Study of biochemical pathway and genes in biosynthesis of silymarin in cell suspension and hairy root cultures of Silybum marianum for increasing the production of silymarin
2011
Hasanloo, Tahereh | Seperifar, Roshanak | Khayam Nekoie, Mojtaba
Silymarin is composed of many flavonolignans, in seed of Silybum marianum L. Gearten (family composite) that have been used to treat liver diseases. Hairy root cultures of S. marianum could be able to produce silymarin. There is evidence that the use of elicitors has been an important strategy for improving the production of secondary metabolites and identifies cell signaling pathways. Therefore in this research different concentration of biotic elicitor (Salicylic acid 1, 2, 4, 6 and 8 mg/50 ml culture media) and Methyl jasmonate (50, 100, 150 and 250 μM) and abiotic elicitor (Ag+ 0.2, 0.4, 0.8, 1 and 2 mM) were used. Treated and non-treated hairy roots were harvested 72h after elicitation. Detection and identification of silymarin was carryied out by High- Performance Liquid chromatograph method. The highest content of SLM was obtained by 0.8 mg/50 ml culture salicylic acid, 2 mM Ag+ and 100 μM methyl jasmonate. Hairy roots were treated by best concentration of elicitors for exposure time (0, 24, 48, 72, 96 and 120 h). The highst content of silymarin was obtained 24, 96 and 96 h after elicitation by salicylic acid, Ag+ and methyl jasmonate, respectively. That were 2.42, 3.68 and 3.49-times of non treated hairy roots respectively. The highest dry weights were obtained by non-treated hairy roots. To obtain further evidence for signaling pathway lipoxigenase, peroxides, ascorbate peroxides activity, hydrogen peroxides and linoleic acid content were detected by spectrophotometer and gas chromatography methods. These observation showed that elicitation cause to increasing linoleic acid content and lipoxigenase activity.
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