Callus Extracts Research Articles

Bio-callus synthesis of silver nanoparticles, characterization, and antibacterial activities via Cinnamomum camphora callus culture

Plant tissue culture is considered as a basic and fundamental component of plant biotechnology and is considered a good approach for green synthesis of nanoparticles (NPs) because it is safe, eco-friendly and clean method. In the current study, callus extract of Cinnamonum camphora was utilized to the synthesis of silver nanoparticles (Ag-NPs) for the first time. Callus culture of C. camphora was propagated in murashige and skoog (MS) medium, and sub-cultured for 3 successive times with 4 week intervals. The propagated callus cultures are incubate at different five light conditions using 1500 Lux., white, green, yellow, red and full dark condition. After that, calli which showed the best callus dry biomass was utilized to synthesis Ag-NPs through formation of callus extract. Green synthesized Ag-NPs was characterized by UV–Vis spectroscopy, TEM, SEM-EDX, DLS, FT-IR, and XRD. Results affirmed the ability of callus extract of C. camphora to fabricate spherical Ag-NPs with average size 5.47–9.48 nm at wavelength 420 nm. Also, the crystallographic structure of nanoparticle are confirmed by XRD analysis. Silver ion was represented by 57.3% of the total weight elements in NPs solutions as showed by EDX analysis. In addition, Ag-NPs exhibited broad spectrum activities against Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Pseudomonas aeruginosa with inhibition zones 19.6 ± 0.8, 19 ± 0.5, 17.1 ± 0.7 and 15.1 ± 0.4 mm respectively.

Qualitative and Quantitative Evaluation of Active Constituents in Callus of Lavandula angustifolia plant in Vitro

This study was conducted to describe a protocol for the callus establishing culture of Lavandula angustifolia plant and estimating their content of volatile oil. The quantity of volatile oil callus tissues was compared with that of leaves production. Callus was induced from leaf explants on Murashige and Skoog medium (MS) supplemented with Naphthalene acetic acid (NAA) and Benzyl adenine (BA) in different concentrations. Maximum callus fresh weight was obtained in the combination of 10 mg/L BA and 3 mg/L NAA which reached 18 g after four weeks. The results of this work showed that the quantity of volatile oil from the highest fresh weight callus was 6 ml compared with quantity of 18g of leaves which gave 0.5 ml. Volatile oil of leaf and callus extracts were analyzed using gas chromatography mass spectrometry method (GC-MS) which showed linoleic acid (56.61%) and oleic acid (57.93%) as main components.

Phytochemicals Screening and Antifungal Activity of Balanites aegyptiaca Seed and Callus Extract against Candida albicans

Balanites aegyptiaca is a medicinal plant that serves as a source of phytochemicals with antimicrobial effect. This work aimed at screening for phytochemical constituents and investigating the antifungal activity of B. aegyptiaca seed and callus extract against Candida albicans. Callus induction from B. aegyptiaca seed kernel explant was done on MS basal nutrient medium supplemented with 0.5 BAP + 1.0 2, 4-D + 1.0 NAA. Cultures were kept under a controlled temperature and light conditions for five weeks. Plant materials were extracted using solvent extraction. 50 ml of the solvents: methanol and n-hexane were mixed with five grams (5 g) each of the grounded plant materials (1:10) w/v. Determination of antimicrobial activity was done using disc diffusion assay. Diffusion discs of approximately 6 mm diameter were prepared from Whatman No. 1 filter paper, then sterilized and autoclaved before drying in an oven. 10 µl of 50 and 100 mg/ml concentration of each crude extracts was impregnated on separate sterile disc using sterile micropipette tips. The diameter of zone of inhibition at 100 and 50 mg/ml showed the methanolic extract of callus had the highest zone of inhibition with 17 ± 0.69 mm and 11 ± 0.94 mm. The lowest diameter of zone inhibition of callus extracts was recorded by n-hexane extract at 100 (15 ± 0.46 mm) and 50 mg/ml (09 ± 0.57 mm) respectively. Also, the MIC ranged between 6.25 and 12.50 mg/ml and MFC recorded value of 12.50 mg/ml. Seed methanolic extract had the highest zones of inhibition of 15 ± 0.34 mm and 10 ± 0.62 mm at 100 and 50 mg/ml respectively, while the lowest value (12 ± 0.51 mm and 09 ± 0.23) was recorded in n-hexane at 100 and 50 mg/ml. From the results, both the MIC and MFC of seed extracts ranged from 12.50 to 25.00 mg/ml. Callus extracts showed stronger antifungal activities compared to the seed extracts. Therefore, from the result, Callus extract from B. aegyptiaca can serve as a good source of therapeutic compounds for fungal related disease.

