Callus Extracts Research Articles

Improving the Production of Secondary Metabolites via the Application of Biogenic Zinc Oxide Nanoparticles in the Calli of Delonix elata: A Potential Medicinal Plant.

The implementation of nanotechnology in the field of plant tissue culture has demonstrated an interesting impact on in vitro plant growth and development. Furthermore, the plant tissue culture accompanying nanoparticles has been showed to be a reliable alternative for the biosynthesis of secondary metabolites. Herein, the effectiveness of zinc oxide nanoparticles (ZnONPs) on the growth of Delonix elata calli, as well as their phytochemical profiles, were investigated. Delonix elata seeds were collected and germinated, and then the plant species was determined based on the PCR product sequence of ITS1 and ITS4 primers. Afterward, the calli derived from Delonix elata seedlings were subjected to 0, 10, 20, 30, 40, and 50 mg/L of ZnONPs. The ZnONPs were biologically synthesized using the Ricinus communis aqueous leaf extract, which acts as a capping and reducing agent, and zinc nitrate solution. The nanostructures of the biogenic ZnONPs were confirmed using different techniques like UV-visible spectroscopy (UV), zeta potential measurement, Fourier transform infrared spectra (FTIR), X-ray diffraction (XRD), and scanning electron microscopy (SEM). Adding 30 mg/L of ZnONPs to the MS media (containing 2.5 µM 2,4-D and 1 µM BAP) resulted in the highest callus fresh weight (5.65 g) compared to the control and other ZnONP treatments. Similarly, more phenolic accumulation (358.85 µg/g DW) and flavonoid (112.88 µg/g DW) contents were achieved at 30 mg/L. Furthermore, the high-performance liquid chromatography (HPLC) analysis showed significant increments in gallic acid, quercetin, hesperidin, and rutin in all treated ZnONP calli compared to the control. On the other hand, the gas chromatography and mass spectroscopy (GC-MS) analysis of the calli extracts revealed that nine phytochemical compounds were common among all extracts. Moreover, the most predominant compound found in calli treated with 20, 30, 40, and 50 mg/L of ZnONPs was bis(2-ethylhexyl) phthalate, with percentage areas of 27.33, 38.68, 22.66, and 17.98%, respectively. The predominant compounds in the control and in calli treated with 10 mg/L of ZnONPs were octadecanoic acid, 2-propenyl ester and heptanoic acid. In conclusion, in this study, green ZnONPs exerted beneficial effects on Delonix elata calli and improved their production of bioactive compounds, especially at a dose of 30 mg/L.

Comparative Analysis of Water Extracts from Roselle (Hibiscus sabdariffa L.) Plants and Callus Cells: Constituents, Effects on Human Skin Cells, and Transcriptome Profiles

Roselle (Hibiscus sabdariffa L.) is a plant that has traditionally been used in various food and beverage products. Here, we investigated the potential of water extracts derived from Roselle leaves and callus cells for cosmetic and pharmaceutical purposes. We generated calluses from Roselle leaves and produced two different water extracts through heat extraction, which we named Hibiscus sabdariffa plant extract (HSPE) and Hibiscus sabdariffa callus extract (HSCE). HPLC analysis showed that the two extracts have different components, with nucleic acids and metabolites such as phenylalanine and tryptophan being the most common components in both extracts. In vitro assays demonstrated that HSCE has strong anti-melanogenic effects and functions for skin barrier and antioxidant activity. Transcriptome profiling of human skin cells treated with HSPE and HSCE showed significant differences, with HSPE having more effects on human skin cells. Up-regulated genes by HSPE function in angiogenesis, the oxidation-reduction process, and glycolysis, while up-regulated genes by HSCE encode ribosome proteins and IFI6, functioning in the healing of radiation-injured skin cells. Therefore, we suggest that the two extracts from Roselle should be applied differently for cosmetics and pharmaceutical purposes. Our findings demonstrate the potential of Roselle extracts as a natural source for skincare products.

Phytochemical Composition and Detection of Novel Bioactives in Anther Callus of Catharanthus roseus L.

