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Gentiana spp. are ornamental as well as valuable medicinal plants. This research was conducted to investigate the effects of different cytokinin and auxin treatments on shoot propagation and rooting of both G. lutea and G. boissieri, and to establish the most appropriate micropropagation protocols for these species. For this purpose, MS media containing benzyl adenine (BA) (1, 2, 3, and 4 mg/l), zeatin (0.5, 1, 2, and 3 mg/l), and kinetin (0.5, 1, 2, and 3 mg/l) were used for shoot propagation, and the optimal cytokine applications were determined by examining shoot number per explant (SN/E) and shoot length (SL). To determine rooting ability, shoots were transferred to MS media containing indole acetic acid (IAA), indole butyric acid (IBA), and naphthalene acetic acid (NAA) at different concentrations (1, 2, 3, and 4 mg/l). The optimum auxin applications for root and plant development were determined by examining the rooting percentage (%, RP), root number per shoot (RN /S), root length (RL), shoot length (SL), and plant fresh weight (FW). As a result of the study, it was found that the media containing 3 mg/l and 2 mg/l BA were the most suitable shoot propagation media for G. lutea and G. boissieri, respectively. The media containing 4 mg/l IBA and 1 mg/l AgNO3 gave the highest rooting and growth parameters for G. lutea, while G. boissieri had the best rooting and growth values in media supplemented with 2 mg/l NAA. In addition, G. boissieri was found to be superior to G. lutea in both shoot propagation and rooting. [ABSTRACT FROM AUTHOR]

Melatonin was discovered in plants in 1995, and so-called phytomelatonin. Its characteristic qualities as an antioxidant molecule and with amphiphilic nature make it an excellent candidate to use in plant biotechnological techniques. In plant physiology, melatonin has demonstrated with multiple studies great possibilities of application in crop improvement in aspects such as germination, rooting, plant growth, leaf-senescence, floration, fruit-set, parthenocarpy, fruit ripening and cut flower conservation, among others. In this work, we reviewed many studies on the possible role of melatonin to improve the different plant tissue culture goals, among them: the role of melatonin in callus culture induction and its effects in plant biomass production; the possible action of melatonin as an inductor or elicitor of different secondary metabolite generation in in vitro cultures; the interesting possibilities of melatonin as a protective agent (safener) against abiotic stressors present in in vitro cultures; the potential of melatonin in regulating plant organogenesis, modulating shooting/rooting responses; at last, the studies on the relevant role of melatonin in plant material cryopreservation and the improvement of cell transformation using A. tumefaciens have been discussed. [ABSTRACT FROM AUTHOR]

Zani WNWM, Sidik NJ, Isa NAM, Awal A, Osman NI, Alias N, Nordin MK. 2024. Influence of media variations and growth regulators on in vitro propagation of Dendrocalamus asper bamboo. Nusantara Bioscience 16: 237-244. This study reports the effects of different types of media and combinations of plant growth regulators on the in vitro propagation of Dendrocalamus asper (Schult.f.) Backer bamboo species, a species known for its economic significance and challenges in traditional cultivation. The experiment was initiated through the cultivation of in vitro nodal segments as explants in various strengths (half-strength and full-strength) of Murashige and Skoog's (MS) and Vacin & Went (VW) media, supplemented with 1.0 mg L-1 6-benzylaminopurine (BAP) hormone. Optimal results were achieved in full-strength MS media with 1.0 mg L-1 BAP, with the highest shoot number (5.6 shoots) and length (2.14±0.18 cm), surpassing outcomes in VW media. Shoot multiplication in full-strength MS media, with varying BAP and indole-3-butyric acid (IBA) hormone combinations (0.5, 1.0, 2.0, and 4.0 mg L-1), was conducted. The combination of 4.0 mg L-1 BAP and 0.5 mg L-1 IBA yielded the highest number of shoots (5.17±3.97), while supplementation with 4.0 mg L-1 IBA alone resulted in the longest shoot with 1.89±1.15 cm. These findings underscore the significance of tailored conditions for optimal in vitro propagation of this species. Further investigations could explore additional factors influencing the propagation process for better refinement of bamboo cultivation techniques. [ABSTRACT FROM AUTHOR]

Plant tissue culture biotechnology offers a sustainable way to adapt to climate change by propagating disease-free plants with the desired traits. This study investigated stakeholders' agronomic, social and economic evaluation of tissue culture plants (TCPs). It unravels micro-level outcomes and indicators that determine TCPs evaluation through a systematic literature review. The 35 articles examined show an imbalance of studies in the three evaluation streams, with social studies on horticultural crops, especially bananas in developing countries, dominating. The results of the TCPs evaluation vary considerably between stakeholders. The main determinants of evaluation are TCPs attributes. TCPs are very knowledge-intensive and costly to establish and manage. Therefore, customising of TCPs to local conditions and closer collaboration between stakeholders are essential for successful implementation. Advisory services, social entrepreneurship and networks are needed to create knowledge-sharing platforms and ecosystems. While single-stream evaluations reveal TCPs superiority over conventionally propagated crops, it is suggested that an integrated assessment be undertaken to combine social, economic, and agronomic indicators and evaluate the technology simultaneously. [ABSTRACT FROM AUTHOR]

Plantlets grown in vitro show different growth responses and photosynthetic pigment production when they are exposed to different light conditions. The aim of the present study is to assess the agronomic and physiological performance of sugarcane (Saccharum spp. hybrid) grown in vitro under different light intensity and spectrum variations. Plantlets measuring approximately 2 cm, grown in vitro, based on somatic embryogenesis, were cultivated in Murashige and Skoog (MS) basic medium and subjected to five light intensities: 26, 70, 100, 113 and 144 µmol m− 2 s− 1, at different light spectrums: 100% white, 100% red, 100% blue and blue/red combinations at %R: %B (30:70; 70:30; and 50:50) ratios obtained by LEDs, for 35 days. The optimal light intensity for maximizing dry weight and photosynthetic pigment production was 113 µmol m⁻² s⁻¹, which contributed to enhanced plantlet vigor. Furthermore, plantlets subjected to white, red, and blue light spectra showed the highest dry weight accumulation, while the 50% blue:50% red spectrum was most effective in promoting biosynthesis of photosynthetic pigments. In conclusion, this study demonstrated that the growth and physiological responses of sugarcane (Saccharum spp. hybrid) plantlets grown in vitro are significantly influenced by variations in light intensity and spectrum. These findings offer valuable insights for optimizing in vitro cultivation conditions to enhance sugarcane growth and development.Key message: Photon flux density and wavelength affect growth and pigments accumulation in Saccharum officinarum L. and the combination 50% blue:50% red enabled photosynthetic pigments to reach greater biosynthesis. [ABSTRACT FROM AUTHOR]

Marigold (Tagetes erecta L.), holds significance for traditional medicine and ornamental gardening in India. This study investigates the optimization of micropropagation in cultivars Pusa Narangi Gainda and Pusa Basanti Gainda, via axillary shoot tip culture through germinated seedlings. It emphasizes the impact of hormone concentrations on shoot multiplication, vitrification, and related physiological responses, including the activities of key enzymes like chitinase and β-glucanase. Elevated cytokinin levels were found to suppress shoot growth and induce oxidative stress, evidenced by increased malondialdehyde and enzyme activity. The research identifies critical factors influencing vitrification through correlation analysis, PCA, and eigen analysis, offering strategies to enhance marigold micropropagation and reduce physiological disorders. Notably, this is the first report documenting chitinase and β-glucanase activity in marigolds.Key message: This work offers the first documentation of the function of biological components as β-1.3 glucanase and chitinase in vitrification. [ABSTRACT FROM AUTHOR]

Zingiberaceae family, also known as the ginger family, comprises 53 genera and 1300 species. The members of this family are widely known for their ethnobotanical properties. Owing to their anti-inflammatory, antimicrobial, antidiabetic, and cardioprotective properties, the rhizomes have been used in traditional medicine to treat a wide range of illnesses like cold, inflammation, gastritis, arthritis, diabetes, and respiratory problems. The medicinal properties of the plants are attributed to the wide array of secondary metabolites present in them. They also find their use in food, spices, ornamentals, perfumes, and cosmetics. The current review focuses on biotechnological techniques like direct regeneration, indirect regeneration via callus induction and somatic embryogenesis, microrhizome induction, and in vitro secondary metabolite production that have been established to meet the increasing commercial and pharmaceutical demands. This review also identifies unrevealed possibilities, knowledge gaps, and considerable challenges that should be taken into account for developing novel concepts pertaining to the biotechnology of Zingiberaceae in the near future. [ABSTRACT FROM AUTHOR]

