Search Results (34)

Lilium, a traditional plant with dual medicinal and ornamental values, is restricted in its industrial development by the low natural propagation rate of bulbs and dependence on imported high-quality germplasm. To address this bottleneck, this study used new lines of LA hybrid lilies and Asiatic hybrid lilies (three cultivars) as experimental materials to establish an efficient and stable tissue culture and rapid propagation system. Key procedures including disinfection of different explants (bulb scales and capsules), adventitious bud induction and proliferation, rooting culture, as well as acclimatization and transplantation were systematically evaluated. The results showed that bulb scales were superior to capsule seeds as explants in tissue culture (contamination rate 9.44%, regeneration rate 11.92%). After disinfection with 75% ethanol combined with 10% sodium hypochlorite, the contamination rate could be controlled at 14.29–21.43%, and the regeneration rate reached 100%. Supplementation with 50 g·L⁻¹ sucrose + 1 mg·L⁻¹ 6-BA + 0.1 mg·L⁻¹ NAA (Treatment TA1), 50 g·L⁻¹ sucrose + 2 mg·L⁻¹ 6-BA + 0.5 mg·L⁻¹ NAA (Treatment TA4) in MS medium, combined with dark culture, could effectively promote adventitious bud induction, proliferation and bulblet enlargement. For the rooting stage, the optimal media were 1/2 MS + 0.5 g·L⁻¹ activated charcoal + 2 mg·L⁻¹ 6-BA + 0.5 mg·L⁻¹ NAA (Treatment TB4) or MS + 0.3 g·L⁻¹ activated charcoal + 2 mg·L⁻¹ 6-BA + 0.5 mg·L⁻¹ NAA (Treatment TB1), and the highest rooting rate of ‘Pink Renault’ reached 100%. When plantlets from all three cultivars were combined and acclimatized and transplanted into sterilized peat soil, the overall survival rate was 89.33%. The TOPSIS method was also adopted for comprehensive evaluation to screen out the optimal culture conditions for different varieties. Based on phenotypic observation and physiological index data, ‘Pink Renault’ showed great potential as an excellent propagation germplasm. The integrated and optimized technical system provides a feasible solution for large-scale and industrialized seedling production of medicinal and ornamental lilies, and is of great practical significance for the efficient utilization of germplasm resources and sustainable development of the lily industry. Full article

Rosa canina L. is a medicinal and nutritionally valuable species with increasing industrial demand, yet its conventional propagation is limited by low rooting capacity and high genetic heterogeneity. In this study, a complete and reproducible in vitro micropropagation protocol was established, from explant introduction to plantlet acclimatization. Axillary buds were disinfected and introduced into Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (BAP). Shoot multiplication was achieved using sequential cytokinin treatments, and shoot elongation was promoted by adding liquid MS medium containing activated charcoal (AC). The highest and fastest root induction percentage (up to 75%) was obtained on WPM with 2 mg·L⁻¹ IBA and under a 16 h light/8 h dark photoperiod. Light promoted adventitious root formation depending on the nutrient formulation. Thereafter, shoots developed well-structured root systems in vitro, and plantlets fully survived to ex vitro acclimatization. This protocol provides an efficient platform for the large-scale propagation of R. canina and demonstrates that its auxin-driven adventitious rooting is strongly conditioned by the interaction between basal medium composition and photoperiod. Full article

Metal and metal oxide nanoparticles (NPs) synthesized through biologically mediated reduction of metal ions using biomolecules derived from microorganisms, algae, or plants are attracting growing attention in plant biotechnology due to their multifunctional properties and environmental advantages compared with conventional physicochemical synthesis. This review provides a comprehensive analysis of biological approaches for NP production using bacteria, fungi, algae, cyanobacteria, whole plants, and in vitro plant cell cultures. The main biosynthetic mechanisms, types of reducing and capping metabolites, metal specificity, and typical NP characteristics are described for each system, with emphasis on their relative productivity, scalability, reproducibility, and biosafety. Special consideration is given to plant cell and tissue cultures as highly promising platforms that combine the metabolite diversity of whole plants with precise control over growth conditions and NP parameters. Recent advances highlight the significance of bioengineering of reductive capacity as a novel strategy to enhance the efficiency and controllability of NP biosynthesis. Since NP formation is driven by key biomolecules, targeted modification of biosynthetic pathways through metabolic and genetic engineering can substantially increase NP yield and allow fine-tuning of their structural and functional properties. The applications of biogenic NPs in plant biotechnology are systematically evaluated, including their use as environmentally safe disinfectants for explants and seed sterilization, modulators of callus induction and morphogenesis, and abiotic elicitors that enhance the accumulation of economically valuable secondary metabolites. Remaining challenges, such as variability in NP characteristics, limited scalability, and insufficient data on phytotoxicity and environmental safety, are discussed to outline future research priorities. The synthesis–function relationships highlighted here provide a foundation for developing sustainable NP-based technologies in modern agriculture. Full article

