Planta Med 2001; 67(3): 249-253
DOI: 10.1055/s-2001-12006
Original Paper
Biochemistry, Physiology, in vitro Cultures
© Georg Thieme Verlag Stuttgart · New York

Decreased Scopolamine Yield in Field-Grown Duboisia Plants Regenerated from Hairy Roots

Carles Roig Celma1 , Javier Palazón1 , Rosa M. Cusidó1 , M. Teresa Piñol1 , Michael Keil2,*
  • 1 Departament de Productes Naturals, Biologia Vegetal i Edafologia, Facultat de Farmàcia, Universitat de Barcelona, Barcelona, Spain
  • 2 Boehringer Ingelheim Pharma KG, A Feinchemikalien/DPL, Ingelheim, Germany
Further Information

Publication History

April 7, 2000

August 9, 2000

Publication Date:
31 December 2001 (online)

Abstract

Hairy root cultures were obtained from hybrid clones of Duboisia myoporoides x D. leichhardtii following transformation by Agrobacterium rhizogenes strain A4. Shoots spontaneously regenerating from the hairy root cultures were rooted and transferred to soil. The plants displayed typical morphological alterations known as hairy root syndrome to varying degrees. PCR analysis confirmed that all transformed plants contained the rolA, rolB and rolC genes, irrespective of the degree of morphological alterations. A field test of the transformed regenerated plants revealed that those plants displaying the strongest hairy root syndrome symptoms had the highest content of the tropane alkaloid scopolamine. However, the overall scopolamine and hyoscyamine yield of all transformed plants was clearly reduced compared to untransformed control plants. These results demonstrate that the A. rhizogenes-transformed plants tested in this study do not provide a viable alternative to agricultural farming of hybrid clones of D. myoporoides x D. leichhardtii obtained by conventional breeding.

References

  • 1 Yeoman M M, Yeoman C L. Transley Review No. 90. Manipulating secondary metabolism in cultured plant cells.  New Phytology. 1996;  134 553-69
  • 2 Wysokinska H, Chmiel A. Transformed root cultures for biotechnology.  Acta Biotechnologia. 1997;  17 131-59
  • 3 Wilson P DG. The pilot-scale cultivation of transformed roots. In: Doran PM, editor Hairy roots - culture and applications. Amsterdam; Harwood Academic Publishers 1997: 179-90
  • 4 Christey M C. Transgenic crop plants using Agrobacterium rhizogenes - mediated transformation. In: Doran PM, editor Hairy roots - culture and applications. Amsterdam; Harwood Academic Publishers 1997: 99-111
  • 5 Sevón N, Dräger B, Hiltunen R, Oksman-Caldentey K M. Characterization of transgenic plants derived from hairy roots of Hyoscyamus muticus .  Plant Cell Reports. 1997;  16 605-11
  • 6 Tanaka N. Genetic transformation of Vinca minor L. In: Bajaj YPS, editor Biotechnology in Agriculture and Forestry 45 - Transgenic Medicinal Plants. Berlin; Springer 1999: 355-65
  • 7 Franklin C I, Dixon R . Initiation and maintenance of callus and cell suspension cultures. In: Dixon RA, Gonzales RA, editors Plant cell culture - A practical approach. Oxford; IRL Press 1994: 1-25
  • 8 Vervliet G, Holsters M, Teuchy H, Van Montagu M, Schell J. Characterization of different plaque-forming and defective temperate phages in Agrobacterium strains.  Journal of Genetics and Virology. 1975;  26 33-48
  • 9 Plank K H, Wagner K G. Determination of hyoscyamine and scopolamine in Datura innoxia plants by high performance liquid chromatography.  Zeitung für Naturforschung. 1986;  41 391-5
  • 10 Muranaka T, Kitamura Y, Ikenaga T. Genetic transformation of Duboisia species. In: Bajaj YPS (ed) Biotechnology in Agriculture and Forestry 45 - Transgenic Medicinal Plants. Berlin; Springer 1999: 117-32
  • 11 Yukimune Y, Hara Y, Yamada Y. Tropane alkaloid production in root cultures of Duboisia myoporoides obtained by repeated selection.  Bioscience, Biotechnology and Biochemistry. 1994;  58 1443-6
  • 12 Sevón N, Oksman-Caldentey K-M, Hiltunen R. Efficient plant regeneration from hairy root-derived protoplasts of Hyoscyamus muticus .  Plant Cell Reports. 1995;  14 738-42
  • 13 Nilsson O, Olsson O. Getting to the root: the role of the Agrobacterium rhizogenes rol genes in the formation of hairy roots.  Physiologia Plantararum. 1997;  100 463-73
  • 14 Matsuda J, Okabe S, Hashimoto T, Yamada Y. Molecular cloning of hyoscyamine 6β-hydroxylase, a 2-oxoglutarate-dependent dioxygenase, from cultured roots of Hyoscyamus niger .  Journal of Biological Chemistry. 1991;  266 9460-4
  • 15 Cusido R M, Palazón J, Piñol M T, Bonfill M, Morales C. Datura metel: In vitro production of tropane alkaloids.  Planta Medica. 1999;  65 144-8
  • 16 Jouhikainen K, Lindgren L, Jokelainen T, Hiltunen R, Teeri T H, Oksman-Caldentey K-M. Enhancement of scopolamine production in Hyoscyamus muticus L. hairy root cultures by genetic engineering.  Planta. 1999;  208 545-51

Dr. Michael Keil

Boehringer Ingelheim Pharma KG

A Feinchemikalien/DPL

55216 Ingelheim

Germany

Email: keilm@ing.boehringer-ingelheim.com

Fax: +49-(0)6132-772095