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Digitalis purpurea L. is one of the important plant species of Nilgiris, Kashmir and Darjeeling regions of India, belonging to the family Plantaginaceae, with well-known pharmacological applications. In the present investigation, an in vitro culture technique of indirect shoot organogenesis of D. purpurea is being explored; the biochemical attributes, the antioxidant activities and the metabolomic analyses were made by utilizing untargeted Gas Chromatography-Mass Spectrometry (GC-MS) and Ultra Performance Liquid Chromatography coupled with electronspray ionization/quadrupole-time-of-flight-mass spectrometry (UPLC-ESI-QTOF-MS) approaches. Initially, the leaf explants were used for callus induction and proliferation and maximum callusing frequency (94.44%) and fresh biomass (4.9 g) were obtained on MS, fortified with 8.8 µM BAP (6-benzyl amino purine) + 0.9 µM 2,4-D (2,4-dichlorophenoxyacetic acid), subsequently shoot formation (indirect organogenesis) was noted on the same MS medium with a shoot induction frequency of 83.33%. Later on, the biochemical and antioxidant potential of in vivo-, in vitro grown leaf and leaf derived callus were assessed. Significantly higher total phenol, flavonoid, DPPH (2,2-diphenyl-1-picrylhydrazyl), POD (peroxidase) and SOD (superoxide dismutase) activities were noticed in in vitro grown callus and leaf tissues compared with field grown leaf. The GC-MS analysis of each methanolic extract (in vivo-, in vitro derived leaf and leaf derived callus) displayed the presence of more than 75 bioactive compounds viz loliolide, stigmasterin, alpha-tocopherol, squalene, palmitic acid, linoleic acid, beta-amyrin, campesterol etc. possessing immense therapeutic importance. The UPLC-MS based metabolite fingerprinting of each methanolic extracts were conducted in both positive and negative ionization mode. The obtained results revealed variation in phytochemical composition in field - and laboratory grown tissues, indicating the impact of in vitro culture conditions on plant tissues. The detected phytocompounds belongs to various classes such as flavonoids, steroids, terpenoids, carbohydrates, tannins, lignans etc. The medicinally important metabolites identified were 20, 22-dihydrodigoxigenin, digoxigenin monodigitoxoside, apigenin, luteolin, kaempferide, rosmarinic acid, nepitrin and others. The results of the present study suggest that in vitro culture of D. purpurea could successfully be utilized for the novel drug discovery by producing such important phytocompounds of commercial interest in shorter duration without harming the plants' natural population. [ABSTRACT FROM AUTHOR]

This work studied the micropropagation of fegra fig (F. palmata Forssk.) during which we experienced the incidence of shoot-tip necrosis (STN). STN was evident during the shoot elongation stage, which was regenerated on Murashige and Skoog (MS) medium supplemented with 2 mg/L 6-benzylaminopurine. To alleviate SNT, we conducted a series of experiments and supplemented the medium with calcium chloride (40, 80, and 120 mg/L), ascorbic acid (50, 100, and 150 mg/L), silver nitrate (1, 2, and 3 mg/L), and boric acid (9.3, 12.4, and 15.5 mg/L). Results showed that all the treatments controlled STN at varying levels, and supplementation of medium with 3 mg/L silver nitrate reduced the incidence of STN from 80% to 24%. The regenerated shoots were rooted on the same medium with incubation of cultures in the dark for 3 weeks and subsequent 4 weeks of incubation under 16/8-hour light/dark photoperiod. The growth parameters (number of shoots and roots, length of the main shoot and root, fresh and dry weights), photosynthetic pigments (chlorophylls and carotenoids), and relative water content of plantlets were restored with the application of 3 mg/L silver nitrate to the medium. Incubation of cultures initially in the dark followed by 16/8-hour light incubation facilitated axillary shoot elongation. On the basis of our findings, it is recommended to culture the regenerated axillary shoots of fegra fig onto MS medium containing 3% sucrose, 1.5 mg/L activated charcoal, and 3 mg/L silver nitrate to manage STN effectively. [ABSTRACT FROM AUTHOR]

