Phytochemistry (extraction, identification and measurement of active components)
Fahimeh Gharaie; Monireh Cheniany; Ali Ganjeali
Abstract
Background and Objectives: Teucrium polium L. is a herbaceous plant from the mint family that has many uses in traditional medicine. In recent years, numerous medicinal effects such as anti-diabetes, anti-cancer, anti-spasm, and reducing fat and blood pressure have been reported. Therefore, cultivation ...
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Background and Objectives: Teucrium polium L. is a herbaceous plant from the mint family that has many uses in traditional medicine. In recent years, numerous medicinal effects such as anti-diabetes, anti-cancer, anti-spasm, and reducing fat and blood pressure have been reported. Therefore, cultivation of this plant under laboratory conditions is necessary to preserve its survival, reproduction, and increased secondary metabolites. Among the solutions, elicitors in the plant tissue culture technique are one of the most widely used methods to increase secondary-medicinal metabolites. Chitosan, as a biopolymer, induces defense responses, increases antioxidant enzyme activity, and accumulates phenolic compounds. In the present study, the effect of chitosan bio-elicitor was studied on some phenolic compounds of T. polium under in vitro culture to optimize secondary metabolites and increase antioxidant capacity.Methodology: Leaf explants were prepared from preserved T. polium in hydroponic conditions and fed with Hoagland's solution. After sterilization with sodium hypochlorite 5% (v/v) and ethanol 70% (v/v), middle slices were prepared from the leaf explant and cultured on Murashige and Skoog (MS) culture medium containing separate and combined concentrations of benzyl amino purine (BAP)(1 and 1.5 mg.L-1) and naphthalene acetic acid (NAA)(0.5 mg.L-1). The samples were kept for four weeks in the dark at 25 degrees Celsius. With callogenesis, the samples were transferred to light conditions of 16 hours of light and 8 hours of darkness. At the end of the sixth week, the induced calli were subcultured. The calli were treated with chitosan at different concentrations (0, 50, 100, and 150 mg.L-1). After eight weeks, green calli were collected. Finally, to prepare the final extract for biochemical assays, extraction was done from green calli. The content of phenolic compounds (phenol, flavonoid, flavone, and phenolic acids), antioxidant activity, and phenylalanine ammonialyase (PAL) activity was measured.Results: According to the results, the application of different concentrations of chitosan (0, 50, 100, and 150 mg.L-1) and all hormonal treatments (BAP1, BAP1.5, and BAP1.5+NAA 0.5 mg.L-1) caused a significant increase in the content of phenolic compounds, antioxidant capacity, and PAL activity compared to the control samples. So the application of the simultaneous and combined treatment of BAP at a concentration of 1.5 mg.L-1 with NAA at a concentration of 0.5 mg.L-1, along with the treatment of 100 mg.L-1 of chitosan induced green calli of T. polium in in vitro culture, results in the maximum content of phenolic derivatives (1884.95 for phenol, 936.65 for o-diphenol, 1462.28 for flavonoid, 631.07 for flavone, and 662.41 for phenolic acids). As compared to the control, the antioxidant capacities (measured by DPPH and FRAP assays) increased by 68.34 percent and 71.92 percent, respectively. A significant increase in PAL activity (65.81%) was observed in induced calli.Conclusion: Considering the importance of T. polium as a medicinal plant, chitosan, a successful elicitor, promotes the synthesis of phenolic secondary metabolites. As a result, the increase of antioxidant power and PAL activity in the callus of T. polium.
Y. Shakoori; B. Kashefi
Abstract
Considering the importance of medicinal properties of tarragon, and its propagation problems, tissue culture techniques can be used for its propagation. The aim of this study was to compare different culture media and chemical plant growth regulators on the growth traits and secondary metabolites of ...
