In Vitro Propagation of "Salsaparrilha" [Herreria salsaparrilha Mart. (Asparagaceae)]: An Antisyphilitic Medicinal Species.

An efficient procedure for in vitro propagation of Herreria salsaparrilha Martius was established from single-node explants (fourth and fifth nodes from apex to the base) derived from donor plants maintained under shading-house conditions. After surface sterilization, explants are inoculated in test tubes containing 15 mL of Murashige and Skoog (MS) medium without growth regulators. Cultures are maintained under 35 µmol m-2 s-1 irradiance, a 16/8-h light/dark light regime, at 26 ± 2 °C. The subcultures are carried out under the same conditions, adding 6-benzyladenine 1.0 mg/L and Phytagel® 2.8 g/L. Shoots are elongated and rooted by transferring individual shoots to half-strength MS medium without growth regulators. After 25-30 days, elongated rooted shoots are transferred to plastic pots containing 25-30 mL of sterile distilled water, covered with a transparent plastic bag, and kept under the same growth room conditions for 2 days. Plants are transferred to cups containing autoclaved and washed sand and kept in a shading house (50% light interception) for acclimatization. True-to-type adult plants were successfully recovered under ex vitro conditions.

Optimizing PGRs for in vitro shoot proliferation of pomegranate with bayesian-tuned ensemble stacking regression and NSGA-II: a comparative evaluation of machine learning models.

BACKGROUND: The process of optimizing in vitro shoot proliferation is a complicated task, as it is influenced by interactions of many factors as well as genotype. This study investigated the role of various concentrations of plant growth regulators (zeatin and gibberellic acid) in the successful in vitro shoot proliferation of three Punica granatum cultivars ('Faroogh', 'Atabaki' and 'Shirineshahvar'). Also, the utility of five Machine Learning (ML) algorithms-Support Vector Regression (SVR), Random Forest (RF), Extreme Gradient Boosting (XGB), Ensemble Stacking Regression (ESR) and Elastic Net Multivariate Linear Regression (ENMLR)-as modeling tools were evaluated on in vitro multiplication of pomegranate. A new automatic hyperparameter optimization method named Adaptive Tree Pazen Estimator (ATPE) was developed to tune the hyperparameters. The performance of the models was evaluated and compared using statistical indicators (MAE, RMSE, RRMSE, MAPE, R and R2), while a specific Global Performance Indicator (GPI) was introduced to rank the models based on a single parameter. Moreover, Non­dominated Sorting Genetic Algorithm­II (NSGA­II) was employed to optimize the selected prediction model. RESULTS: The results demonstrated that the ESR algorithm exhibited higher predictive accuracy in comparison to other ML algorithms. The ESR model was subsequently introduced for optimization by NSGA­II. ESR-NSGA­II revealed that the highest proliferation rate (3.47, 3.84, and 3.22), shoot length (2.74, 3.32, and 1.86 cm), leave number (18.18, 19.76, and 18.77), and explant survival (84.21%, 85.49%, and 56.39%) could be achieved with a medium containing 0.750, 0.654, and 0.705 mg/L zeatin, and 0.50, 0.329, and 0.347 mg/L gibberellic acid in the 'Atabaki', 'Faroogh', and 'Shirineshahvar' cultivars, respectively. CONCLUSIONS: This study demonstrates that the 'Shirineshahvar' cultivar exhibited lower shoot proliferation success compared to the other cultivars. The results indicated the good performance of ESR-NSGA-II in modeling and optimizing in vitro propagation. ESR-NSGA-II can be applied as an up-to-date and reliable computational tool for future studies in plant in vitro culture.

Variations in Cold Resistance and Contents of Bioactive Compounds among Dendrobium officinale Kimura et Migo Strains.

