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.
Improvement and breeding
S. Jafari; S Mousavi-Fard; A. Rezaei Nejad; H. Mumivand; K. Sorkheh
Abstract
Milk thistle (Silybum marianum (L. Gaertn.)) is a plant with various medicinal uses which is adapted to different climates. To investigate the effects of different concentrations of chitosan (CS) and titanium dioxide (TiO2) on increasing yield and biochemical indices of milk thistle ecotypes, a farm ...
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Milk thistle (Silybum marianum (L. Gaertn.)) is a plant with various medicinal uses which is adapted to different climates. To investigate the effects of different concentrations of chitosan (CS) and titanium dioxide (TiO2) on increasing yield and biochemical indices of milk thistle ecotypes, a farm factorial experiment was conducted in a randomized complete block design with three replications during two crop years. Experimental treatments included milk thistle ecotype at five levels (Budakalazi, Sari, Khomein, Khorramabad, and Ahvaz) and type and elicitor concentration at nine levels (foliar application of distilled water (control), bulk CS (50 and 100 mg L-1), nano CS (50 and 100 mg L-1), bulk TiO2 (50 and 100 mg L-1), and nano TiO2 (50 and 100 mg L-1)). The results showed that foliar application of CS and TiO2 (bulk and nano) effectively increased yield and defense enzymes. In all these traits, foliar application of CS and TiO2 in nano form was more effective compared to the bulk form and control. Based on the mean comparisons results, the plant yield increased with foliar application of nano CS at the concentration of 50 mg L-1. Also, foliar application of nano CS at the concentration of 100 mg L-1 increased the activity of defense enzymes. In general, foliar application of nano CS increased the yield as well as improved the biochemical traits of milk thistle ecotypes, especially the Budakalazi one, which could be considered as a suitable way to increase the yield of this plant. Considering the higher yield of Budakalazi ecotype, it could be also recommended to be used in breeding programs.
S. Fabriki-Ourang; H.S. Shahabzadeh
Abstract
In this study, the effects of salicylic acid and methyl jasmonate as two inducers were studied on antioxidant activity, photosynthetic pigments and flavonoids under salinity stress in Chelidonium majus L. This experiment was conducted as a factorial in a completely randomized design with four replications ...
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In this study, the effects of salicylic acid and methyl jasmonate as two inducers were studied on antioxidant activity, photosynthetic pigments and flavonoids under salinity stress in Chelidonium majus L. This experiment was conducted as a factorial in a completely randomized design with four replications at Imam Khomeini International University during 2017. The elicitors were treated as foliar and mixed with irrigation. The factors studied included elicitors at three levels (distilled water as control, 2 mM salicylic acid, and 100 µM methyl jasmonate), salinity stress at two levels, urban water as control (EC=0.62 ds/m), salinity at 30 mM NaCl (EC=3.36 ds/m), and plant organs at three levels (leaf, root, stem). The analysis of variance showed that the main effects of salinity, elicitors, organs and their interactions were significant for most of the traits. The results of mean comparison for elicitors showed that methyl jasmonate increased carotenoid, flavonoids and anthocyanin. Both salicylic acid and methyl-jasmonate treatments reduced the amount and activity of catalase, superoxide dismutase and guaiacol peroxidase enzymes under salinity stress. In mean comparison of the triple interaction effects, salinity+salicylic acid+leaf was the best combination of factors for chlorophyll (1.68 mg.g-1 FW) and carotenoid (0.54 mg.g-1 FW) contents. Also, the maximum content for anthocyanin (5.7 OD.gr-1 FW) was observed in non-salinity+methyl jasmonate+leaf and for flavonoids in salinity+non-hormone (3.42 OD.gr-1 FW), non-salinity+salicylic acid (3.27 OD.gr-1 FW) and salinity+methyl jasmonate (3.2 OD.gr-1 FW) in leaf, respectively. The amount of total protein increased in salinity+salicylic acid (56.72 mg.g-1 FW) and salinity+methyl jasmonate (53.27 mg.g-1 FW) in contrast to reduction in salinity+non-hormone (43.35 mg.g-1 FW). In conclusion, by applying the two mentioned elicitors, greater celandine does not need to increase the antioxidant defense system and will potentially improve the yield of biomass and some secondary metabolites under salinity stress.
A. Kheiry; H. Tori; N. Mortazavi
Abstract
Peppermint (Mentha piperita L.) from Lamiaceae family is one of the most widely used medicinal plants. In addition to therapeutic effects, it is used as flavoring in the production of various foods and medicinal products. In addition to genotype, the quality and quantity of active ingredients in medicinal ...
