy roots (HRs) induced by Agrobacterium rhizogenes are a commonly used chassis in synthetic biology to enhance the production of active compounds in medicinal plants. Methods: A biosynthesis system to acquire the active ingredients of G. glabra was established using an HR culture system. It employed a transcriptome analysis to identify the change in gene expression following treatment with methyl jasmonate (MeJA). Results: After 28 days of suspension culture, the biomass of HRs increased by approximately 34.5-fold and reached 1.83 g/100 mL flask. Treatment with MeJA significantly increased the contents of liquiritigenin, liquiritin, and glabridin in the HRs. The transcriptome data indicated that MeJA activated the flavonoid biosynthetic pathway genes in the HRs, which was largely consistent with the qRT-PCR results. Furthermore, the overexpression of the GgCHS6 gene substantially increased the content of flavonoids in HRs. Conclusions: Collectively, this study established an HR system to biosynthesize the active ingredients of G. glabra using metabolic engineering and genetic engineering techniques and provides several valuable candidate genes for further functional study. Full article

Woody plants, comprising forest and fruit tree species, provide essential ecological and economic benefits to society. Their genetic improvement is challenging due to long generation intervals and high heterozygosity. Genetic transformation, which combines targeted DNA delivery with plant regeneration from transformed cells, offers a powerful alternative to accelerating their domestication and improvement. Agrobacterium tumefaciens, Rhizobium rhizogenes, and particle bombardment have been widely used for DNA delivery into a wide variety of explants, including leaves, stems, hypocotyls, roots, and embryos, with regeneration occurring via direct organogenesis, callus-mediated organogenesis, somatic embryogenesis, or hairy root formation. Despite successes, conventional approaches are hampered by low efficiency, genotype dependency, and a reliance on challenging tissue culture. This review provides a critical analysis of the current landscape in woody plant transformation, moving beyond a simple summary of techniques to evaluate the co-evolution of established platforms with disruptive technologies. Key advances among these include the use of developmental regulators to engineer regeneration, the rise in in planta systems to bypass tissue culture, and the imperative for DNA-free genome editing to meet regulatory and public expectations. By examining species-specific breakthroughs in key genera, including Populus, Malus, Citrus, and Pinus, this review highlights a paradigm shift from empirical optimization towards rational, predictable engineering of woody plants for a sustainable future. Full article

The aim of the present study was to evaluate the morphogenetic efficiency of the new flax (Linum usitatissimum L.) cultivar Silesia. The plant material consisted of five ecotypes of Silesia seeds selected by determining the fatty acid composition in individual plants. Thus, five ecotypes with different fatty acid compositions were applied for germination, and the resulting seedlings were used for callus induction. The observed in vitro morphogenic response of explants to the used callus induction medium was 100%, followed by varying development of shoots, with the highest value being 17 shoots per callus, with an average length of 7.15 cm (for ecotype 1). The biochemical analyses showed photosynthetic pigments were significantly affected by the tested ecotypes. The main conclusion is that the selection of plant material as a source of tissue cultures is crucial for efficient regeneration and organogenesis. One factor potentially influencing these processes is fatty acid composition and the ratio of linoleic to α-linolenic acid. A ratio ranging from 1.36 to 1.68 in the source material (seeds) used for initiation of tissue cultures resulted in the highest efficiency of shoot regeneration and number of obtained shoots per callus. A strong negative correlation (−0.78) was observed between shoot regeneration efficiency and the ratio of linoleic to α-linolenic acid in seeds from which the explants were obtained. Moreover, an efficient protocol of micropropagation from callus tissue was established for the new flax cultivar Silesia. An interesting insight into the metabolism of the obtained regenerants allowed us to determine the relationship between the content of chlorophyll and rhizogenesis efficiency. For these two parameters, the calculated correlation coefficient was 0.66. A strong relationship (high correlation coefficient: 0.79) was also established between flavonoid content and length of obtained regenerants, pointing to the developmental role of flavonoids. Full article

Despite the great structural diversity, plant lignans, coumarins, and xanthones share numerous biological activities, ranging from antimicrobial, anti-inflammatory, and antioxidant to antineoplastic and neuroprotective. The compounds, products of the shikimic acid biosynthetic pathway, also play an important role in plant–environment interactions. In a search for sustainable and renewable sources of these valuable plant products, numerous in vitro culture systems were investigated, including hairy root cultures. The Rhizobium rhizogenes-transformed root cultures of over 40 plant species representing 17 families of the plant kingdom were studied in this respect. The present review focuses on the hairy roots that may be efficient producers of valuable plant products with the prospect of use in the pharmaceutical, food, or cosmetics industry. In vitro culture systems based on hairy roots, which were used to elucidate the biosynthesis pathways of the high-added-value plant compounds, were also considered. Full article

