Enhanced production of withaferin-A in shoot cultures of Withania somnifera (L) Dunal

Mir, Bilal Ahmad; Khazir, Jabeena; Hakeem, Khalid Rehman; Koul, Sushma; Cowan, Don A.

doi:10.1007/s13562-014-0264-8

Cited by 18 publications

(21 citation statements)

References 11 publications

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“…The medicinal properties of W. somnifera have been attributed to withanolides synthesized in the leaves and roots. In recent years, there is an increasing demand of these bioactive withanolides in pharmaceutical industry (Mir et al, 2012(Mir et al, , 2014a. However, the composition and quality of withanolides shows great variability in plants growing in different environmental conditions (Kumar et al, 2011;Mir et al, 2014a).…”

Section: Introductionmentioning

confidence: 99%

Cold stress affects antioxidative response and accumulation of medicinally important withanolides in Withania somnifera (L.) Dunal

Mir

Khazir

et al. 2015

Industrial Crops and Products

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Withania somnifera (L.) Dunal (Indian ginseng) is a high value medicinal plant. It synthesizes a large array of biologically active withanolides. In this study, two month old seedlings of AGB002 (wild genotype) and AGB025 (cultivated genotype) of W. somnifera were subjected to cold stress (4 • C) under controlled envi-ronment. Plants were analyzed for three medicinally important secondary metabolites (withanolide A, withanone and withaferin A), lipid peroxidation (MDA), cell injury, superoxide radical (O 2 •− ) accumula-tion and anti-oxidative enzymes activities such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR). Increases in the titers of superoxide anion and MDA were observed from day 1 to day 7 in both genotypes, although the increase on the first day of exposure was significantly higher. Enzymatic activities of SOD, CAT, APX and GR also showed an increasing trend in both genotypes and reached a maximum on day 7 of the cold temperature exposure; however, this increase was higher in AGB002 than AGB025. Withanolide A (WS-1) in the roots of both genotypes sig-nificantly decreased on the first day of cold exposure and then showed a recovery until day 7. WS-1 was not detected in the leaves of either genotype. Withanone (WS-2) content in the leaves also decreased towards the end of the cold period in both genotypes. Cold stress also elicited the accumulation of WS-2 in AGB025, but was not detectable in control seedlings. At maturity, WS-2 was also detected in control plants. Furthermore, a significant increase in the leaf withaferin A (WS-3) content was recorded from day 1 to day 7 of the cold exposure in both the genotypes, suggesting the possible involvement of with-anolides in cold-protection. AGB002 showed comparatively higher accumulation of antioxidant enzymes and selected marker withanolides than AGB025, indicating that AGB002 is better adapted to cold than AGB025. It could be inferred from these observations that cold stress induces bioactive withanolide accu-mulation in W. somnifera as a mechanism for scavenging reactive oxygen species (ROS). These studies also provide an impetus for enhancing the withanolide accumulation in W. somnifera using controlled environment technology.

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Section: Introductionmentioning

confidence: 99%

Cold stress affects antioxidative response and accumulation of medicinally important withanolides in Withania somnifera (L.) Dunal

Mir

Khazir

et al. 2015

Industrial Crops and Products

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“…Thereafter, chloroform was distilled off under reduced pressure. The residue was weighed and dissolved in HPLC grade methanol (Ranbaxy Fine Chemicals) and subjected to HPLC for detection and quantification of WA as described previously (Mir et al 2014a). HPLC was carried out on a Shimadzu HPLC system consisting of a Pump LC-10ATVP, an automatic sampling unit (auto-sampler), SIL-10ADVP, a column oven CTO-10ASVP, a diode array detector, SPD-M10AVP and SCL-10AVP Version 5.40 (Shimadzu).…”

Section: Withaferin a Extraction And Quantificationmentioning

confidence: 99%

“…Cytokinins are a class of plant growth substances that are active in promoting cell division as well as being active in numerous physiological processes, including plant cell growth and differentiation (Mahesh and Jeyachandran, 2013). The stimulating effect of BA alone or in combination with Kn on bud breaking and in vitro multiple shoot formation has been reported in W. somnifera (Ray and Jha, 2001;Sharada et al 2007;Ahuja et al 2009;Dewir et al 2010;Mir et al 2014a), Trichodesma indicum (Mahesh and Jeyachandran, 2013) and Drimia robusta (Baskaran et al 2013) and many other species (Kumar et al 2005;Baskaran et al 2012;Thiem et al 2013). The positive effect of BA was observed only unto a threshold of 5.0 μM as the further increase in the concentration of BA alone or in combination with Kn did not elicit any greater response in the parameters studied.…”