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Considering the importance of medicinal properties of tarragon, and its propagation problems, tissue culture techniques can be used for its propagation. The aim of this study was to compare different culture media and chemical plant growth regulators on the growth traits and secondary metabolites of tarragon (Artemisia dracunculus L.). Treatments were culture medium at three levels of Murashige and Skoog (MS), Gamborg (B5) and Schenk & Hildebrandt (SH), and growth regulators at five levels of Kinetin (KIN), NAA, and 2,4-D alone (each with a concentration of 0.5 mg l-1), and two combinations of NAA+KIN and 2,4-D+KIN. The present study was conducted as a factorial based on completely randomized design with four replications. The results showed that culture media and growth regulators treatments had significant effects on callus characteristics, regenerated plantlets, callus secondary metabolites, and plant essential oil and its composition. MS medium caused a higher callus induction percentage and more increase in its fresh and dry weight. MS with NAA+KIN and 2,4-D+KIN caused the highest regeneration percentage (35.5), root length (1.28 cm), root (6.5 g) and stem (2.6 g) dry weight, the amount of callus phenolics, and also the amount of essential oil (2.42%) and ocimene (5.78%). Estragol percentage of essential oil increased by 2,4-D, while NAA enhanced ocimene, limonene, and linalool. The percentage of limonene in treatment with NAA alone and linalool in treatment with 2,4-D+ KIN was higher than those of other treatments. The results showed that the highest growth index, callus secondary metabolites, and plant essential oil were observed in MS medium containing growth regulators. Also, the application of kinetin with NAA and 2, 4-D resulted in improved callus growth, regeneration, essential oil, and some of its compounds production in tarragon.
M. Modarres
Abstract
Salvia leriifolia Benth., belonging to Lamiaceae, has antinociceptive, anti-inflammatory, antidiabetic and antioxidant properties. In order to callus induction from apical meristem and embryo of salvia leriifolia to produce phenolic acids, an experiment was carried out in a completely randomized design. ...
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Salvia leriifolia Benth., belonging to Lamiaceae, has antinociceptive, anti-inflammatory, antidiabetic and antioxidant properties. In order to callus induction from apical meristem and embryo of salvia leriifolia to produce phenolic acids, an experiment was carried out in a completely randomized design. The apical meristem and embryo explants were cultured in MS medium supplemented with 2,4-D (0, 1, 2, 3mg/L), KIN (0, 0.3, 1mg/L), BAP (0, 0.5, 1, 2, 3mg/L) and NAA (0, 0.5, 1, 1.5mg/L). The fresh and dry weights were evaluated and accumulation of phenolic acids was measured after four weeks of culture by HPLC. The best treatments for callus induction and growth was at the 2,4-D 2mg/L and KIN 1mg/L. Accumulation of caffeic acid in embryo callus was higher than that of leaves of Salvia leriifolias and concentration of rosmarinic acid was comparable to the leaves. The highest salvianolic acid B concentration was found in callus of apical meristem that was about four times higher than that of leaves.
M. Movahedi; V. Ghasemiomran; S. Torabi
Abstract
The present study was carried out to investigate the possibility of micro-propagation and to determine the optimal medium composition and combination of Cannabis sativa L. growth regulators under in vitro conditions. Seeds were surface-sterilized and then cultured on MS basal medium. One month later, ...
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The present study was carried out to investigate the possibility of micro-propagation and to determine the optimal medium composition and combination of Cannabis sativa L. growth regulators under in vitro conditions. Seeds were surface-sterilized and then cultured on MS basal medium. One month later, leaf and hypocotyl explants, obtained from the seedlings grown at in vitro condition, were used in MS culture medium containing NAA hormone (0.5, 1, 2 and 3 mg/l) either alone or in combination with 0.5mg/l BA; and 2,4-D (0.1, 0.2, 0.5 and 1 mg/l) alone or in combination with 0.5mg/l BA. Callus formation was the response of explants in most media. Direct shoot regeneration from explants was not observed but shoot induction from callus was seen only in 0.1 mg/L 2,4-D+ 0.5 mg/L BA. The highest volume of induced callus was formed on MS medium 0.1 mg/L 2,4-D+ 0.5 mg/L BA using leaf as explant. Root induction from some explants was observed in different treatments. The highest fresh weight of calli belonged to the leaf explant cultured on MS medium containing 1 mg/L 2,4-D+ 0.5 mg/L BA. Callus induction and rooting occurred easily and the explants did not respond well to regeneration.
M. Movahedi; V. Ghasemiomran; S. Torabi
Abstract
Cannabis sativa L., belonging to cannabinaceae family, is a medicinal plant that produces a diverse array of secondary metabolites. The present study aimed to optimize in vitro direct and indirect regeneration in this valuable plant. For this reason, leaf and hypocotyl explants from in vitro grown plantlets ...
