Tropane alkaloid production by hairy roots of Atropa belladonna obtained after transformation with Agrobacterium rhizogenes 15834 and Agrobacterium tumefaciens containing rol A, B, C genes only

J Biotechnol. 2000 Aug 25;81(2-3):151-8. doi: 10.1016/s0168-1656(00)00287-x.

Abstract

Atropa belladonna leaf disks were infected by a wild strain Agrobacterium rhizogenes 15834 harboring the Ri-TL-DNA and by a disarmed Agrobacterium tumefaciens strain harboring a construction with only rol ABC and npt II genes. Thirteen root lines were established and examined for their growth rate and alkaloid productivity to evaluate the possible role of rol genes in morphological differentiation and in tropane alkaloid formation. A great diversity has been observed in the growth rate of these 13 root lines. The root biomass increased up to 75 times. The total alkaloid contents were similar in the root lines obtained by infection with A. rhizogenes 15834 and A. tumefaciens rol ABC. The last ones accumulated between 4 (1.1 mg g(-1) DW) and 27 (8 mg g(-1) DW) times more alkaloids than the intact roots (0.3 mg g(-1) DW). This work has shown that the rol ABC genes were sufficient to increase tropane alkaloid production in A. belladonna hairy root cultures.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agrobacterium tumefaciens / genetics*
  • Atropa belladonna / cytology
  • Atropa belladonna / genetics
  • Atropa belladonna / metabolism*
  • Atropa belladonna / microbiology
  • Bacterial Proteins / genetics
  • Belladonna Alkaloids / metabolism*
  • Cell Division
  • Cell Line, Transformed
  • DNA, Bacterial / analysis
  • Kinetics
  • Plant Roots / chemistry
  • Plant Roots / metabolism
  • Plants, Medicinal*
  • Plants, Toxic*
  • Transfection
  • Tropanes / metabolism*
  • beta-Glucosidase / genetics

Substances

  • Bacterial Proteins
  • Belladonna Alkaloids
  • DNA, Bacterial
  • Tropanes
  • rolA protein, Agrobacterium rhizogenes
  • RolB protein, Agrobacterium rhizogenes
  • cytokinin-beta-glucosidase
  • beta-Glucosidase