Sugar beet (Beta vulgaris L.), a biennial sugar crop, provides about 16% of the world’s sucrose production. PEG and Agrobacterium tumefaciens-mediated transformation have been established for sugar beet. However, the traditional transformation of sugar beet is time-consuming, low efficiency, and dependent on tissue regeneration. Recently, the use of Agrobacterium rhizogenes for genetic transformation without tissue culture has become a new possibility. Here, we describe an optimized A. rhizogenes-mediated transformation for the generation of composite sugar beet without tissue culture. By dipping A. rhizogenes K599 colonies onto a wound of hypocotyl and petiole, about 81.7% and 51.1% of shoots and leaves could be induced to produce adventitious roots. Of these, more than 60% of the explants contained transformed adventitious roots. Specifically, we discovered that the transformation efficiency was significantly improved when the MAS promoter was employed instead of the CaMV35S promoter. The transformation in adventitious roots was also validated by qRT-PCR and Western blot at the transcriptional and translational levels. The transformed adventitious roots have great potential for the study of taproot development, sugar accumulation, and resistance to root diseases, which is closely related to sugar beet yield and quality. Full article

In the present study, the propagation ability of rootstock 1103 Paulsen in a hydroponic system was investigated. In the first part of the experiment, the effects of indolebutyric acid (IBA), dopamine (L-DOPA), and their combination on rhizogenesis were examined. The experiment was conducted under controlled conditions in a hydroponic system. Key parameters evaluated included rooting percentage, average root diameter, average number of roots per cutting, total root area, total root length, and moisture content of the cuttings. Results showed that L-DOPA treatment, followed by the IBA + L-DOPA combination, exhibited the most favorable outcomes across these parameters. The hydroponic system proved highly effective for root formation compared to other substrates (e.g., peat, perlite, sand, or their combinations), provided that continuous aeration of the water was ensured for adequate oxygenation. The second part of the experiment focused on the response of phenolic compounds, antioxidants, sugars, and starch in woody cuttings subjected to different treatments (control, IBA, L-DOPA, and IBA + L-DOPA) and how these compounds varied over time. The objective was to assess the influence of the treatments on both the rhizogenesis process and the biochemical profile of the cuttings throughout the experiment. This research aims to contribute to the understanding of the rooting behavior of 1103 Paulsen in hydroponic systems and to evaluate the physiological and biochemical responses of cuttings under different treatments. Full article

The southern region of Kazakhstan represents the northernmost boundary of the natural habitat of five wild almond species, among which Amygdalus communis L. is of particular interest due to a range of favorable traits for use in breeding programs and cultivation in the region. The current distribution range of common almond growth was clarified using GPS to determine precise coordinates, and a schematic map was developed. Monitoring revealed a significant reduction in population size. In the surveyed areas, 54 trees were selected and described. Seed material was collected from 34 genotypes and characterized according to a descriptor. Genotypes A3, A8, and A15 were identified as having favorable trait combinations. To restore populations and preserve the gene pool of Amygdalus communis L., a method of clonal micropropagation was employed. The composition of the nutrient medium was optimized for establishment, multiplication, and rhizogenesis. It was determined that Murashige and Skoog (MS) medium without phytohormones is effective for in vitro establishment (70% regeneration rate). For multiplication, MS medium with 0.5 mg/L BAP (6-benzylaminopurine) was used (with a multiplication rate of 3.5 per explant). For rhizogenesis, MS medium with 0.5 mg/L BAP, 0.02 mg/L gibberellic acid (GA), and 0.1 mg/L IBA (indole-3-butyric acid) was used. A total of 340 clonal Amygdalus communis L. plants with closed root systems were grown for field collection. The research results can be applied for the restoration, propagation, and conservation of populations both in vitro and in situ, as well as for the inclusion of selected high-performing genotypes in breeding programs. Full article

Halophytes are salt-tolerant plants with ethnomedicinal value and growing potential in food and cosmetics; their adaptability to extreme conditions makes them promising candidates for sustainable agriculture and crop development in salt-affected areas. In vitro plant tissue culture further supports this by enabling resilient plant production in the face of climate and food security challenges. In this study, in vitro cultures of two medicinal halophytes from the genus Plantago (P. coronopus and P. crassifolia) were established to optimize their micropropagation protocol. Seed germination percentages, growth parameters, micropropagation rates, rooting efficiency, and physiological condition were evaluated. Growth media (modified MS medium) differed in the type of cytokinin. The seed germination efficiency was monitored at weekly intervals for 8 weeks, and other growth parameters were evaluated in 6- and 12-week cultures. Differences in both the rate and efficiency of in vitro germination between the two species were observed, with approximately 73% germination reached by P. coronopus and 47% by P. crassifolia after 4 weeks, and 80% and 53% after 8 weeks, respectively. The addition of 0.5 mg dm⁻³ kinetin plus 0.5 mg dm⁻³ IAA (indole acetic acid) proved to be effective in promoting growth in P. coronopus, resulting in longer plantlets and higher multiplication rates, while the addition of meta-topolin (mT) was a better stimulator of shoot and root growth in P. crassifolia. The highest multiplication coefficient, 6.22 for P. coronopus and 4.90 for P. crassifolia, was obtained on the P1 medium for both species. Importantly, medium with mT also had a stimulating effect on rooting in both species over the long term (12-week culture). The developed PTC enables efficient propagation and trait selection in halophytes, supporting sustainable large-scale production of the studied Plantago species, and facilitating future research on salt stress tolerance. Full article

