Enhanced production of flavonolignans in hairy root cultures of Silybum marianum by over-expression of chalcone synthase gene

Rahnama, Hassan; Razi, Zahra; Dadgar, Morteza Najafi; Hasanloo, Tahereh

doi:10.1007/s13562-012-0122-5

Cited by 29 publications

(18 citation statements)

References 20 publications

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“…Schijlen et al (2007) found that the RNA interference silencing of CHS gene in tomatoes could cause the reduction of total flavonoid levels to a few percent of those in wild-type ones. Rahnama et al (2013) reported that the overexpression of CHS gene could result in the transgenic Silybum marianum hairy roots containing a 10-fold increase in silybin (an active flavonolignan) content. Moreover, CHS gene expression can be induced by environmental stresses such as fungal pathogen attack, wounding, or UV irradiation, which is capable of triggering the accumulation of flavonoid phytoalexins to improve plant resistance (Dao et al, 2011).…”

Section: Biosynthesis Gene Expression In Ithrcs Underlying Chitosan Ementioning

confidence: 99%

Chitosan elicitation of Isatis tinctoria L. hairy root cultures for enhancing flavonoid productivity and gene expression and related antioxidant activity

Jiao

Gai

Wang

et al. 2018

Industrial Crops and Products

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Elicitation for phytochemical enhancement via cost-effective elicitors can overcome the limitation of commercial application faced by plant cell and organ culture technology. Chitosan is a natural, low-cost, and nontoxic elicitor that can trigger plant defense responses with the concomitant enhancement in phytochemical biosynthesis. In this work, the elicitation of Isatis tinctoria L. hairy root cultures by chitosan was conducted to enhance the production of pharmacologically active flavonoids. In comparison with control (2.31 ± 0.29 mg/g DW), a 7.08-fold enhancement of total flavonoids (16.35 ± 0.88 mg/g DW) was achieved in 24 day-old I. tinctoria hairy root cultures elicited by 150 mg/L chitosan for 36 h. Interestingly, the multiple hydroxyl-substituted flavonoids (rutin, quercetin, isorhamnetin, and isoliquiritigenin) were noticed to increase significantly in chitosan-elicited I. tinctoria hairy root cultures. Moreover, the transcription of associated genes involved in flavonoid biosynthesis pathway was significantly up-regulated underlying chitosan elicitation, among which chalcone synthase and flavonoid 3′-hydroxylase might play an important role in flavonoid enhancement. Additionally, extracts from chitosan-elicited I. tinctoria hairy root cultures exhibited higher antioxidant activities with lower IC 50 values as compared with control. Overall, a cost-effective strategy via the simple chitosan elicitation is provided here to enhance the production of high-added value flavonoids in I. tinctoria hairy root cultures, which paves the way toward the successful commercialization of this in vitro culture system in the future.

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Section: Biosynthesis Gene Expression In Ithrcs Underlying Chitosan Ementioning

confidence: 99%

Chitosan elicitation of Isatis tinctoria L. hairy root cultures for enhancing flavonoid productivity and gene expression and related antioxidant activity

Jiao

Gai

Wang

et al. 2018

Industrial Crops and Products

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“…In addition to verifying the hairy roots based on their morphology, a molecular method such as PCR was performed using rol gene-specific primers. The production of hairy roots in G. glabra, Gmelina arborea, S. marianum, and Plumbago rosea plants has been confirmed using rolB gene-specific primers [20,23,38,39]. The integration of CYP88D6 gene into the hairy root was determined by PCR using CYP88D6-specific primers, showing a 1482-bp band in putative transgenic hairy roots, but not in control samples (Fig.…”

Section: Confirmation Of Co-transformed Hairy Rootsmentioning

confidence: 77%

“…Genomic DNA of hairy roots was extracted by CTAB method [29]. PCR was performed using A. rhizogenes VirD1 F 5′-ATG TCG CAA GGC AGT AAG -3′ and R 5′-CAA GGA GTC TTT CAG CAT G-3′ as negative control to ensure the absence of bacteria in the results [30], rolB F 5′-TTA GGC TTC TTT CAT TCG GTT TAC TGC AGC -3′ and R 5′-ATG GAT CCC AAA TTG CTA TTC CCC ACGA-3′ to determine the integration of T-DNA from Ri plasmid into the roots [20], and CYP88D6 F 5′-ATG GAA GTA CAC TGG GTT TGC-3′ and R 5′-CTA CGC ACA TGA AAC CTT TATC-3′ to confirm the presence of CYP88D6 gene in transgenic hairy roots [25].…”