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Peppermint (Mentha piperita L.) from Lamiaceae family is one of the most widely used medicinal plants. In addition to therapeutic effects, it is used as flavoring in the production of various foods and medicinal products. In addition to genotype, the quality and quantity of active ingredients in medicinal plants are influenced by environmental factors. This research was aimed to study the effects of drought stress and jasmonic acid on the quality and quantity of peppermint as a factorial experiment in a completely randomized design at University of Zanjan in 2015. The treatments consisted of three irrigation levels [50%, 75%, and 100% (control) of field capacity] as main factor and three concentrations of jasmonic acid (0, 50, and 100 mg l-1) as sub-factor. Rhizomes of plants were used for plant production. The plant samples were collected at flowering stage and after drying in the shade, the essential oil was extracted with water distillation method by Clevenger apparatus. The essential oil components were identified by GC and GC-MS. The results showed that the highest fresh and dry weight, height, total chlorophyll and relative leaf water content were obtained in control with 100 mg l-1 jasmonic acid. The highest peroxidase enzyme content was obtained at 50% field capacity drought stress with 100 mg l-1 jasmonic acid. The highest essential oil content was obtained at 75% field capacity drought stress with application of 50 mg l-1 jasmonic acid. The main essential oil components were menthol, menthon, 1,8-cineole, cyclohexanol, and methyl acetate. The highest amounts of menthol, menton, and 1,8-cineole were obtained from 100 mg l-1 jasmonic acid and the highest amount of cyclohexanol was obtained from 50 mg l-1 jasmonic acid. However, application of jasmonic acid decreased the amount of methyl acetate from 7.24% in control to 0.56% in 100 mg l-1 of JA treatment.
R. Jafari Hajati; V. Payamnoor; K. Ghasemi Bezdi; N. Ahmadian Chashmi
Abstract
This study aimed to enhance the production of betulin and betulinic acid using suspension cultures of birch (Betula pendula Roth) and elicitation of the cell cultures by methyl jasmonate (MeJA) and salicylic acid (SA). To do this, at the first step, the cell growth curve was investigated in a 16-day ...
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This study aimed to enhance the production of betulin and betulinic acid using suspension cultures of birch (Betula pendula Roth) and elicitation of the cell cultures by methyl jasmonate (MeJA) and salicylic acid (SA). To do this, at the first step, the cell growth curve was investigated in a 16-day period. Then, two elicitors, namely, MeJA (at final concentration of 0, 50, 100, 150 and 200 µM) and SA (at final concentration of 0, 100, 200, 300 and 400 µM) were separately supplemented to 8-day-old cell cultures and the cells were harvested 1, 2, 3, 5 and 7 days after elicitations. Fresh weight (FW), dry weight (DW) and cell viability were measured. In addition, betulin and betulinic acid content were analyzed using HPLC. The results showed the significant effects of different concentrations of SA and MeJA on metabolites content and FW and DW. Maximum amount of betulin was observed about 4-fold (2.5 mg g-1 DW) higher than the control treatment by addition of 100 µM SA, two days after elicitation. Moreover, betulinic acid content was enhanced about 5 mg g-1 DW, 4.5-fold compared to control, one day after addition of 200 µM SA. Furthermore, the high accumulation of betulin (2.3 mg g-1 DW) was obtained in the elicited cell by 50 µM MeJA, seven days after elicitation. Also, the maximum amount of betulinic acid, about 3 mg g-1 DW, was observed in the cells elicited by 100 µM MeJA, two days after elicitation. Overall, the effect of SA on the production of betulin and betulinic acid was significantly more than the effect of MeJA.
M. Parsa; A. Zeinali
Abstract
Tropane alkaloids such as atropine and scopolamine have wide application in the treatment of diseases such as asthma and antispasmodic due to anticholinergic agents. In the present study, the effects of salicylic acid (SA) on the production of two alkaloids, atropine and scopolamine ,were studied ...
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Tropane alkaloids such as atropine and scopolamine have wide application in the treatment of diseases such as asthma and antispasmodic due to anticholinergic agents. In the present study, the effects of salicylic acid (SA) on the production of two alkaloids, atropine and scopolamine ,were studied in hairy root and in vitro grown root cultures of Hyoscyamus niger L. The roots were cultured in liquid B5 medium containing different concentrations of SA (0, 0. 1, 1, 2 and 4 mM) in various exposure times (24, 96 and 168 hours). Eventually, root growth index, and atropine and scopolamine content were assayed after 30 days. In in vitro grown roots, treatment with 1mM SA resulted in the highest production of atropine after 168 hours, while the highest amount of scopolamine (649.53 µg/g D. W) was obtained in 2mM SA (after 96 hours), showing more than 13-fold increase compared to the control. In hairy root cultures, the most significant contents of atropine were observed in the medium containing 2mM SA after 96 h. Moreover, the highest content of scopolamine was achieved in medium treated with 0.1 mM SA after 96 hours. In general, atropine content in hairy roots was considerably higher than that of in vitro grown roots. In contrast, scopolamine content in in vitro grown roots was significantly more than that of hairy roots. Moreover, the rate of root growth declined as a result of increasing of elicitor concentration in the medium.