The study of the reproduction characteristics of roses of the garden class Shrub, the definition of the dependence of the hormonal determination of explant rhizogenesis on the concentrations of phytohormones that are part of the nutrient medium, are relevant and has both scientific and practical interest. This study presents the results of studies of hormonal determination of rhizogenesis in explants of cultivars of roses of the garden class Shrub: Gärtnerfreude, Lavender Dream, Pomponella, Red Cascade, Sommerabend cultivated in vitro on nutrient medium containing growth regulators. It has been established that of the nutrient medium modified by the addition of 0.2-1.0 mg/l α-naphthylacetic acid (α-NAA), the most effective was the medium with the content of α-NAA 0.5 mg/l, the content of macro- and microelements half of the Murashige and Skoog prescriptions, and a decrease in the sucrose content to 2.0%. On this medium, the frequency of rhizogenesis averaged 61.2% for the studied cultivars. Hormonal determination of rhizogenesis and efficiency of root formation in vitro in the Shrub rose regenerants depended on the genotype of the plant: cv. Lavender Dream (66.0%) and cv. Sommerabend (67.0%) had the highest rhizogenesis ability. The use of the universal growth regulator Humifield in combination with 0.5 mg/l α-NAA contributed to an increase in the rooting rate of the studied rose cultivars up to 70.0-86.0%. [ABSTRACT FROM AUTHOR]

This paper reports an efficient in vitro rooting and acclimatization method for Pennisetum 'Vertigo®'. For that purpose, the influence of the IAA (indole-3-acetic acid) in MS (Murashige and Skoog) media, and different types of LED light (100% blue, 100% red, a combination of red and blue (70% + 30%), a combination of red, blue with yellow, green, or far-red (35% + 15% + 50%), and white LED) were investigated. A fluorescent lamp and a medium without auxin were used as controls. Subsequently, the plants were transferred ex vitro in trays to the greenhouse and later transplanted into pots. In all tested combinations, the shoots rooted at a high rate of 70–100% (except under blue light). The best results were obtained under white LED or the fluorescent lamp, where the highest number of roots was formed; however, IAA did not increase the rooting rate, the number, or the weight of roots. Shoots rooted under LEDs and on media containing IAA generally exhibited higher soluble sugar content compared to those rooted on auxin-free media. Consistent with the rooting results, the best acclimatization was observed in plants rooted under white LED or fluorescent light on auxin-free media. [ABSTRACT FROM AUTHOR]

• Putrescine enhances microshoot regeneration in Turiaçu pineapple. • Nodular cluster induction boosts propagation efficiency. • Morphoanatomical insights shed light on cluster development. • Nodule clusters offer a promising method for pineapple propagation. Nodular clusters are a morphogenetic pathway used in the in vitro propagation of Bromeliaceae due to their high responsiveness in the regeneration of microshoots. Additionally, endogenous polyamines, especially putrescine, may contribute to the induction of these nodular clusters. However, the potential effects of this regulator on the regeneration of microshoots when applied to explants are unclear, especially in traditional pineapple cultivars such as "Turiaçu". Therefore, our objective here was to induce and characterize the morphoanatomy of nodular clusters in combinations of 6-benzylaminopurine (BAP) and putrescine (PUT) in "Turiaçu" pineapple. Leaf segments induced in culture medium supplemented with 4 μM naphthaleneacetic acid (NAA), 8 μM BAP, and 100 μM PUT resulted in an induction rate of 56 %, with the formation of microshoots via nodular clusters, increased length, and the number of leaves per microshoot. Morphoanatomical studies revealed that in the medium with 4 μM NAA, 8 μM BAP, and 100 μM PUT, the clusters presented a voluminous size, microshoots with leaves, and the development of new leaves along the base of the explant, and the formation of meristemoids with small cells arranged in a spiral. The results indicate that auxins and cytokinins combined with exogenous putrescine are efficient for inducing nodule clusters. Thus, exogenous putrescine improves the efficiency of microshoot regeneration by nodular clusters. [ABSTRACT FROM AUTHOR]

Date palm(Phoenix dactylifera L.) holds substantial agricultural, nutritional, and economic significance, particularly in arid and semi-arid regions. Traditional propagation methods, such as offshoot and seed propagation, face limitations including slow growth rates, limited availability of offshoots, and genetic instability, making the adoption of in vitro culture techniques essential for the mass propagation and genetic improvement of Date palm. This comprehensive review delves into the intricacies of in vitro culture protocols for Date palm, focusing on the development of multiple shoot, root, and organ cultures. The review meticulously examines the critical factors influencing each stage of in vitro propagation, including the selection of explants, optimization of culture media, and control of environmental conditions. The efficacy of different plant growth regulators (PGRs), such as cytokinins and auxins, in promoting shoot proliferation, root induction, and organogenesis is compared and analyzed, highlighting the optimal concentrations and combinations for successful tissue culture outcomes. For multiple shoot induction, the review discusses the use of shoot tips, axillary buds, and immature inflorescences, with media formulations like MS, B5, and WPM supplemented with specific PGRs such as BAP and kinetin. The process of root induction is examined, emphasizing the role of auxins like IBA and NAA in developing robust root systems and the influence of environmental factors like light and temperature. Organ culture techniques, particularly embryogenic callus induction and somatic embryogenesis, are explored, detailing the use of high auxin concentrations for callus formation and the transition to media with balanced PGRs for embryo maturation. Additionally, the review addresses the challenges of somaclonal variation and genetic stability in tissue culture, proposing strategies for monitoring and mitigating these issues through genetic and epigenetic analyses. The potential of integrating advanced biotechnological tools, such as CRISPR/Cas9, for precise genetic modifications is discussed, alongside the need for improved acclimatization protocols to enhance the survival rates of in vitro propagated plantlets. Future research directions are identified, focusing on minimizing somaclonal variation, optimizing acclimatization, exploring diverse explant sources, and scaling up production through automated culture systems. By synthesizing the latest research findings and offering detailed comparisons of various in vitro protocols, this review aims to provide a valuable resource for researchers and practitioners seeking to enhance the efficiency and success of Date palm micropropagation, ultimately contributing to the sustainable cultivation and genetic improvement of this vital crop. [ABSTRACT FROM AUTHOR]

Tissue culture is used to multiply Al-Taif rose (Rosa damascena f. trigintipetala (Diek) R. Keller) plants in order to meet the demands of the fragrance, cosmetic, and floriculture industries. The use of arbuscular mycorrhizal fungus (AMF) could potentially improve plant growth and acclimatization performance to ex vitro conditions. Thus, in the current study, we investigated how AMF Rhizophagus fasciculatus influences the growth, establishment, and physiological performance of micropropagated Al-Taif rose plants during the acclimatization stage. The growth and physiological parameters of the AMF-treated plants were evaluated after a 12 week growth period in the growth chambers. The plants treated with AMF exhibited greater height (25.53 cm) and biomass growth values for both shoot fresh weight (0.93 g/plant) and dry weight (0.030 g/plant), more leaves (11.3/plant), more leaf area (66.15 cm2), longer main roots (15.05 cm/plant), total root length (172.16 cm/plant), total root area (64.36 cm2/plant), and biomass from both fresh weight (383 mg/plant) and dry weight (80.00 mg/plant) of the plants. The plants treated with AMF also exhibited increased rates of net CO2 assimilation, stomatal conductance, and transpiration compared to the control plants. The proline content in the leaves and roots was significantly lower in the AMF-treated plants than untreated plants. The Fv/Fm ratio, which serves as an indicator of the intrinsic or maximal efficacy of Photosystem II (PSII) demonstrated a notable decline in the untreated Al-Taif rose plants. These results elucidate the advantageous impact of AMF colonization on micropropagated Al-Taif rose plants, thereby enhancing their resilience against adverse ex vitro conditions. [ABSTRACT FROM AUTHOR]

Salinity is one of the most important abiotic stress factors that affect plant growth and limit agricultural productivity. In this study, the effects of iron (FeNP), silver (AgNP), and silicon dioxide (SiNP) nanoparticles on the morphological and physiological parameters of in vitro boysenberry plants grown under salinity stress (NaCl) were investigated. According to our study results, higher values were obtained from SiNP application in terms of shoot development parameters; FeNP application was found to be more successful for root development; AgNP application was effective in terms of SPAD, leaf relative water content (LRWC), and relative growth rate (RGR); and FeNP application increased superoxide dismutase (SOD) and catalase (CAT) enzyme activities. Salt stress significantly affected root development, SPAD values, LRWC and RGR, and SOD and CAT enzyme activities. As a result, under salt stress conditions, SiNP, FeNP, and AgNP applications can significantly reduce the negative effects of stress and promote the vegetative development of the plant compared to control conditions. [ABSTRACT FROM AUTHOR]

