Enhanced production of tropane alkaloids in transgenic Scopolia parviflora hairy root cultures over-expressing putrescine N-methyl transferase (PMT) and hyoscyamine-6β-hydroxylase (H6H)

Kang, Young Min; Park, D. J.; Jiang, Min; Song, Hye Young; Jeong, Mi Jin; Kim, Y. D.; Kang, Shin-Min; Karigar, Chandrakant S.; Choi, Myung-Suk

doi:10.1007/s11627-011-9367-2

Cited by 20 publications

(5 citation statements)

References 23 publications

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“…Overexpression of PMT and H6H in Hyoscyamus niger and Atropa belladonna in transgenic hairy root cultures [65,66] and expressional analysis of H6H gene from Hyoscyamus niger and Hyoscyamus tenuicaulis [68] has been reported. Our study is also in agreement with the work carried out regarding the expressional analysis of PMT and H6H genes in Scopolia parviflora [67].…”

Section: Plos Onesupporting

confidence: 92%

“…and scopolamine concentrations in Hyoscyamus niger [65], Atropa belladonna [66], Scopolia parviflora [67]. Our study showed a maximum increase of 10.3 and 4.8 fold in the expression of H6H and PMT, respectively, at 0.03% EMS to control, suggesting that EMS at low concentration leads to an increase in expression, whereas the decrease in expression is seen with a higher concentration of EMS.…”

Section: Plos Onementioning

confidence: 43%

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Promoting the accumulation of scopolamine and hyoscyamine in Hyoscyamus niger L. through EMS based mutagenesis

et al. 2020

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The overexploitation of medicinal plants is depleting gene pool at an alarming rate. In this scenario inducing the genetic variability through targeted mutations could be beneficial in generating varieties with increased content of active compounds. The present study aimed to develop a reproducible protocol for in vitro multiplication and mutagenesis of Hyoscyamus niger targeting putrescine N-methyltransferase (PMT) and 6β-hydroxy hyoscyamine (H6H) genes of alkaloid biosynthetic pathway. In vitro raised callus were treated with different concentrations (0.01%-0.1%) of Ethyl Methane Sulfonate (EMS). Emerging multiple shoots and roots were obtained on the MS media supplemented with cytokinins and auxins. Significant effects on morphological characteristics were observed following exposure to different concentrations of EMS. EMS at a concentration of 0.03% was seen to be effective in enhancing the average shoot and root number from 14.5±0.30 to 22.2 ±0.77 and 7.2±0.12 to 8.8±0.72, respectively. The lethal dose (LD 50) dose was calculated at 0.08% EMS. The results depicted that EMS has an intense effect on PMT and H6H gene expression and metabolite accumulation. The transcripts of PMT and H6H were significantly upregulated at 0.03-0.05% EMS compared to control. EMS treated explants showed increased accumulation of scopolamine (0.639 μg/g) and hyoscyamine (0.0344μg/g) compared to untreated.

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Section: Plos Onesupporting

confidence: 92%

Section: Plos Onementioning

confidence: 43%

Promoting the accumulation of scopolamine and hyoscyamine in Hyoscyamus niger L. through EMS based mutagenesis

et al. 2020

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“…Combining the overexpression of both PMT and H6H in A. belladonna has resulted in approximately a 2.5-fold increase of scopolamine above wild-type levels (Liu et al, 2010). This growth increase could be attributed to the role of polyamines in the processes of growth and cell differentiation (Kang et al, 2011). Using the plant Scopolia parviflora, a similar construct using PMT and H6H was used to make transgenic hairy roots.…”

Section: Metabolic Engineering Of Tropane Alkaloidsmentioning

confidence: 99%

Increasing the Pace of New Discoveries in Tropane Alkaloid Biosynthesis

Jirschitzka

Dolke

D’Auria

2013

New Light on Alkaloid Biosynthesis and Future Prospects

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“…Overexpression of geraniol 10-hydroxylase (G10H) and a jasmonate-responsive transcript factor (ORCA3) in the HR of Catharanthus roseus improves catharanthine production, another valuable TA [60]. More recently, Kang and coworkers [61] using the same strategy, co-expression of pmt and h6h, obtained transgenic HR lines of Scopolia parviflora. Consequently to the introduction of these key enzyme genes, production of the alkaloids hyoscyamine and scopolamine was enhanced.…”

Section: Overexpression or Co-expression Of Enzymes Involved In The B...mentioning

confidence: 99%

Hairy Roots, their Multiple Applications and Recent Patents

Talano

Oller²,

González³

et al. 2012

BIOT

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In the last years, hairy root (HR) cultures are gaining attention in the biotechnology industry. This particular plant cell culture derives from explants infected with Agrobacterium rhizogenes. They constitute a relatively new approach to in vitro plant biotechnology and modern HR cultures are far away from the valuables findings performed by Philip R. White in the 1930's, who obtained indefinite growth of excised root tips. HR cultures are characterized by genetic and biochemical stability and high growth rate without expensive exogenous hormones source. HR cultures have allowed a deep study of plant metabolic pathways and the production of valuable secondary metabolites and enzymes, with therapeutic or industrial application. Furthermore, the potential of HR cultures is increasing continuously since different biotechnological strategies such as genetic engineering, elicitation and metabolic traps are currently being explored for discovery of new metabolites and pathways, as well as for increasing metabolites biosynthesis and/or secretion. Advances in design of proper bioreactors for HR growth are being of great interest, since scale up of metabolite production will allow the integration of this technology to industrial processes. Another application of HR cultures is related to their capabilities to biotransform and to degrade different xenobiotics. In this context, removal assays using this plant model system are useful tools for phytoremediation assays, previous to the application in the field. This review highlights the more recent application of HRs and those new patents which show their multiple utilities.

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Enhanced production of tropane alkaloids in transgenic Scopolia parviflora hairy root cultures over-expressing putrescine N-methyl transferase (PMT) and hyoscyamine-6β-hydroxylase (H6H)

Abstract: Scopolia parviflora adventitious roots were metabolically engineered by co-expression of the two gene putrescine N-methyl transferase (PMT) and hyoscyamine-

Cited by 20 publications

References 23 publications

Promoting the accumulation of scopolamine and hyoscyamine in Hyoscyamus niger L. through EMS based mutagenesis

Promoting the accumulation of scopolamine and hyoscyamine in Hyoscyamus niger L. through EMS based mutagenesis

Increasing the Pace of New Discoveries in Tropane Alkaloid Biosynthesis

Hairy Roots, their Multiple Applications and Recent Patents

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