Corylus avellana L. is a rare and endangered species in Kazakhstan, included in the national Red Book. The results of morphological and genetic characterization of the sole known natural population of C. avellana in the Western Kazakhstan region are presented in this study. Sixty wild accessions were evaluated based on tree and leaf morphological traits using standard descriptors in accordance with Bioversity International guidelines. Genetic diversity was assessed using ten nuclear simple sequence repeat (SSR) markers. A total of 120 alleles were detected across the nuclear loci, with the number of alleles per locus ranging from 9 to 16 and an average of 12. The mean effective number of alleles (Ne) per locus was 3.862. A high level of intraspecific polymorphism was observed, with an average observed heterozygosity (Ho) of 0.70. The population showed considerable genetic diversity, as highlighted by a mean Shannon’s diversity index of 1.526. STRUCTURE, PCoA, and phylogenetic analyses confirmed strong differentiation between the wild Kazakh population and the cultivated hazelnut germplasm. Due to the lack of viable seeds, in vitro conservation was initiated using vegetative shoots. A two-step disinfection protocol, involving Plant Preservative Mixture and mercuric chloride, significantly improved explant survival, enabling successful establishment of an aseptic in vitro collection. These findings highlight the urgent need for targeted conservation strategies and show the potential of biotechnological approaches for safeguarding Kazakhstan’s only natural C. avellana population. Full article

For its rapid growth, high yield, and broad adaptability, Arundo donax is widely used in various applications, yielding considerable economic and ecological benefits. However, widespread cultivation is challenging because A. donax can only be propagated asexually. In this study, a tissue culture method was developed using the clustered bud proliferation pathway. The explant type, disinfection method, induction medium, proliferation medium, and rooting medium were optimized to efficiently harvest high-quality A. donax seedlings. Using axillary buds with whole cane fragments as the most suitable explants, they were first sterilized with 75% alcohol for 30 s and then disinfected with 0.1% mercuric chloride for 5 min. 97.8% of explants could successfully form clustered buds on Murashige and Skoog (MS) medium supplemented with 5.0 mg/L 6-benzylaminopurine (6-BA) and 1.0 mg/L 3-indolebutyric acid (IBA). Each individual bud achieved efficient propagation with a proliferation coefficient as high as 33.3 on MS medium supplemented with 4.0 mg/L 6-BA and 1.0 mg/L IBA. In addition, all buds were capable of rooting on 1/2 MS medium supplemented with 0.5 mg/L 1-naphthaleneacetic acid (NAA). The resultant rooted seedlings survived and developed into plantlets, averaging 44.84 cm in height and 2.54 mm in thickness, following a 30-day acclimation period. This protocol provides a robust foundation for the large-scale, high-quality propagation of A. donax, supporting its broader application in ecological restoration and bioresource industries. 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

Ilex paraguariensis (yerba mate), a culturally and economically important South American species, faces significant challenges in vitro, including contamination, phenolic oxidation, and low regeneration rates. Nanoparticles have recently emerged as promising tools to overcome such limitations. This study evaluated silver (AgNPs) and chitosan nanoparticles (ChNPs) in eight experiments using nodal, leaf, and internodal explants. Surface disinfection with 1% colloidal silver solution 20 ppm significantly reduced contamination (17.2% and 15%) while maintaining viability (62.1%). However, supplementation of culture media with AgNPs (4–75 mg·L⁻¹) or ChNPs (5–120 mg·L⁻¹) did not improve nodal segment responses. In leaf explants, 4 mg·L⁻¹ AgNPs proved most effective, reducing contamination and markedly decreasing callus oxidation from 63.3% to 10.0%. Callogenesis was enhanced when AgNPs were combined with growth regulators, with the highest induction at 6 mg·L⁻¹ AgNPs + zeatin (38.1%) and 4 mg·L⁻¹ AgNPs + BAP (42.9%). Conversely, in internodal segments, AgNPs combined with BAP completely inhibiting callus formation. The resulting calli exhibited compact and friable morphologies but no signs of somatic embryogenesis. Overall, the effectiveness of AgNPs depends on their formulation, explant type, and interaction with cytokinins. Optimization of nanoparticle formulation and hormonal balance remains essential to maximize efficacy while minimizing toxicity. Full article