Cannabis has been cultivated for thousands of years for a variety of purposes, including fiber, seeds, oil, and medicinal compounds. The cannabis industry is growing rapidly because several countries have recently legalized the use of cannabis. In these countries, the industry related to cannabinoid ingredients such as cannabidiol (CBD) and Δ9-tetrahydrocannabinol (THC) is steadily increasing every year. High concentrations of cannabinoids are mainly produced in unfertilized female flowers. Maintaining plants with high cannabinoid content is essential for producing uniform substances in large quantities. This study established an in vitro micropropagation protocol that can maintain the mother plant characteristics of Cannabis sativa 'Cheungsam'. As a result of this experiment, the shoot tips and nodes of Cannabis sativa 'Cheungsam' at various concentrations (0, 0.25, 0.5, 1.0 mg/L) of 2iP, BA, and mT plant growth regulators (PGRs), and all concentrations of 2iP showed better results compared to two other hormones. However, the cut surfaces turned black, and excessive hyperhydricity occurred. Based on these symptoms, activated charcoal was added to the medium with the assumption that necrosis and hyperhydricity occur due to the accumulation of reactive oxygen species (ROS). When treated with 0.5 g/L charcoal, hyperhydricity was not overcome, and there was no difference compared to the control. As a new alternative, we divided the experiments into MS (Murashige and Skoog) and DKW (Driver and Kuniyuki Walnut) medium, which were commercially available. As a result, the rate of hyperhydricity was reduced, the cut surface did not turn black, and the growth conditions were also improved. Subsequently, ½ MS medium and ½ DKW medium were treated with various concentrations of IBA alone and with combinations of IBA and NAA for rooting. As a result, ½ DKW with IBA 0.5 mg/L showed the highest rooting rate and the best root condition for Cheungsam. After 4 weeks, when considering rooted plants with a height above 5 cm that were acclimatized, the acclimatization rate reached 100%. In conclusion, the Cannabis sativa 'Cheungsam' plants used in this study produced healthy shoots on DKW medium containing 1.0 mg/L 2iP and 0.5 mg/L of IBA in ½ DKW medium showed the best rooting rate. [ABSTRACT FROM AUTHOR]

Aquilaria crassna Pierre ex Lecomte is a principal species renowned for its production of agarwood. However, the active components of agarwood are not universally in compliance with the standards set by the Chinese Pharmacopoeia. We have identified an elite A. crassna tree with agarotetrol and alcohol extract levels that exceed these standards and have successfully established a stable in vitro micropropagation system using stem segments from this elite tree. The effects of auxins and minerals on axillary-bud induction, shoot multiplication, and rooting were investigated. The most effective medium for axillary-bud induction was a half-strength Murashige and Skoog (1/2MS) medium supplemented with 0.50 mg/L 6-benzylaminopurine (6-BA), achieving an induction rate of 53.33% with minimal hyperhydricity. The optimal shoot proliferation medium was an MS medium with 0.40 mg/L 6-BA, yielding a propagation coefficient of 2.96 without hyperhydricity. The best rooting medium comprised quarter-strength MS (1/4MS) macroelements and 1/2MS microelements with 0.10 mg/L naphthaleneacetic acid (NAA), resulting in an 82.54% rooting rate. Substrate effects on transplant survival and growth were also evaluated, and peat soil was identified as the best substrate, achieving a survival rate of 96.67%. This study introduces a straightforward and efficient in vitro micropropagation system utilizing mature A. crassna as explants. It holds significant importance for the consistent production of agarwood that complies with the standards of the Chinese Pharmacopoeia and provides a model for the targeted breeding of medicinal plants. [ABSTRACT FROM AUTHOR]

Producing uniform Cannabis sativa (Cannabis) for medicinal/recreational flower production through sexual propagation has been problematic, leading to dominance of clonal propagation from "mother plants" in the cannabinoid industry, which also faces significant limitations. Cannabis tissue culture (TC) methods have been developed to overcome these challenges, but the long-term health and maintenance of Cannabis explants in TC have been largely overlooked in previous studies. The current study focused on the development of an efficient and optimized micropropagation protocol covering the entire process, with a specific focus on the health and performance in the multiplication stage. Multiplication media were formulated hormone-free to avoid longer-term vitrification issues, resulting in single-main-shoot cultures rather than multiple-shoot cultures. This instigated the use of stage II explant types different from the standard shoot tips previously used for multiple shoot cultures. Multiplication media were further improved from the basal salt composition via nitrogen and calcium additives. The optimized protocol was used on eight diverse Cannabis cultivars to test its applicability across various genetic backgrounds. Results indicated that the protocol was effective for conservation purposes across all cultivars and achieved good long-term multiplication rates for some but not all. The outcomes of this study mark a significant stride towards an efficient Cannabis TC methodology ready for more comprehensive industrial applications. [ABSTRACT FROM AUTHOR]

Momordica cochinchinensis (Lour.) Spreng. is a tropical dioecious plant belonging to the family, Cucurbitaceae also known as Gac, baby jack fruit, cochinchin gourd and sweet gourd. The plant is noted for its densely orange fruit, rich in nutrients, especially with 70 times more lycopene content than tomatoes. It possesses medicinal properties and the seeds are used in traditional Chinese medicine called Mubiezhi to treat fluxes, liver and spleen disorders, wounds and hemorrhoids. This study evaluated the effect of combination of pH, sucrose and BAP concentration on in vitro multiple shoot induction response of M. cochinchinensis according to guidelines of design of experiment. For those three factors, Response surface methodology (RSM) using the Box-Behnken design (BBD) was chosen and applied to the design matrix. The response shoot number/explant was evaluated by in vitro culture under variable conditions given in the design matrix. The response shoot number/explant was evaluated using in vitro culture under variable conditions given in the design matrix. The outcome of this method is an equation that calculates the shoot number as a function of pH, sucrose, and BAP concentration This study created response surface plots and contour plots for each factor, by analysing each factor's level of influence on the response shoot induction. [ABSTRACT FROM AUTHOR]

Viruses and viroids pose a significant challenge in citriculture, and their control is crucial for plant health. This study evaluated the effectiveness of in vitro thermotherapy combined with a meristem tip culture for eliminating citrus exocortis viroid (CEVd) and hop stunt viroid (HSVd) from a new limonime hybrid (Citrus x limon var. limon x Citrus latifolia var. latifolia). The elimination success was confirmed by RT-PCR assays. The in vitro elimination rate for CEVd during the shoot proliferation stage (43%) was higher than for HSVd (21%). Accordingly, in the subsequent rooting stage, the in vitro elimination rate for CEVd (50%) was higher than for HSVd (33%). Successful CEVd and HSVd eradication at a 100% rate was confirmed in the ex vitro acclimatized plants in the greenhouse. The study also established an efficient micropropagation protocol. The optimal treatment for in vitro shoot induction was 0.5–2 mg L−1 benzyladenine (BA) + 0.5 mg L−1 gibberellic acid (GA3) + 0.25 mg L−1 naphthalene acetic acid (NAA), while for shoot elongation, it was 0.5 mg L−1 BA + 0.5 mg L−1 kinetin (KIN) + 0.5 mg L−1 GA3 + 0.25 mg L−1 NAA. Rooting was best promoted by 1 mg L−1 NAA. This study provides valuable insights for the mass production of viroid-free propagation material in this new lemon x lime hybrid, contributing to the conservation of genetic resources in citrus breeding programs through the combined application of in vitro thermotherapy and an in vitro meristem tip culture, a novel and highlighted achievement reported for the first time in this study. [ABSTRACT FROM AUTHOR]

This study focuses on the endangered neo-endemic Baltic dunes species Linaria loeselii Schweigg. (Plantaginaceae), also known as Linaria odora (M. Bieb.). By utilizing in vitro cultures, we successfully germinated seeds collected in situ. Our method, which involved using media supplemented with 5 µmol/L 6-benzylaminopurine, led to the indirect regeneration of shoots after 60 days of culture in the dark, significantly increasing the number of progeny plants. Additionally, the medium supplemented with 2.85 μmol/L indole-3-acetic acid and 10.2 μmol/L paclobutrazol allowed rooting after 30 days of shoot fragments. This research provides a potential basis for developing Linaria loeselii introduction programs into the environment, thereby contributing to the conservation of this endangered species. [ABSTRACT FROM AUTHOR]

