Biotechnology
ebrahim dorani; sanaz hemmati asl
Abstract
Background and objectives: Anthraquinones are secondary plant metabolites that play a significant role in the treatment of certain diseases, such as hepatitis and cancer. Anthraquinones are utilized not only in textiles and food production but also in the cosmetics, perfume, and pharmaceutical ...
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Background and objectives: Anthraquinones are secondary plant metabolites that play a significant role in the treatment of certain diseases, such as hepatitis and cancer. Anthraquinones are utilized not only in textiles and food production but also in the cosmetics, perfume, and pharmaceutical industries due to their high antioxidant levels. In vitro culture of plants has provided a suitable alternative method for producing medicinal compounds under controlled conditions, not affected by environmental factors. The hairy root culture system, based on inoculation with Agrobacterium rhizogenes, has become an influential approach for producing secondary metabolites in plant roots, such as ginseng, over the last two decades. One of the advantages of in vitro production of these compounds is the possibility of increasing their yield through the use of influential nutrient elements, as well as inducing the production process and biosynthesis of secondary substances with various biological and non-biological elicitors. The objective of this research was to investigate the effect of salt concentration in the culture medium and sucrose levels on increasing hairy root biomass, and to examine the influence of different abiotic elicitors on enhancing anthraquinone production in madder hairy roots.Methodology: For this purpose, in the first experiment, after inducing hairy roots, the effect of sucrose concentration (30, 45, and 60 g/L) in combination with basal medium salt strength (1/2 MS and MS) on root biomass enhancement was studied in a factorial experiment with a completely randomized design and four replications. In the second experiment, the effects of different elicitors, including titanium dioxide (50 and 100 mg/L), chitosan (100 and 150 mg/L), silver nitrate (50 and 100 mg/L), proline (250 and 500 mg/L), gibberellic acid (1 and 2 mg/L), and naphthalene acetic acid (1 and 2 mg/L), were investigated for their role in increasing anthraquinone (alizarin) production in hairy roots.Results: In the first experiment, the 1/2 MS medium containing 45 g/L sucrose produced the highest biomass, followed by MS containing 45 and 60 g/L sucrose, MS with 30 g/L sucrose, and 1/2 MS containing 60 g/L sucrose. The best medium for biomass production from this experiment was selected for the subsequent experiment, in which the effect of different elicitor treatments for two days on the accumulation of the secondary metabolite (alizarin) was assessed. The results showed that the accumulation of the secondary metabolite in the medium containing 250 mg/L of proline exhibited a significant increase compared to other elicitors, followed by 50 mg/L of silver nitrate and 2 mg/L of gibberellic acid, respectively. The secondary metabolite content in media supplemented with two different levels of naphthalene acetic acid did not differ significantly from that of the control.Conclusion: Overall, this study demonstrated that the optimal root biomass production was achieved in 1/2 MS culture medium containing 45 g/L of sucrose, while the highest anthraquinone content was obtained in the presence of proline.
R. Ghodsi; B. Hosseini; A. Hedayati
Abstract
Hyoscyamus species such as H. reticulatus L. and H. pusillus L. are rich sources of tropane alkaloids, mainly hyoscyamine and scopolamine, which are used for their mydriatic, antispasmodic, anticholinergic, analgesic and sedative properties. In this study, in the first experiment, the effects of four ...
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Hyoscyamus species such as H. reticulatus L. and H. pusillus L. are rich sources of tropane alkaloids, mainly hyoscyamine and scopolamine, which are used for their mydriatic, antispasmodic, anticholinergic, analgesic and sedative properties. In this study, in the first experiment, the effects of four strains of Agrobacterium rhizogenes (ATCC 15834, A4, A7 and A13) and three explants (leaf, hypocotyls, and cotyledon) on hairy root induction were investigated in both H. reticulatus and H. pusillus species. In the second experiment, the effects of various culture media (MS, 1/2 MS, 1/4 MS, and B5) were evaluated on some morphological and phytochemical traits of hairy roots. The results showed that in H. reticulatus, maximum hairy root induction (78%) was obtained in the cotyledon explant inoculated with A7 strain and maximum hairy roots fresh weight (6.25 g) and dry weight (0.5 g) were recorded in MS medium. In this species, the highest total phenol (3.82 mg GAE g-1 FW) and total flavonoid (7 mg QUE g-1 FW) content were observed in MS medium; however, no significant changes were observed in the activity of antioxidant enzymes of hairy roots cultured in different media. In H. pusillus, the leaf explant inoculated with A13 strain showed maximum hairy root induction (90%), and the highest hairy root fresh weight (5.25 g) and dry weight (0.43 g) were observed in MS medium and the highest total phenol (2.26 mg GAE g-1 FW) and flavonoid (9.22 mg QUE g-1 FW) content were obtained in B5 medium. This study showed that basal culture medium type and plant species had a significant impact on the biomass and phytochemical characteristics of hairy roots.
