Plant Cell, Tissue and Organ Culture

, Volume 35, Issue 3, pp 253–257 | Cite as

Agrobacterium rhizogens mediated transformation of Rauvolfia serpentina: Regeneration and alkaloid synthesis

  • B. D. Benjamin
  • G. Roja
  • M. R. Heble
Original Research Paper


Shoot cultures of Rauvolfia serpentina infected with Agrobacterium rhizogenes 15834 produced tumourous tissue at the site of injection, which eventually developed callus with hairy roots. Sporadic shoot formation occurred from the hairy roots. The shoots were grown to maturity in the green house. The mature transformed plants (RST) showed distinct variations in their physiological characteristics. The flowers of the transformed plants were more delicate and less pigmented when compared to the flowers of the mature normal plants. The roots of the transformed plants were hairy with a number of lateral branches, whereas the roots of the normal plant had very few lateral branches. The biomass of the transformed plant was 86.39 g/plant (fresh wt), significantly higher than the normal plant which was 77.3 g/plant (fresh wt). The total alkaloid content in the mature transformed plant (0.073 g per plant) was similar to the normal plant (0.078 g per plant), although the hairy roots contained little alkaloid.

Key words

Agrobacterlum rhizogenes indole alkaloids Rauvolfia serpentina transgenic plants 



Murashige & Skoog's basal medium


modified Linsmaier & Skoog's basal medium




naphthalene acetic acid


thin layer chromatography


high performance liquid chromatography


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Flores HE (1986) Use of plant cell and organ culture in the production of biological chemicals. in: Lebaron H, Mumma RO, Honeycutt RC & Duesing JH (Eds) Application of Biotechnology to Agricultural Chemistry. Proc. 190 Amer. Chem. Soc. Symposium series 190Google Scholar
  2. Habib SM & Court WE (1974) The separation and identification of microquantities of Rauwolfia alkaloids. Planta Med. 25: 331–341Google Scholar
  3. Hamill JD, Parr AJ, Robins RJ & Rhodes MJC (1986) Secondary product formation by cultures of Beta vulgaris and Nicotiana rustica transformed with Agrobacterium rhizogenes. Plant Cell Rep. 5: 111–114Google Scholar
  4. Hatamoto H, Boulter ME, Shirsat AH, Croy EJ & Ellis JR (1990) Recovery of morphologically normal transgenic tobacco plants from hairy roots co-transformed with Agrobacterium rhizogenes and a binary vector plasmid. Plant Cell Rep. 9: 88–92Google Scholar
  5. Kamada H, Okamura M, Satak M, Harada EH & Shimomura K (1986) Alkaloid production by hairy root cultures in Atropa belladonna (1986). Plant Cell Rep. 5: 239–242Google Scholar
  6. Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–479Google Scholar
  7. Ondrej M & Protiva J (1987) In vitro culture of crown gall and hairy root tumours of Atropa belladonna. Differentiation and alkaloid production. Biol. Plant. (Praha). 29: 241–246Google Scholar
  8. Ondrej M, Hrouda M & Kostrica P (1989) Potato transformation of Agrobacterium rhizogenes Ri plasmid. Biol. Plant. (Praha). 31: 312–314Google Scholar
  9. Petit AC, David C, Dahl GA, Ellis JG, Guyon P, Casse-Delbart F & Tempe J (1983) Further extension of the opine concept: plasmids in Agrobacterium rhizogenes cooperate for opine degradation. Mol. Gen. Genet. 190: 204–214Google Scholar
  10. Roja PC, Benjamin BD, Heble MR & Chadha MS (1984) Indole alkaloids from multiple shoot cultures of Rauwolfia serpentina. Planta Med. 1: 73–74Google Scholar
  11. White FF & Nester EW (1980) Hairy root plasmid encodes virulence traits in Agrobacterium rhizogenes. J. Bact. 141: 1134–1141Google Scholar
  12. Yamamoto O & Yamada Y (1986) Production of reserpine and its optimisation in cultured Rauvolfia serpentina Benth. cells. Plant. Cell Rep. 5: 50–53Google Scholar

Copyright information

© Kluwer Academic Publishers 1993

Authors and Affiliations

  • B. D. Benjamin
    • 1
  • G. Roja
    • 1
  • M. R. Heble
    • 1
  1. 1.Plant Biotechnology SectionBhabha Atomic Research CentreBombayIndia

Personalised recommendations