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Background. The problem of obtaining certified planting material with biotechnological methods is important for expanding commercial and homestead plantations of hazelnut cultivars in Belarus. Materials and methods. Regenerated plants of cv. ‘Akademyk Yablokov’, representing the genus Corylus L., were a model object for studying in vitro morphogenesis and effectiveness of antibiotics against bacterial contamination, so that a protocol could be developed to obtain healthy planting material of hazelnut cultivars. The plants produced during this study were included in the duplicate ex situ collection of nut crops preserved in vitro in the active growth state. Results. Single exposure to the antibiotic kanamycin monosulfate at a concentration of 100 mg/L during antibiotic therapy in the stage of in vitro micropropagation eliminated bacterial infection in 83.3% of regenerated plants, and twofold exposure in 100%. Further cultivation revealed its phytotoxic aftereffect manifested in the form of necrosis on most of the regenerated plants and a decrease in the activity of gemmogenesis and growth. Neither single nor twofold exposure to cefotaxime sodium salt at a concentration of 90 mg/L caused elimination of bacterial infection, but gemmogenesis and regenerated plant growth retained their activity during subsequent cultivation on antibiotic-free media. The best development parameters were observed on a modified Murashige–Skoog medium with 6 mg/L 6-BA, 0.01 mg/L IBA, and 0.1 mg/L GA3 (average number of shoots: 2.2; number of microcuttings: 2.3), and a modified DKW medium with 6 mg/L 6-BA, 0.01 mg/L IBA, and 0.1 mg/L GA3 (average number of shoots: 2.05; microcuttings: 2.9). The use of zeatin as a cytokinin to stimulate adventitious morphogenesis or activate the growth of axillary meristems at a concentration of 5 or 6 mg/L was not as effective as 6-BA.

The study of the reproduction characteristics of roses of the garden class Shrub, the defini-tion of the dependence of the hormonal determination of explant rhizogenesis on the con-centrations of phytohormones that are part of the nutrient medium, are relevant and has both scientific and practical interest. This study presents the results of studies of hormonal de-termination of rhizogenesis in explants of cultivars of roses of the garden class Shrub: Gärt-nerfreude, Lavender Dream, Pomponella, Red Cascade, Sommerabend cultivated in vitro on nutrient medium containing growth regulators. It has been established that of the nutrient medium modified by the addition of 0.2-1.0 mg/l α-naphthylacetic acid (α-NAA), the most effective was the medium with the content of α-NAA 0.5 mg/l, the content of macro- and microelements half of the Murashige and Skoog prescriptions, and a decrease in the sucrose content to 2.0%. On this medium, the frequency of rhizogenesis averaged 61.2% for the studied cultivars. Hormonal determination of rhizogenesis and efficiency of root formation in vitro in the Shrub rose regenerants depended on the genotype of the plant: cv. Lavender Dream (66.0%) and cv. Sommerabend (67.0%) had the highest rhizogenesis ability. The use of the universal growth regulator Humifield in combination with 0.5 mg/l α-NAA contribut-ed to an increase in the rooting rate of the studied rose cultivars up to 70.0-86.0%.

Eucheuma denticulatum and Kappaphycus alvarezii are two of the most commercially important, cultivated carrageenophytes in Southeast Asia. Their large-scale farming through repeated vegetative propagation and virtual monocropping has resulted in a variety of phyconomic issues. The efficacy of two European, commercial liquid extracts from the brown seaweeds, i.e., Ascophyllum nodosum (ANE), a fucoid, and Laminaria digitata (LDE), a kelp, on the direct formation of axes and survival of these two eucheumatoids was examined using tissue culture techniques. Seaweed apical sections (2–3 mm long) were cultured for 45 days, with different concentrations of ANE and LDE (i.e., 0.005, 0.05, 0.5, 1.5, and 2.5 mL L−1) and compared with a control (UV-filtered seawater). Lower concentrations (< 1.5 mL L−1) of both liquid extracts promoted morphogenesis and growth of E. denticulatum and K. alvarezii in tissue culture, with the dilution rate of 0.05 mL LDE L−1 resulting in a high percentage survival (i.e., 95.3 ± 0.9%), formation of direct axes (i.e., 95.3 ± 0.9%), and the longest length (i.e., 10.0 ± 0.3 mm) for E. denticulatum after a typical 45-day culture period. A 0.5 mL ANE L−1 concentration was found to be beneficial for the tissue culture of K. alvarezii, given the relatively high final survival and direct axes formation (i.e., 96.2 ± 2.2%) and axes length (i.e., 8.2 ± 0.1 mm). This study supports the application of these temperate, brown seaweed-derived extracts as a phycobiostimulant enrichment in eucheumatoid micropropagation for mass production of plantlets for out-planting purposes. [ABSTRACT FROM AUTHOR]

Abstract Background Kale, a versatile cruciferous crop, valued for its pro-health benefits, stress resistance, and potential applications in forage and cosmetics, holds promise for further enhancement of its bioactive compounds through in vitro cultivation methods. Micropropagation techniques use cytokinins (CKs) which are characterized by various proliferative efficiency. Despite the extensive knowledge regarding CKs, there remains a gap in understanding their role in the physiological mechanisms. That is why, here we investigated the effects of three CKs – kinetin (Kin), 6-benzylaminopurine (BAP), and 2-isopentenyladenine (2iP) – on kale physiology, antioxidant status, steroidal metabolism, and membrane integrity under in vitro cultivation. Results Our study revealed that while BAP and 2iP stimulated shoot proliferation, they concurrently diminished pigment levels and photosynthetic efficiency. Heightened metabolic activity in response to all CKs was reflected by increased respiratory rate. Despite the differential burst of ROS, the antioxidant properties of kale were associated with the upregulation of guaiacol peroxidase and the scavenging properties of ascorbate rather than glutathione. Notably, CKs fostered the synthesis of sterols, particularly sitosterol, pivotal for cell proliferation and structure of membranes which are strongly disrupted under the action of BAP and 2iP possibly via pathway related to phospholipase D and lipoxygenase which were upregulated. Intriguingly, both CKs treatment spurred the accumulation of sitostenone, known for its ROS scavenging and therapeutic potential. The differential effects of CKs on brassicasterol levels and brassinosteroid (BRs) receptor suggest potential interactions between CKs and BRs. Conclusion Based on the presented results we conclude that the effect evoked by BAP and 2iP in vitro can improve the industrial significance of kale because this treatment makes possible to control proliferation and/or biosynthesis routes of valuable beneficial compounds. Our work offers significant insights into the nuanced effects of CKs on kale physiology and metabolism, illuminating potential avenues for their application in plant biotechnology and medicinal research.

Light-emitting diodes (LEDs) have become an alternative light source to the fluorescent lamp (FL) for the maintenance of plant tissue cultures due to their low heat emission, low power consumption and the ability to fine-tune the light spectrum. In this study, the effect of LEDs on the growth and leaf stomata features of in vitro cultivated raspberries (Rubus idaeus L. 'Lloyd George') was examined. The plantlets were grown in vitro under a lighting system based on the Philips GreenPower LED research module. Four groups of LEDs emitting white (W), red (R), blue (B), and mixed (W:R:B: far red = 1:1:1:1) lights and FL (control) were used. As a second control (marked as EV), plants grown in the multiplication and rooting stage under fluorescent lamps and then acclimatized ex vitro in a greenhouse for 90 days in natural light were included. Relative growth rate (RGR), protein content as well as stomata morphology and density of the plantlets were analysed after three passages under corresponding light treatment. The results show that different LEDs specifically affect the growth and size of leaf stomata and density of in vitro cultured raspberry plants and can be applied as an effective modulator of morphogenesis during micropropagation. The combination of white, blue, red and far red LED stimulated the accumulation of biomass and proteins, as well as the formation of a higher number of stomata on the lower surface of the leaves, which would be a prerequisite for more effective control of water loss from plantlets during the ex vitro acclimatization.