Catharanthus roseus L. (G.) Don is the most widely studied plant because of its high pharmacological value. In vitro culture uses various plant parts such as leaves, nodes, internodes and roots for inducing callus and subsequent plant regeneration in C. roseus. However, till now, little work has been conducted on anther tissue using plant tissue culture techniques. Therefore, the aim of this work is to establish a protocol for in vitro induction of callus by utilizing anthers as explants in MS (Murashige and Skoog) medium fortified with different concentrations and combinations of PGRs. The best callusing medium contains high α-naphthalene acetic acid (NAA) and low kinetin (Kn) concentrations showing a callusing frequency of 86.6%. SEM-EDX analysis was carried out to compare the elemental distribution on the surfaces of anther and anther-derived calli, and the two were noted to be nearly identical in their elemental composition. Gas chromatography-mass spectrometry (GC-MS) analysis of methanol extracts of anther and anther-derived calli was conducted, which revealed the presence of a wide range of phytocompounds. Some of them are ajmalicine, vindolinine, coronaridine, squalene, pleiocarpamine, stigmasterol, etc. More importantly, about 17 compounds are exclusively present in anther-derived callus (not in anther) of Catharanthus. The ploidy status of anther-derived callus was examined via flow cytometry (FCM), and it was estimated to be 0.76 pg, showing the haploid nature of callus. The present work therefore represents an efficient way to produce high-value medicinal compounds from anther callus in a lesser period of time on a larger scale.

In vitro effects of undifferentiated callus extracts from Plantago major L, Rhodiola rosea L and Silybum marianum L in normal and malignant human skin cells

Background and objectivesThe occurrence of non-melanoma and melanoma skin cancers is currently increasing rapidly with one in every three cancers diagnosed as a skin cancer. A useful strategy to control the progression of skin cancer could be the use of plant flavonoids that suppress pro-inflammatory cytokines involved in tumor initiation and progression. In this study, the anti-inflammatory and antioxidant activity of undifferentiated callus extracts from Plantago major L, Silybum marianum L and Rhodiola rosea L was investigated both in normal and malignant skin cells. MethodsAntioxidant activity of the extracts was analyzed by using the Trolox Equivalent Antioxidant Capacity (TEAC) assay. High-Performance Thin-Layer Chromatography (HPTLC) was performed to demonstrate the phytochemical profile, and the total flavonoid content was analyzed with an aluminum chloride colorimetric method. The anti-inflammatory effect was investigated by cell treatments using the plant extracts. Thereafter, the possible suppression of induced IL-6 response was measured from the cultured skin cancer cell lines A2058 and A431, and normal primary keratinocytes with Enzyme-Linked Immunosorbent Assay (ELISA). ResultsThe HPTLC analysis assessed that the extracts contained a complex phytochemical profile that was rich in phenolic and flavonoid compounds. Dose response assays showed that concentrations between 15 and 125 μg/mL of all three plant extracts could be used to investigate an effect on the IL-6 production. The S. marianum extract had the most pronounced anti-inflammatory effect, which significantly inhibited induced IL-6 production in both normal keratinocytes and skin cells derived from epidermal carcinoma. The extract from S. marianum also had the highest flavonoid content and showed the highest antioxidant activity of the three extracts tested. ConclusionAll in all, we have confirmed that undifferentiated callus extracts of S. marianum possess properties such as antioxidant and anti-inflammatory activities in both normal and malignant keratinocytes, and thus could be a promising agent controlling the pro-inflammatory IL-6 production.

Cydonia oblonga Mill. Pulp Callus Inhibits Oxidative Stress and Inflammation in Injured Cells.

The pharmacological activity of a callus extract from the pulp of Cydonia oblonga Mill., also known as quince, was investigated in murine macrophage (RAW 264.7) and human keratinocyte (HaCaT) cell lines. In particular, the anti-inflammatory activity of C. oblonga Mill. pulp callus extract was assessed in lipopolysaccharides (LPS)-treated RAW 264.7 by the Griess test and in LPS-treated HaCaT human keratinocytes by examining the expression of genes involved in the inflammatory process, including nitric oxide synthase (iNOS), interleukin-6 (IL-6), interleukin-1β (IL-1β), nuclear factor-kappa-B inhibitor alfa (ikBα), and intercellular adhesion molecule (ICAM). The antioxidant activity was evaluated by quantizing the reactive oxygen species (ROS) production in the hydrogen peroxide and tert-butyl hydroperoxide-injured HaCaT cell line. The obtained results indicate that C. oblonga callus from fruit pulp extract has anti-inflammatory and antioxidant activities, suggesting its possible application in delaying and preventing acute or chronic diseases associated with aging or in the treatment of wound dressing.