Dendrobium officinale is a valuable traditional Chinese herbal plant that is both medicinal and edible. However, the yield of wild Dendrobium officinale is limited. Adverse stress affects the growth, development, and yield of plants, among which low temperature is the primary limiting factor for introducing Dendrobium officinale to high-latitude areas and expanding the planting area. Therefore, this study aims to explore the variations in growth ability, cold resistance, and contents of bioactive compounds among different Dendrobium officinale strains. Four strains of Dendrobium officinale were selected as experimental materials and were subjected to low-temperature stress (4 °C). The agronomic traits, physiological indices, as well as the expressions of cold resistance-related genes (HSP70, DcPP2C5, DoCDPK1, and DoCDPK6) in the roots and leaves of Dendrobium officinale, were determined. The contents of bioactive compounds, including polysaccharides, flavonoids, and phenols were also measured. Compared with the other strains, Xianju had the highest seed germination and transplantation-related survival rates. Under low-temperature stress, Xianju exhibited the strongest cold resistance ability, as revealed by the changes in water contents, chlorophyll levels, electrical conductivities, enzyme activities, and expressions of the cold resistance-related genes. Additionally, the polysaccharide content of Xianju increased the most, while the stem flavonoid and leaf phenol contents were elevated in all four strains under cold treatment. Therefore, selecting excellent performing strains is expected to expand the planting area, improve the yield, and increase the economic benefits of Dendrobium officinale in high latitude areas with lower temperatures.

Micropropagation and Genetic Fidelity of Fegra Fig (Ficus palmata Forssk.) and Grafting Compatibility of the Regenerated Plants with Ficus carica.

Ficus palmata is an important fig species that produces edible and nutritious fruit and possesses several therapeutic uses. This study reports an effective method for the micropropagation of F. palmata using nodal explants. In vitro shoots were cultured for 7 weeks onto MS medium fortified with different concentrations of cytokinins, light intensities, sucrose concentrations, and light/dark incubation treatments. Optimal axillary shoot proliferation (10.9 shoots per explant) was obtained on a medium containing 30 g/L sucrose and supplemented with 2 mg/L 6-benzylaminopurine (BAP) under 35 µmol/m2/s light intensity. Dark incubation limited the foliage growth but favored shoot elongation and rooting compared with light incubation. Elongated shoots, under dark conditions, were rooted (100%; 6.67 roots per explant) onto MS medium containing 1 mg/L indole-3-acetic acid (IAA) and 1.5 g/L activated charcoal. The micropropagated plantlets were acclimatized with a 95% survival rate. In this study, the genetic fidelity of micropropagated F. palmata clones along with their mother plant was tested using randomly amplified polymorphic DNA (RAPD), inter-simple sequence repeats (ISSR), and start codon targeted (SCoT) molecular markers. The genetic similarity between the micropropagated plantlets and the mother plant of F. palmata was nearly 95.9%, assuring high uniformity and true-to-type regenerated plants. Using micropropagated F. palmata plantlets as a rootstock proved appropriate for the grafting F. carica 'Brown Turkey'. These findings contribute to the commercial propagation and production of the fig crop.

Effectiveness of light-emitting diodes for arsenic and mercury accumulation by Ceratophyllum demersum L.: An innovative advancement in phytoremediation technology.

Light Emitting Diodes (LEDs) have emerged as a tool with great potential in the field of phytoremediation, offering a novel approach to enhance the efficiency of plant-based remediation techniques. In this work investigated the influence of LEDs on the phytoremediation of arsenic (As) and mercury (Hg) by Ceratophyllum demersum L., propagated using tissue culture methods. In addition, the biochemical properties of the plants exposed to metal toxicity were examined. Phytoremediation experiments employed concentrations of As (0.01-1.0 mg/L) and Hg (0.002-0.2 mg/L), with application periods set at 1, 7, 14, and 21 days. In addition to white, red and blue LEDs, white fluorescent light was used for control purposes in the investigations. A positive correlation was observed between higher metal concentrations, extended exposure times, and increased metal accumulation in the plants. Red LED light yielded the highest level of heavy metal accumulation, while white fluorescent light resulted in the lowest accumulation level. Examination of the biochemical parameters of the plants, including photosynthetic pigment levels, protein quantities, and lipid peroxidation, revealed a pronouncedly enhanced performance in specimens subjected to red and blue LED illumination, surpassing outcomes observed in other light treatments. The findings of this study introduce innovative avenues for the effective utilization of red and blue LED lights in the realm of phytoremediation research. Thus, the interaction between LEDs, tissue culture, and the phytoremediation process could lead to synergistic effects that contribute to more effective and sustainable remediation strategies.