Ultraviolet-B-induced changes on phenolic compounds, antioxidant capacity and HPLC profile of in vitro-grown plant materials in Echium orientale L

The application of UV-B radiation is effective stress factor for the biosynthesis and accumulation of valuable plant secondary metabolites. The effect of UV-B irradiation on phenolic compounds, total flavonoid and phenolic contents and antioxidant activity of in vitro grown Echium orientale L. shoots and calli were investigated in current study. For this purpose, phenolic compounds (rosmarinic acid, chlorogenic acid, ferulic acid and quercetin) were determined by High Performance Liquid Chromatography (HPLC) analysis while antioxidant activity, total phenolic and flavonoid contents were evaluated by UV–vis spectrophotometric measurements. The results showed that callus extracts contained higher amount of total phenolics and flavonoids and the phenolic compounds than shoot extracts of E. orientale. Besides, the results revealed that the phenolic compounds, especially rosmarinic acid and quercetin, increased in shoot and callus extracts of E. orientale which were harvested one week later after UV-B treatment when compared with the control. The results also showed that UV-B irradiation increased the antioxidant activity and total flavonoid content in both plant samples which were harvested one week later after UV-B treatment. However, the total phenolic content increased in callus extracts which were harvested immediately after UV-B irradiation.

Induction and Growth Kinetics Callus of Tomato (Solanum lycopersicum)

Plant callus extracts are potential to be developed as ingredient in skincare products. Tomato callus is supposed to contain protein-derivatives and or other components such as secondary metabolites that play a role in skin regeneration. Therefore the production of calli is important to be studied for callus sustainable supply. This research aims to obtain optimum medium for callus induction and to analyze tomato callus development anatomically. In vitro culture response was assessed in tomato plant (Solanum lycopersicum L. ‘Permata’) for optimum callus induction. Seeds were grown on ¼ MS medium for 10-15 days. Hypocotyl was excised and cultured on MS medium + 2 mg/l 2,4-D for 15 days as the explants for callus induction. Callus was transferred to MS medium with 8 variations of PGRs including the combination of BAP + NAA, and 2,4-D. Both fresh and dry weight was measured every 5 days over 60 days to establish the growth kinetics and growth efficiency of callus. Anatomic characters of calli were examined through paraffin-embedded method. The result showed of MS medium supplemented with 2.0 mg/l NAA and 0.2 mg/l BAP is optimum for tomato callus induction, based on highest number of the absolute growth rate on fresh weight (73.77% per day), dry weight (3.84% per day), and callus initiation time (5.56 days) achieved by the medium. Cells in the ground tissue of tomato hypocotyl are competent to be dedifferentiated into a callus. This research results were expected to find out suitable methods for tomato callus production in preparation for skincare uses.

A Comparative Study of in vivo Plant and in vitro Callus Extracts of Centratherum punctatum Cass

