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Qualitative and quantitative variations in withanolides and expression of some pathway genes during different stages of morphogenesis in Withania somnifera Dunal

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Abstract

Withania somnifera Dunal is an important and extensively studied medicinal plant; however, there is no report available that relates withanolide content and its profile in relation to the expression of pathway genes during different morphogenic stages. In this study, withanolide A, withaferin A, and withanone, the major withanolides of W. somnifera, were measured in different in vitro stages during organogenesis, viz., shoot to root (direct rhizogenesis)/root to shoot (indirect via callus phase) transition vis-à-vis expression levels of key pathway genes involved in withanolide biosynthetic pathways. The morphogenic transitions were found to be tightly linked to the pattern of accumulation of withanolides. The high expression levels of most of the pathway genes in in vitro shoots in comparison to in vitro root and callus tissues exhibited a direct co-relation with the maximum withanolide content (>2.7 mg/gDW). The biogenesis of withaferin A, a major constituent of the leaves, was however found to be tightly linked to shoots/green tissue. In addition, we were also able to establish an efficient regeneration system from roots for their further utilization in biotechnological applications.

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Abbreviations

IAA:

Indole-3-acetic acid

IBA:

Indole-3-butyric acid

NAA:

α-Naphthalene acetic acid

2, 4-D:

2,4-Dichlorophenoxy acetic acid

TDZ:

Thidiazuran

HPLC:

High-performance liquid chromatography

DW:

Dry weight

FW:

Fresh weight

MS:

Murashige and Skoog (1962)

HMGR:

Hydroxymethylglutaryl Co-A reductase

FPPS:

Farnesyldiphosphate synthase

SS:

Squalene synthase

SE:

Squalene epoxidase

CAS:

Cycloartenol synthase

GT:

Glucosyl transferase

References

  • Bhojwani SS, Razdan MK (2004) Plant tissue culture: theory and practice, a revised edition. North Holland, Amsterdam

    Google Scholar 

  • Chatterjee S, Srivastava S, Khalid A, Singh N, Sangwan RS, Sidhu OP, Roy R, Khetrapal CL, Tuli R (2010) Comprehensive metabolic fingerprinting of Withania somnifera leaf and root extracts. Phytochemistry 71:1085–1094

    Article  PubMed  CAS  Google Scholar 

  • Chaurasiya ND, Gupta VK, Sangwan RS (2007) Leaf ontogenic phase-related dynamics of withaferin A and withanone biogenesis in Ashwagandha (Withania somnifera Dunal) an important medicinal herb. J Plant Biol 50:508–513

    Article  Google Scholar 

  • Chaurasiya ND, Sangwan RS, Mishra LN, Tuli R, Sangwan NS (2009) Metabolic clustering of a core collection of Indian ginseng Withania somnifera Dunal through DNA, isoenzyme, polypeptide and withanolide profile diversity. Fitoterapia 80:496–505

    Article  PubMed  CAS  Google Scholar 

  • Chaurasiya ND, Sangwan NS, Sabir F, Misra LN, Sangwan R S (2012) Withanolide biosynthesis recruits both mevalonate and DOXP pathways of isoprenogenesis in Ashwagandha Withania somnifera L. (Dunal). Plant Cell Reports. doi:10.1007/s00299-012-1302-4

  • Choi D, Ward BL, Bostock RM (1992) Differential induction and suppression of potato 3-hydroxy-3-methylglutaryl coenzyme A reductase genes in response to Phytophthora infestans and to its elicitor. Plant Cell 4(10):1333–1344

    PubMed  CAS  Google Scholar 

  • Dale S, Arro M, Becerra B, Morrice NG, Boronat A, Hardie DG, Ferrer A (1995) Bacterial expression of the catalytic domain of 3-hydroxy-3-methylglutaryl-CoA reductase (isoform HMGR1) from Arabidopsis thaliana, and its inactivation by phosphorylation at Ser577 by Brassica oleracea 3-hydroxy-3-methylglutaryl-CoA reductase kinase. Eur J Biochem 233(2):506–513