Cyanobacterial elicitor enhances the biomass of Mentha piperita L. and improves the production of high-value rosmarinic acid under in vitro culture of apical meristem.

BACKGROUND: Rosmarinic acid (RA), like other phenolic compounds, is sources of antioxidants and anti-inflammatory agents in medicinal plants. In vitro culture of plants can improve the medicinal plants' metabolite profile and phenolic compound quantity. To date, various methods have been proposed to increase this medicinal metabolite in plants, among which the use of bioelicitors can be mentioned. In the present study, a native isolate of heterocystous cyanobacteria, Nostoc spongiaeforme var. tenue ISB65, was used to stimulate the production of biomass and content of RA in Mentha piperita L. (peppermint) grown in vitro from apical meristem. Mentha piperita L. explants were inoculated in half strength Murashige and Skoog (1/2 MS) medium containing cyanobacterial lysate (CL). After 50 days of culturing, the growth indices, the content of photosynthetic pigments, and RA in control and treated plants were measured. RESULTS: CL inoculation resulted in a significant enhancement in the vegetative growth indices of peppermint, including root and shoot length, plant biomass and leaf number. The content of photosynthetic pigments also increased in cyanobacteria-treated plants. Inoculation with CL increased the RA content by 2.3-fold, meaning that the plants treated with CL had the highest RA content (7.68 mg. g- 1 dry weight) compared to the control (3.42 mg. g- 1 dry weight). Additionally, HPLC analysis revealed the presence of several auxins in CL. CONCLUSIONS: The presence of auxins and the chemical content of CL such as K+ and Ca2+, as regulators of metabolic pathways and molecular activities of cells, may be responsible for the enhanced growth and phenolic compounds of plants under tissue culture conditions. An improvement in RA content in the tissue culture of medicinal plants treated with CL was reported for the first time in this investigation.

Micropropagation of Stevia (Stevia rebaudiana Bert.) in RITA®.

Stevia rebaudiana Bert. is a plant that contains noncaloric sweeteners highly appreciated in the food industry. However, there is a high demand for propagules to establish commercial plantations, and the conventional reproduction types for this species are inefficient. Micropropagation is a technique that allows obtaining a large number of plants and can be used to meet the demand in the field. However, it requires in vitro propagation techniques such as temporary immersion systems (SIT) to increase yield and reduce production costs. This chapter describes an effective protocol for the large-scale micropropagation of S. rebaudiana using a TIS.

Temporary Immersion Systems in Plant Micropropagation.

Temporary immersion systems (TIS) are technological tools that support plant micropropagation. Given their high efficiency in the in vitro propagation of shoots, a current goal is to update the protocols addressing micropropagation in semisolid culture systems to protocols involving TIS. To this end, different parameters have been evaluated, including TIS types and designs, immersion times, immersion frequencies, and volume of medium per explant, among other characteristics. This has resulted in the improved production of propagules of plants of economic interest and the production of physiologically upgraded plants with high percent survival during acclimatization. TIS are specialized culture flasks that provide countless advantages during the commercial micropropagation of plants.

Direct Shoot From Root and True-to-Type Micropropagation of Limonium "Misty Blue" in Partially Immersed Culture on an Aluminum Mesh Raft.