The present study aims at comparative study between plant and callus extract, with respect to analysis of phytochemical constituents, antioxidant, antibacterial activity and cytotoxicity properties of Centratherum punctatum using aqueous medium and different solvents such as, methanol and ethyl acetate. In vitro studies in MS media supplemented with BAP 4.5 mg/L + Kn 4.0 mg/L has shown the high callus induction percentage of 92.33% with a maximum callus weight of 1.08 g. The phytochemical analysis of aqueous, methanol and ethyl acetate extract of C. punctatum in vivo plant and in vitro callus showed the presence of alkaloids, flavonoids, phenols and carbohydrates. The aqueous extract of both plant and callus showed the presence of tannins, proteins and steroids whereas the methanol extract showed the presence of tannins, amino acids and terpenoids. The ethyl acetate extract showed terpenoids and protein. FTIR analysis of plant and callus aqueous extract had a maximum characteristic band at 3399.87 cm-1 and 3412.73 cm-1 respectively indicating the presence of N-H stretching. GC-MS analysis revealed the presence of 11 different compounds in ethyl acetate extracts of plant and the callus extract revealed the presence of 15 different compounds which was absent in the plant extract. Plant extract exhibited maximum total phenol content than callus extract. The in vitro callus extract showed higher DPPH radical scavenging activity with lower inhibition percentage than in vivo plant extract. A maximum zone of inhibition was observed in methanol extract of in vivo plant and in vitro callus (15 mm and 14 mm respectively) against Bacillus subtilis. The ethyl acetate extract of in vivo plant and in vitro callus had a zone of 14 mm and 12 mm against E. coli. A maximum zone of inhibition (12 mm and 11 mm respectively) was observed in both methanol and ethyl acetate of in vivo plant and in vitro callus against Staphylococcus aureus. Antiproliferative analysis revealed that in vivo plant has inhibitory percentage of 23.6 whereas callus exhibited 28.5% against HeLa cells.

A Comparison of Chemical Profiles of Callus, Plant and Agarwood Extracts of Gyrinops walla using Thin Layer Chromatography

Agarwood produced by Gyrinops walla Gaertner, an endemic plant species of Sri Lanka, arguably the costliest wood, is used as a major constituent in cosmetic industries. Application of plant cell culture technique serves as the best alternative for the production of fragrance compounds of G. walla while ensuring the sustainability of the species. Using the advantages of plant cell culture and Thin Layer Chromatography (TLC), a preliminary study was carried out to compare the phytochemical constituents of callus and plant extracts (i.e stem, leaves, bark and agarwood) of G. walla. Different callus lines were developed on MS and modified MS media with reduced level of NH4NO3, supplemented with different combinations and concentrations of plant growth regulators and multiplied. Freeze dried samples were extracted with hexane, ethyl acetate and methanol and run on a TLC plate under different eluting solvent systems. A higher number of TLC spots were observed in the ethyl acetate extracts of all samples, under 15 percent methanol: chloroform solvent system. Based on TLC chemical profiles of all callus lines, MS media added with 1.0 mg/L BAP, 0.5 mg/L kinetin and 0.1 mg/L 2,4-D and ½ MS contained 1.0 mg/L BAP and 0.1 mg/L 2,4-D were selected as best lines. The extracts of the selected two callus lines matched with the fingerprint profiles of best plant samples i.e stem, bark and agarwood. Compounds present in the extracts of callus samples, plant samples and agarwood, appearing at Rf=0.56, 0.74 and 0.78, could be used as possible markers for G. walla. The chromatography profiling by TLC has proven the presence of similar phytochemical constituents in callus, plant samples and agarwood. Therefore, it can be concluded that plant cell culture technique guarantees the possibility to produce same compounds that are present in plant and agarwood of G. walla.

ENHANCEMENTS OF GROWTH AND METABOLITES OF INDICA RICE CALLUS (ORYZA SATIVA L. CV. PATHUMTHANI1) USING TiO2 NANOPARTICLES (NANO-TiO2)

In this research, the effects of a various in concentration of TiO2 nanoparticles (NanoTiO2) were applied to study the cell growth, metabolites and antioxidant responses of callus in indica rice plants. Dose-dependent changes in growth performances, metabolite accumulations and antioxidant activities of callus were found. Mature seeds of indica rice cv. Pathumthani1 were cultured in callus induction medium with different concentrations of Nano-TiO2 (0-600 mg L-1 ). After five weeks of cultivation, the callus formations were measured in fresh weights and dry weights, and the contents of phenolic compounds, flavonoids, and the antioxidant activities of callus extracts were assayed. The Nano-TiO2 induced of fresh weights and dry weights in callus formations when compared with the control treatment (0 mg L-1 Nano-TiO2) were also observed. The flavonoid and phenolic compound contents in rice callus exposed to 400 mg L-1 Nano-TiO2 were 118% and 139%, respectively, higher than rice callus unexposed to Nano-TiO2. Moreover, the antioxidant activities of the callus metabolites studied shows that the percentages of inhibition were up-regulated (84%) when exposed to 400 mg L-1 Nano-TiO2, as a response to the highlevel depletion of the free radicals in metabolites of rice callus. The results obtained from this research showed that the addition of Nano-TiO2 had impacts on the callus growth and metabolite accumulations in rice callus. The results suggested that addition of Nano-TiO2 at appropriate levels could role as an elicitor for biosynthesis of valuable metabolites and antioxidant properties for pharmaceutical applications in further research.