    Article  PubMed  CAS  Google Scholar 

  • Facchini P (2001) Alkaloid biosynthesis in plants: biochemistry, cell biology, molecular egulation and metabolic engineering applications. Annu Rev Plant Mol Biol 52:29–66

    Article  CAS  Google Scholar 

  • Franklin G, Sheeba CJ, Lakshmisita G (2004) Regeneration of eggplant (Solanum Melongena L.) from root explants. In Vitro Cell Dev Biol Plant 40:188–191

    Article  Google Scholar 

  • George EF (ed) (1993) Plant propagation by tissue culture, 2nd edn. Part 1. The technology. Exegetics, Wilts

  • Gupta P, Akhtar N, Tewari SK, Sangwan RS, Trivedi PK (2011) Differential expression of farnesyl diphosphate synthase gene from Withania somnifera in different chemotypes and in response to elicitors. Plant Growth Regul 65:93–100

    Article  CAS  Google Scholar 

  • Gupta P, Agarwal AV, Akhtar N, Sangwan RS, Singh SP, Trivedi PK (2012) Cloning and characterization of 2-C-methyl-D-erythritol-4-phosphate pathway genes for isoprenoid biosynthesis from Indian ginseng. Protoplasma. doi:10.1007/s00709-012-0410-x

  • Hosseini-Nasr M, Rashid A (2002) Thidiazuron-induced shoot-bud formation on root segments of Albizzia julibrissin is an apex-controlled, light-independent and calcium-mediated Response. Plant Growth Regu 36:81–85

    Article  CAS  Google Scholar 

  • Howell SH (1998) Molecular genetics of plant development. Cambridge University Press, Cambridge, UK

  • Huetteman CA, Preece JE (1993) Thidiazuron:a potent cytokinins for woody plant tissue culture. Plant Cell Tiss Org Cult 33:105–119

    Article  CAS  Google Scholar 

  • Manickam VS, Mathavan RE, Antonisamy R (2000) Regeneration of Indian ginseng plantlets from stem callus. Plant Cell, Tissue Org Cult 62:181–185

    Article  Google Scholar 

  • Martin KP, Zhang CL, Hembrom ME, Slater A, Madassery J (2008) Adventitious root induction in Ophiorrhiza prostrata: a tool for the production of camptothecin (an anticancer drug) and rapid propagation. Plant Biotech Rep 2:163–169

    Article  Google Scholar 

  • Mirjalili MH, Moyano E, Bonfill M, Cusido RM, Palazon J (2009) Streoidal lactones from Withania somnifera, an ancient plant for novel medicine. Molecules 14:2373–2393

    Article  PubMed  CAS  Google Scholar 

  • Mirjalili MH, Moyano E, Bonfill M, Cusido RM, Palazón J (2011) Overexpression of the Arabidopsis thaliana squalene synthase gene in Withania coagulans hairy root cultures. Biol Plant 55:357–360

    Article  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Nagella P, Murthy HN (2010) Production of withanolide-A from adventitious root cultures of Withania somnifera. Acta Physiol Plant 32:1017–1022

    Article  Google Scholar 

  • Rani G, Virk GS, Nagpal A (2003) Callus induction and plantlet regeneration in Winthania somnifera (L.) Dunal. In Vitro Cell Dev Biol Plant 39:468–474

    Google Scholar 

  • Reed RC, Brady SR, Muday GK (1998) Inhibition of auxin movement from the shoot into the root inhibits lateral root development in Arabidopsis. Plant Physiol 118:1369–1378

    Article  PubMed  CAS  Google Scholar 

  • Sabir F, Sangwan NS, Chaurasiya ND, Misra LN, Sangwan RS (2008) In vitro withanolide production by Withania somnifera L. cultures. Z Naturfor 63c:409–413

    Google Scholar 

  • Sabir F, Sangwan RS, Singh J, Misra LN, Pathak N, Sangwan NS (2011) Biotransformation of withanolides by cell suspension cultures of Withania somnifera (Dunal). Plant Biotech Rep 5:127–134