Commercial plant tissue culture now primarily serves the ornamental horticulture industry. The main pillars of the commercial tissue culture business are scalability of production, cost reduction, limited labor involvement, high quality, and genetic homogeneity of propagated plants. Based on these requirements, the current protocol employs a partially immersed liquid culture medium supported by a flexible aluminum mesh raft with a wire stand to facilitate shoot organogenesis from the horizontally placed root explants and hold the plants upright for shoot multiplication and rooting of Limonium Misty Blue. It is a florist crop that is in high demand as both dried and fresh flower fillers in various floral decorations. The majority of cultivated Limonium or statice cultivars are heterozygous in nature and propagate commercially through in vitro propagation to cater to the huge demand for planting materials needed for flower production. This is the first protocol to describe direct shoot organogenesis from the roots in a liquid half-component of Murashige and Skoog's (1962) (MS) basal medium supplemented with 1.6 µM NAA and 1.1 µM BA. The regenerated shoots are multiplied and rooted at the same time on the raft in a MS-based liquid culture medium that included 0.44 µM BA and 1.07 µM NAA. In comparison to agar-gelled medium, plants cultured in liquid medium grow more quickly without any signs of hyperhydricity. In liquid medium, a clump of 4-5 shoots is formed from a single shoot explant within 4 weeks and are rooted simultaneously within 6 weeks. On average, seven explants may fit on each raft, so on average, 25 healthy plants are produced from a single bottle. The regenerated plants are easily hardened in the greenhouse, and using ISSR-based molecular markers, the genetic homogeneity of the randomly selected hardened plants can be determined.

Tissue Culture via Protocorm-like Bodies in an Orchids Hybrids Paphiopedilum SCBG Huihuang90.

This study successfully established an efficient in vitro propagation system for Paphiopedilum SCBG Huihuang90 via protocorm-like body (PLB) formation from seed-derived calluses, PLB proliferation and differentiation, root induction and greenhouse acclimatization. Furthermore, 1/2 Murashige and Skoog (MS) + 0.025 mg/L 2,4-Dichlorophenoxyacetic acid (2,4-D) was suitable for the proliferation of PLBs, and 1/2MS + 10% coconut water (CW, v/v) + 0.5 g/L activated carbon (AC) was suitable for PLB differentiation. PLBs at different developmental stages required different kinds of sugars. This study provided a reference for research on the propagation techniques of other Paphiopedilum.

Aloe monticola Reynolds: A refugee of the mountains - contributing towards its conservation through in vitro propagation.

In vitro micropropagation study of Aloe monticola Reynolds was conducted using offshoots in Murashige and Skoog (MS) media enriched with plant growth regulators (PGRs). Initiation experiment, carried out by culturing sterilized explants in full-strength MS media enriched with 0.0-1.0 mg/L benzyl amino purine (BAP) alone and in combination with 0.10 mg/L indole-3-butyric acid (IBA), resulted in 89 - 100 % healthy and live (i.e., initiated) explants in 9-29 days. Regeneration study, conducted by culturing initiated explants in full-strength MS media supplemented with 0.0-3.0 mg/L BAP alone and in combination with 0.50 mg/L IBA, showed that treatments enriched with 1.0-3.0 mg/L BAP in combination with 0.50 mg/L IBA yielded better shooting responses than the rest of the treatments. The rooting responses of the shoots were also tested by culturing in half-strength MS media enriched with 0.0-4.0 mg/L NAA alone and in combination with 0.25 mg/L IBA. Better rooting responses were observed in treatments supplemented with 1.0-4.0 mg/L NAA in combination with 0.25 mg/L IBA with two exceptions. The responses of A. monticola plantlets to primary and secondary acclimatization in greenhouse, nursery shade, and direct sunlight in coco peat, composted soil, and manured soil were excellent - with survival percentages of 90-98 %. The findings of this empirical research are important for developing refined protocol of in vitro micropropagation of this ecologically important but endangered plant.

Influence of the Abiotic Elicitors Ag Salts of Aspartic Acid Derivatives, Self-Organized in Nanofibers with Monomeric and Dimeric Molecular Structures, on the Antioxidant Activity and Stevioside Content in Micropropagated Stevia rebaudiana Bert.