Anticancer Potential of Calli Versus Seedling Extracts Derived from Rosmarinus officinalis and Coleus hybridus.

In Saudi Arabia, the incidence and mortality rates of breast cancer are high. Although current treatments are effective, breast cancer cells develop resistance to these treatments. Numerous studies have demonstrated that active compounds in plant extracts, such as the phenolic compound Rosmarinic Acid (RA), exert anti-cancer effects. We investigated the anticancer properties of methanolic crude extracts of seedlings and calli of Rosmarinus officinalis and Coleus hybridus, two Lamiaceae species. MCF-7 human breast cancer cells were treated with methanolic crude extracts obtained from plant calli and seedlings generated in vitro, and cell proliferation was evaluated. Transcriptional profiling of the seedling and callus tissues was also conducted. The mRNA expression levels of RA genes were higher in C. hybridus seedlings than in R. officinalis seedlings, as well as in C. hybridus calli than in R. officinalis calli, except for TAT and C4H. In addition, seedling and callus extracts of both R. officinalis and C. hybridus showed anti-proliferative effects against MCF-7 cells after 24 or 48 h of treatment. At a low concentration of 10 μg/mL, C. hybridus calli and seedling extracts showed the most significant anti-proliferative effects after 24 and 48 h of exposure (p < 0.01); controls (doxorubicin) also showed significant inhibition, but lesser than that observed with C. hybridus (p < 0.05). Results with R. officinalis callus and seedling extracts did not significantly differ from those with untreated cells. Methanolic extracts of R. officinalis and C. hybridus are potentially valuable options for breast cancer treatment.

Box-Behnken supported development and validation of robust HPTLC method: an application in estimation of punarnavine in leaf, stem, and their callus of Boerhavia diffusa Linn.

Boerhavia diffusa (BD) Linn. (Nyctaginaceae) is one of the most commonly used herbs in the Indian traditional system of medicine for the urinary disorders. The aim of the current investigation was to carry out initiation, development, and maintenance of BD callus cultures and quantitative estimation of punarnavine in plant and callus extracts. Leaves and stem of BD were used as explant for the tissue culture studies using Murashige and Skoog (MS) basal medium. MS Media comprising 2,4-Dichlorophenoxy acetic acid (2,4-D) (1ppm) and 2,4-D (1ppm) + Indole-3-acetic acid (IAA) (1.0ppm) were found to yield friable callus from leaf explant; similarly, 2,4-D (0.3ppm) + IAA (0.75ppm) + Kinetin (0.3ppm) and 2,4-D (0.5ppm) + Naphthalene acetic acid (NAA) (1.5ppm) + Kinetin (0.3ppm) were found to yield friable callus from the stem explant. High-performance thin-layer chromatography method was been developed for the quantitative estimation of punarnavine (R f = 0.73) using mobile phase containing toluene: ethyl acetate: formic acid in the ratio (7.0:2.5:0.7, v/v/v) at 262nm. The validated method was found linear (r 2 = 0.9971) in a wide range (100-1000ng spot-1), precise, accurate, and robust. The values of limit of detection, LOD = 30.3ng spot-1, and limit of quantification, LOQ = 100.0ng spot-1. The robustness of the method was proved by applying the Box-Behnken design (BBD). The developed method found appropriate for the quality control of medicinal plants containing punarnavine as a constituent.