Anti-Inflammatory and Anti-Quorum Sensing Effect of Camellia sinensis Callus Lysate for Treatment of Acne.

Cutibacterium acnes (C. acnes) is involved in the pathogenesis of acne by inducing inflammation and biofilm formation, along with other virulence factors. A Camellia sinensis (C. sinensis) callus lysate is proposed to reduce these effects. The aim of the present work is to study the anti-inflammatory properties of a callus extract from C. sinensis on C. acnes-stimulated human keratinocytes and the quorum-quenching activities. Keratinocytes were stimulated with thermo-inactivated pathogenic C. acnes and were treated with the herbal lysate (0.25% w/w) to evaluate its anti-inflammatory effect. C. acnes biofilm was developed in vitro and treated with 2.5 and 5% w/w of the lysate to evaluate quorum sensing and the lipase activity. The results showed that the lysate was able to reduce the production of interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), and C-X-C motif chemokine ligand 1 (CXCL1), and decrease the nuclear translocation of nuclear factor kappa light chain enhancer of activated B cells (NF-κB). The lysate did not show bactericidal activity but showed diminished biofilm formation, the lipase activity, and the production of autoinducer 2 (AI-2), a member of a family of signaling molecules used in quorum sensing. Therefore, the proposed callus lysate could have the potential to reduce acne-related symptoms without the eradication of C. acnes, which is part of the natural skin microbiome.

Elicitor-induced phytochemicals production in Berberis lycium Royle

Medicinally important alkaloids (i.e., berberine, palmatine) and polyphenols compounds in-vitro callus cultures of Berberis lycium Royle were quantified. The maximum callus biomass and phytochemical concentrations were observed after five weeks of culture in the medium containing methyl jasmonate (MeJA) and salicylic acid (SA). The cell cultures established with lower elicitation (i.e., 50 µM SA and 100 µM MeJA) concentration showed significantly (p < 0.05) higher biomass accumulation and highest alkaloids and polyphenol production compared to the control. Furthermore, upon 100 μM MeJA treatment, the elicited callus extract showed significantly higher (56.67%) antimutagenic activity than SA treatment (41.71%), and the result was comparable with that of reference compounds (gallic acid; 59.43%). Besides, the antimicrobial potential of the callus extract was investigated in different solvent mediums (i.e., methanol, acetone, ethyl acetate) and against four microorganisms, i.e., Bacillus subtilis, B. megaterium, Escherichia coli, Serratia marcescens. Of the different solvent mediums, the maximum zone of inhibition (11.5 ± 0.51 mm) was observed for E. coli in methanolic extract. High-performance liquid chromatography equipped with a photodiode detector (HPLC-PDA) analyses revealed the presence of polyphenols compounds such as gallic acid, catechin, m-coumaric acid, vanillin, phloridzin, p-coumaric acid, salicylic acid, naringin, o-coumaric acid and alkaloids (berberine, palmatine) in the callus culture. This is the first report of in-vitro elicitor-mediated production of different phytochemicals, polyphenolics, and alkaloids in B. lycium callus cultures. The optimized protocol developed can be used efficiently in large-scale production of pharmaceutically important metabolites, thus opening the door for industrial exploration and the conservation of this important medicinal plants.

Secondary Metabolites Profiling, Antimicrobial and Cytotoxic Properties of Commiphora gileadensis L. Leaves, Seeds, Callus, and Cell Suspension Extracts.