Euonymus koopmannii is a rare and protected species in Kazakhstan, valued for its ecological role in soil stabilization and its ornamental properties. This study presents the first use of micropropagation and phylogenetic analysis for the endemic plant E. koopmannii. Seedlings of E. koopmannii proved to be more effective than internodes as primary explants for plant micropropagation of in vitro culture, with a multiplication coefficient of 28.5 from seedlings and 6.1 from internodes. On MSR I medium supplemented with 0.5 mg/L IBA and 0.05 mg/L IAA, a higher success rate of 67% was achieved for root formation of test tube-grown E. koopmannii plants. Using mannitol as an osmotic agent at a concentration of 8 mg/L prolonged the storage time of E. koopmannii under slow growth conditions when compared to CCC and abscisic acid. Phylogenetic relationships and species identification were analyzed using four DNA-barcoding markers, comparing E. koopmannii with species from NCBI. All candidate barcoding markers showed sufficient levels of interspecific genetic variation among Euonymus species. In addition, ITS region and rbcL gene sequences effectively distinguished E. koopmannii from other species. These results provide fundamental information that will be valuable for future biotechnological and molecular studies. [ABSTRACT FROM AUTHOR]

WUSCHEL-related homeobox (WOX) genes are plant-specific homeobox transcription factors and are known to play a key role during in vitro embryogenesis and organogenesis. To decipher the structural basis and genetic diversification of WOX family genes, this study performed a genome-wide analysis in Plantaginaceae. In the current study, Picrorhiza kurrooa and Antirrhinum majus (snapdragon; a model organism) of the Plantaginaceae family were used for in silico analysis of WOX gene family members. The physiochemical properties, subcellular localization, and phylogenetic analysis in P. kurrooa were evaluated. Further, to understand gene structure, chromosomal localization, and synteny, A. majus along with Arabidopsis thaliana was used due to unavailability of chromosome-level resolved genome in P. kurrooa. Five WOX and one WUS members were identified in the P. kurrooa genome while eight WOX and eight WUS members were identified in A. majus distributed in five clades. Phylogenetic analysis revealed that PkWOX and AmWOX showed greater similarity to each other, as do PkWUS and AmWUS, compared to AtWOX and AtWUS. Further, the tree was separated in three different clades, including ancient, intermediate, and WUS clade. Chromosomal localization of WOX and WUS genes in A. majus revealed their distribution between 2 and 7 chromosomes while chromosome 1 and 8 did not contain any member. Expression analysis of P. kurrooa WOX and WUS genes showed differential expression during the different stages of in vitro organogenesis (mother leaf, callus initiation, callus proliferation, and regeneration) and different temperature gradients (25°C, 15°C, and 10°C). Taken together, this study provided significant insight into the developmental and stress responsiveness of WOX gene family members in P. kurrooa, which can be further explored as promising candidates. [ABSTRACT FROM AUTHOR]

Chinese artichoke (Stachys sieboldii [Miq.]) is a popular healthcare food owing to its high contents of stachyose. Viral infections have caused the severe degeneration of germplasm in this plant species. An efficient virus elimination and micropropagation system is required to produce Chinese artichoke plants that are free of viruses. A protocol for virus elimination in Chinese artichoke was established by comparing the potential for growth of the terminal buds using shoot apices as starting materials, in media with or without plant growth regulators (PGRs) and various concentrations of sucrose. Murashige and Skoog (MS) medium without any PGR performed better than that supplemented with PGRs as indicated by a higher rate of survival of shoot apices, higher plant height, more root numbers and adventitious buds, and increased biomass (P < 0.05). An experiment to further optimize the sucrose concentration demonstrated that MS medium with 5.0% sucrose (w/v) efficiently induced the growth of both roots and shoots, thus, achieving the efficient micropropagation of Chinese artichoke within 4 wk. Compared with the mother plants in which viruses had not been eliminated, the virus-free plants had significantly higher numbers of tubers, increased yield, and a higher content of stachyose (P < 0.05). Higher contents of endogenous hormones were detected in Chinese artichoke, which may explain the efficient micropropagation without the use of exogenous PGRs. This simple protocol enabled the production of virus-free Chinese artichoke to enhance the yield of tubers and high content of stachyose. [ABSTRACT FROM AUTHOR]

Petiole segments of an economically ornamental plant Oxalis triangularis A.st.-Hil have been used in the present study for rapid, reliable and systematic in vitro propagation. The morphogenic effect of a-Naphthalene acetic acid (NAA) at various concentrations (0.5 - 3.0 mg/L) was studied individually or in combination with 6-benzylaminopurine (BAP). Superior multiplication rates were achieved on Murashige and Skoog (MS) medium supplemented with 0.5 mg/L NAA and 1.0 mg/L BAP. Of the tested combinations, maximum shoot regeneration (100.00 %), mean shoot number (14.60 ± 0.37 shoots per segment), and shoot length (18.11 ± 0.30 mm) were recorded on MS medium supplemented with 0.5 mg/L NAA and 1.0 mg/L BAP after 6 weeks of incubation. Half-strength MS medium augmented with 1.0 mg/L indole-3-butyric acid (IBA) was the optimal medium for in vitro root induction, (24.60 ± 0.37 roots per shoot with 9.10 ± 0.32 mm in length) after 6 weeks of incubation. The regenerated plantlets showed 100 % survival success during acclimatization and exhibited normal growth characteristics and morphology. This optimized protocol is cost-effective and highly efficient for the rapid mass propagation of Oxalis triangularis A.st.-Hil. [ABSTRACT FROM AUTHOR]

Aquatic plants are grown in aquaria for their beauty and to maintain the quality of water. The explants of Lobelia cardinalis L. were obtained from the Aquatic Plant Center Co., Ltd., Thailand. In vitro clonal propagation offers alternative strategy for its rapid clonal multiplication. In this study, multiple and rapid propagations of Lobelia cardinalis L. with tissue culture techniques were aimed. Highly efficient and repeatable in vitro regeneration protocol was established from nodal segments of Lobelia cardinalis L. Various concentrations and combinations of various plant growth regulators (PGRs) were employed to induce multiple shoots, shoot elongation, number of leaves and rooting of shoots to obtain complete plantlets of Lobelia cardinalis L. Among 2 PGRs such as 6-benzylaminopurine (BAP) and a-Naphthalene acetic acid (NAA) tested for multiple shoot induction. Regenerated shoots had a higher number of shoots on MS medium contained cytokinins with auxins. The maximum shoot proliferation (100 %), number of shoots per explant (6.20 ± 0.51 shoots), shoot length (11.67 ± 0.69 mm) and number of leaves per explant (58.10 ± 0.38 leaves per explant) were observed in MS medium supplemented with 1.0 mg/L BAP and 2.0 mg/L NAA. Rooting was achieved within 3 to 4 weeks on all the rooting media, but MS medium augmented with 1.0 mg/L indole-3-butyric acid (IBA) produced the maximum number of strong and healthy roots (11.70 ± 0.27 roots). The regenerated complete plantlets with healthy roots and shoot system were transferred to 3.5 cm diameter black net plastic pots containing rockwool medium and successfully acclimatized with a survival rate of 100 % in the greenhouse and no phenotypic variations were observed among regenerated plants. The success of this protocol offers highly efficient method of the plant multiplication, which would be beneficial for plant tissue culturist, and nursery people where regular supply of plants is a large number required. This protocol can be utilized for commercial scale propagation. [ABSTRACT FROM AUTHOR]

Use of meta-Topolin (mT) has significantly affected tissue culture results in several plant species. In this study, a fast and efficient micropropagation protocol for Withania somnifera was standardized. Also, a comparison was made between the effect of using M.S. media supplemented with mT and M.S. media supplemented with different concentrations of B.A.P. + AdSO4 on the in vitro regeneration of W. somnifera. M.S. media supplemented with I.B.A. was used to obtain a healthy and advantageous root system. The resulting plantlets were analyzed for their total chlorophyll and protein content. The comparison revealed that 2.5 mg/l mT supplemented M.S. media showed better results with 88.9 ± 0.42% bud breaking percentage, a shoot multiplication rate of 22.4 ± 4.16 shootlets per explants, total chlorophyll content of 0.887 ± 0.004 mg/g and protein content of 25.67 ± 0.25%. Healthy adventitious root systems were observed in regenerated shootlets inoculated in M.S. media supplemented with 2.0 mg/l I.B.A. During hardening, 80% of plants treated initially with mT survived. Whereas treatment with M.S. media supplemented with 1.0 mg/l B.A.P. + 50 mg/l AdSO4 resulted in bud breaking percentage of 61.1 ± 0.09%, a shoot multiplication rate of 15.8 ± 7.81 shootlets per explant, total chlorophyll content of 0.7194 ± 0.0055 mg/g and protein content of 23.33 ± 0.25%. During hardening, 78% of plantlets treated initially with B.A.P. + AdSO4 survived. Therefore, the study concludes that mT may be used as an alternative source of cytokinin for obtaining healthy plantlets with a higher rate of plant regeneration in W. somnifera.Key message: The study carried out to standardize a viable protocol for meta-Topolin as alternative source of cytokinins in micropropagation for Withania somnifera and to compare the effects using different phytohormones. [ABSTRACT FROM AUTHOR]