Biosynthesized silver nanoparticles from aqueous extracts of sweet lime fruit and callus tissues possess variable antioxidant and antimicrobial potentials

Different plant extracts have been utilized for biological synthesis of metal nanoparticles. However, no published literature compares the antimicrobial potential of silver nanoparticles (AgNPs) derived from extracts of mature fruit juice (SLF-AgNPs) versus callus (SLC-AgNPs) tissue of same plant. These SLF- and SLC-AgNPs exhibited variable physical and biological properties. Distinct UV-Vis absorption peak (at 250 nm) for SLF-AgNPs indicated formation of small-sized monodisperse particles. Both SLF-AgNPs (5 to 35 nm) and SLC-AgNPs (3 to 270 nm) exhibited quasi-spherical shapes. The FT-IR spectra showed occurrence of shared peaks at 3335, 1647, and 1080 cm−1 while SLC specific peaks at 1065, 1042 and 1017 cm−1. The SLF-AgNPs possessed higher DPPH free radical scavenging potential. These nanoparticles also showed better antibacterial and antimycotic action epitomized as higher inhibition zone diameters compared to SLC-AgNPs. This work can be furthered to identify predominant mechanism for improved antimicrobial potential of the synthesized AgNPs.

Interactive Effect of Melatonin and UV-C on Phenylpropanoid Metabolite Production and Antioxidant Potential in Callus Cultures of Purple Basil (Ocimum basilicum L. var.s purpurascens).

The present study evaluated the interactive effect of melatonin and UV-C on phenylpropanoid metabolites profile and antioxidant potential of Ocimum basilicum L. Callus was treated with varying concentrations of melatonin and UV-C radiations for different time durations, either alone and/or in combination. Individual treatments of both UV-C and melatonin proved to be more effective than combine treatments. Results indicated that UV-C (10 min) exposure increased rosmarinic acid (134.5 mg/g dry weight (DW)), which was 2.3-fold greater than control. Chichoric acid (51.52 mg/g DW) and anthocyanin (cyanide 0.50 mg/g DW) were almost 4.1-fold, while peonidin was found 2.7-fold higher in UV-C (50 min) exposure. In the case of melatonin, 1.0 mg/L concentrations showed maximum rosmarinic acid (79.4 mg/g DW) accumulation; i.e., 1.4-fold more, as compared to the control. However, 2 mg/L melatonin accumulate chichoric acid (39.99 mg/g DW) and anthocyanin (cyanide: 0.45 mg/g DW and peonidin: 0.22 mg/g DW); i.e., 3.2, 3.7 and 2.0-fold increase, as compared to the control, respectively. On the other hand, melatonin-combined treatment (melatonin (Mel) (4 mg/L) + UV-C (20 min)) was proved to be effective in caffeic acid elicitation, which was 1.9-fold greater than the control. Furthermore, antioxidant potential was evaluated by both in vitro (DPPH, ABTS and FRAP assays) and in cellulo methods. Maximum in vitro antioxidant activity (DPPH: 90.6% and ABTS: 1909.5 µM) was observed for UV-C (50 min)-treated cultures. The highest in vitro antioxidant activity measured with the ABTS assay as compared to the FRAP assay, suggesting the main contribution of antioxidants from basil callus extracts acting through a hydrogen atom transfer (HAT) over an electron transfer (ET)-based mechanism. Cellular antioxidant assay was evaluated by production of ROS/RNS species using yeast cell cultures and further confirmed the protective action of the corresponding callus extracts against oxidative stress. Overall, both melatonin and UV-C are here proved to be effective elicitors since a positive correlation between the induced production of phenolic compounds, and in cellulo antioxidant action of basil callus extracts were observed.

