Methyl jasmonate increases silymarin production in Silybum marianum (L.) Gaernt cell cultures treated with β-cyclodextrins (original) (raw)
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Journal of Biotechnology, 2005
The biosynthesis of the flavonolignan silymarin, a constitutive compound of the fruits of Silybum marianum with strong antihepatotoxic and hepatoprotective activities, is severely reduced in cell cultures of this species. It is well known that elicitation is one of the strategies employed to increase accumulation of secondary metabolites. Our study here reports on the effect of several compounds on the production of silymarin in S. marianum cultures. Yeast extract (YE), chitin and chitosan were compared with respect to their effects on silymarin accumulation in S. marianum suspensions and only yeast extract stimulated production. Jasmonic acid (JA) potentiated the yeast extract effect. One of the jasmonic acid derivatives, methyl jasmonate (MeJA), strongly promoted the accumulation of silymarin. Methyl jasmonate acted in a number of steps of the metabolic pathway of flavonolignans and its stimulating effect was totally dependent of "de novo" protein synthesis. Chalcone synthase (CHS) activity was enhanced by methyl jasmonate; however there did not appear to be a temporal relationship between silymarin accumulation and increase in enzyme activity. Also, this increase was not blocked by the protein synthesis inhibitor cycloheximide (CH). This study indicates that elicitor treatment promotes secondary metabolite production in S. marianum cultures and that jasmonic acid and its functional analogue plays a critical role in elicitation.
Enhanced accumulation of flavonolignans in Silybum marianum cultured roots by methyl jasmonate
Phytochemistry Letters, 2012
Root cultures of Silybum marianum (L.) Gaertn. (Asteraceae) were established from in vitro germinated sterile plantlets. The cultures grew in hormone-free Murashige and Skoog medium. The flavonolignan content in the cultured roots was determined by HPLC using 30% acetonitrile in acidified water (0.5% phosphoric acid). The major flavonolignans produced by the cultured roots were silychristin (74.2 mg g À1 fresh weight (FW)) and silydianin (8.1 mg g À1 FW). The flavonolignan precursor taxifolin was also detected in the cultured roots (40.8 mg g À1 FW). Addition of methyl jasmonate to 7-days-old root cultures for 48 h increased the content of the produced flavonolignans and taxifolin to approximately 300% of the control cultures. Methyl jasmonate also enhanced about sixfold the accumulation of a compound identified as 3,3 0 ,5,5 0 ,7-pentahydroxyflavanone.
2016
Milk thistle (Silybum marianum L. Gaertn., Asteraceae) is an annual or biennial, broadleaf plant native in North African and Mediterranean with highly valued medicinal properties. The active principle in S. marianumis silymarin; which is an isomeric mixture of flavonolignans is used in pharmaceutical industries. Silymarin is the most commonly used herbal product for chronic liver disease and may also be beneficial for reducing the chances for developing certain cancers. Due to a growing demand for silymarin, it is justifiable to investigate ways to optimized production of it. Spiny leaves and flowers of the Milk thistle make difficult traditional agriculture of this plant; also the total average daily air temperature from the formation of inflorescence shoots to the milk thistle harvest has a significant effect on silymarin content. Thus ‘In vitro’ culture of cells and tissues may offer an alternative for the production of silymarin. Efforts were carried out to isolate flavanolignan...
Plant Science, 1999
Flavonolignan production was examined in callus and cell suspension cultures derived from cotyledons of Silybum marianum (L.) Gaertn. The flavonolignan content in 2-month-old calluses was lower than the values obtained for fruits (1.5%) and progressively decreased over repeated subcultures. Higher accumulation was observed in cell suspension cultures than in callus but continued to be very low with respect to the values obtained for fruits (14%). The growth and flavonolignan production of suspensions were tested using different concentrations of KNO 3 , KH 2 PO 4 , iron and calcium. Only the removal of calcium ions promoted flavonolignan accumulation, although under this condition, growth was significantly reduced.