The large-leaved Homalomena (LLH, Homalomena pendula) represents an endangered medicinal plant species within Vietnam, primarily attributed to its recognized tonic properties. Despite its imminent threat of extinction within Vietnamese ecosystems, the development of a robust protocol for molecular species identification and large-scale propagation of LLH remains absent. Consequently, we present the first conservation endeavor for LLH based on plant micropropagation techniques, with plant materials validated through anatomical observations and DNA barcoding (rbcL). Our investigation yielded five rbcL sequences specific to LLH, serving as the current best barcode for LLH identification and thereby facilitating forthcoming taxonomic endeavors. Optimization of in vitro culture conditions revealed that the Murashige and Skoog (MS) medium supplemented with 2 mg/L 6-benzylaminopurine, 0.5 mg/L α-naphthaleneacetic acid, and 60 g/L mashed potato, alongside the incorporation of 0.5 mg/L indole-3-butyric acid to the basal MS medium, yielded optimal outcomes for shoot proliferation and root development, respectively. After successful micropropagation, acclimatization of rooted plantlets to a substrate comprising soil, coconut coir, and rice husk (in a 1:1:1 ratio) culminated in a 100% survival rate among the plants.Key message: The first established protocol for species identification and micropropagation of H. pendula. [ABSTRACT FROM AUTHOR]

The safe management of sewage waste is a current concern due to population growth and waste production. Biosolids, derived from sewage sludge treatment, are globally used as organic fertilizers, aligning with Sustainable Development Goal 6 for resource recycling. However, biosafety concerns arise due to the presence of metals and microplastics in biosolids, potentially impacting soil and water. This study investigated biosolids’ use for in vitro cultivation of Bowdichia virgilioides Kunth. Results indicate that while biosolids can replace traditional nutritional media, balancing their concentration is crucial for optimizing plant growth. The WPM (Wood Plat Medium) remains essential for in vitro cultivation, but substituting it with biosolids at concentrations of up to 2 g L− 1 is feasible, providing similar plant development compared to the WPM medium. However, when combined, there is a complex and challenging interaction between biosolids and the culture medium. [ABSTRACT FROM AUTHOR]

Lavandula stoechas subsp. luisieri and Pterospartum tridentatum are two valuable aromatic and medicinal plants. Their biometric and morphological parameters, such as the number of new shoots, length of the longest shoot, multiplication rate, and fresh weight, were evaluated using the multiplication MS medium protocol. The rooting protocols involved immersing the explants in IBA (1 g L−1) and a commercial IBA (3.3 g L−1) preparation (Clonex®). Slow-growth conservation assays were carried out using two different sucrose concentrations (15 g L−1 and 30 g L−1), and a control, with the cultures kept at 4 °C for 12 months. The multiplication rate for L. stoechas subsp. luisieri was 6.8, and that of P. tridentatum was 13.3, achieved using the MS medium supplemented with 0.2 mg L−1 BAP, 1 mg L−1 BAP, and 0.5 mg L−1 IBA. The application of Clonex® showed the best ex vitro rooting results in L. stoechas subsp. luisieri (77%) and P. tridentatum (90%). In the slow-growth conservation assays, at 4 °C, in darkness for 12 months, an excellent survival rate was achieved in L. stoechas subsp. luisieri (>80%) and P. tridentatum (>90%), even at the reduced sucrose concentration. This study demonstrates the effectiveness of in vitro multiplication and ex vitro rooting protocols for two valuable aromatic and medicinal plants. These findings are significant for the ex situ conservation of these species, as they provide effective long-term preservation and utilization strategies. [ABSTRACT FROM AUTHOR]

Podophyllum hexandrum Royle, also known as Podophyllum emodi Wall, holds significant ecological, ornamental, and medicinal values. However, it has become endangered due to overexploitation, prolonged seed dormancy, slow natural regeneration, and climate change. This study developed an efficient in vitro protocol for callogenesis and micropropagation of P. hexandrum to conserve germplasm in in vitro conditions. Callus formation from various plant parts, including the leaf, stem, rhizome, radicle, and cotyledon, was induced using Murashige and Skoog (MS) medium supplemented with different plant growth regulators. The combination of benzyladenine at 1 mg L−1 and 4-dichlorophenoxy acetic acid at 3 mg L−1 was optimal for biomass production, yielding 215.88 ± 0.31 mg, with growth per gram at 8.32 ± 0.32 and a growth rate of 13.62 ± 0.25 mg/day on MS medium. For shoot proliferation, benzyladenine (3.5 mg L−1) and naphthalene acetic acid (0.5 mg L−1) combined with activated charcoal showed the highest shoot induction percentage per explant. For shoot regeneration from calluses, 6-benzylaminopurine (0.5 mg L−1) and thidiazuron (2 mg L−1) were most effective, producing superior shoot length, number of regenerations, and regeneration percentage. Root induction was successful with α-naphthalene acetic acid supplementation (0.5 to 1.5 mg L−1) in MS medium, resulting in the highest number per explant (4.08 ± 0.08), length (5.45 ± 0.15 cm), and rooting rate (87.00 ± 1.66%) of roots in plantlets. Subculturing for callus culture was performed every 28 days for up to four subcultures to prevent nutrient depletion and toxic metabolite accumulation, ensuring tissue health and viability. Continuous subculturing of callus on MS medium maintained healthy P. hexandrum germplasm in vitro. Overall, this micropropagation protocol provides a rapid system for conserving P. hexandrum germplasm. [ABSTRACT FROM AUTHOR]

The demand for bioactive secondary metabolites of natural origin is increasing every day. Micropropagation could be a strategy to respond more quickly to market demands, regardless of seasonality. This research aims to evaluate in vitro-grown plants of two hop varieties, namely Columbus and Magnum, as a potential source of bioactive compounds. The extracts were characterized in terms of total phenolic content by a Folin–Ciocalteu assay and antioxidant capacity by DPPH•, ABTS+, and FRAP assays. The bioactive compound profile of the extracts from both varieties was determined by using UPLC-ESI-QqQ-MS/MS. The results confirmed richness in (poly)phenols and other secondary metabolites of the in vitro-grown hop plantlets. Thirty-two compounds belonging to the major families of phytochemicals characteristic of the species were identified, and twenty-six were quantified, mainly flavonoids, including xanthohumol and isoxanthohumol, phenolic acids, as well as α- and β-acids. This study confirms the validity of in vitro-derived hop plantlets as source of bioactive compounds to be used in the nutraceutical, pharmaceutical, and food industries. [ABSTRACT FROM AUTHOR]

Background: Kale, a versatile cruciferous crop, valued for its pro-health benefits, stress resistance, and potential applications in forage and cosmetics, holds promise for further enhancement of its bioactive compounds through in vitro cultivation methods. Micropropagation techniques use cytokinins (CKs) which are characterized by various proliferative efficiency. Despite the extensive knowledge regarding CKs, there remains a gap in understanding their role in the physiological mechanisms. That is why, here we investigated the effects of three CKs – kinetin (Kin), 6-benzylaminopurine (BAP), and 2-isopentenyladenine (2iP) – on kale physiology, antioxidant status, steroidal metabolism, and membrane integrity under in vitro cultivation. Results: Our study revealed that while BAP and 2iP stimulated shoot proliferation, they concurrently diminished pigment levels and photosynthetic efficiency. Heightened metabolic activity in response to all CKs was reflected by increased respiratory rate. Despite the differential burst of ROS, the antioxidant properties of kale were associated with the upregulation of guaiacol peroxidase and the scavenging properties of ascorbate rather than glutathione. Notably, CKs fostered the synthesis of sterols, particularly sitosterol, pivotal for cell proliferation and structure of membranes which are strongly disrupted under the action of BAP and 2iP possibly via pathway related to phospholipase D and lipoxygenase which were upregulated. Intriguingly, both CKs treatment spurred the accumulation of sitostenone, known for its ROS scavenging and therapeutic potential. The differential effects of CKs on brassicasterol levels and brassinosteroid (BRs) receptor suggest potential interactions between CKs and BRs. Conclusion: Based on the presented results we conclude that the effect evoked by BAP and 2iP in vitro can improve the industrial significance of kale because this treatment makes possible to control proliferation and/or biosynthesis routes of valuable beneficial compounds. Our work offers significant insights into the nuanced effects of CKs on kale physiology and metabolism, illuminating potential avenues for their application in plant biotechnology and medicinal research. [ABSTRACT FROM AUTHOR]