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Cannabis sativa L., belonging to cannabinaceae family, is a medicinal plant that produces a diverse array of secondary metabolites. The present study aimed to optimize in vitro direct and indirect regeneration in this valuable plant. For this reason, leaf and hypocotyl explants from in vitro grown plantlets were excised and cultured on MS media containing different concentrations of BA (0.1, 0.2, 0.5, 1, 2 and 3mg/l) and TDZ (0.1, 0.2, 0.5, 1, 2 and 3mg/l) alone or in combination with 0.5 mg/l IBA. The explants reaction in all media led to formation of calli. Direct shoot regeneration from explants was not observed but shoot induction from callus was seen on MS media containing different concentrations of BA in hypocotyl explants. The highest volume of callus was formed on MS medium containing 2 mg/l TDZ+ 0.5mg/l IBA using leaf as explant. The highest wet and dry weight of callus belonged to leaf explant cultured on MS medium containing 1 mg/l TDZ+ 0.5 mg/l IBA. MS medium containing 0.5 mg/l TDZ+ 0.5 mg/l IBA produced no callus in both explants. The addition of IBA in different concentration of BA had significant effects on volume, wet and dry weight of calli in both explants whereas it had no significant effects in different concentrations of TDZ.
K. Pirian; Kh. Piri
Abstract
Portulaca oleracea L. as a medicinal plant, having valuable secondary metabolites, such as noradrenaline, dopamine and omega-3, is used as anti-cancer, antioxidant, and blood purifier factor. Callus inducted from hairy roots of some medicinal plants are used to increase the production of secondary metabolites, ...
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Portulaca oleracea L. as a medicinal plant, having valuable secondary metabolites, such as noradrenaline, dopamine and omega-3, is used as anti-cancer, antioxidant, and blood purifier factor. Callus inducted from hairy roots of some medicinal plants are used to increase the production of secondary metabolites, cell suspension culture, protoplast culture, induction of embryonic callus and gene transfer agents. In this experiment, various concentrations of two hormones of BA and 2,4-D were examined on transgenic hairy roots of P. oleracea to produce callus. Hairy roots transgenic of P. oleracea were induced by Agrobacterium rhizogene, 15834 strain. Hairy roots produced by Agrobacterium rhizogenes15834 strain, were transferred to 1/2MS medium containing different concentrations of BA and 2,4-D hormones. BA and 2,4-D hormones were used, both at three levels of 0, 0.5 and 1 mg per liter and in three replications. Our results indicated that BA and 2,4-D hormones alone as well as without hormone treatment (control) did not show any callus formation. The treatments containing different ratios of BA and 2,4-D hormones, caused callus formation in varying degrees. The medium containing 1mg/L BA and 1 mg/L of 2,4-D hormones showed the higher production of callus.
M. Omidi; B. Behjat Sasan; M.R. Naghavi; S. Kalate Jari; A.R. Etminan
Abstract
Taxus baccata L. is an endangered forest tree species. Taxol is recognized as a highly effective anticancer drug. It is mainly extracted from the bark and needles of taxus. A factorial experiment was carried out on the base of completely randomized design to find the best combination of media, explants ...
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Taxus baccata L. is an endangered forest tree species. Taxol is recognized as a highly effective anticancer drug. It is mainly extracted from the bark and needles of taxus. A factorial experiment was carried out on the base of completely randomized design to find the best combination of media, explants and plant regulators for callus induction in order to produce biomass taxol. In this research three factors were used as follows: MS medium and 4-modified MS media, which differ in salts and nitrogen construction and glutamine, two plant regulators (NAA, 2, 4-D and Kin), and two explants (stem and leaf). Following two months of callus, the tissue was transferred to ½ MS supplemented with 0.4 mg/l 2, 4-D+ 3mg/l Kin. The maximum percentage of callus induction (96.97%) was obtained from stem segments on ½ MS (475 mg/l KNO3, 412.5 mg/l NH4NO3) medium in combination with glutamine (100 mg/l) and containing 2,4-D (1mg/l) + Kin (1mg/l). The best response (80.67 mm2) was observed from leaf on ½ MS in combination with glutamine (100 mg/l) and containing NAA (2mg/l)+ Kin (0.2 mg/l) for callus size. Stem callus showed meristematic cells. According to the results, glutamine showed significant effects on callus induction and growth. The effect of low concentrations of growth regulators on callus induction and callus size was also more than that of high concentrations of growth regulators.