Antioxidant activity of in vitro plantlets and callus cultures of Randia echinocarpa, a medicinal plant from northwestern Mexico

Randia echinocarpa, an endemic plant to Northwest Mexico, is used as food and in traditional medicine, and several of its biological activities have been demonstrated (antioxidant, antimutagenic, antidiabetic, and immunomodulatory). Plant tissue culture is a safe and scalable system for plant propagation and production of bioactive compounds. Therefore, this study aims to establish protocols for seed germination and callus culture of R. echinocarpa and to evaluate the antioxidant activity of methanol extracts (ME) of plantlets and calli via the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) methods. Seeds were cultured in media with different concentrations of Murashige and Skoog (MS) salts and sucrose, and a higher germination rate and plantlet growth was observed in half-strength MS medium with 15 g L−1 of sucrose. Calli were obtained from cotyledon and hypocotyl explants cultured in MS media with different concentrations of benzyl aminopurine (BAP) and indole-3-acetic acid (IAA). All treatments induced callus formation in 100% of explants; however, the medium containing 1 mg L−1 BAP + 1 mg L−1 IAA was selected because it produced calli with higher biomass and friable texture. The ME of cotyledons showed the highest antioxidant activity values (μmol Trolox per 100 g dry weight) in DPPH (345.5) and ABTS (1166.4) assays, whereas the ME of calli from hypocotyls showed a higher antioxidant activity than the ME of calli from cotyledons in both antioxidant assays. The tissue culture protocols established here will be useful for R. echinocarpa germplasm conservation and propagation, as well as for the production of bioactive compounds.

Identification of DNA variation in callus derived from Zingiber officinale and anticoagulation activities of ginger rhizome and callus

BackgroundThe medicinal plants have been used as alternative treatments for many diseases in many countries. Thus, the possibility of the alteration of some naturally relatively cheap sources into highly valuable products for pharmaceutical and biological importance via tissue culture is investigated in this study. Special attention is needed to estimate the molecular genetic variation between the studied plant and the callus.ResultsGinger (Zingiber officinale Roscoe) rhizome was affected by various concentrations of cytokinin and auxin for the induction of callus. The highest percentage of callus induction and maximal callus fresh weight was achieved when Murashige and Skoog (MS) medium was supplemented with 2 mg/l 2, 4-D + 1mg/l BA. The genetic variations accompanied with in vitro conditions of callus induction was evaluated by four primers of inter-simple sequence repeat (ISSR) that amplified 36 bands. The highest readings for clotting times were found by using the sulfated neutral extract of ginger rhizome at a concentration of 400 μg/ml, and the extract of sulfated alkaline plant callus had an anticoagulation activity at (200 μg/ml) comparable to that of a standard preparation of heparin sodium.ConclusionThese results showed that ginger (Zingiber officinale Roscoe) rhizome was affected by various concentrations of cytokinin and auxin for induction of callus especially when MS was supplemented with 2 mg/l 2, 4-D + 1 mg/l BA. Also, the results of ISSR markers confirmed the occurrence of genetic variations during callus induction process. The results indicated that the sulfated alkaline of ginger rhizome and sulfated aqueous extracts of ginger rhizome and callus exhibited anticoagulant activity. So, it was clear that the addition of sulfate group into the investigated extracts enhances the anticoagulation activities.

Enhancing bioactive potential by growth regulators in callus of Mentha longifolia L. leaves for anti-inflammatory and analgesic activities

Mentha longifolia L., popularly known as Mint, is a medicinal herb against inflammation and analgesic pains caused by venom of snakes and scorpions. Its leaves are commonly used by nomads of Ahaggar to treat external wounds. Unfortunately, this medicinal plant is less accessible in its natural zone. Hence, here, we propose a biotechnological approach to induce callus yielding the same effects. Callus was induced from young leaves cultivated on modified Murashige and Skoog solid medium supplemented with 30 g L-1 sucrose, 100 mg glutamine and growth regulators under dark conditions. Aqueous and C-glycoside extracts of plant and callus, respectively were prepared and analyzed by HPLC-DAD/LC-ESI-MS. The study of acute toxicity at 2000 mg kg-1 followed by the anti-inflammatory activity assessment using carrageenan-induced paw edema and the analgesic activity of acetic acid induced writhing were performed on mice. Compared to the control, the trials showed 79% higher rate of friable callus induced on 2,4-D and BAP. No mortality or signs of toxicity were observed in mice. Unexpected higher anti-inflammatory and analgesic activities by callus extracts can be attributed to their phenolic composition.