Commiphora gileadensis L. is an important endangered medicinal plant that belongs to the family Burseraceae. In this study, C. gileadensis callus culture was established successfully using mature leaves as explants cultured on Murashige and Skoog (MS) media supplemented with 24.50 μM of indole butyric acid (IBA) and 2.22 μM 6-Benzylaminopurine (BAP) (callus induction media). The obtained callus was maintained on MS medium supplemented with 16.11 μM naphthalene acetic acid (NAA) in combination with 6.66 μM BAP, which resulted in a substantial increase in callus fresh and dry weights. The cell suspension culture was established successfully using liquid callus induction media supplemented with 3.0 mg·L-1 proline. Thereafter, the chemical constituents of different C. gileadensis methanolic extracts (callus, cell suspension, leaves, and seeds) were profiled, and their cytotoxic and antimicrobial properties were investigated. The LC-MS GNPS analyses were applied for chemical profiling of the methanolic plant extracts, and several natural products were identified, including flavonols, flavanones, and flavonoids glycosides, with two unusual families that included puromycin, 10-hydroxycamptothecin, and justicidin B. The methanolic extracts have shown selective antimicrobial and cytotoxic properties against different microbes and cancer cell lines. For instance, leaf extract showed the highest zone of inhibition for Staphylococcus aureus, while cell suspension culture was effective against Staphylococcus epidermidis and Staphylococcus aureus. All extracts showed selective activity against A549 cell lines for the cytotoxicity assay, while the leaf extract had a broad cytotoxic effect against all tested cell lines. This study revealed that C. gileadensis callus and cell suspension cultures can be employed to increase the in vitro formation of biologically active compounds that may have cytotoxicity and antibacterial action against different cancer cell lines and bacterial species. Further studies are required to isolate and identify such constituents that corroborate the observed activities.

Farm or Lab? A Comparative Study of Oregano's Leaf and Callus Volatile Isolates Chemistry and Cytotoxicity.

Oregano (Origanum vulgare, Lamiaceae plant family) is a well-known aromatic herb with great commercial value, thoroughly utilized by food and pharmaceutical industries. The present work regards the comparative assessment of in vitro propagated and commercially available oregano tissue natural products. This study includes their secondary metabolites' biosynthesis, antioxidant properties, and anticancer activities. The optimization of callus induction from derived oregano leaf explants and excessive oxidative browning was performed using various plant growth regulators, light conditions, and antioxidant compounds. The determination of oregano callus volatiles against the respective molecules in maternal herbal material was performed using gas chromatography-mass spectrometry (GC/MS) analysis. In total, the presence of twenty-seven phytochemicals was revealed in both leaf and callus extracts, from which thirteen molecules were biosynthesized in both tissues studied, seven compounds were present only in callus extracts, and seven metabolites only in leaf extracts. Carvacrol and sabinene hydrate were the prevailing volatiles in all tissues exploited, along with alkanes octacosane and triacontane and the trimethylsilyl (TMS) derivative of carvacrol that were detected in significant amounts only in callus extracts. The MTT assay was employed to assess the in vitro cytotoxic properties of oregano extracts against the epithelial human breast cancer MDA-MB-231 and the human neuroblastoma SK-N-SH cell lines. The extracts displayed concentration and time-dependent responses in cell proliferation rates.

Biologically active compounds in Scutellaria baicalensis L. callus extract: Phytochemical analysis and isolation

Plant cells and tissue cultures are sources of secondary plant metabolites. Substances produced by callus cultures can expand the raw material base in pharmacy and food production. However, isolating biologically active substances from medicinal plants is a labor- and time-consuming process. As a result, new and efficient technological processes adapted for extraction from callus cultures are in high demand, and new algorithms of isolation and purification of biologically active substances remain a relevant task.&#x0D; This research featured callus cultures of Scutellaria baicalensis. The procedures for phytochemical analysis and isolation of biologically active substances involved such physicochemical research methods as high-performance chromatography (HPLC), thin-layer chromatography (TLC), UV spectrometry, and IR spectrometry.&#x0D; The high performance liquid chromatography confirmed the presence of flavonoids represented by baicalein (5,6,7-trioxyflavone), baicalin (baicalein 7-O-glucuronide), scutellarein (5,6,7,4-tetraoxyflavone), scutellarin (7-O-glucuronide scutellarein), vagonin, and oroxylin. The spectral analyses also detected skutebaicalin. The highest total content of diterpene belonged to the samples extracted with 70% ethanol at 70°C. The content of diterpene was 0.09 mg/cm3 in terms of betulin. The biologically active substances were isolated from the callus extracts of S. baicalensis with a recovery rate of ≥ 80%. The purification scheme made it possible to obtain highly-pure individual biologically active compounds: trans-cinnamic acid, baicalin, and oroxylin A had a purity of ≥ 95%; baicalein had a purity of ≥ 97%; scutellarin and luteolin reached ≥ 96%.&#x0D; The new technological extraction method made it possible to obtain extracts from S. baicalensis callus cultures, which were tested for the component composition. The developed isolation algorithm and purification scheme yielded biologically active substances with a purification degree of ≥ 95%.