Regenerating sweet potato from field-derived plant material requires careful management of several critical factors, including the effectiveness of the disinfectant, the age of the explant, and the genotype used. In this context, establishing a reliable aseptic protocol is essential for successful in vitro culture. This study aimed to assess the effects of two disinfectants (sodium hypochlorite and mercuric chloride), three sweet potato genotypes (Nakabo, Boyapleu, and Irene), and three explant ages (2, 3, and 4 weeks) on clean culture establishment and regeneration efficiency from nodal explants. The findings revealed that regeneration success is significantly influenced by the type and concentration of disinfectant, explant age, and genotype. Treatment with 10% sodium hypochlorite markedly reduced contamination, achieving clean culture and regeneration rates of 75.72 ± 3.36% and 86.83 ± 3.02%, respectively, regardless of explant age. In contrast, higher concentrations of mercuric chloride induced necrosis in the explants. The highest clean culture rate (93.82 ± 1.16%) was observed in 3-week-old explants, which also showed a regeneration rate of 54.93 ± 3.19%. Furthermore, the Boyapleu and Irene genotypes demonstrated good suitability for in vitro culture, whereas the Nakabo genotype performed poorly under the tested conditions. Full article

Roses are one of the most essential ornamental flowers in the world. At present, traditional techniques such as cross breeding are mainly used in rose breeding. The inefficiency of the in vitro regeneration system has become the limiting step for the innovation and genetic improvement of rose germplasm resources. A tissue culture rapid propagation system of Rosa ‘Pompon Veranda’ was established using the stem segments with shoots as the initial experimental material. The results showed that the best disinfection method was to soak the explants in 75% ethanol for 1 min, and then soak them in 15% sodium hypochlorite solution for 15 min. The contamination rate was only about 6%. The best rooting medium for tissue culture seedlings was 1/2MS with 0.1 mg∙L⁻¹ NAA, and the rooting rate can reach around 95%. On this basis, calluses were induced by using leaflets of tissue-cultured seedlings as explants. The results showed that the optimal medium for inducing callus tissue was MS + 5.0 mg∙L⁻¹ 2,4-D, with an induction rate of 100%. The calluses were cultured in the medium of MS with 0.01 mg∙L⁻¹ NAA, 1.5 mg∙L⁻¹ TDZ and 0.1 mg∙L⁻¹ GA₃ for 12 days in the dark and then transferred to light conditions. The differentiation rate of callus was 10.87%. On the medium of MS with 0.5 mg∙L⁻¹ 6-BA, 0.004 mg∙L⁻¹ NAA and 0.1 mg∙L⁻¹ GA₃, the shoots could regenerate into whole plants. This study has established an in vitro regeneration system of R. ‘Pompon Veranda’, which is a key perquisite for the subsequent establishment of its genetic transformation system. Moreover, this method will also be an important reference for studies on quality traits such as floral scent and prickles of Rosa plants. Full article

In vitro micropropagation is used to rapidly shorten the breeding process of crops, such as kale, an internationally widespread winter vegetable. The aim of this study is to develop optimised micropropagation protocols for three kale varieties. First, it was determined which seed surface disinfection method resulted in the highest germination rate and the lowest infection rate. Secondly, it was investigated which of several existing Brassica protocols and one modified protocol from the literature provided the highest regeneration efficiency of kale explant types (cotyledons, hypocotyl, root, and intact seedlings as the control) after eight weeks of cultivation. Germination was highest and fastest after disinfection with 10% NaClO for 10 min for “Frostara” and at 5% for 2.5 min for “Schatteburg”. The infection rate and speed were lowest in treatments with 10% NaClO. The regeneration efficiency and number of newly formed leaves, roots, shoots, and stems varied between media, explant type, and kale variety. Most new leaves and shoots were formed when hypocotyls were used as explant type. Roots regenerated mostly more roots than shoots, stems, and leaves. A higher ratio of auxin to cytokinin in the culture medium partially increased leaf regeneration. The addition of AgNO₃ increased shoot regeneration and reduced yellowing and leaf drop. Phenotypic anomalies occurred less frequently in media with lower hormone concentrations. All tested protocols are suitable for kale micropropagation, but regeneration was highly dependent on the medium for different varieties and explant types. Therefore, this study builds a basis for future micropropagation of kale and the development of variety-specific protocols for maximum commercial success. Full article

