Document Type : Research Paper
Authors
1 tabriz university
2 biotechnology, university of tabriz, tabriz,iran
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 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.
Keywords
Main Subjects
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