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Indirect regeneration of Withania somnifera and comparative analysis of withanolides in in vitro and greenhouse grown plants

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Biologia Plantarum

Abstract

The present study reports an efficient protocol for indirect shoot organogenesis and plantlets regeneration of Withania somnifera (L.) Dunal. Leaf explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of 6-benzylaminopurine (BAP) and indole-3-acetic acid (IAA). The highest callus induction rate (89.5 %) and shoot regeneration rate (92 %) were obtained when 2 mg dm−3 BAP was combined with 0.5 mg dm−3 IAA. Three major withanolides (withaferine A, 12-deoxywithastramonolide and withanolide A) were investigated in different plant organs from in vitro and greenhouse grown plants. Leaves contained higher contents of withanolides and phenolics than roots or stems, whereas roots contained the highest contents of flavonoids and polysacharides. In vitro grown plants contained greater contents of phenolics, flavonoids and polysaccharides while lower contents of withanolides than greenhouse grown plants.

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Abbreviations

BAP:

6-benzylaminopurine

DPPH:

1,1-diphenyl-2-picrylhydrazyl

HPLC:

high-pressure liquid chromatography

IAA:

indole-3-acetic acid

LC-MS:

liquid chromatography-mass spectrometry

TLC:

thin-layer chromatography

References

  • Banerjee, S., Naqvi, A.A., Mandal, S., Ahuja, P.S.: Transformation of Withania somnifera (L.) Dunal by Agrobacterium rhizogenes: infectivity and phytochemical studies. — Phytother. Res. 8: 452–455, 1994.

    Article  CAS  Google Scholar 

  • Bessalle, R., Lavie, D.: Semi-quantitative reversed-phase high-performance liquid chromatographic analysis of the ecotypes of Withania somnifera chemotype III. — J. Chromatogr. 389: 195–210, 1987.

    Article  CAS  Google Scholar 

  • Carlo, G.D., Mascolo, N., Izzo, A.A., Capasso, F.: Flavonoids: old and new aspects of a class of natural therapeutic drugs. — Life Sci. 65: 337–353, 1999.

    Article  PubMed  Google Scholar 

  • Folin, O., Ciocalteu, V.: On tyrosine and tryptophane determination in proteins. — J. biol. Chem. 27: 627–650, 1927.

    Google Scholar 

  • Ganzera, M., Choudhary, M.I., Khan, I.A.: Quantitative HPLC analysis of withanolides in Withania somnifera. — Fitoterapia 74: 68–76, 2003.

    Article  CAS  PubMed  Google Scholar 

  • Gupta, A.P., Verma, R.K., Misra, H.O., Gupta, M.M.: Quantitative determination of withaferin A in different plant parts of Withania somnifera by TLC densitometry. — J. med. aromat. Plant. Sci. 18: 788–790, 1996.

    CAS  Google Scholar 

  • Hatano, T., Kagawa, H., Yasuhara, T., Okuda, T.: Two new flavonoids and other constituents in licorice: their relative astringency and radical scavenging effects. — Chem. Pharm. Bull. 36: 2090–2097, 1988.

    CAS  PubMed  Google Scholar 

  • Hunter, I.R., Walden, M.K., Heftmann, E., Glotter, E., Kirson, I.: Separation of withanolides by high-pressure liquid chromatography with coiled columns. — J. Chromatogr. 170: 437–442, 1979.

    Article  CAS  Google Scholar 

  • Jia, Z.-S., Tang, M.-C., Wu, J.-M.: The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. — Food Chem. 64: 555–559, 1999.

    Article  Google Scholar 

  • Jiang, Y.H., Jiang, X.L., Wang, P., Hu, X.K.: In vitro antioxidant activities of water-soluble polysaccharides extracted from Isaria farinosa B05. — J. Food Biochem. 29: 323–335, 2005.

    Article  CAS  Google Scholar 

  • Kulkarni, A.A., Thengane, S.R., Krishnamurthy, K.V.: Direct in vitro regeneration of leaf explants of Withania somnifera (L.) Dunal. — Plant Sci. 119: 163–168, 1996.

    Article  CAS  Google Scholar 

  • Kulkarni, A.A., Thengane, S.R., Krishnamurthy, K.V.: Direct shoot regeneration from node, internode, hypocotyl and embryo explants of Withania somnifera. — Plant Cell Tissue Organ Cult. 62: 203–209, 2000.

    Article  Google Scholar 

  • Manickam, V.S., Elango-Mathavan, R., Antonisamy, R.: Regeneration of Indian ginseng plantlets from stem callus. — Plant Cell Tissue Organ Cult. 62: 181–185, 2000.

    Article  Google Scholar 

  • Murashige, T., Skoog, F.: A revised medium for rapid growth and bioassay with tobacco tissue culture. — Physiol. Plant. 15: 473–495, 1962.

    Article  CAS  Google Scholar 

  • Rice-Evans, C.A., Miller, N.J., Pagana, G.: Structure-antioxidant activity relationship of flavonoids and phenolic acids. — Free Radical Biol. Med. 20: 933–956, 1996.

    Article  CAS  Google Scholar 

  • Roja, C.l., Heble, M.R., Sipahimalani, A.T.: Tissue cultures of Withania somnifera: morphogenesis and withanolide synthesis. — Phytother. Res. 5: 185–187, 1991.

    Article  CAS  Google Scholar 

  • Sen, J., Sharma, A.K.: Micropropagation of Withania somnifera from germinating seeds and shoot tip. — Plant Cell Tissue Organ Cult. 26: 71–73, 1991.

    Article  CAS  Google Scholar 

  • Vishnoi, A., Babbar, S.B., Gupta, S.C.: Induction of androgenesis in anther cultures of Withania somnifera. — Z. Pflanzenphysiol 94: 169–171, 1979.

    CAS  Google Scholar 

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Acknowledgements

This work was supported by the Korea Science and Engineering Foundation (KOSEF) grant funded by the Korea government (MOST) (R01-2007-000-10543-0).

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Authors and Affiliations

  1. Department of Horticulture, Faculty of Agriculture, Kafrelsheikh University, Kafr El-Sheikh, 33516, Egypt

    Y. H. Dewir

  2. Floriculture Section, National Botanical Research Institute, Lucknow, India

    D. Chakrabarty

  3. Research Center for The Development of Advanced Horticultural Technology, Chungbuk National University, Cheong-Ju, 361-763, Korea

    S. -H. Lee, E. -J. Hahn & K. -Y. Paek

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  1. Y. H. Dewir
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  2. D. Chakrabarty
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  3. S. -H. Lee
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  4. E. -J. Hahn
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  5. K. -Y. Paek
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Correspondence to Y. H. Dewir.

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Dewir, Y.H., Chakrabarty, D., Lee, S.H. et al. Indirect regeneration of Withania somnifera and comparative analysis of withanolides in in vitro and greenhouse grown plants. Biol Plant 54, 357–360 (2010). https://doi.org/10.1007/s10535-010-0063-6

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  • DOI: https://doi.org/10.1007/s10535-010-0063-6

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