Phytochemistry (extraction, identification and measurement of active components)
L. Moradipour; V. Payam Noor
Abstract
In biotechnology processes, increasing the production of biomass and secondary metabolites is one of the economic factors of such projects. The present study was conducted to investigate the effects of magnetic field as an abiotic elicitor on increase of biomass and secondary metabolites of Ganoderma ...
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In biotechnology processes, increasing the production of biomass and secondary metabolites is one of the economic factors of such projects. The present study was conducted to investigate the effects of magnetic field as an abiotic elicitor on increase of biomass and secondary metabolites of Ganoderma lucidum. Mycelium with a diameter of 1 cm of pured fungi was exposed to magnetism at four levels including 0, 20, 40, and 60 mT for 0, 30, 60, and 90 min with three replications. The best treatment was selected based on growth rate and antioxidant activity. The amounts of secondary compounds in mycelium under selected magnetic field (60 mT for 90 min) including betulin, betulinic acid, ascorbic acid, astaxanthin, total polysaccharide, and antioxidants were measured and compared with control and natural fungi. The results showed that magnetization increased betulin (a potent anticancer triterpene) more than 2.5 times compared to natural fungi and 3.7 times compared to control mycelium. Polysaccharides concentration in the treated mycelium and control media was obtained 5.05 and 5.17 times more than natural fungi, respectively. Ganoderic acid content was measured in the treated mycelium (intracellular) and PDB medium (extracellular). The highest amount of ganoderic acid (1.9 mg.ml-1) was obtained in the treated mycelium. Natural fungi and PBD medium had almost the same amount of this compound (1.87 and 1.86 mg.ml-1). The amount of betulinic acid (an anticancer compound derived from betulin), ascorbic acid, and astaxanthin was obtained high in natural fungus followed by 90 min magnetization at 60mT level treatment and control, respectively. Antioxidants percentage was about 85% in control and about 75% in natural fungi and treated mycelium. According to the results, there seems to be a good prospect to use in vitro cultures of Ganoderma instead of using natural fungi and also constructing costly farms.
V. Payamnoor; J. Nazari; R. Jafari Hajati
Abstract
Betulin and betulinic acid are from the most important anticancer and anti-HIV metabolites, and the birch species (Betula spp.) bark is considered as the primary source of these metabolites. Due to the extinction of these tree species in Iran, it is necessary to replace the metabolites extraction from ...
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Betulin and betulinic acid are from the most important anticancer and anti-HIV metabolites, and the birch species (Betula spp.) bark is considered as the primary source of these metabolites. Due to the extinction of these tree species in Iran, it is necessary to replace the metabolites extraction from the birch bark with modern methods such as cell and tissue culture to produce the metabolites. The aim of this study was to determine the effect of explant type on the amount of betulin and betulinic acid produced in calli of two birch species B. pendula and B. litwinowii under in vitro conditions compared to the amount of metabolites extracted from the tree bark. Bark and leaf explants of two mentioned species were cultured in WPM medium containing 1 mg/L BAP and 0.1 mg/L 2,4-D for callogenesis. The amount of betulin and betulinic acid in three-month calli was measured using the HPLC technique and compared with the amount of these metabolites in one-centimeter stem bark samples taken from nature. The bark explant was more successful in callogenesis, and calli derived from this explant had more active ingredients. The amount of betulin and betulinic acid from the extract of bark sample taken from nature was respectively obtained to be 5.23 and 2.91 percent for B. pendula, and 5.65 and 2.52 percent for B. litwinowii. Moreover, calli derived from the bark explant of B. pendula and B. litwinowii contained 0.023 and 0.016 percent of betulin and 0.053 and 0.057 percent of betulinic acid, respectively. Generally, the results indicated that the bark explant was more capable of callogenesis and secondary metabolite induction than the leaf explant in both birch species under in vitro conditions.
J. Nazari; V. Payamnoor; M.R. Kavosi; J. Asadi
Abstract
Lichens are small environments containing two fungus and algae symbiosis. They are one of the richest sources of natural ingredients with anti-tumor, antibiotic, and antioxidant properties, some of which are used as drugs and for the treatment of certain diseases. This research was aimed to identify ...
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Lichens are small environments containing two fungus and algae symbiosis. They are one of the richest sources of natural ingredients with anti-tumor, antibiotic, and antioxidant properties, some of which are used as drugs and for the treatment of certain diseases. This research was aimed to identify and determine the amount of betolin and betulinic acid as well as evaluation of the antioxidant activity of lichen Betula pendula Roth. Roth.and its symbiotic fungus. It was found that the lichen isolated from the bark of birch contained bioactive compounds entitled Raminalina sinensis. The symbiotic fungus (Arthrinium arundinis) was detected by the molecular method. This fungus belongs to the ascomycetes, registered for the first time in the NCBI website with MG198621 code. In the extract of lichen and its symbiotic fungus, the amount of betulinic acid and betulinic acid was determined using HPLC. Betulinic acid and betulin, belonging to the terpenes, are known as a strong anticancer agent. The mentioned ingredientswere significantly found in the lichen tissue(2.17 and 0.075 percent, respectively) and in its symbiotic fungus A. arundinis (1.6 and 0.025 percent, respectively). In this way, the lichen and its symbiotic fungus are introduced for the first time as new sources containing these two active ingredients. Measurement of antioxidant properties of lichen extract and its symbiotic fungus was examined by methanol and ethanol solvents. It was found that the lichen extract and the symbiotic fungus, besides having alternative metabolites, have antioxidant properties, which are able to remove toxic free radicals. These results could have valuable effects in medicine and industry.
V. Payamnoor; R. Jafari Hajati
Abstract
Birch (Betula pendula Roth.) is one of the most important species exposed to extinction due to the limited natural habitat and lack of regeneration. Plant tissue culture technique is an appropriate solution for asexual reproduction of this species and to protect it in controlled conditions. In ...
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Birch (Betula pendula Roth.) is one of the most important species exposed to extinction due to the limited natural habitat and lack of regeneration. Plant tissue culture technique is an appropriate solution for asexual reproduction of this species and to protect it in controlled conditions. In this research, callogenesis of B. pendula was investigated using five types of explants (leave, stem, nodal, petiole and bark) in WPM and NT media enriched with two hormonal combinations including A) 1mg/l BAP, 0.1 mg/l 2,4-D and B) 0.1 mg/l BAP, 0.01mg/l TDZ). The results indicated that the minimum callogenesis was observed in nodal and petiole explants in NT medium enriched with A hormonal combination. Callogenesis, fresh and dry weight of bark, stem and leave explants were significantly decreased in WPM and NT media containing B hormonal combination as compared with A hormonal combination. The maximum callogenesis was observed in bark explant in WPM (81.14) and NT (70.36) media with A hormonal combination as compared with other explants. The average fresh and dry weight of this explant in NT medium was more than that of WPM medium with the same hormonal combination.
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.