Determination of antioxidant activity, total phenolics and fatty acids in essential oils and other extracts from callus culture, seeds and leaves of Argania spinosa (L.) Skeels

Argan (Argania spinosa (L.) Skeels) is an endangered and endemic agroforestry species of Morocco highly appreciated for its nutraceutical properties. Herein, the antioxidant activity, total phenolic content and fatty acids were evaluated in different extracts obtained from callus culture, seeds and leaves of four argan genotypes: G25, G36, G41 and G84. Callus induction, proliferation and morphology varied depending on genotype and explant type. The highest callus induction rate (97.5%) was observed in the cotyledon explants of genotype G84. The radical scavenging activity values ranged from 76.5 to 98.1%. The essential oils extracted from calli induced from seedling-derived leaves exhibited a slightly higher radical scavenging activity (91.7%) than those extracted from field-grown leaves (90–91.1%). The total phenolic content ranged from 0.72 mg/g dry weight gallic acid equivalent in the methanolic extracts of G84 callus obtained from seedling-derived leaves to 198.26 mg/g dry weight gallic acid equivalent in the essential oils of G84 seeds. The fatty acid composition varied significantly among the different samples. The essential oils extracted from seeds and callus obtained from cotyledon explants have high contents in oleic and linoleic acids (26–37.9% and 25–36.8%, respectively), while the major fatty acid found in the essential oils of leaves and callus obtained from seedling-derived leaves was eicosenoic acid (18.8–45.4%). The present study showed that argan callus culture could be envisaged for sustainable and continuous production of bioactive compounds, and that each extract analyzed had unique and distinct characteristics. The essential oils and other extracts obtained from argan callus induced in vitro were characterized and were compared with those obtained from seeds and field-grown leaves

Bioactive Compounds Isolated From In Vitro Callus and Wild Plant of Hibiscus Sabdariffa Linn. And Its Anticancer Activities

Bioactive compounds were isolated from in vitro callus as well as wild plant of Hibiscus sabdariffa Linn. is an annual shrub belonging to the family Malvaceae. The bioactive compounds were isolated from two different organic extracts are methanolic crude and acidified aqueous acetone. These extracts were analyzed antioxidant potential by free radical scavenging activity of DPPH and Total antioxidant activities. The strong DPPH free radical scavenging activity of the acidified aqueous acetone and methanolic extracts of wild plant and in vitro callus of H. sabdariffa showed maximum of 94.03 and 94.50% of inhibition, respectively. IC50 value of acidified aqueous acetone and methanolic extracts of wild plant and in vitro callus was 35.77 and 30.47 µg/mL concentrations, respectively. Based on the antioxidant potential, the isolated compounds were studied the antiviral, anticancer activities. The antiviral activity of H. sabdariffa show evidence of antiviral activity for H1N1 and FLU B viruses by using Vero cell line. The good antiviral activity was observed both extracts as well as isolated compounds. In vitro cytotoxicity study was carried out for the extract and isolated compounds of extracts of plant and callus by used human cervical carcinoma cells (HeLa) and human colon carcinoma cells (HT 29). Apoptosis cell death percentage was observed in and human carcinoma cancer cells (HT 29) with H. sabdariffa and its isolated compounds by DNA fragmentation method. We could observe distinct ladder pattern in the compounds treated cells. The DNA fragmentation induced by the compounds indicating programmed cell death. The present work strongly justifies that the isolated compounds of Hibiscus sabdariffa is a good candidate for anticancer.

Biomass and terpenoids profile of callus extract of Piper betle L. var. Nigra with abiotic elicitor cobalt (II) chloride