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

Galanthamine is an immensely valuable alkaloid exhibiting anti-cancer and antiviral activity. The cultivation of plant tissues in in vitro conditions is a good source for the synthesis and enrichment of secondary metabolites of commercial interest. In this study, the Amaryllidaceae alkaloid galanthamine was quantified in three Zephyranthes species, such as Zephyranthes candida, Zephyranthes grandiflora, and Zephyranthes citrina, and the impact of the methyl jasmonate (MJ) signaling molecule on galanthamine accumulation was monitored in in vitro-derived plant tissues. This is the first ever study of the MJ-regulated accumulation of galanthamine in in vitro-grown Zephyranthes tissues. Shoot regeneration was obtained in all three Zephyranthes species on Murashige and Skoog (MS) medium containing 2.0 mgL−1 benzylaminopurine (BAP) + 0.5 mgL−1 naphthalene acetic acid (NAA). The regenerated shoots were rooted on a medium containing 2.0 mgL−1 indole butyric acid (IBA). A GC-MS study of Zephyranthes extracts revealed the presence of 34 phyto-compounds of varied levels with therapeutic activities against diseases. The galanthamine content was quantified in plant parts of the three Zephyranthes species using high-performance thin layer chromatography (HPTLC); the maximum was found in Z. candida bulb (2.41 µg g−1 dry wt.), followed by Z. grandiflora (2.13 µg g−1 dry wt.), and then Z. citrina (2.02 µg g−1 dry wt.). The galanthamine content showed bulb > leaf > root source order. The in vitro-generated plantlets were treated with different MJ concentrations, and the galanthamine yield was measured in bulb, leaf, and root tissues. The highest galanthamine content was recorded in bulbs of Z. candida (3.97 µg g−1 dry wt.) treated with 150 µM MJ, showing an increase of 64.73% compared to the control. This accumulation may be attributed to MJ-induced stress, highlighting the potential commercial synthesis of galanthamine in vitro. [ABSTRACT FROM AUTHOR]

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

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. [ABSTRACT FROM AUTHOR]

KIGELIA AFRICANA belongs to the family Bignoniaceae, and this species has been exploited for many years for its medicinal properties caused by the secondary metabolites found in its fruit. Thus, this study aimed to determine the foliar micromorphology and histophyto-chemistry of K. africana leaf extracts using microscopy, histochemistry, and phytochemical screening. Microscopy revealed glandular peltate trichomes on the adaxial and abaxial surfaces of young and mature leaflets. Histochemical analysis showed that the peltate trichomes secreted lipids and alkaloids, and histo-phytochemical analysis indicated that the leaf extracts contained various secondary metabolites, including alkaloids, saponins, tannins, and phenols. The presence of glandular trichomes is a characteristic feature of species belonging to the family Bignoniaceae, and the phytochemicals present in the leaf extracts were similar to those found in fruit extracts. This study revealed the presence of peltate trichomes on the surfaces of young and mature leaflets, and histo-phytochemistry showed that the leaves contained various classes of phytochemical compounds. However, further work must be conducted to investigate the fine ultrastructure of the leaves and peltate trichomes as well as the biological activity of various phytochemicals. [ABSTRACT FROM AUTHOR]

Red radish is a nutritious root vegetable crop that has a short production cycle. Water deficit limits plant productivity, affecting its quantity and quality. Compost amendment offers a potential solution to mitigate water deficit effects. This study assessed the impact of compost manure rates (0%, 50%, 75%, and 100%) and irrigation treatments (40%, 60%, 80%, and 100% of evapotranspiration) on 'Crimson Giant' red radish production. Significant differences in growth and quality were observed among these treatments. Compost rates of 75% and 100% improved leaf gas exchange, plant growth (leaf count, fresh weight, dry weight, and area; stem length), root development, total yield (root fresh weight, dry weight, diameter, and length), and root quality (vitamin C and total soluble solid and titratable acidity). The 100% compost and 100% irrigation combination achieved the highest yields. Under water deficit, applying 75% or 100% compost with 80% irrigation conserved 20% of water while maintaining radish output. Overall, compost amendment effectively enhanced red radish growth and production under water deficit. [ABSTRACT FROM AUTHOR]