Is acute low-dose gamma irradiation an effective elicitor for secondary metabolism in Leontopodium alpinum (Cass.) callus culture?

Leontopodium alpinum Cass. (Edelweiss) is a protected species in some European countries and has become of growing interest for exploitation by the food supplements and cosmetics industry, due to its valuable composition in bioactive and biocompatible compounds. Small scale field cultivation and in vitro biotechnological approaches are economically viable alternatives to the production via chemical synthesis. In this study, we aimed to compare two performant L. alpinum callus culture lines and to investigate the effect of acute low dose gamma irradiation on biomass accumulation (Gi), total phenolic content (TPC), total flavonoid content (TFC), antioxidant activity (DPPH) and secondary metabolites’ profile. A purple, pigmented line (PC) was obtained under lightning conditions and in the absence of light, an unpigmented greenish line (GC) was established. Both lines displayed impressive proliferative capacity after 14 days of culture, with an 18.31 ( ± 1.83) fold mass increase for PC and a 13.27 ( ± 2.42) for GC. In respect to Gi, TPC, TFC and DPPH, the purple callus line significantly outperformed the unpigmented line. According to the UPLC-HRMS analysis of the methanolic extracts, both callus lines displayed similar metabolic profiles, having similar quantities of 3,5-dicaffeoylquinic acid, leontopodic acids A and B, but 60% higher concentration of chlorogenic acid in the GC line. In the methanolic callus extracts, 9 economically important phenylpropanoid derivatives were identified. The two callus lines responded differently to low dose gamma irradiation. Although the control values measured for Gi, TPC, TFC and DPPH were significantly higher in the PC line, this line was generally negatively impacted by the gamma treatments. On the contrary, higher, but not significant, values were registered in some GC variants post irradiation with 25, 30 and 40 Gy. UPLC-HRMS investigations emphasized the overproduction of 7 compounds, post irradiation with all doses, for the PC line, while for the unpigmented line only one compound was stimulated by all irradiation treatments. The highest stimulative effect was obtained for the 35 and 40 Gy experimental variants, in the GC line, where dihydroxybenzoic acid glucoside accumulation was increased by 160%. The results of this study showed, for the first time, the different responses of two distinct L. alpinum callus lines using gamma irradiation elicitation method, which can be successfully applied to stimulate the production of certain industrially important compounds as chlorogenic acid, dicaffeoylquinic acid isomers or characteristic glucaric acids.

Extract of Bletilla formosana callus elevates cellular antioxidative activity via Nrf2/HO-1 signaling pathway and inhibits melanogenesis in zebrafish

BackgroundBletilla species are endangered terrestrial orchids used in natural skin care formulas in Asia for a long history. In order to explore the bioactivity potential of Bletilla species as a cosmetic ingredient in a sustainable resource manner, the callus of Bletilla formosana (Hayata) Schltr. was established and extracted by an eco-friendly supercritical fluid CO2 extraction (SFE-CO2) method. The intracellular reactive oxygen species (ROS) scavenging activity and antioxidation-related gene expression of the callus extract were evaluated in both Hs68 fibroblast cells and HaCaT keratinocytes. The melanogenesis-inhibitory effect was investigated in B16F10 melanoma cells and in an in vivo zebrafish model. ResultsThe calli of B. formosana were propagated for 10–15 generations with a consistent yellow friable appearance and then subjected to SFE-CO2 extraction to obtain a yellow pasty extract. Obvious intracellular ROS scavenging activity of the extract was detected in both Hs68 and HaCaT cells with 64.30 ± 8.27% and 32.50 ± 4.05% reduction at the concentration of 250 μg/mL. Moreover, marked expression levels of heme oxygenase-1 (HO-1) and (NAD(P)H) quinone oxidoreductase-1 (NQO1) genes were detected after 6-h and 24-h treatments. These results indicate the cellular antioxidative activity of B. formosana callus extract was probably activated via the nuclear factor erythroid 2-related factor 2 (Nrf2)/HO-1 signaling pathway. Melanogenesis-inhibitory effect of the extract was observed in α-MSH stimuli-inducing B16F10 cells with 28.46% inhibition of intracellular melanin content at the concentration of 50 μg/ml. The effect was confirmed with in vivo zebrafish embryos that showed a relative pigmentation density of 80.27 ± 7.98% at the concentration of 100 μg/mL without toxicity. ConclusionOur results shed light on a sustainable utilization of Bletilla species as a potential ingredient for skin.

