Optimization of Callus Induction and Cell Suspension Culture of Betula pendula Roth for Improved Production of Betulin, Betulinic Acid, and Antioxidant Activity (original) (raw)
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Data in Brief, 2016
Plant tissue or cell culture keeps a significant role in micropropagation in the plant production industry. Combination of 6-Benzylaminopurine (BAP) and other plant growth regulators like 1-Naphthaleneacetic acid (NAA) or Indole-3-acetic acid (IAA) or indole-3-butyric acid (IBA) was used in the most of the research in tissue culture. The study was carried out to investigate the optimization of the concentration of IBA and BAP combination (0, 0.25, 0.50, 1.0, 1.50, 2.0, 2.5, 3.0 and 3.5 mg/l) for the root, callus and leaf proliferation from the leaf cutting slice. The highest number (6.75) of root proliferation was observed in the concentration of 2.0 mg/l IBAþ 0.25 mg/l BAP combination. The callus initiation was found in the concentration of IBA 1.0-3.5 mg/l þ BAP 1.0-2.0 mg/l. However, the highest callus weight was observed at the concentration of IBA 1.5 mg/l þBAP 1.0 mg/l combination than other combination of concentrations. Positively leaf initiation and formation was better in the concentration of IBA 1-3.5 mg/l þBAP 1.0-2.0 mg/l combination. In addition, the 2,2-diphenyl-2-picrylhydarzyl (DPPH) free radical scavenging potential was higher (70.1%) in leaves extract than in callus extracts (46.3%) at the concentration of 10 mg/ml though both extracts had lower DPPH free radical scavenging activity compared to the positive control, vitamin C and BHT. Theresults conclude that the optimum concentration was IBA 1.5 mg/l þBAP 1.0 mg/l combination to produce callus cell Contents lists available at ScienceDirect
Preparative Biochemistry & Biotechnology, 2018
Betulin (B) and betulinic acid (BA) are two triterpenes with diverse pharmacological and physiological actions. Elicitation of Betula pendula Roth cell cultures by elicitors is an excellent strategy to increase B and BA levels. Six abiotic and biotic elicitors were studied to improve accumulation of B and BA in the cell culture of B. pendula. The B and BA production in treated cells was verified by HPLC. The results showed the maximum growth index (7) on day 3 in cells treated with 0.5 mg L À1 chlorocholine chloride (CCC). The increased accumulation of BA in the cells treated with 200 mg L À1 of chitosan was found to be 5.9 Â (6.5 mg g À1 DW) higher over control cells. Treating the cells with 2 mg L À1 of CCC, after 7 days, led to 149.3Â enhancement of B content (19.4 mg g À1 DW) over the controls. Production of this triterpenoid at a much shorter time with a much higher growth rate can be economic and lead to producing large amounts of B and BA for anticancer and HIV drugs preparation.
Antioxidant Activity of Dry Birch (Betula Pendula) Leaves Extract
Folia Medica
Background: Betula pendula is widespread in Europe and Asia. It has been used in traditional medicine since ancient times. Birch leaf extracts are known to exhibit a number of pharmacological activities. Antioxidant activity has also been reported. Aim: The aim of this work was to investigate the antioxidant activity of a dry leaf extract from Betula pendula Roth. Materials and methods: The total flavonoid content was determined. Some of the most commonly used methods were applied to evaluate the antioxidant capacity of the extract in vitro and in vivo. The ability of the extract to scavenge DPPH free radicals was evaluated by the method described by Brand-Williams with suitable modifications. ABTS decolorization assay was also applied. The in vivo assay was performed after acute and chronic administration of the extract into white albino rats, in a dose of 100 and 500 mg/kg bw. The antioxidant potential of the plasma was determined using FRAP reagent. Results: A total flavonoid con...
Evidence-Based Complementary and Alternative Medicine, 2016
Leaf, seed, and tuber explants ofC. latifoliawere inoculated on MS medium supplemented with various concentrations of BAP and IBA, alone or in combinations, to achievein vitroplant regeneration. Subsequently, antioxidant and antibacterial activities were determined fromin vitroandin vivoplant developed. No response was observed from seed culture on MS media with various concentrations of PGRs. The highest percentage of callus was observed on tuber explants (94%) and leaf explants (89%) when cultured on MS media supplemented with IBA in combination with BAP. A maximum of 88% shoots per tuber explant, with a mean number of shoots (8.8±1.0), were obtained on MS medium supplemented with combinations of BAP and IBA (2.5 mg L−1). The best root induction (92%) and mean number (7.6±0.5) from tuber explants were recorded on 2.5 mg L−1IBA alone supplemented to MS medium. The higher antioxidant content (80%) was observed fromin vivotuber. However, tuber part from the intact plant showed higher...
Optimizing vitamin B12 biosynthesis by mycelial cultures of Lentinula edodes (Berk.) Pegl
Enzyme and Microbial Technology, 2008
Preparations derived from Lentinula edodes (Berk.) Pegl. are widely used as dietary supplements, they contain compounds with immune system enhancing and chemopreventive properties. These preparations contain a blend of minerals and vitamins, including vitamins A, D, B 1 , B 2 , C, and niacin, but only scant vitamin B 12. Our goal was to optimize the growth conditions of submerged mycelial cultures of L. edodes in order to obtain a new dietary supplement enriched in vitamin B 12. We designed a biotechnological process in which cobalamin precursors, cobalt chloride and the methyl donors betaine, methionine, and choline, were added to the culture medium. The vitamin B 12 content in mycelial extracts was determined by RP-HPLC, while total Co 2+ content was determined by HPIC. At the optimal Co 2+ concentration (40 g/mL), the vitamin B 12 level in L. edodes mycelium reached 95 g/g of dry weight. All methyl donors had a positive effect on cobalamin biosynthesis, at least doubling its concentration in mycelia as compared mycelia grown in non-enriched medium. The amount of cobalamin in submerged cultivated mycelia of L. edodes was 10,000-fold higher than that recorded for fruiting bodies. Our results suggest that these optimized culture conditions could be applied to obtain a new cobalamin-enriched dietary supplement derived from L. edodes.