Tylophora indica (Burm. f.) Merrill is an endangered medicinal plant that possesses various active agents, such as tylophorinine, kaempferol, quercetin, α-amyrin and beta-sitosterol, with multiple medicinal benefits. α-amyrin, a triterpenoid, is widely known for its antimicrobial, anti-inflammatory, gastroprotective and hepatoprotective properties. In this study, we investigated the metabolite profiling of tissues and the effects of cadmium chloride and chitosan on in vitro accumulation of alkaloids in T. indica. First, the callus was induced from the leaf in 2,4-D-, NAA- and/or BAP-fortified MS medium. Subsequent shoot formation through organogenesis and in vitro roots was later induced. Gas chromatography–mass spectrometry (GC–MS)-based phytochemical profiling of methanolic extracts of in vivo and in vitro regenerated plants was conducted, revealing the presence of the important phytocompounds α-amyrin, lupeol, beta-sitosterol, septicine, tocopherol and several others. Different in vitro grown tissues, like callus, leaf and root, were elicited with cadmium chloride (0.1–0.4 mg L−1) and chitosan (1–50 mg L−1) to evaluate the effect of elicitation on α-amyrin accumulation, measured with high-performance thin layer chromatography (HPTLC). CdCl2 and chitosan showed improved sugar (17.24 and 15.04 mg g−1 FW, respectively), protein (10.76 and 9.99 mg g−1 FW, respectively) and proline (7.46 and 7.12 mg g−1 FW), especially at T3 (0.3 and 25 mg L−1), in the leaf as compared to those of the control and other tissues. The antioxidant enzyme activities were also evaluated under an elicitated stress situation, wherein catalase (CAT), superoxide dismutase (SOD) and ascorbate peroxidase (APX) displayed the highest activities in the leaf at T4 of both of the two elicitors. The α-amyrin yield was quantified with HPTLC in all tested tissues (leaf, callus and root) and had an Rf = 0.62 at 510 nm wavelength. Among all the concentrations tested, the T3 treatment (0.3 mg L−1 of cadmium chloride and 25 mg L−1 of chitosan) had the best influence on accumulation, irrespective of the tissues, with the maximum being in the leaf (2.72 and 2.64 μg g−1 DW, respectively), followed by the callus and root. Therefore, these results suggest future opportunities of elicitors in scaling up the production of important secondary metabolites to meet the requirements of the pharmaceutical industry. [ABSTRACT FROM AUTHOR]

As a result of its domestication, breeding and subsequent commercialization, African violet (Saintpaulia ionantha H. Wendl.) has become the most famous and popular Saintpaulia species. There is interest in producing cultivars that have increased resistance to pests and low temperature, in the introduction of novel horticultural characteristics such as leaf shape, flower colour, size and form, and in improved productivity and enhanced flower duration in planta. In African violet, techniques such as the application of chemical mutagens (ethylmethanesulfonate, N-nitroso-N-methylurea), radiation (gamma (γ)-rays, X-rays, carbon ion beams) and colchicine have been successfully applied to induce mutants. Among these techniques, γ radiation and colchicine have been the most commonly applied mutagens. This review offers a short synthesis of the advances made in African violet breeding, including studies on mutation and somaclonal variation caused by physical and chemical factors, as well as transgenic strategies using Agrobacterium-mediated transformation and particle bombardment. In African violet, Agrobacterium-mediated transformation is affected by the Agrobacterium strain, selection marker, and cutting-induced wounding stress. Somaclonal variation, which arises in tissue cultures, can be problematic in maintaining true-to-type clonal material, but may be a useful tool for obtaining variation in flower colour. The only transgenic African violet plants generated to date with horticulturally useful traits are tolerant to boron (heavy metal) stress, or bear a glucanase-chitinase gene.

An effective and reproducible micropropagation protocol was developed for Centratherum punctatum Cass. Successful in vitro initiation of callus and subsequent plant regeneration were obtained on nodal explants cultured on MS medium supplemented with plant growth regulators (PGRs). The maximum frequency of callus formation (98.3%) was noted on MS containing 4.0 mg/L 6-Benzylaminopurine (BAP) and 3.5 mg/L Kinetin with a maximum callus weight of 2.02 g. The best shoot induction frequency (100%) with an average of 30.2 shoots per explant was achieved when 4.5 mg/L BAP and 4.0 mg/L Kinetin were added to the MS. The same PGR combination resulted in the best callus-mediated shoot formation (8.3 shoots/callus mass). The highest rhizogenic response (95.3%) with an average 26.1 roots per shoot and root length of 6.2 cm was obtained with 1.0 mg/L Indole-3-acetic acid (IAA)-supplemented MS medium. The gas chromatography–mass spectrometry (GC-MS) technique was applied in the present study to analyze the methanolic extracts of the leaf, callus, and root of regenerated C. punctatum shoots to detect the different phytochemical constituents. The leaf extract of the regenerated C. punctatum showed 37 phytocompounds; some important bioactive compounds were the Phytol,1,6-Octadien 3,5-Dimethyl-Cis, 4,8-Dimethylnona-3,8-dien-2-one, 2,6-Octadiene, Stigmasterol, Chondrillasterol, Lanosteryl acetate, etc. In the callus, the extract had a total of 57 phytocompounds; among them, the Stigmasterol, Guanosine, and Tri-decanoic acid were the major ones. In the root extract, the GC-MS revealed a low number of 23 phytocompounds, the important compounds of which were Stigmasterol, Trimethylsilyl (TMS) derivative, Chrysantenyl 2-methuylbutanoate, 4-tert-Butoxybutan-1-ol, etc. The order in terms of numbers of phytocompounds present in tissue sources are callus > leaf > root. [ABSTRACT FROM AUTHOR]