Green Synthesis of Silver Nanoparticles Using Nigella sativa Callus and Cellular Suspension Cultures with Different pH Values

The current study included an extensive protocol for the first time globally. This protocol was concerned with the green synthesis of silver nanoparticles (AgNPs) from Nigella sativa callus and cellular suspended cultures. The results confirmed the success of the process of green synthesis of AgNPs from Nigella sativa callus by the color change in the reaction solution from colorless to brown and reddish-brown as a result of the reduction of Ag ions in the extract after addition of AgNO3 solution (1:1 v/v). UV-Vis spectroscopy was used to initially prove the existence of AgNPs, which showed an absorption peak ranging between (420-440 nm) caused by the phenomenon of Surface Plasmon Resonance (SPR) of AgNPs. (SEM) showed nanoparticle objects ranging from (18.341 - 40.932) nm for standard extracts and (17.261 - 29.13) nm for suspended cultures extracts. (EDX) proved the success of producing AgNPs in different proportions depending on the type of tissue cultures used and the difference in the pH value of the extracts (6, 9, 12), as the highest % of AgNPs production reached 4.46% in suspended cultures extracts from pH 12, followed by callus extracts, which reached 4.12% at pH 12. biologically prepared AgNPs proved their effectiveness in inhibiting the growth of Gram+ and Gram- bacteria, as the inhibition zone ranged (19– 35 mm) depending on the type of extract and the degree of acidity (pH) applied to the extract.

Production of Boeravinone-B, total phenolic, flavonoid content and antioxidant activity from callus cultures of Punarnava (Boerhavia diffusa L.)

Boerhavia diffusa L. (Punarnava) is a medicinal herb, rich in diversified plant secondary metabolites used in curing various health ailments. Boeravinone-B is one of the important phytochemicals reported in Punarnava, possessing various pharmacological activities. It belongs to the family of rotenoids, belonging to the isoflavone group. Production of Boeravinone-B from the Punarnava through conventional propagation is comparatively very low, and alternative interventions are of utmost importance to meet the growing demand. In view of this, the present study aims to develop biotechnological approaches like cell/tissue culture as a substitute strategy for the accumulation of biomass and Boeravinone-B biosynthesis. Callus was established from leaf explants of Boerhavia diffusa L. when cultured on MS semi solid medium fortified with varied concentrations and combinations of auxins and cytokinins. The callus induced on Murashige and Skoog medium (MS medium) supplemented with 5.0 ppm 2,4-Dichlorophenoxyacetic acid (2,4-D) favored the highest production of Boeravinone-B analyzed through High-performance Liquid chromatography (HPLC) and it was found to be 673.95 ?g g-1 Dry weight (DW). The total phenolic and flavonoid content were determined for the callus extracts and the results showed that callus induced on 5.0 ppm 2,4-D medium showed the highest phenolic and flavonoid content, which was 63.48 mg g-1 Gallic acid equivalent (GAE) Dry weight (DW), and 30.22 mg g-1 Quercetin equivalent (QE) DW. Similarly, antioxidant activities (radical scavenging, metal chelating, and reducing power) were performed, and it was found that callus induced on 5.0 ppm 2,4-D showed the highest anti-oxidant potential. Radical scavenging activity was found to be 91.1%, and 74% of metal chelating activity was recorded, and a similar trend was observed with respect to reducing power as well. The results of the present study lay foundation for optimization and subsequent large-scale production of Boeravinone-B from callus/cell suspension cultures.

