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

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Abstract

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.

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Abbreviations

MS:

Murashige & Skoog's basal medium

MLS:

modified Linsmaier & Skoog's basal medium

BA:

benzyladenine

NAA:

naphthalene acetic acid

TLC:

thin layer chromatography

HPLC:

high performance liquid chromatography

References

  • 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 190

  • Habib SM & Court WE (1974) The separation and identification of microquantities of Rauwolfia alkaloids. Planta Med. 25: 331–341

    Google Scholar 

  • 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: 111–114

    Google Scholar 

  • 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: 88–92

    Google Scholar 

  • 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: 239–242

    Google Scholar 

  • Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–479

    Google Scholar 

  • 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: 241–246

    Google Scholar 

  • Ondrej M, Hrouda M & Kostrica P (1989) Potato transformation of Agrobacterium rhizogenes Ri plasmid. Biol. Plant. (Praha). 31: 312–314

    Google Scholar 

  • 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: 204–214

    Google Scholar 

  • Roja PC, Benjamin BD, Heble MR & Chadha MS (1984) Indole alkaloids from multiple shoot cultures of Rauwolfia serpentina. Planta Med. 1: 73–74

    Google Scholar 

  • White FF & Nester EW (1980) Hairy root plasmid encodes virulence traits in Agrobacterium rhizogenes. J. Bact. 141: 1134–1141

    Google Scholar 

  • Yamamoto O & Yamada Y (1986) Production of reserpine and its optimisation in cultured Rauvolfia serpentina Benth. cells. Plant. Cell Rep. 5: 50–53

    Google Scholar 

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Benjamin, B.D., Roja, G. & Heble, M.R. Agrobacterium rhizogens mediated transformation of Rauvolfia serpentina: Regeneration and alkaloid synthesis. Plant Cell Tiss Organ Cult 35, 253–257 (1993). https://doi.org/10.1007/BF00037278

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  • DOI: https://doi.org/10.1007/BF00037278

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