Black betel ( Piper betle L. var. Nigra) is traditional medicinal plant potentially produce secondary metabolites. Its secondary metabolite content can be promoted using callus culture with abiotic elicitor. This study was aimed to determine the effect of cobalt (II) chloride (CoCl 2 ) concentration towards biomass and terpenoids profile of leaf callus of Piper betle L. var. Nigra. Leaf explants of Piper betle L. var. Nigra were cultured in solid Murashige and Skoog (MS) medium supplemented with growth regulators 2,4-D (0.5 mg/L) and BAP (2.0 mg/L) to induce callus formation. Elicitation was performed after callus reached five weeks of age by performing subculture on MS medium added with CoCl 2 (0.5 mg/L; 1.0 mg/L; 2.5 mg/L). Calli of Piper betle L. var. Nigra were harvested after one, two, or three weeks before they were extracted using methanol and analyzed of its terpenoids content using Gas Chromatography-Mass Spectrometry (GC-MS) method. Results showed that calli had compact and crumb texture with green, white, brown, and white color. Callus added 1.0 mg/L CoCl 2 at two weeks age produce terpenoids at 5.95%. Callus with 1.0 mg/L CoCl 2 at three weeks of age produced higher fresh and dry weight at 916.4±162.3 mg and 93.3±11.7 mg respectively. Callus supplemented with 1.0 mg/L CoCl 2 at two weeks of age had higher terpenoids percentage compared to control. Two of the highest terpenoid types found in the callus of black betel leaf explant were 1,2-epoxy-1-vinylcyclododecene and hexadecanoic acid.

CALLUS INDUCTION OF Sonchus arvensis L. AND ITS ANTIPLASMODIAL ACTIVITY.

Background:Malaria is a global health problem that requires urgent need for new drugs. Tempuyung (Sonchus arvensis L.) possesses many potential medicinal compounds. As the plant is originally found wild, it is important to reproduce its secondary metabolites by tissue culture. The objectives of this study were to look for effective methods to induce callus from leaf explants of Sonchus arvensis L. and to test its in vitro antiplasmodial activity.Materials and Methods:The leaves and petioles of the plant were cultured on Murashige and Skoog (MS) solid medium supplemented with indole acetic-3-acid (IAA), indole-3-butyric acid (IBA), naphthalene acetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D) and benzyl amino purine (BAP), in light and dark incubations. The best results obtained from callus induction were then treated by with several concentrations of sucrose (1- 5%). The best results from callus induction were then extracted with methanol for antiplasmodial test by Trager and Jensen’s method. It was also tested against 3D7 strain of Plasmodium falciparum.Results:The combination of 1mg/L 2,4-D and 0.5 mg/L BAP in dark incubation was the best treatment for callus induction of tempuyung. It produced the best quality of callus and the shortest period for callusing. Sucrose treatment had various effects on leaves callusing, but had no effect on petioles callusing, whereby 4% sucrose was the best treatment for leaves callusing in dark incubation. The methanol extract of the best callus had anti-plasmodial activity with IC50=0.343 µg/mL.Conclusion:Methanol extract of tempuyung callus shows potential as an antimalarial drug but more studies would be required.

Cytoprotective activity of carrot and tomato callus extracts and the ex‐ pression of cytokines in UV‐B irradiated fibroblast cells

Studies have suggested that both carrot (Daucus carota L.) and tomato (Solanum lycopersicum L.) callus extracts contain antoxidant compounds that might have the potental to protect cells from free radicals such as H2O2 that contribute to cell damage. The other sources of free radical exposure in human cells, such as UV‐B, should also be examined. UV‐B exposure can trigger increased expression of inflammatory cytokines such as cyclooxygenase‐2 (COX‐2) and tumor necrosis factor‐α (TNF‐α) and the antinflammatory cytokine interleukin‐10 (IL‐10), which causes photoaging. This study was conducted to investgate the cytoprotectve actvity of carrot and tomato callus aqueous extracts by observing cell viability using the MTT assay. Immunocytochemistry methods were used to examine the effects of carrot and tomato callus aqueousextracts on the expression of COX‐2, TNF‐α, and IL‐10 in human dermal fbroblast adult (HDFa) cells exposed to UV‐B light. Carrot and tomato callus aqueous extracts were obtained by the maceraton method using aqua bidistlled solvent. Results showed that both carrot and tomato callus aqueous extracts at 0.5 mg/mL exhibited the highest cytoprotectve effect in HDFa cells compared to that at other concentratons. Both carrot and tomato callus aqueous extracts could also decrease the expression of COX‐2 and TNF‐α, whereas carrot callus aqueous extract increased the expression of the ant‐inflammatory cytokine IL‐10 in HDFa cells.