Safety Assessment of Apple-Derived Ingredients as Used in Cosmetics.

The Expert Panel for Cosmetic Ingredient Safety (Panel) reviewed the safety of 26 apple-derived ingredients, which reportedly function mostly as skin conditioning agents in cosmetic products. Because apple-derived ingredients may be obtained from different apple cultivars, the composition of ingredients derived from different cultivars should be similar to that of ingredients reviewed in this safety assessment. Additionally, industry should continue to use good manufacturing practices to limit impurities that could be present in botanical ingredients. The Panel reviewed the available data to determine the safety of these ingredients and concluded that 21 of these ingredients are safe in cosmetics in the present practices of use and concentrations described in this safety assessment. However, the Panel also determined that the available data are insufficient to determine the safety of Pyrus Malus (Apple) Root Extract, Pyrus Malus (or Malus Domestica) (Apple) Stem Extract, Malus Domestica (Apple) Callus Extract, and Malus Domestica (Apple) Oil.

Chemical Composition and Biological Activities of Lavandula coronopifolia Poir Extracts: A Comparison between Callus Culture and Native Plant

Nowadays, according to the development and high functions of in vitro cultivation, one of the questions that researchers are looking for is whether the characteristics and compositions of the obtained product meet the needs of the society or not. Therefore, this research was done aimed to compare the bioactive characteristics of callus and native plant aqueous extracts of Lavandula coronopifolia Poir. Thus, two different culture media were utilized for calluses’ growth. Extracts from these calluses and native plants were obtained using the sequential ultrasound-microwave assisted extraction (SUMAE) method. The extraction conditions were a constant temperature of 70°C ultrasound in 20 min at 250 W and 8 minutes at 800 W for microwave at 85°C. At first, results revealed that callus cultures were not able to produce any significant amounts of essential oil. Therefore, aqueous extracts obtained from calluses as well as the natural plant were examined in terms of antioxidant activity, total phenolic content, total flavonoid content, and antimicrobial activity. Moreover, their volatile components were detected with GC-MS. Both DPPH and FRAP techniques demonstrated that the callus on MS + Auxin had stronger antioxidant activity than the other callus and native plant. It also had the greatest total phenolic content. In terms of total flavonoid content, the native plant has the highest value. Generally, it can be concluded that the extract of callus on MS + Auxin has shown significant activity against the tested microorganisms than the other samples.

Micropropagation and GC–MS analysis of bioactive compounds in bulbs and callus of white squill

Medicinal herbs are the main source of bioactive compounds used in the medical industry. White squill (Urginea maritima) is an important medicinal and ornamental plant cultivated in the Mediterranean region. This study reports an efficient protocol for in vitro propagation of Urginea and investigates important bioactive compounds present in the bulbs and in vitro–produced callus. The least number of days for callus induction and shoot regeneration was achieved with Murashige and Skoog (MS) basal media supplemented with 1.0 mg L−1 1-naphthaleneacetic acid (NAA) plus 0.1 mg L−1 6-benzylaminopurine (BAP) and 1.0 mg L−1 NAA plus 0.4 mg L−1 BAP, respectively, while the highest number of shoots and fresh weight were obtained at medium supplemented with 1.0 mg L−1 NAA plus 0.5 mg L−1 BAP. Gas chromatography-mass spectrometry (GC–MS) analysis of Urginea bulb methanol extract showed the existence of important secondary metabolites, such as palmitic acid (C16H32O2), 9-hexadecenoic acid (C16H30O2), phthalic acid 2-ethylhexyl propyl ester (C19H28O4), tetradecanoic acid (C14H28O2), undecanoic acid (C11H22O2), and oleic acid (C18H34O2), in addition to other important compounds, such as 13-heptadecyn-1-ol, 9,12-octadecadienoic acid, 1-monolinoleoylglycerol trimethylsilyl ether, 2-methyl-1-hexadecanol, and octadecanoic acid. Callus methanol extracts showed a reduction in the percentages of most phyto-components compared to bulb extract except for oleic acid, 3-(octadecyloxy) propyl ester and 3-hydroxydodecanoic acid; on the other hand, some important compounds were detected only in callus extract possessing anti-cancer, antiviral, and anti-inflammatory effects, such as farnesol (C15H26O), 7-methyl-Z-tetradecen-1-ol acetate (C17H32O2), ethyl iso-allocholate (C26H44O5), 4-trifluoroacetoxypentadecane (C17H31F3O2), and 2-hydroxyhexadecanoic acid (C16H32O3).