The only biocidal iodine species in topical iodine disinfectants is molecular iodine (I2). I2, a biomolecule, has broad-spectrum antimicrobial activity and does not generate resistance. Physicians, regulatory agencies, and scientists have assumed that I2 is responsible for the skin staining and irritation associated with the clinical use of iodine disinfectants; this assumption is deeply embedded in the medical community but is not supported with empirical data. This study provides the first experimental data that measure the biocompatibility of I2 with human skin. Human skin explants in tissue culture were evaluated at 3, 7, and 24 h after being exposed to I2 (500 to 15,000 ppm). Cell viability was measured relative to phosphate-buffered saline using 3-[4,5-dimethylthiazol-2yl]-2,5-diphenyl-tetrazolium bromide (MTT). The biocidal activity of I2 vapor emitted from silicone was demonstrated against bacteria growing on agar to confirm I2 off-gassing from skin was an active biocide. Additionally, statistically significant bacterial reductions with both gas and solution phase I2 were observed in a static and dynamic five-species wound biofilm. The data suggest that high, e.g., 50–5000 ppm, levels of I2 should be incorporated into topical iodine disinfectants instead of the very low (0.2–10 ppm) levels found in 10% povidone iodine products currently in use. Full article

Nandina domestica ‘Firepower’ is one of the most popular colorful foliage species in landscaping. However, it is currently propagated mainly by seeding and cuttings, with a low reproduction coefficient, hindering the cultivation of this species. Therefore, establishing an in vitro regeneration system would be beneficial for the industrialized production of Nandina domestica ‘Firepower’. In this study, an ex vivo regeneration system was established using the direct organogenesis pathway. In early April, the new shoots of Nandina domestica ‘Firepower’ were selected, and the stem segments of 1~2 cm were cut as the disinfection materials for the explants. The optimal formulation for inducing axillary shoots was 1/2 MS + 1.5 mg L⁻¹ 6-benzylaminopurine (BA) + 0.3 mg L⁻¹ indole-3-butric acid (IBA). The optimal formulation for the differentiation and proliferation of axillary shoots was 1/2 MS + 1.5 mg L⁻¹ BA + 0.01 mg L⁻¹ IBA with a multiplicity of proliferation of 9.22. We determined that the rooting of axillary shoots required a combination of IBA, naphthalene acetic acid (NAA), and activated carbon (AC). The optimal formulation for rooting was 1/2 MS + 0.2 mg L⁻¹ NAA + 0.3 mg L⁻¹ IBA + 0.2 mg L⁻¹ AC. After a two-day hardening period for tissue-cultured plantlets, a substrate consisting of peat soil, vermiculite, and perlite at a ratio of 2:2:1 was determined to be the optimal cultivation formulation. This system provides a framework for the industrialized production of Nandina domestica ‘Firepower’. Full article

Viburnum opulus L. ‘Roseum’ is a highly valuable ornamental plant for landscaping, but it has a long propagation cycle and low propagation coefficient. In this study, stem segments with axillary buds from Viburnum opulus L. ‘Roseum’ were used as explants. We systematically analyzed the use of sodium hypochlorite for the sterilization of explants, as well as the effects of different plant growth regulator combinations and concentrations on shoot bud induction, shoot proliferation, the rooting of tissue-cultured shoots, and the transplanting of the tissue-cultured shoots. A complete rapid propagation technology system for Viburnum opulus L. ‘Roseum’ was established. The results showed that a disinfection method using 75% ethanol for 30 s and soaking in 5% sodium hypochlorite for 5 min was the most suitable for disinfecting the stem segments of Viburnum opulus L. ‘Roseum’, which showed low contamination and a 73.33% survival rate. The ideal medium for primary bud induction was WPM (Woody Plant Basal Medium) + 2.0 mg·L⁻¹ 6-benzylaminopurine (6-BA) + 0.15 mg·L⁻¹ indole-3-butyric acid solution (IBA) + 25 g·L⁻¹ sucrose. The optimal medium for shoot proliferation was WPM + 1.0 mg·L⁻¹ 6-BA + 0.15 mg·L⁻¹ IBA + 25 g·L⁻¹ sucrose, achieving an induction rate of 7.17. For the rooting of tissue-cultured shoots, the most suitable formulation was 1/2 WPM + 0.3 mg·L⁻¹ naphthaleneacetic acid (NAA) + 0.3 mg·L⁻¹ activated charcoal (AC) + 25 g·L⁻¹ sucrose, which induced robust and developed root systems. This study provides a technical basis for the establishment of a fast propagation system for the industrial production of Viburnum opulus L. ‘Roseum’. Full article