The use of nanomaterials in biotechnology for the in vitro propagation of medical plants and the accumulation of certain biologically active metabolites is becoming an efficient strategy. This study aimed to evaluate the influence of the concentration (0, 1, 10, 50, and 100 mg L-1) of two types of nanofibers on the growth characteristics, the antioxidant status, and the production of steviol glycosides in micropropagated Stevia rebaudiana Bert. plantlets. The nanofibers were synthesized by aspartic acid derivatives (L-Asp) Ag salts self-organized into nanofibers with two different molecular structures: monomeric, containing one residue of L-Asp with one hydrophilic head which bonds one Ag ion (NF1-Ag salt); and dimeric, containing two residues of L-Asp with two hydrophilic heads which bond two Ag ions (NF2-Ag salt). An increase in the shoots from the explants' number and length, biomass accumulation, and micropropagation rate was achieved in the plants treated with the NF1-Ag salt in concentrations from 1 to 50 mg L-1 after 30 days of in vitro proliferation compared to the NF2-Ag salt. In contrast, the plants grown on MS media supplemented with NF2-Ag salt exhibited an increase in the level of stevioside, rebaudioside A, and mono- (CQA) and dicaffeoylquinic (DCQA) acids as compared to the NF1-Ag salt.

Quantitative regeneration: Skoog and Miller revisited.

In 1957, Skoog and Miller published their seminal work on the effects of hormones upon plant growth. By varying the concentrations of auxin and cytokinin, they observed dramatic differences in shoot and root growth from tobacco stem cultures. Their finding that quantitative differences in hormone concentrations could dramatically alter the fate of developing organs provided a foundation for understanding organ formation and tissue regeneration. Their in vitro assays established plant propagation techniques that were critical for regenerating transgenic plants. Here, I discuss their original paper, what led to their findings and its impact on our understanding of hormone interactions, how plants regenerate and in vitro tissue culture techniques.

Zingiber zerumbet (L.) Roscoe ex Sm.: biotechnological advancements and perspectives.

Shampoo ginger (Zingiber zerumbet) is a multipurpose ginger that has confirmed their role as food, medicine, and for decorative purposes. The rhizome possesses zerumbone, curcuminoids, and other bioactive molecules that play crucial roles in treating several human diseases. To date, several reports are existing on the in vitro biotechnology of Z. zerumbet. The present review highlights the consolidated clarification and comprehensive explanation of in vitro biotechnological implications based on plant tissue culture for the improvement of Z. zerumbet. Studies on biotechnological involvement in shampoo ginger were primarily emphasized in the study of the last 3 decades, for instance, in vitro regeneration, micro-rhizome production, callus culture, somatic embryogenesis, ex vitro establishment, and molecular assessment of in vitro-raised clones. Moreover, this review provides insights into different in vitro culture systems and endophytes involvement in the production of secondary metabolites. This review will assist for advanced research areas related to in vitro manipulation of shampoo ginger, especially for the commercial cultivation of secondary metabolites rich clones of Z. zerumbet. Moreover, it will provide an insight into crop upgrading and breeding programs of this underutilized, aromatic, and medicinal plant for amended yield and quality. KEY POINTS: • Z. zerumbet is an aromatic spice and an ornamental • This review comprehensively assesses Z. zerumbet tissue culture • Key shortcomings and future directions of Z. zerumbet biotechnology.

In Vitro-Based Production of Virus-Tested Babaco (Vasconcellea x heilbornii, syn. Carica pentagona) in Ecuador: An Integrated Approach to an Endangered Crop.

