Gene expression and flavonolignan production in fruits and cell cultures of Silybum marianum

Torres, María Mercedes; Corchete, Purificación

doi:10.1016/j.jplph.2016.02.004

Cited by 25 publications

(33 citation statements)

References 28 publications

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“…The results indicate that S. miltiorrhiza flavonoid biosynthesis-related genes were MeJA-responsive and the responses were in a tissue- and time-specific manner. It is consistent with the results from other plant species showing the regulation of MeJA in flavonoid (particularly anthocyanin) accumulation [ 78 ] and the stimulation of anthocyanin biosynthetic genes, including CHS [ 74 , 79 ], CHI [ 80 ], F3H [ 80 , 81 ], F3′H [ 80 ], DFR [ 78 ] and ANS [ 78 ].The expression levels of SmCHS1 , SmCHS3 , SmCHS5 , SmCHI3 , SmCHI4 , SmF3′H2 , SmF3′H3 , SmF3′H4 , SmFLS2 , SmANS , and SmDFR in roots reached the peak at the time-point of 24 h or 36 h treatment, and then gradually decreased. Similar results were observed for CHS expression in hairy roots of Scutellaria viscidula [ 79 ].…”

Section: Resultssupporting

confidence: 91%

Identification and Characterization of Flavonoid Biosynthetic Enzyme Genes in Salvia miltiorrhiza (Lamiaceae)

Deng

et al. 2018

Molecules

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Flavonoids are a class of important secondary metabolites with a broad spectrum of pharmacological functions. Salvia miltiorrhiza Bunge (Danshen) is a well-known traditional Chinese medicinal herb with a broad diversity of flavonoids. However, flavonoid biosynthetic enzyme genes have not been systematically and comprehensively analyzed in S. miltiorrhiza. Through genome-wide prediction and molecular cloning, twenty six flavonoid biosynthesis-related gene candidates were identified, of which twenty are novel. They belong to nine families potentially encoding chalcone synthase (CHS), chalcone isomerase (CHI), flavone synthase (FNS), flavanone 3-hydroxylase (F3H), flavonoid 3′-hydroxylase (F3′H), flavonoid 3′,5′-hydroxylase (F3′5′H), flavonol synthase (FLS), dihydroflavonol 4-reductase (DFR), and anthocyanidin synthase (ANS), respectively. Analysis of intron/exon structures, features of deduced proteins and phylogenetic relationships revealed the conservation and divergence of S. miltiorrhiza flavonoid biosynthesis-related proteins and their homologs from other plant species. These genes showed tissue-specific expression patterns and differentially responded to MeJA treatment. Through comprehensive and systematic analysis, fourteen genes most likely to encode flavonoid biosynthetic enzymes were identified. The results provide valuable information for understanding the biosynthetic pathway of flavonoids in medicinal plants.

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Section: Resultssupporting

confidence: 91%

Identification and Characterization of Flavonoid Biosynthetic Enzyme Genes in Salvia miltiorrhiza (Lamiaceae)

Deng

et al. 2018

Molecules

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“…Chalcone synthase (CHS) is a vital enzyme of the flavonoid and flavonolignans biosynthesis pathway [17,57]. Silymarin is synthesized in milk thistle plants via the phenylpropanoid pathway converting phenylalanine to 4-coumaroyl-CoA ( Figure S2).…”

Section: Discussionmentioning

confidence: 99%

“…Silymarin is synthesized in milk thistle plants via the phenylpropanoid pathway converting phenylalanine to 4-coumaroyl-CoA ( Figure S2). CHS and chalcone isomerase (CHI) are the enzymes that catalyze the condensation of three malonyl-CoA molecules and 4-hydroxycin-namoyl-CoA to produce the chalcone derivative, naringenin, which is the first dedicated step in the pathway of phenylpropanoid in plants [57,58]. This latter compound is hydroxylated, giving at the end taxifolin, which is the precursor of the flavonoid part of silymarin [58].…”

Section: Discussionmentioning

confidence: 99%

“…Overall, our findings revealed that drought stress improved the expression of the three genes (CHS1, CHS2, and CHS3), which was coordinated by a definite increase in silymarin content (23.76 mg•g −1 DM) in milk thistle. Torres and Corchete [57] observed a relationship between the expression of SmCHS genes engaged in the silymarin biosynthesis pathway and the production of these metabolites in milk thistle. Sanjari, Shobbar, Ebrahimi, Hasanloo, Sadat-Noori and Tirnaz [17] identified CHS1, CHS2, and CHS3 genes in different parts of milk thistle plant (different leaves, petals, flower heads, and diverse parts of the stem), which are perhaps involved in the silymarin biosynthetic pathway in milk thistle.…”

Section: Discussionmentioning

confidence: 99%

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The Impact of Drought Stress on Antioxidant Responses and Accumulation of Flavonolignans in Milk Thistle (Silybum marianum (L.) Gaertn)