Preliminary phytochemical analysis and antibacterial activity of leaf and leaf derived callus extracts of Passiflora caerulea: An important medicinal plant

In the current study the Preliminary phytochemical analysis and antibacterial activity of leaf and leaf derived callus extracts of Passiflora caerulea were carried out in different solvent extracts.MS medium showed a better response than B5 medium and White’s medium for callus induction. The plant and callus extracts showed the presence of carbohydrates, tannins and glycosides and leaf extracts showed the presence of carbohydrates, whereas stem extracts showed the presence of carbohydrates and phlobatannins. . The methanol, chloroform, leaf extract showed an inhibition zone of 8.00 mm against B. cereus and E. coli. Methanol and hexane Callus extract showed an inhibition zone of 7.00mm against E. coli and P. aeruginosa. The presence of antibacterial property of the solvent extracts against selected microorganisms might be due to the presence of some compounds. Further phytochemical studies are required to explore compounds responsible for the activity.

Preliminary phytochemical analysis and antibacterial activity of leaf and leaf derived callus extracts of Passiflora caerulea: An important medicinal plant

Preliminary phytochemical analysis and antibacterial activity of leaf and leaf derived callus extracts of Passiflora caerulea: An important medicinal plant

Cytotoxicity, Phytochemical Screening and Genetic analysis of Ginger (Zingiber officinale Rosc.) Callus and Rhizome

• Ginger rhizome extracts displayed strong cytotoxic effect against HT29, HT116 and MCF-7 cancer cells lines with highest activity against HT29. • Ginger callus extracts recorded non-cytotoxic effect against all examined cancer cells with IC50 >100 µg/mL. • Ginger rhizome extracts contained more phenols and terpenes than the callus ones. • The principal components of ginger; 6-gingerol and 6-shogaol were not detected in callus extracts. • Incidence of genetic changes in DNA of ginger callus during in vitro culture in 2,4-D containing medium. • Most polymorphic bands (11 out of 12) were generated in callus during in vitro culture with total polymorphism 23.07% between callus and plant. Ginger ( Zingiber officinale Rosc.) is well known as distinctive spice with considered therapeutic values. In the current work we aimed to investigate cytotoxic effect, phytochemical screening and genetic analysis of ginger callus and their rhizome. Cytotoxicity of ginger rhizome and callus extracts was examined against some human cancer cell lines HT29, HT116 and MCF-7 by MTT method. Phytochemicals were screened by thin layer chromatography (TLC) and genetic analysis was performed by ISSR markers. Ginger rhizome principle components, 6-gingerol and 6-shogoal were used as standards to detect their presence in callus. Results showed that, rhizome extracts displayed a strong cytotoxicity towards all tested cancer cells with the highest activity (IC 50 20.4 ± 3.0 µg/mL; P< 0.05) against HT29. IC 50 of ginger callus extracts were >100 µg/mL in all examined cancer cell lines. TLC chromatograms of rhizome and callus extracts revealed the presence of phenols and terpenes, whereas 6-gingerol and 6-shogaol were only detected in rhizome extracts. Genetic analysis by ISSR markers revealed occurrence some genetic changes in DNA of callus compared to that of rhizome with total polymorphism 23.07%. In conclusion, ginger cytotoxicity to HT29, HT116 and MCF-7 cancer cells was mainly associated with the presence of 6-shogaol and 6-gingerol in extracts, which appeared as strong cytotoxic effect in rhizome compared to non-cytotoxic effect was evaluated in the callus extracts. Therefore, further studies on the callus elicitation and precursors feeding to induce biosynthesis of ginger bioactive molecules are recommended.