Effect of Abiotic and Biotic Elicitors on Callus and Suspension from Piper Betle L. Var. Nigra
Syntax Idea
Piper betle L. var. Nigra (black betel) contains secondary metabolites and has biological activity as antibacterial, antifungal, antioxidant, etc. To increase the production of secondary metabolites, an alternative method is needed, namely cell suspension culture. This study aims to determine the effect of abiotic and biotic elicitor on callus biomass produced from cell suspension culture. Leaf explants were grown on Murasige and Skoog (MS) medium with the addition of growth regulator 2.4-D 0.5 mg/L and BAP 2.0 mg/L with abiotic elicitors CuSO4, ZnSO4, HgCl2 and CoCl2 with a concentration of 0.5; 1.0 and 2.5 mg/L. The biotic elicitor used was Aspergillus niger with a concentration of 0.025%; 0.050% and 0.1%. The cultures were incubated for 8 weeks. 0.5 g callus was subcultured on 25 mL cell suspension medium. The suspension culture was shaken at 110 rpm. In this suspension culture, the callus was incubated for 3 weeks. After 3 weeks of age callus on suspension medium was harvested a...
Plants
Bryonolic acid is a triterpenoid compound found in cucurbitaceous roots. Due to its biological activities, this compound gets more attention to improve production. Herein, we carried out efficient ways with high bryonolic acid productions from Trichosanthes cucumerina L., a Thai medicinal plant utilizing plant cell cultures. The results showed that calli (24.65 ± 1.97 mg/g dry weight) and cell suspensions (15.69 ± 0.78 mg/g dry weight) exhibited the highest bryonolic acid productions compared with natural roots (approximately 2 mg/g dry weight). In the presence of three elicitors (methyl jasmonate, yeast extract, and chitosan), cell suspensions treated with 1 mg/mL of chitosan for eight days led to higher bryonolic acid contents (23.56 ± 1.68 mg/g dry weight). Interestingly, cell culture and root extracts with high bryonolic acid contents resulted in significantly higher percent cell viabilities than those observed under control (1% v/v DMSO) treatment in Saos-2 and MCF-7 cells. The...
Musa balbisiana is a wild banana genotype with important traits such as drought tolerance and disease resistance. Uniform and clean plants are often required to study these traits in different laboratories but plants can only be generated through a tissue culture process yet for a long time a protocol for regeneration of the same has not been available. Here, we demonstrate that modification of the anti-oxidant content of the in-vitro plant proliferation medium through adjusting the concentration of ascorbic acid and thiamine HCl in the basal MS medium together with subjecting the explants to dark culturing conditions improved proliferation of M. balbsiana by over 10 fold. These treatments resulted in 40 shoots per initial explant material at the best performance.
Artificial Cells, Nanomedicine, and Biotechnology
Prunella vulgaris L. is one of the therapeutic herbs containing various polyphenolics, which is used for multiple medicinal purposes. In this study, plant growth regulators (PGRs)-induced calli cultures from seed-derived leaf explants were exploited for the production of stress enzymes and polyphenolics. A growth curve was plotted for each PGR for 49 days period, which showed a distinct lag, log and decline phases. Here, the combination of naphthalene acetic acid (NAA) and 6-benzyleadenine (BA; 0.5 and 2.0 mg l À1 ) produced maximum fresh (6.32 FW-g/100 ml) and dry biomass (0.75 DW-g/100 ml) in contrast to control. The maximum synthesis of SOD (0.0154 FW-nM/min/mg) was detected on media comprising mixture of NAA and BA (1.5 mg l À1 ), while POD enzyme (0.366 FW-nM/min/mg) was higher at 0.5 mg l À1 NAA and 2, 4-dichlorophenoxy acetic acid. Further, NAA and BA (1.5 and 2.0 mg l À1 ) boosted up the synthesis of phenolics (18.83 GAE-mg/g-DW) and flavonoids content (18.05 RE-mg/g-DW) than control. Moreover, NAA of 1.0 and 2.0 mg l À1 were found supportive for maximum antioxidant activity (87.4%) and total protein (716 mg BSAE/mg-DW). This study will contribute in the development of cell culture in fermenter and synthesis of antioxidant secondary metabolites for commercial uses.
Effect of Vitamins on In Vitro Organogenesis of Plant
Vitamins are necessary compounds synthesized and utilized in plants. In tissue culture media, vitamin addition is not always common; since the amount needed by plants is relatively unknown and varies. Vitamins, in combination with other media constituents, have been shown to have direct and indirect effects on callus growth, somatic growth, rooting, and embryonic development. For example, different studies have shown that thiamine is associated with cytokinin and has a role in inducing callus growth and rooting. Moreover, thiamine was essential in facilitating the production of more secondary metabolites such as proteases in pineapple. Both biotin and riboflavin play a role in callus development as well. Specifically, riboflavin exerts different effects on plant rooting either positively and negatively. Vitamin D known to cause uptake of calcium in animal tissue, exerts a similar effect in plants. In addition, vitamin D causes cell elongation and meristematic cell division. Vitamin C, known for its anti-oxidative properties, has also enhanced shoot growth and rooting.