A. Norozi; B. Hosseini; M. Jafari; M. Farjaminezhad
Abstract
Henabne(Hyoscyamus reticulatus L.) is a rich source of tropane alkaloids including hyoscyamine (Hyos) and scopolamine (SCP), widely used in pharmaceutical field. The aim of the present study was to assess the effect of different concentrations (0, 50, 100 and 200 µM) of methyl jasmonate (MeJA) ...
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Henabne(Hyoscyamus reticulatus L.) is a rich source of tropane alkaloids including hyoscyamine (Hyos) and scopolamine (SCP), widely used in pharmaceutical field. The aim of the present study was to assess the effect of different concentrations (0, 50, 100 and 200 µM) of methyl jasmonate (MeJA) at two different times of exposure (24 and 48 h) on growth, antioxidant enzymes activity, tropane alkaloids content, and the expression level of hyoscyamine-6-beta-hydroxylase (h6h) gene in H. reticulatus hairy root cultures. The results showed that MeJA at high concentration (200 µM) led to the decline of fresh and dry weight of hairy roots. Catalase (CAT), guaiacol peroxidase (GPX) and ascorbate peroxidase (APX) activities were significantly increased by MeJA, especially at 200 µM in elicited hairy roots. As well, CAT and GPX activities, but not of APX, were also affected by exposure time of MeJA. According to the GC/MS analysis, the highest percentage of SCP (13.96%) and Hyos (21.9%) were respectively obtained when hairy roots were exposed to MeJA at concentrations of 100 and 200 µM for 48 and 24 h, which were, respectively, 1.6- and 1.25-fold higher than those in the control roots. According to the results of semi-quantitative RT-PCR analysis, the highest expression of h6h gene (6-fold higher than in the control) was obtained after 24 h exposure to 100 µM MeJA. These results indicate that MeJA, due to stimulation of the expression of key genes involved in the biosynthetic pathway of tropane alkaloids, could be used as an effective elicitor for increased production of Hyos and SCP.
M. Noori; Sh. Gharanjik; A. Safipour Afshar; F. Saeid Nematpur
Abstract
The hairy roots induced by Agrobacterium rhizogenes due to the stability and mass production in hormone free culture provide an appropriate tissue for secondary metabolite production, and the content of metabolites produced could be affected by bacterial strain. In the current study, the effect of different ...
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The hairy roots induced by Agrobacterium rhizogenes due to the stability and mass production in hormone free culture provide an appropriate tissue for secondary metabolite production, and the content of metabolites produced could be affected by bacterial strain. In the current study, the effect of different strains of A. rhizogenes including A4, A13 and 15834 on the percentage of hairy root induction of Echinacea purpurea (L.) moench. in the leaf and hypocotyl explants were studied. In another experiment, total phenolics and polysaccharides content of hairy roots were measured compared to the control (untransformed) roots. The results showed that the percentage of hairy root induction was significantly affected by bacterial strains, so that in E. purpurea leaves incubated by strains A13, 15834 and A4, the hairy root induction was 60%, 40% and 0, and in hypocotyl explants was 85%, 45% and 70%, respectively. Maximum total phenolic compounds were observed in A4 strain. As well, in comparison to control, the polysaccharide content in hairy root induced by strains 15834 and A13 increased and by A4 strain decreased significantly. Differences observed in this study about hairy root induction and secondary metabolites production could be due to different ability of A. rhizogenes strains in T-DNA transferring to plant cells.
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.