Advancements in micropropagation techniques have made it easier to produce large numbers of cannabis clones, but these methods may also introduce genetic instability over successive generations. This instability often manifests as somaclonal variation, characterized by the progressive accumulation of genetic mutations or epigenetic alterations with each subculture. In this study, we examined how mutations accumulate in cannabis clones subjected to 6–11 subcultures. Using genotyping-by-sequencing, we identified 9405 polymorphic variants across 70 clones. The analysis revealed a correlation between the number of subcultures and the frequency of these mutations, revealing that genetic changes accumulate over successive subcultures despite clones sharing the same chronological age. Furthermore, we evaluated the functional impacts of accumulated mutations, with particular attention to implications on gene function and overall plant health. While rare, 14 high-impact variants were identified in genes that are important for plant development. Notably, six variants were also found in genes related to cannabinoid and terpene synthesis pathways, potentially affecting the plant's biochemical composition. These findings highlight the need for genetic assessments in micropropagation protocols, impacting plant breeding and conservation. Understanding genetic variations in clonally propagated plants optimizes practices for stability. Crucial for cannabis and horticultural plants, it emphasizes techniques to prevent genetic decay and ensure viability. [ABSTRACT FROM AUTHOR]

Plant biotechnology has become a viable strategy for agriculture, its applications include strategies aimed at obtaining and maintaining pathogen-free cultivars, genetic improvement (anther cultivation, embryo rescue, etc.) and micropropagation, used to obtain a greater number of individuals in less time and reduced spaces. Each of its phases depends largely on an adequate culture medium, since the growth, development and morphogenesis of the explant is mainly due to its chemical composition. For the production of garlic "seed" (Allium sativum L.), all phases are carried out in a growth chamber under automatically controlled aseptic and artificial conditions and at a temperature ranging between 22-25 °C. Somatic embryogenesis allows complete plants to be regenerated from a single cell. Garlic is an annual plant that only reproduces asexually, which is why bacteria, fungi and mainly viruses, which contribute to a 50-80 % decrease in its yield, affect it. An alternative to avoid these problems is the use of chemotherapy, thermotherapy or the combined use of one of these techniques with the cultivation of meristems, to obtain plants of high biological quality. Depending on the behavior of the culture during its in vitro stage, its response will be in the acclimatization phase and later in field conditions. This work was carried out with the objective of deepening the knowledge of some biotechnological techniques that are applied in the cultivation of garlic. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the impact of varying gamma irradiation doses on banana scalps' survival and regeneration potential, specifically focusing on their interaction with different culture media components. Banana scalps, derived from a selected cultivar (Calcutta-4), were exposed to gamma irradiation doses ranging from 10 Gy to 70 Gy. Following irradiation, the scalps were transferred to fresh culture media, and their survival rates and regeneration potential were assessed. The findings revealed a clear inverse relationship between radiation dose and scalp survival. Lower doses of 10 Gy and 20 Gy exhibited higher survival rates when cultured on a modified MS medium. In comparison, higher doses, such as 50 Gy and 60 Gy, led to a significant decrease in survival. The optimal gamma irradiation dose for both scalp survival and successful regeneration fell within the range of 30 Gy to 40 Gy when using a medium supplemented with lower concentrations of BAP and Ads (1.5 mg/l to 2.0 mg/l BAP + 200 mg/l to 300 mg/l Ads). Furthermore, the study demonstrated that the surviving irradiated scalps exhibited a high potential for shoot regeneration when cultured on an MS medium with 3.0 mg/l BAP + 400 mg/l Ads. Gamma irradiation facilitated the initiation of new shoots, and the specific concentration of plant growth regulators and media composition influenced the response. These findings have significant implications for optimizing tissue culture techniques and improving crop production in banana cultivars and are crucial for advancing banana farming practices. [ABSTRACT FROM AUTHOR]

• Investigation of strategies for enhanced biosynthesis of bioactive compounds. • Pathway for the biosynthesis of glucosinolates, isothiocyanates, thiocarbamates, and omoringone. • Commercialization challenges addressing, contamination, propagation and factors impacting yield and quality. • Role in green synthesis of nanoparticles with antimicrobial, antioxidant, and other potential applications. Moringa oleifera Lam., also known as the "drumstick tree" or "miracle tree,'' has a wide range of applications in both traditional and modern medicine due to presence of specific bioactive compounds. This review investigates the chemical composition of the plant, the mechanisms involved in the biosynthesis of bioactive compounds, and strategies for increasing their production. Glucosinolates, isothiocyanates, and other secondary metabolites are among the important bioactive compounds found in Moringa , which have shown anti-inflammatory, antimicrobial, antioxidant, and anticancer properties. These findings are gaining popularity for their potential applications in medicine, nutrition, and industry. This paper discusses the challenges of commercializing Moringa while focusing on issues related to contamination, propagation methods, environmental factors that influence yield and novel use of M. oleifera for the green synthesis of metallic and nonmetallic nanoparticles such as silver, zinc, copper, iron, gold, chitosan and others. These nanoparticles have antimicrobial, antioxidant, and photocatalytic properties, making them useful for applications in healthcare, environmental remediation, and other fields. Finally, M. oleifera is a versatile and resourceful plant with the ability to resolve a wide range of health concerns and provide long-term viable alternatives to a wide range of medicinal and therapeutic applications. [ABSTRACT FROM AUTHOR]

This research focuses on the seasonal patterns of the ecophysiological recovery of four olive cultivars (Arbequina, Coratina, Frantoio, and Urano), both micropropagated and self-rooted, grown in a mature, irrigated, super high-density (SHD) orchard under Mediterranean conditions (Southern of Italy). The aim was to observe the impact of the micropropagation method on the ecophysiological responses. Ecophysiological parameters, including leaf water potential (LWP), stomatal conductance (gs), net photosynthetic rate (Pn), and transpiration rate (E) were assessed. Self-rooted trees consistently exhibited superior gs, E, and Pn recovery compared to the micropropagated ones. 'Arbequina' maintained elevated levels of Pn under water-deficit conditions. 'Coratina' exhibited increases in gs and E after irrigation. 'Frantoio' demonstrated recovery capabilities, with lower LWP and higher Pn under stress. 'Urano' micropropagated trees achieved higher gs values in mid-summer, while self-rooted trees sustained higher Pn later in the season. This field research highlighted the important role of the propagation method in optimizing the physiological performance of olive cultivars in SHD orchards. Furthermore, it highlighted the necessity of long-term studies on the effects of propagation methods and their interactions with other farming practices. [ABSTRACT FROM AUTHOR]

Zuccagnia punctata Cav. (“jarilla poposa”) is a monotypic argentine native medicinal plant used by indigenous communities for the treatment of several pathologies. The pharmacological properties have been scientifically validated and its potential use in phytocosmetic and pharmaceutic industry as antiaged, hypoglycemic, anti-inflammatory, antitumoral, antioxidant, antibacterial and antimicotic has been previously demonstrated. However, the species has a low propagation rate in natural conditions and has been included in the preliminary red list of endangered plants in category 3. Therefore, it is necessary to develop methodologies leading to its conservation and propagation to achieve its sustainable use. The objective of this work is to perform a micropropagation protocol for Z. punctata species for the sustainable use and conservation of this species. Furthermore, the study evaluated also the impact of “in vitro” propagation on the content of bioactive compounds of ethanolic extracts obtained from Z. punctata plants obtained “in vitro” with collected wild plants. Seeds have been used as mother material. The seed germination to obtain explants were performed on Murashige and Skoog medium (MS) without plant growth regulators (PGRs) containing sucrose, agar, and ascorbic acid. The same medium was employed for the multiplication of shoots. “In vitro” rooting was achieved in liquid and semisolid medium, and a higher yield was obtained in the latter. The plants were transferred into commercial substrate and acclimatized in a greenhouse. The micropropagated plants (MP) had a high content of total phenolic compounds similar to those obtained from wild adult plants (WAP) collected in the Monte Desert. The flavonoid content of MP was higher than that of WAP. The main chalcones identified in WAP, 2′,4′ dihydroxychalcone (DHC) and 2′,4′-dihydroxy-3′-methoxychalcone (DHMC) were also present in the MP with a rate 1/1, and 1/3 in WAP. The extracts obtained from the MP showed antioxidant, antiinflammatory and antimicrobial activity, similar to WAP. The “in vitro” propagation and acclimatization of Z. punctata constitutes a fundamental tool to achieve large-scale multiplication of this species in the future and the sustainable production of bioactive compounds with commercial applications. Reimplantation in the natural habitat could prevent the loss of this medicinal genetic resource. [ABSTRACT FROM AUTHOR]