Babaco (Vasconcellea x heilbornii), a fruit-bearing vegetatively propagated crop native to Ecuador, is appreciated for its distinctive flavor and nutritional properties. The aim of this research was to determine a functional protocol for tissue culture propagation of virus-free babaco plants including in vitro establishment, multiplication, rooting, and acclimation. First, symptomless babaco plants from a single commercial nursery were analyzed for virus detection and cared for using different disinfection treatments in the greenhouse to reduce contamination during the in vitro establishing step, and three cytokinins, 6-(γ,γ-Dimethylallylamino) purine (2IP), 6-Benzylaminopurine (BAP), and Thidiazuron (TDZ), were used to determine the best hormone for multiplication. The best treatment for plant disinfection was the weekly application of copper sulfate at the greenhouse and a laboratory disinfection using ethanol (EtOH) (70%), Clorox (2%), and a solution of povidone iodine (2.5%), with an 80% survival during in vitro plant establishment. TDZ showed a better multiplication rate when compared with other hormones, and 70% of the rooted plants were successfully acclimated at the greenhouse. Generated plants were virus-free when tested against babaco mosaic virus (BabMV) and papaya ringspot virus (PRSV), two of the most important viruses that can affect babaco. An efficient protocol to produce virus-free babaco plants was elaborated with an integrated use of viral diagnostic tools to ensure the production of healthy start material to farmers.

How Scarification, GA3 and Graphene Oxide Influence the In Vitro Establishment and Development of Strelitzia.

The propagation of strelitzia plants can be carried out in vitro as an alternative to combine the aseptic conditions of the culture medium with the use of strategies to promote germination and controlled abiotic conditions. However, this technique is still limited by the prolonged time and low percentage of seed germination, which is the most viable explant source, due to dormancy. Thus, the objective of this study was to evaluate the influence of chemical and physical scarification processes of seeds combined with gibberellic acid (GA3), as well as the effect of graphene oxide in the in vitro cultivation of strelitzia plants. Seeds were subjected to chemical scarification with sulfuric acid for different periods (10 to 60 min) and physical scarification (sandpaper), in addition to a control treatment without scarification. After disinfection, the seeds were inoculated in MS (Murashige and Skoog) medium with 30 g L-1 sucrose, 0.4 g L-1 PVPP (polyvinylpyrrolidone), 2.5 g L-1 Phytagel®, and GA3 at different concentrations. Growth data and antioxidant system responses were measured from the formed seedlings. In another experiment, the seeds were cultivated in vitro in the presence of graphene oxide at different concentrations. The results showed that the highest germination was observed in seeds scarified with sulfuric acid for 30 and 40 min, regardless of the addition of GA3. After 60 days of in vitro cultivation, physical scarification and scarification time with sulfuric acid promoted greater shoot and root length. The highest seedling survival was observed when the seeds were immersed for 30 min (86.66%) and 40 min (80%) in sulfuric acid without GA3. The concentration of 50 mg L-1 graphene oxide favored rhizome growth, while the concentration of 100 mg L-1 favored shoot growth. Regarding the biochemical data, the different concentrations did not influence MDA (Malondialdehyde) levels, but caused fluctuations in antioxidant enzyme activities.

Leaves micromorphology, chemical profile, and bioactivity of in vitro-propagated Nepeta cyrenaica (Lamiaceae).

INTRODUCTION: The endemic species Nepeta cyrenaica Quézel & Zaffran, native to northeastern Libya, is valued as an important honey-bearing plant. OBJECTIVES: This study was aimed to examine the micromorphology, phytochemistry, and bioactivity of in vitro-propagated N. cyrenaica for the first time. MATERIALS AND METHODS: The leaf indumentum was examined using light and scanning electron microscopy and further characterised for histochemistry. The chemical composition of essential oil (EO) was performed using GC-MS analysis, while dichloromethane (DCM), methanol (ME), ethanol (ET), and aqueous (AQ) extracts were analysed using qualitative and quantitative LC/MS analyses. The antioxidant activities of EO and extracts were assessed using three parallel assays, while enzyme-inhibiting effects were evaluated against four enzymes. RESULTS: The leaves bear various types of glandular trichomes, with lipophilic secretion predominating. The main EO component of EO was 1,8-cineole. A considerable number of phenolics and iridoids were tentatively identified in the ME extract. Quantitative LC/MS analysis confirmed that ferulic acid, rosmarinic acid, and epigallocatechin gallate were present in the highest amount in the extracts, in which three iridoids were also quantified. Although the ME extract contained the highest amount of polyphenolics and iridoids, the DCM extract showed the best overall biological potential. Additionally, EO exerted the strongest acetylcholinesterase and tyrosinase inhibition. CONCLUSION: This study demonstrated that the endemic N. cyrenaica can be efficiently grown under in vitro conditions, where it develops various glandular trichomes that are thought to secrete and/or accumulate bioactive compounds with valuable medicinal potential.