Section: Shoot Proliferation and Establishmentmentioning

confidence: 99%

“…The quality and quantity of withanolide constituents are highly affected by genotype and environmental conditions. In this scenario, plant cell and organ culture of an elite genotype is an attractive alternative for large-scale propagation and for the production of bioactive withanolide (Mir et al 2014a). It is the most suited approach for the conservation and commercial propagation of rare and endangered medicinal species to reduce the dependence on the natural habitat for the supply of raw drugs (Baskaran et al 2013;Mir et al 2014a).…”

Section: Introductionmentioning

confidence: 99%

“…Plant tissue culture could also provide a means of disease free healthy clones for extraction of pure drugs molecules. It also offer an elegant alternative for homogeneous, controlled production of metabolites, throughout the year, especially when we take commercial demand into account (Gawde and Paratkar, 2012;Mir et al 2014a). Further in vitro propagation through shoot tip and nodal explants is an easy and economic way for obtaining large number of consistently uniform and true-to-type plants.…”

Section: Introductionmentioning

confidence: 99%

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In vitro propagation and withaferin A production in Withania ashwagandha, a rare medicinal plant of India

Mir¹,

Mir²,

Koul³

2014

Physiol Mol Biol Plants

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Withania ashwagandha, belonging to the family Solanaceae, is an important medicinal herb of India with restricted geographic distribution. It is a rich source of withaferin A (WA) and other bioactive withanolides. In the present study a rapid in vitro mass propagation protocol of W. ashwagandha was developed from nodal explants. Nodal explants were cultured on MS medium supplemented with various concentrations and combinations of plant growth regulators (PGRs). The highest number of regenerated shoots per ex-plant (33±2.7) and highest WA (13.4±1.15 mg/g of DW) production was obtained on MS medium supplemented with 5.0 μM 6-benzyladenine (BA) and 1.0 μM Kinetin (Kn). In vitro raised shoots were further rooted on half-strength MS medium containing 2.0 μM Indole-3-butyric acid (IBA) and analyzed for WA production. The rooted plantlets when transferred to poly bags in the greenhouse showed 90 % survival frequency. Levels of WA were higher in the in vitro and ex vitro derived shoot and root tissues as compared to field grown mother plants. In an attempt to further maximize WA production, shoot cultures were further grown in liquid MS medium supplemented with 5.0 μM 6-benzyladenine (BA) and 1.0 μM Kinetin (Kn). Root cultures were grown on half strength MS liquid medium fortified with 2.0 μM of IBA. WA production in the liquid cultures was significantly higher compared to the static composition of the same media. This protocol, first of its kind in this plant, can be successfully employed for conservation, proliferation and large-scale production of WA. The regenerated plants can also be used in traditional medicine as an alternative to naturally collected plants.

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Metabolite Profiling in Withania somnifera Roots Hydroalcoholic Extract Using LC/MS, GC/MS and NMR Spectroscopy

Trivedi¹,

Panda²,

Sethi³

et al. 2017

Chemistry & Biodiversity

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Ashwagandha (Withania somnifera) is a very well-known herbal medicine and it was well studied for its active metabolites throughout the World. Although, nearly 40 withanolides were isolated from W. somnifera root extract, still there is remaining unidentified metabolites due to very low abundance and geographical variation. Advanced separation technology with online identification by mass and nuclear magnetic resonance (NMR) are nowadays used to find out the new compounds in the crude herbal extract. This article described the metabolite profiling of ashwagandha root hydroalcoholic extract using ultra-performance liquid chromatography coupled with a positive ion electrospray ionization tandem mass spectrometry through gas chromatography mass spectrometry (GC/MS) and NMR spectroscopy. A total of 43 possible withanolides was identified and proposed their structures based on the mass of molecular and fragment ions. GC/MS and NMR analysis indicated the presence of several known withanolides including withaferin A, withanolide D, withanoside IV or VI, withanolide sulfoxide, etc. To the best of our knowledge, dihydrowithanolide D at m/z 473 (t 7.86 min) and ixocarpalactone A at m/z 505 (t 8.43 min) were first time identified in the ashwagandha root hydroalcoholic extract. The current study that described the identification of withanolides with summarized literature review might be helpful for designing the experiment to identify of the new chemical constituents in Withania species.

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Enhanced production of withaferin-A in shoot cultures of Withania somnifera (L) Dunal

Cited by 18 publications

References 11 publications

Cold stress affects antioxidative response and accumulation of medicinally important withanolides in Withania somnifera (L.) Dunal

Cold stress affects antioxidative response and accumulation of medicinally important withanolides in Withania somnifera (L.) Dunal

In vitro propagation and withaferin A production in Withania ashwagandha, a rare medicinal plant of India

Metabolite Profiling in Withania somnifera Roots Hydroalcoholic Extract Using LC/MS, GC/MS and NMR Spectroscopy

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