Rose is a widely favored floriculture crop that is commercially propagated through the application of tissue culture techniques. Here, we report an effective method for axillary shoot proliferation in Al-Taif rose, an important cultivar for rose oil industry. Stem nodes were excised from an adult donor Al-Taif rose shrub and cultured for 4 weeks on Murashige and Skoog's (MS) medium supplemented with 6-benzylaminopurine (BAP) or gibberellic acid (GA3) at 0 and 3 mg·L−1 to induce the sprouting of axillary shoots. Al-Taif rose shoots were cultured in vitro for 6 weeks on MS medium fortified with different concentrations of cytokinins, light/dark incubation and different culture types (gelled and liquid/bioreactor culture). The culture conditions that were applied had a noteworthy impact on the responses of Al-Taif rose shoot proliferation. The supplementation of the medium with 6-benzylaminopurine (BAP) resulted in an augmented rate of shoot proliferation in comparison to other cytokinins. Additionally, dark incubation limited foliage growth, leaf yellowing and abscission and favored shoot proliferation compared with light incubation. Liquid culture using bioreactors provided higher axillary shoot proliferation and growth as compared with gelled culture. A continuous immersion system with a net provided the highest axillary shoots (four shoots per explant) and shoot length (16.5 cm), whereas an immersion system without a net provided the highest fresh weight of axillary shoots (499 mg per explant). These findings will improve commercial propagation and contribute to the rose oil industry of Al-Taif rose. [ABSTRACT FROM AUTHOR]

Lycorine is an important Amaryllidaceae alkaloid showing anti-cancerous activity on numerous cell lines; and it also demonstrates antiviral activity against several viruses including SARS-CoV-2 responsible for COVID-19. The in vitro cultivation of plant parts is a good alternative for elicitor mediated secondary metabolite synthesis, mass propagation, and cloning of elite genotypes. In this study, the quantification of lycorine from Zephyranthes candida, Zephyranthes grandiflora, and Zephyranthes citrina was evaluated, and the influence of methyl jasmonate (MJ) treatments on accumulation of lycorine yield was observed in the in vitro cultivated tissues. The influence of methyl jasmonate (MJ) on accumulation of lycorine was investigated for the first time in in vitro regenerated bulb, leaf, and root tissues of Zephyranthues. The efficient in vitro shoot regeneration was achieved in Z. candida (73.9%), Z. grandiflora (85.5%), and Z. citrina (76.5%) on Murashige and Skoog (MS) medium amended with 2.0 mg·L−1 Naphthalene acetic acid + 0.5 mg·L−1 Benzylaminopurine. The regenerants had the highest i.e., 7.46 mean numbers of bulblets per shoot. The best root induction (80.66%) was achieved on 2.0 mg/L Indole butyric acid (IBA). The lycorine content was quantified through High performance thin layer chromatography (HPTLC) in three field grown Zephyranthes species, was highest in Z. candida (1.93 µg g−1 dry wt.), followed by Z. grandiflora (1.87 µg g−1 dry wt.) and Z. citrina (1.62 µg g−1 dry wt.). The yield of lycorine was observed in this order i.e., bulb > leaf > root. The regenerated plants were supplemented with MJ and maximum lycorine yield was noted at 100 µM in bulb tissues of Z. candida (2.74 µg g−1 dry wt.) with an increase in percentage of 39.08. The enhancement could be due to MJ-induced stress as the biochemical attributes and anti-oxidant enzyme activity were high with elevated level of MJ. This enrichment may auger commercial manufacturing and utilization of lycorine in future. [ABSTRACT FROM AUTHOR]