The study aimed to find an accurate propagation program for the rootstock of peach Prunus persica cv. Garnem. The apical shoots and nodes of this rootstock were used as explants for the stages of initiation and multiplication, and they were planted on MS medium after adding different concentrations of Active Charcoal (AC), Polyvinyl Pyrrolidone (PVP), and Benzyl adenine (BA). The shoots were planted for the purpose of rooting on MS medium containing different concentrations of salts (1/4 MS, 1/2 MS and full MS salts) with different concentrations of IBA. In the establishment phase, the apical shoots grown on medium supplied with AC were given the highest response rate, the highest average length of plantlet, and the highest number of leaves. The best multiplication of explants in terms of the number of shoots and their length were obtained when cultivated on a medium supplemented with BA at 1 and 2 mg.L-1, whereas the application of BA at 1 mg. L-1 to the cultivated nodes gave the highest number of leaves. The best rooting percent was obtained when plantlets were s treated with 2 mg. L-1 was applied to 1/4 MS medium. The highest number of roots was obtained from plantlets grown in 1/2 MS medium treated with IBA at 3 mg.L-1, whereas the longest root was obtained from plantlets grown in 1/2 MS medium treated with 1 mg.L-1 IBA. [ABSTRACT FROM AUTHOR]

One of the important sources of human selenium intake is vegetables, so increasing of selenium in edible parts of plants, biofortification, provides an effective approach to reduce selenium deficiency. Origanum vulgare L. is an important medicinal plant which is also used as a valuable spice. The present study was done to evaluate the ability of selenium accumulation of micropropagated O. vulgare under sodium selenate and/or salicylic acid treatment. Micropropagation of O. vulgare was done from shoot-tips as explants on modified Murashige & Skoog (MS) medium. After 1 month, rooted plantlets were transferred to modified Murashige & Skoog solidified medium containing sodium selenate (0, 25 and 50 µM) and/or salicylic acid (0 and 100 mgL−1). After 4 wk, growth, selenium accumulation, biochemical parameters and oxidative stress indicators were determined. The results showed that the plantlets were able to accumulate 16.1 µg g−1 DW and 28.27 µg g−1 DW selenium in shoots at 25 and 50 µM selenate sodium concentrations respectively. Both concentrations of selenium increased shoot dry weight, length of shoot and root, soluble sugars and total protein content, peroxidase and superoxide dismutase activity. On the contrary, malondialdehyde content decreased under selenium treatment. It was found that by increasing selenium concentration the content of total protein, H2O2 and antioxidant enzymes activity increased significantly. Application of salicylic acid in combination with selenium improved growth but did not affect on selenium accumulation. The obtained results can be considered as the first report of biofortification of O. vulgare under tissue culture technique. [ABSTRACT FROM AUTHOR]

This study aimed to develop a suitable micropropagation protocol for black mulberry (Morus nigra L.), which cannot be easily clonally propagated like the other mulberry species. Clonal propagation of this species is gaining importance due to its health benefits and utilization in pharmacology. The effects of the culture media and cytokinin concentrations on the micropropagation of black mulberry were investigated. Single-axillary bud explants grown on Murashige and Skoog medium, Driver and Kuniyuki walnut medium, and Nas and Read medium (NRM) containing 0.25 mg L-1 of benzyladenine (BA) plus 0.01 mg L-1 of indole-3-butyric acid (IBA) were compared to cultures grown on NRM with thidiazuron (TDZ), meta-Topolin (mT), or BA at concentrations of 1.25, 2.5, 5.0, or 7.5 mg L-1 plus 0.01 mg L-1 of IBA. The culture medium had a significant effect on the shoot length, which was greater on the NRM (3.3 shoots per explant) compared to the other media. Shoot multiplication was similar on all of the tested media. TDZ negatively affected shoot formation and triggered rosette shoots and/or large callus formation. BA and mT promoted both shoot proliferation and elongation. Around 84% of the micro shoots rooted and the rooted shoots were successfully acclimatized to ex vitro conditions. [ABSTRACT FROM AUTHOR]

Genetic variability induced by cell and tissue culture techniques provides a new source of diversity for improvement of ornamental plants species. For this purpose, the effect of Thidiazuron (TDZ) on callus induction and number of subcultures of Salvia splendens was studied. The results showed that the MS medium supplemented with TDZ (0.5 mg/l) and Indole Acetic Acid IAA (0.1 mg/l) produced the highest percentage for explants producing callus (60%). For shoot multiplication, the best results were achieved when shoot tips were cultured on MS medium supplemented with TDZ (0.2 mg/l) alone. For subculture the same concentration of TDZ (0.2 mg/l) in media of eleven subcultures were performed at 21-day intervals over a total period of 231 days. For molecular variation detection, SCoT molecular markers were employed. Five SCoT primers were used and a total of 24 bands were obtained. The results showed high level of polymorphism between parent and their regenerated plants (subculture No. 11). These markers revealed distinctly different patterns of DNA fragments in all somaclones, and some distinctive bands were linked to the regenerated plants. For rooting, the best rooting percentage (100.00%), highest number of roots/shoot (7.00) and root length (2.58 cm) were observed when the shoots were cultured on MS hormonefree medium supplemented with 15 g sucrose/l. Plantlets were successfully acclimatized on Peat moss + perlite (1:1) with a survival rate of 100 %. [ABSTRACT FROM AUTHOR]

Plants are the sources of valuable biomass that are being currently used in many areas. It is important to produce high biomass for efficient commercial production. Amongst the many factors that affect in vitro propagation of plants, changing or enriching the media composition is one of the commonly used techniques in micropropagation of plants. Biochar is a solid product obtained from organic wastes and because of its rich composition, it has many beneficial effects on plants. In our study, Lavandula officinalis plantlets were subjected to two types of biochars (Geocharged biochar and Biorfe biochar) at 0.5 and 2 g/L concentrations and their effects were investigated by means of plant growth, biomass accumulation and biochemical composition. The results showed that 0.5 g/L concentration of biochar had better effects than 2 g/L concentration and except for biochemical composition, biochar type had no significant effect on plant growth and biomass accumulation. Mean root dry weights and multiple shoot formations/explant enhanced up to 3.7 and 4.17 times higher than the control at 0.5 g/L concentration. Explant browning was also detected lower in biochar-applied media. The differences between biochemical accumulations of different media were also found statistically significant. The total concentrations of phenolics and flavonoids and radical scavenging activities were detected lower when biochars were applied. The total antioxidant concentration was higher in the control group. These findings showed that biochars lowered the negative effects of the culture conditions for L. officinalis plantlets. [ABSTRACT FROM AUTHOR]

The influence of the temperature and different light qualities emitted by light-emitting diodes (LEDs) and fluorescent lamps (Fl) on the micropropagation of the tree fern Cyathea delgadii was evaluated. The most efficient somatic embryo production was obtained on internode explants at 22 °C. The optimal temperature range for obtaining well-developed plants was 24–28 °C. This stimulated the elongation and development of the first leaf and the formation of the next leaf and roots primordia. Temperatures lower than 24 °C and higher than 28 °C inhibited the formation of young sporophyte organs and delayed their development. The RBUV (35% red, 15% blue, and 50% UV) and B (100% blue) lights and Fl light were beneficial for the sporophyte production on internode explants. However, plants obtained under RBUV light were undeveloped. The white LED light stimulated the number of explants capable of gametophyte production and development. The RB light (70% red and 30% blue) enhanced the number of roots of newly-formed plants. Most of the LED lights tested had a good impact on root elongation compared to Fl light and constant darkness. The R light (100% red) benefits leaf development and elongation. Research shows that temperature and LED lightning play a significant role in the process of morphogenesis in C. delgadii, significantly affecting the embryogenic competence of somatic cells and the development of sporophytes.Key message: A wide range of temperatures and light qualities have been investigated for optimizing the efficiency of in vitro morphogenesis of the tree fern Cyathea delgadii. [ABSTRACT FROM AUTHOR]