Production of β-β,Dimethylacrylshikonin in Callus Cultures of Onosma echioides Var hispidum Clarke

  • Surrinder K Lattoo
  • Sushma Koul
  • Manoj K Dhar
  • Ravi K Khajuria
  • Devinder K Gupta
  • Autar K Dhar
  • Ghulam N Qazi


Callus tissues derived from leaf segments of Onosma echioides var hispidum on three basal media viz. Murashige & Skoog (MS), Gamborg et al (B5) and White’s containing 3% sucrose produce napthaquinone pigments in presence of and NAA. However, β-β,dimethylacrylshikonin synthesis was triggered in dark in undifferentiated parenchyma cells on B5 agar medium containing 1 x10−5 M Kn and 2×10−6 M IBA when proliferated calli after 16 weeks (4th generation) were transferred to it and incubated at 23 ± 1 °C. The pigment biosynthesis increased linearly from 4th to 6th week after a lag of first 3 weeks. Callus grew exponentially after a lag of 2 weeks and diminished from 6th week onward. During 8 weeks of growth, callus grew from 0.8 to 8.2 g and the β-β, dimethylacrylshikonin showed the highest level of 25.41 μg g−1 of fresh tissue. Light microscopic examination of semi-thin sections of pigmented tissue revealed pigment accumlation between the plasma membrane and cell wall and also in the intercellular spaces. Exposure of cultures to white fluorescent light for more than 2 h resulted in complete repression of pigment bosynthesis. The investigations suggest that the regulatory mechanism for the biosynthesis and accumulation of napthaquinone pigment(s) may be similar to that of Lithospermum erythrorhizon. Pigment producing capability of callus cultures of O. echioides var hispidum can be exploited as an alternative raw source for the production of shikonin derivatives.

Key words

Onosma echioides var hispidum Boraginaceae callus cultures shikonin β-β,dimethylacrylshikonin 





indole-3-butyric acid




□-napthaleneacetic acid


Murashige and Skoog


Gamborg et al medium


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  1. 1.
    Anonymous, Wealth of India, CSIR, New Delhi, 7 (1966) 95.Google Scholar
  2. 2.
    Khajuria RK & Jain SM, Indian J Chern, 32B (1993) 390.Google Scholar
  3. 3.
    Wagner H, Kreher B & Juric K, Arzneim Forsch, 38 (1988) 273.Google Scholar
  4. 4.
    Sankawa U, Ebizuka Y, Miyazaki T, Isomura Y, Otsuka H, Shibta S. Inomata M & Fukuoka F, Chern Pharm Bul, 25 (1977) 2392.CrossRefGoogle Scholar
  5. 5.
    Tabata M & Fujita Y, In, Biotechnology in plant science (M Zaitlin, Editor), Academic Press, New York, (1985) pp 207–307.CrossRefGoogle Scholar
  6. 6.
    Ozaki Y, Sakaguchi I, Tsukada M & Tabata M, J Nat Prod, 49 (1986) 466.CrossRefGoogle Scholar
  7. 7.
    Koul S, Sambyal M, Khajuria RK & Jain SM, Fitoterapea, LXIV (1993) 552.Google Scholar
  8. 8.
    Murashige T & Skoog F, Physiol Plant, 15 (1962) 473.CrossRefGoogle Scholar
  9. 9.
    Gamborg OL, Miller RA & Ojima K, Exp Cell Res, 50 (1968) 151.CrossRefPubMedGoogle Scholar
  10. 10.
    White PR, The cultivation of animal and plant cells, The Ronald Press, New York (1963) p 228.Google Scholar
  11. 11.
    Morimoto I, Ikegani TKS & Hirala Y, Tetrahadron Lett, 52 (1965) 4737.CrossRefGoogle Scholar
  12. 12.
    Tsukada M & Tabata M, Planta Medica, 50 (1984) 338.CrossRefPubMedGoogle Scholar
  13. 13.
    Tabata M, Mizukami H, Hiraoka N & Konoshima M, Phytochemistry, 13 (1974) 927.CrossRefGoogle Scholar
  14. 14.
    Inouye H, Veda S, Inoue K & Matsumura H, Phytochemistry, 18 (1979) 1301.CrossRefGoogle Scholar
  15. 15.
    Fujita Y, In Applications of plant cell and tissue culture, Wiley Chichester, Ciba Found Symp (1988) 228.Google Scholar
  16. 16.
    Couladouros EA, Plyta ZF, Strongilos AT & Papageorgiou VP, Tetrahedron Lett, 387 (1997) 263Google Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  • Surrinder K Lattoo
    • 1
  • Sushma Koul
    • 1
  • Manoj K Dhar
    • 2
  • Ravi K Khajuria
    • 1
  • Devinder K Gupta
    • 1
  • Autar K Dhar
    • 1
  • Ghulam N Qazi
    • 1
  1. 1.Regional Research Laboratory (CSIR)JammuIndia
  2. 2.Department of BiotechnologyJammu UniversityJammuIndia

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