Section: Pcr Analysis Of Putative Transgenicsmentioning

confidence: 99%

“…A lot of researches have shown the increase in production of secondary metabolites using gene over-expression in transgenic hairy roots. The metabolic engineering approach increased production of flavonolignans through over-expressing petunia chalcone synthase (chsA) gene in hairy root cultures of Silybum marianum [20]. Over-expression of 7-O-acetyltransferase (SalAT) gene was employed as the target for metabolic engineering in Papaver bracteatum hairy root cultures, which led to the enhancement of morphinan alkaloid production [21].…”

Section: Introductionmentioning

confidence: 99%

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Metabolic Engineering of Glycyrrhizin Pathway by Over-Expression of Beta-amyrin 11-Oxidase in Transgenic Roots of Glycyrrhiza glabra

et al. 2018

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Glycyrrhiza glabra is one of the most important and well-known medicinal plants which produces various triterpene saponins such as glycyrrhizin. Beta-amyrin 11-oxidase (CYP88D6) plays a key role in engineering pathway of glycyrrhizin production and converts an intermediated beta-amyrin compound to glycyrrhizin. In this study, pBI121:CYP88D6 construct was transferred to G. glabra using Agrobacterium rhizogene ATCC 15834. The quantitation of transgene was measured in putative transgenic hairy roots using qRT-PCR. The amount of glycyrrhizin production was measured by HPLC in transgenic hairy root lines. Gene expression analysis demonstrated that CYP88D6 was over-expressed only in one of transgenic hairy root lines and was reduced in two others. Beta-amyrin 24-hydroxylase (CYP93E6) was significantly expressed in one of the control hairy root lines. The amount of glycyrrhizin metabolite in over-expressed line was more than or similar to that of control hairy root lines. According to the obtained results, it would be recommended that multi-genes of glycyrrhizin biosynthetic pathway be transferred simultaneously to the hairy root in order to increase glycyrrhizin content.

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“…Although the number of identified diverse type III PKS family of enzymes is growing exponentially, the role of only a few like CHS and STS were extensively studied in transgenic plants. CHS gene was over‐expressed for increasing the production of lignin content in Linumus itatissimum [10], phenolic acids and anthocyanin in potato tubers [11] and flavonolignans in hairy root cultures of Silybum marianum [12]. Moreover, over‐expression of CHS gene in tobacco plants resulted in white flowers [13].…”

Section: Introductionmentioning

confidence: 99%

Over‐expression of bael quinolone synthase in tobacco improves plant vigor under favorable conditions, drought, or salt stress

2014

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a b s t r a c tType III polyketide synthases (PKSs) catalyze the biosynthesis of various medicinally important secondary metabolites in plants, but their role in growth and stress response is unclear. Here, we overexpressed quinolone synthase (QNS) from bael in tobacco. QNS-overexpressing plants showed an overall increase in growth, photosynthetic efficiency and chlorophyll content compared to wild type plants. Second-generation (T 2 ) transgenic plants grew to maturity, flowered early and set viable seeds under favorable conditions without yield penalty. An increased accumulation of flavonoids, phenols and alkaloids was associated with higher tolerance to drought and salinity stress in transgenic plants. Thus, bael QNS seems to function as a positive regulator of plant growth and stress response, and could be potentially used for engineering plants tolerant to abiotic stress.

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Enhanced production of flavonolignans in hairy root cultures of Silybum marianum by over-expression of chalcone synthase gene

Cited by 29 publications

References 20 publications

Chitosan elicitation of Isatis tinctoria L. hairy root cultures for enhancing flavonoid productivity and gene expression and related antioxidant activity

Chitosan elicitation of Isatis tinctoria L. hairy root cultures for enhancing flavonoid productivity and gene expression and related antioxidant activity

Metabolic Engineering of Glycyrrhizin Pathway by Over-Expression of Beta-amyrin 11-Oxidase in Transgenic Roots of Glycyrrhiza glabra

Over‐expression of bael quinolone synthase in tobacco improves plant vigor under favorable conditions, drought, or salt stress

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