Journal of Proteomics, 2013
Elicitation with methyl jasmonate (MeJA) or/and cyclodextrin (CD) strongly induced silymarin (Sm) accumulation in suspensions of Silybum marianum, with most of Sm isomers being detected in the culture medium. This induction provides a model platform to characterize the regulation of flavonolignan accumulation and release in response to elicitors and, with this aim, changes in the S. marianum cell proteome were investigated. The DIGE technique was used to detect statistically significant changes in the cell's proteome. A total number of 1269 unique spots were detected, 67 of which were de-regulated upon elicitation. Nineteen spots were identified by nLC-MS/MS database search analysis. Identified proteins belong to a few categories, including metabolism, stress and defense responses and transport processes. The most abundant group was represented by pathogenesis-related (PR) proteins and heat shock proteins. Two proteins related to transport process were identified and both were upregulated by elicitation. One was identified as Ras-related protein Rab11C of the Rab family of small ATPase superfamily. A second protein was identified as an ABC transporter. Some of the identified proteins are discussed with respect to their putative role in the extracellular flavonolignan accumulation in S. marianum cultures. Biological significance Most approaches to increase secondary metabolite yields using plant cell cultures have been focused on the optimization of its biosynthesis. The study of other post biosynthetic events, like chemical or enzymatic modifications, transport, storage/secretion and catabolism/degradation are also biotechnologically relevant. Secretion is of particular interest since if cell cultures are to be used routinely for the commercial production, they must release the targeted metabolites into the extracellular medium. Elicitor-induced silymarin accumulation and release in S. marianum cell cultures provide a responsive model system to profile both alterations in proteins related to monolignol/ flavonoid biosynthesis and to identify potential systems involved in secretion of secondary metabolites. The proteomic approach undertaken in this work has permitted identify some of the events occurring in elicited S. marianum cell cultures. One attainment of this study is that a
Fitoterapia, 2017
Flavonolignans constitute an important class of plant secondary metabolites formed by oxidative coupling of one flavonoid and one phenylpropanoid moiety. The standardized flavonolignan-rich extract prepared from the fruits of Silybum marianum is known as silymarin and has long been used medicinally, prominently as an antihepatotoxic and as a chemopreventive agent. Principal component analysis of the variation in flavonolignan content in S. marianum samples collected from different locations in Egypt revealed biosynthetic relationships between the flavonolignans. Silybin A, silybin B, and silychristin are positively correlated as are silydianin, isosilychristin, and isosilybin B. The detection of silyamandin in the extracts of S. marianum correlates with isosilychristin and silydianin content. The positive correlation between silydianin, isosilychristin, and silyamandin was demonstrated using quantitative (1)H nuclear magnetic resonance spectroscopy (qHNMR). These correlations can be...
Studies on flavanolignans from cultured cells of Silybum marianum
Acta Physiologiae Plantarum, 2012
Callus and cell suspension cultures of Silybum marianum (L.) Gaertn. (Asteraceae) were established from in vitro germinated sterile plantlets. The cultures grew in Murashige and Skoog medium containing 1 mg l-1 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.1 mg l-1 kinetin. A reversed phase-high performance liquid chromatography method for determination of flavanolignans in plant material was developed using an isocratic solvent system comprising acetonitrile and water containing 0.5% (v/v) phosphoric acid. Silychristin was the major flavanolignan produced by the cultured cells followed by silydianin. Elicitation of cultured S. marianum cells with 100 lg ml-1 yeast extract increased silychristin production from 0.11 to 0.23 mg g-1 fresh weight. Free radical scavenging activity was tested for the cultured cells using 1,1-diphenyl picryl hydrazyl (DPPH) radical. Extract prepared from the cultured cells of S. marianum showed 48% inhibition compared to 55% inhibition of the extract prepared from the fruits. Cytotoxic activity was tested using liver carcinoma cell line (HEPG2). Cultured cells and fruit extracts showed a significant cytotoxic activity of IC 50 = 1.01 and 0.47 lg, respectively. Extract of S. marianum cultured cells ameliorated the adverse effects of carbon tetrachloride-induced hepatic injury in rats and returned the altered levels of biochemical markers to near normal levels.