Bioreactor systems for micropropagation of plants: present scenario and future prospects.

Plant micropropagation has been adapted in the fields of agriculture, horticulture, forestry, and other related fields for large-scale production of elite plants. The use of liquid media and adoption of bioreactors have escalated the production of healthy plants. Several liquid-phase, gas-phase, temporary immersion, and other modified bioreactors have been used for plant propagation. The design, principle, operational mode, merits, and demerits of various bioreactors used for the regeneration of propagules, such as bulblets, cormlets, rhizomes, microtubers, shoots (subsequent rooting), and somatic embryos, are discussed here. In addition, various parameters that affect plant regeneration are discussed with suitable examples.

Approaches for in vitro propagation and production of plumbagin in Plumbago spp.

The genus Plumbago (family Plumbaginaceae), commonly known as leadwort, is a sub-tropical shrub that produces secondary metabolite plumbagin, which is employed by pharmaceutical companies and in clinical research. Plumbagin is a potent pharmaceutical because of its anti-microbial, anti-malarial, antifungal, anti-inflammatory, anti-carcinogenic, anti-fertility, anti-plasmodium, antioxidant, anti-diabetic, and other effects. This review documents the biotechnological innovations used to produce plumbagin. The use of modern biotechnological techniques can lead to a variety of benefits, including better yield, increased extraction efficiency, mass production of plantlets, genetic stability, increased biomass, and more. Large-scale in vitro propagation is necessary to minimize over-exploitation of the natural population and allow the use of various biotechnological techniques to improve the plant species and secondary metabolite production. During in vitro culture, optimum conditions are requisites for explant inoculation and plant regeneration. In this review, we provide information on various aspects of plumbagin, depicting its structure, biosynthesis, and biotechnological aspects (both conventional and advanced) along with the future prospects. KEY POINTS: • Critical assessment on in vitro biotechnology in Plumbago species • In vitro propagation of Plumbago and elicitation of plumbagin • Biosynthesis and sustainable production of plumbagin.

Effect of Salinity Stress on Phenolic Compounds and Antioxidant Activity in Halophytes Spergularia marina (L.) Griseb. and Glaux maritima L. Cultured In Vitro.

The study of halophytes as sources of phenolic compounds, as well as conditions that further enhance the accumulation of biologically active compounds in them, is of particular interest. In this paper, the effect of different salinity levels (25-500 mM in the form of NaCl) on the content of phenolic compounds and the antioxidant activity of two rare halophyte species Spergularia marina (L.) Griseb. and Glaux maritima L. cultured in vitro was investigated. A species-specific reaction of plants to salinization was established. In G. maritima, the maximum total content of phenolic compounds was observed at 50-100 mM, flavonoids 75-400 mM, and hydroxycinnamic acids 200-300 mM, as well as individual phenolics (protocatechuic acid, catechin, astragalin, hyperoside, rutin, isoquercitrin, and apigenin derivative) at 100-300 mM NaCl. For S. marina, on the contrary, there was a slight decrease in the content of phenolic compounds when NaCl was added to the nutrient medium compared to the control. The content of protocatechuic acid, rosmarinic acid, and apigenin derivative significantly decreased with increased salt stress. The change in antioxidant activity at different salinity levels was also species specific. The maximum values of different groups of phenolic compounds in G. maritima were observed at 50-300 mM NaCl. The cultivation of S. marina without the addition of NaCl and at 500 mM NaCl allowed the production of plants with the highest content of phenolic compounds. The obtained results can be further used in the development of protocols for the cultivation of these plants in vitro in order to induce the biosynthesis of phenolic compounds in them.