Passion fruit (Passiflora edulis Sims), belonging to the Passifloraceae family, is an economically important plant in tropical and subtropical regions. The advances in functional genomics research of passion fruit have been significantly hindered by its recalcitrance to regeneration and stable transformation. This study establishes the first efficient Agrobacterium rhizogenes-mediated hairy root transformation system for passion fruit. Utilizing the eGFP marker gene, transformation efficiencies of 11.3% were initially achieved with strains K599, MSU440, and C58C1, with K599 proving most effective. Key transformation parameters were systematically optimized to achieve the following: OD₆₀₀ = 0.6, infection duration 30 min, acetosyringone concentration 100 μM, and a dark co-cultivation period of 2 days. The system’s utility was further enhanced by incorporating the red visual marker RUBY, enabling direct, instrument-free identification of transgenic roots via betaxanthin accumulation. Finally, this system was applied for functional analysis using PeMYB123, which may be involved in proanthocyanidin accumulation. Overexpression of PeMYB123 produced a higher content of proanthocyanidin in hairy roots. Additionally, the PeANR gene involved in the proanthocyanidin pathway was strongly activated in the transgenic hairy roots. This rapid and efficient visually simplified hairy root transformation system provides a powerful tool for functional gene studies in passion fruit. Full article

Hairy root cultures induced by Agrobacterium rhizogenes (Rhizobium rhizogenes) provide a sustainable approach to meet the growing demand for economically valuable plant-derived compounds in the face of depleting natural resources. These cultures exhibit rapid, hormone-independent growth and genetic stability, making them viable for producing bioactive compounds, plant-specialized metabolites, and recombinant proteins. However, challenges remain in optimizing large-scale production, improving bioreactor efficiency, and enhancing metabolite synthesis across different plant species. This review addresses these challenges by exploring the mechanisms behind the induction of hairy root cultures, their applications in genetic and metabolic engineering, and their potential in environmental remediation. The review further highlights recent advances in biotechnology and illustrates how the hairy root system can sustainably meet industrial, pharmaceutical, and agricultural needs. In addition, by pointing out essential research areas such as optimizing culture conditions, increasing metabolite yields, and scaling up production, this work strengthens the significance of hairy root cultures in meeting the demand for high-value products while ensuring sustainable resource utilization. In particular, the integration of hairy root systems with advanced genomic tools such as transcriptomics and CRISPR technology holds immense potential for accelerating pathway-specific metabolic engineering, enhancing biosynthetic flux, and expanding their applications in sustainable agriculture and pharmaceutical innovation. This convergence is expected to drive substantial economic value by optimizing the production of high-value bioactive compounds, improving crop resilience, and facilitating precision medicine. Future work involving systems and synthetic biology will be instrumental in unlocking novel functions and ensuring broader deployment of hairy root cultures across industrial biotechnological platforms. Full article

This study examined the bioproductivity of two poplar genotypes propagated by single-tree stem cuttings. The experiment compared variants using cuttings of different lengths (10–22 cm) and containers with volumes from 1 to 3 L. It was found that the best growth performance of seedlings in height according to the traditional container technology (70.6 ± 5.5–111.5 ± 5.0 cm) was observed in the intersectional hybrid of poplar ‘E.s.-38’ (genotype 1). The predominance of the genotype factor over the technology of rooting cuttings was established. The fast-growing genotype 1, E.s.-38, had higher productivity and plant height indices, suggesting it as a variety that can allow for the growth of standard planting material in containers in one season. For genotype 1, the length of cuttings was 10–14 cm when the container volume was increased to 3 L, which could increase the number of cuttings from one mother plant by 2–3 times. The revealed correlations between the height of the seedling and the diameter of the increment, as well as the analysis of the proportions of plant organs, showed that biological features of the rooting of stem cuttings depended on the genotype of poplars. The natural type G2 was characterized by the prevalence of root system growth over the growth of other organs; in the case of short cuttings, the proportion of leaves increased in plants to enhance photosynthesis and ensure rhizogenesis. Full article