ElSayed

El-hamahmy

Rafudeen

et al. 2019

Plants

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Biosynthesis and accumulation of flavonolignans in plants are influenced by different environmental conditions. Biosynthesis and accumulation of silymarin in milk thistle (Silybum marianum L.) were studied under drought stress with respect to the antioxidant defense system at the physiological and gene expression level. The results revealed a reduction in leaf chlorophyll, ascorbic acid, and glutathione contents. In contrast, H2O2, proline, and antioxidative enzyme activities, such as superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), ascorbate peroxidase (APX), and glutathione reductase (GR), were increased. These results confirmed that milk thistle undergoes oxidative stress under drought stress. Furthermore, transcription levels of APX, SOD, CAT, 1-Cys-Prx, and PrxQ were significantly increased in milk thistle under drought stress. Overall this suggests that protection against reactive oxygen species and peroxidation reactions in milk thistle are provided by enzymatic and non-enzymatic antioxidants. Flavonolignans from milk thistle seeds after different drought treatments were quantified by high-performance liquid chromatography (HPLC) and showed that severe drought stress enhanced the accumulation of silymarin and its components compared with seeds from the control (100% water capacity). Silybin is the major silymarin component and the most bioactive ingredient of the milk thistle extract. Silybin accumulation was the highest among all silymarin components in seeds obtained from drought-stressed plants. The expression of the chalcone synthase (CHS) genes (CHS1, CHS2, and CHS3), which are associated with the silybin biosynthetic pathway, was also increased during drought stress. These results indicated that milk thistle exhibits tolerance to drought stress and that seed derived from severe drought-stressed plants had higher levels of silymarin.

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“…In our studies, it was determined that methyljasmonate (MJ) and random methylated β‐cyclodextrins (Cd) induced a massive release of CA, and some of the isomers of the Sm mixture were also detected in the extracellular medium . Expression studies of genes related to the flavonolignan pathway revealed that elicitation with MJ or Cd up‐regulated transcription; the change in their expression partially overcame rate‐limiting steps for Sm accumulation .…”

Section: Introductionmentioning

confidence: 94%

Silybum marianum cell cultures stably transformed with Vitis vinifera stilbene synthase accumulate t‐resveratrol in the extracellular medium after elicitation with methyl jasmonate or methylated β‐cyclodextrins

Hidalgo

Martínez-Márquez

Cusidó

et al. 2017

Engineering in Life Sciences

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Silybum marianum cell cultures stably transformed with Vitis vinifera stilbene synthase accumulate t-resveratrol in the extracellular medium after elicitation with methyl jasmonate or methylated β-cyclodextrinsThe growing demand for t-resveratrol for industrial uses has generated considerable interest in its production. Heterologous resveratrol production in plant cell suspensions, apart from requiring the introduction of only one or two genes, has the advantage of high biomass yield and a short cultivation time, and thus could be an option for large-scale production. Silybum marianum is the source of the flavonolignan silymarin. Phenylpropanoid synthesis in cultures of this species can be activated by elicitation with methyl jasmonate and methylated β-cyclodextrins, with products of the pathway (coniferyl alcohol and some isomers of the silymarin complex) being released into the medium. Given that stilbene synthase shares the same key precursors involved in flavonoid and /or monolignol biosynthesis, we explored the potential of metabolically engineered S. marianum cultures for t-resveratrol production. Cell suspensions were stably transformed with Vitis vinifera stilbene synthase 3 and the expression of the transgene led to extracellular t-resveratrol accumulation at the level of milligrams per litre under elicitation. Resveratrol synthesis occurred at the expense of coniferyl alcohol. Production of silymarin was less affected in the transgenic cultures, since the flavonoid pathway is limiting for its synthesis, due to the preferred supply of precursors for the monolignol branch. The fact that the expressed STS gene took excessively produced precursors of non-bioactive compounds (coniferyl alcohol), while keeping the metabolic flow for target secondary compounds (i.e. silymarin) unaltered, opens a way to extend the applications of plant cell cultures for the simultaneous production of both constitutive and foreign valuable metabolites.

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Gene expression and flavonolignan production in fruits and cell cultures of Silybum marianum

Cited by 25 publications

References 28 publications

Identification and Characterization of Flavonoid Biosynthetic Enzyme Genes in Salvia miltiorrhiza (Lamiaceae)

Identification and Characterization of Flavonoid Biosynthetic Enzyme Genes in Salvia miltiorrhiza (Lamiaceae)

The Impact of Drought Stress on Antioxidant Responses and Accumulation of Flavonolignans in Milk Thistle (Silybum marianum (L.) Gaertn)

Silybum marianum cell cultures stably transformed with Vitis vinifera stilbene synthase accumulate t‐resveratrol in the extracellular medium after elicitation with methyl jasmonate or methylated β‐cyclodextrins

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