    Article  Google Scholar 

  • Sangwan RS, Chaurasia ND, Mishra LN, Lal P, Uniyal GC, Sharma R, Sangwan NS, Suri KA, Quazi GN, Tuli R (2004) Phytochemical variability in commercial herabal products and preparation of Withania somnifera (Ashwagandha). Curr Sci 86:461–465

    CAS  Google Scholar 

  • Sangwan RS, Chaurasiya ND, Misra LN, Lal P, Uniyal GC, Sangwan NS, Srivastava AK, Suri KA, Qazi GN, Tuli R (2005) An improved process for isolation of withaferin-A from plant materials and products therefrom. US Patent 7,108,870

  • Sangwan RS, Chaurasiya ND, Lal P, Misra L, Uniyal GC, Tuli R, Sangwan NS (2007) Withanolide A biogeneration in in vitro shoot cultures of Ashwagandha (Withania somnifera Dunal), a main medicinal plant in Ayurveda. Chem Pharm Bull 55:1371–1375

    Article  PubMed  CAS  Google Scholar 

  • Sangwan RS, Chaurasiya ND, Sangwan PL, Misra LN, Tuli R, Sangwan NS (2008) Withanolide A is inherently de novo biosynthesized in roots of the medicinal plant Ashwagandha (Withania somnifera). Physiol Plant 133:278–287

    Article  PubMed  CAS  Google Scholar 

  • Sugimoto K, Jiao Y, Meyerowitz EM (2010) Arabidopsis regeneration from multiple tissues occurs via a root development pathway. Develop Cell 18:3463–3471

    Article  Google Scholar 

  • Tuli R., Sangwan R S, Kumar S, Bhattacharya S, Misra LN, Trivedi PK, Tewari SK, Misra P, Chaturvedi P, Sangwan NS, Nair KN, Ojha SK, Mehrotra S, Khajuria A, Suri KA (2009) Ashwagandha (Withania somnifera) A model Indian Medicinal Plant. CSIR Publications pp 294.

  • Verdeil JL, Alemanno L, Niemenak N, Tranbarger TJ (2007) Pluripotent versus totipotent plant stem cells: dependence versus autonomy? Trends Plant Sci 12:245–252

    Article  PubMed  CAS  Google Scholar 

  • Wadegaonkar PA, Bhagwat KA, Rai MK (2006) Direct rhizogenesis and establishment of fast growing normal root organ culture of Withania somnifera Dunal. Plant Cell Tissue Org Cult 84:223–225

    Article  Google Scholar 

  • Widodo N, Kaur K, Shrestha BG, Takagi Y, Ishii T, Wadhwa R, Kaul SC (2007) Selective killing of cancer cells by leaf extract of Ashwgandha: identification of a tumor inhibitory factor and the first molecular insights to its effect. Clin Cancer Res 13:2298–2306

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The financial grant under NMITLI Program of Council of Scientific and Industrial Research, Government of India, New Delhi, is gratefully acknowledged. FS is thankful to CSIR for the award of senior research fellowship.

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Authors and Affiliations

  1. Central Institute of Medicinal and Aromatic Plants (CIMAP-CSIR), P.O. CIMAP, Lucknow, 226015, India

    Farzana Sabir, Smrati Mishra, Rajender S. Sangwan, Jyoti S. Jadaun & Neelam S. Sangwan

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  1. Farzana Sabir
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  2. Smrati Mishra
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  3. Rajender S. Sangwan
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  4. Jyoti S. Jadaun
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  5. Neelam S. Sangwan
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Correspondence to Neelam S. Sangwan.

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Sabir, F., Mishra, S., Sangwan, R.S. et al. Qualitative and quantitative variations in withanolides and expression of some pathway genes during different stages of morphogenesis in Withania somnifera Dunal. Protoplasma 250, 539–549 (2013). https://doi.org/10.1007/s00709-012-0438-y

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  • DOI: https://doi.org/10.1007/s00709-012-0438-y

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