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Planta Med 2001; 67(5): 432-436
DOI: 10.1055/s-2001-15811
Original Paper
Biochemistry, Physiology, in vitro cultures
© Georg Thieme Verlag Stuttgart · New York

Production of Withaferin A in Shoot Cultures of Withania somnifera

Swagata Ray, Sumita Jha*
  • Centre of Advanced Study, Department of Botany, Calcutta University, Calcutta, India
Further Information

Publication History

June 30, 2000

October 29, 2000

Publication Date:
31 December 2001 (online)

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Abstract

Multiple shoot cultures of Withania somnifera were established from single shoot tip explants and their potential for the production of two principle withanolides, withaferin A and withanolide D was investigated. Shoot tips grown on MS medium supplemented with BA (1 mg l-1) induced 10.0 ± 1.15 microshoots per explants and shoot cultures accumulated both withanolides (withaferin A = 0.04 %, withanolide D = 0.06 %). Supplementation of MSSM (solid) agar medium with 4 % sucrose enhanced accumulation of both withaferin A (0.16 %) and withanolide D (0.08 %). Reduction of the agar concentration to 0.16 % increased the number of microshoots induced per explant to 25.5. MSSM liquid medium containing 10 % coconut milk favoured a maximum increase in biomass (27 fold); number of microshoots induced (37.6 ± 1.45) as well as accumulation of withaferin A (0.14 %).

Abbreviations

BA:N 6-Benzyladenine

Kn:Kinetin

2iP:N 6-[2-Isopentenyl]adenine

MS:Murashige and Skoog (1962)

MSSM:Murashige and Skoog's basal medium + BA (1 mg l-1)

Key words

Withania somnifera - Solanaceae - shoot cultures - withaferin A - withanolide D

References

  • 1 Chopra R N, Chopra I C, Handa K L, Kapur L D. In: Indigenous Drugs of India. Calcutta, India; U N Dhar and Sons 1958: 436-7
    Search in Google Scholar
  • 2 Ray A B, Gupta M. Withasteroids, a growing group of naturally occurring steroidal lactones. In: Herz W, Kirby GW, Moore RE, Steglich WT, editors Progress in the Chemistry of Organic Natural Products. 1994 63: 1-106
    Search in Google Scholar
  • 3 Chakraborti S K, De B K, Bandyopadhyay T. Variations in the antitumor constituents of Withania somnifera .  Experientia. 1974;  30 852-3
    PubMedSearch in Google Scholar
  • 4 Menssen H G, Stapel G. C28-steroidal lactone from the roots of Withania somnifera .  Planta Medica. 1973;  24 8
    PubMedSearch in Google Scholar
  • 5 Das H, Dutta S K, Bhattacharya B, Chakraborti S. Antineoplastic agents from plants; antitumor activity of withanolide D.  Indian Journal of Cancer Chemotherapy. 1985;  7 59-65
    PubMedSearch in Google Scholar
  • 6 Roja G, Heble M R, Sipahimalani A T. Tissue cultures of Withania somnifera: morphogenesis and withanolide synthesis.  Phytotherapy Research. 1991;  5 185-7
    PubMedSearch in Google Scholar
  • 7 Yu P LC, El-Olemy M M, Stohs S J. A phytochemical investigation of Withania somnifera tissue cultures.  Lloydia. 1974;  37 (4) 593-7
    PubMedSearch in Google Scholar
  • 8 Sen J, Sharma A K. Micropropagation of Withania somnifera from germinated seeds and shoot tip.  Plant Cell, Tissue and Organ Culture. 1991;  26 71-3
    CrossrefPubMedSearch in Google Scholar
  • 9 Kulkarni A A, Thengane S R, Krishnamurthy K V. Direct in vitro regeneration of leaf explants of Withania somnifera (L) Dunal.  Plant Science. 1996;  119 163-8
    CrossrefPubMedSearch in Google Scholar
  • 10 Ray S, Jha S. Withanolide synthesis in cultures of Withania somnifera transformed with Agrobacterium tumefaciens .  Plant Science. 1999;  146 1-7
    CrossrefPubMedSearch in Google Scholar
  • 11 Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures.  Physiologia Plantarum. 1962;  15 473-97
    PubMedSearch in Google Scholar
  • 12 Gamborg O L, Miller R A, Ojima K. Nutrient requirements of suspension cultures of soybean root cells.  Experimental Cell Research. 1968;  50 151-8
    CrossrefPubMedSearch in Google Scholar
  • 13 Schenk R U, Hildebrandt A C. Medium and techniques for induction and growth of monocotyledonous and dicotyledonous plant cell cultures.  Canadian Journal of Botany. 1972;  50 199-204
    CrossrefPubMedSearch in Google Scholar
  • 14 Lloyd G, McCown B. Commercially-feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot-tip culture.  Int. Plant Prop Soc. Proc.. 1980;  30 421-7
    PubMedSearch in Google Scholar
  • 15 Ray S, Ghosh B, Sen S, Jha S. Withanolide production by root cultures of Withania somnifera transformed with Agrobacterium rhizogenes .  Planta Medica. 1996;  62 571-3
    PubMedSearch in Google Scholar
  • 16 Miura Y, Hirata K, Kurano N, Miyatnoto K, Uchida K. Formation of vinblastine in multiple shoot cultures of Catharanthus roseus .  Planta Medica. 1988;  54 18-20
    PubMedSearch in Google Scholar
  • 17 Indrayanto G, Tristiana E, Mulija H S. Effect of L arginine, casein hydrolysate, banana powder and sucrose on growth and solasodine production in shoot cultures of Solanum laciniatum .  Plant Cell, Tissue and Organ Culture. 1995;  43 237-40
    PubMedSearch in Google Scholar
  • 18 Ali A M, Hughs H G, Safadi F, Dami I. Liquid medium overlays enhanced growth and multiplication of in vitro grapes and apples.  Fruit Varieties Journal. 1996;  50 86-7
    PubMedSearch in Google Scholar
  • 19 Eisenbeiss M, Kreis W, Reinhard E. Cardenolide biosynthesis in light and dark grown Digitalis lanata shoot cultures.  Plant Physiology and Biochemistry. 1999;  37 13-24
    CrossrefPubMedSearch in Google Scholar
  • 20 Anderson L A, Keene A T, Phillipson J D. Alkaloid production by leaf organ, root organ and cell suspension cultures of Cinchona ledgeriana .  Planta Medica. 1982;  46 25-7
    PubMedSearch in Google Scholar

Dr. Sumita Jha

Centre of Advanced Study

Department of Botany

Calcutta University

35, Ballygunge Circular Road

Calcutta 700 019

India