Quercus suber L. (Q. suber) is an evergreen tree species known for producing high-quality cork. Traditional seed propagation of Q. suber has low viability and is time-consuming. Therefore, we used young stem segments of 2~3-year-old seedlings as explants, and optimized protocols for tissue culture and rapid micropropagation of Q. suber. The best disinfection method was 0.10% HgCl₂ (v/v) for 5 min. 0.50 g·L⁻¹ Poly Vinyl Pyrrolidone (PVP) is the best anti-browning agent with a significant reduction in browning by nearly 1.76-fold (58.89% → 33.33%). Woody Plant Medium supplemented with micronutrients and vitamins from Murashige and Skoog Medium (WPMS) was found to be the most suitable for shoot formation. The optimal hormone ratio for development of shoots from axillary buds was 0.60 mg·L⁻¹ 6-benzyladenine (6-BA). Among the cytokinins tested, 0.50 mg·L⁻¹ 6-BA was the most suitable for development of shoots from axillary buds. In additon, the highest percentage of rooting explants (66.67%) and rooting number (3.03) was obtained on WPM basal medium supplemented 0.20 mg·L⁻¹ IBA and 0.20 mg·L⁻¹ NAA. In summary, we have established a set of protocols for tissue culture and rapid micropropagation of Q. suber. These findings lay the foundation for rapid micropropagation and genetic improvement. Full article

Using Panax japonicus as research material, callus induction and culture were carried out, and high-yielding cell lines were screened to establish a suspension culture system that promotes callus growth and the accumulation of the “total saponins” (total content of triterpenoid glycosides or ginsenosides). Using the root as an explant, the medium for callus induction and proliferation was optimized by adjusting culture conditions (initial inoculation amount, carbon source, shaking speed, hormone concentration, culture time) and a high-yielding cell line with efficient proliferation and high total saponins content was screened out. The conditions of suspension culture were refined to find out the most suitable conditions for the suspension culture of callus, and finally, the suspension culture system was established. We found that the lowest (5%) contamination rate was achieved by disinfecting the fresh roots with 75% alcohol for 60 s, followed by soaking in 10% NaClO for 15 min. The highest induction rate (88.17%) of callus was obtained using the medium MS + 16.11 μmol·L⁻¹ NAA + 13.32 μmol·L⁻¹ 6-BA + 30.0 g·L⁻¹ sucrose + 7.5 g·L⁻¹ agar. The callus was loose when the callus subcultured on the proliferation medium (MS + 5.37 μmol·L⁻¹ NAA + 13.32 μmol·L⁻¹ 6-BA + 30.0 g·L⁻¹ sucrose + 3.8 g·L⁻¹ gellan gum) for 21 days. The callus growth was cultured in a liquid growth medium (MS + 5.37 μmol·L⁻¹ NAA + 13.32 μmol·L⁻¹ 6-BA + 30.0 g·L⁻¹ sucrose) with an initial inoculation amount of 40 g·L⁻¹, a shaking speed of 110 r/min and darkness. Cell growth was fastest with a culture period of 21 days. We replaced the growth medium with the production medium (MS + 5.37 μmol·L⁻¹ NAA + 13.32 μmol·L⁻¹ 6-BA + 30.0 g·L⁻¹ glucose) for maximum accumulation of total saponins. [Conclusion] A callus induction and suspension culture system for the root of P. japonicus was established. In this way, we can promote the accumulation of total saponins in callus cells and provide a basis for large-scale cell culture and industrial production of medicinal total saponins. Full article