Effects of the rol C gene on hairy root: induction development and tropane alkaloid production by Atropa belladonna.
Two series of Atropa belladonna hairy root lines were obtained, the first one transformed via Agrobacterium tumefaciens harboring rol C and npt II genes, and the other transformed with rol ABC and npt II genes. Thirteen hairy root lines were obtained and selected on hormone-free medium. The transformation was confirmed by PCR analysis, and these root lines were first examinated for their growth rate. Then hyoscyamine and scopolamine production was measured after 3 and 4 weeks of culture to evaluate the possible role of rol C gene in tropane alkaloid formation. The rol C gene alone played a significant role in the hairy root growth rate (17-fold increase). However this effect was much lower than that induced by the rol ABC genes together (75-fold increase). In contrast, the rol C gene alone was as efficient as the rol ABC genes together (mean value of total alkaloids: 0.36% dry weight, i.e., 12-fold times more than in untransformed roots) to stimulate the biosynthesis of tropane alkaloids in A. belladonna hairy root cultures.
Bonhomme
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V
Laboratoire de Pharmacognosie et Phytotechnologie, Faculté de Pharmacie, 1 Rue des Louvels, 80037 AMIENS, Cédex, France. Valerie.Bonhomme@sa.u-picardie.fr
Laurain-Mattar
D
D
Fliniaux
M A
MA
D001532:Atropa belladonna / Q000235:genetics / Q000378:metabolism*; D001285:Atropine / Q000096:biosynthesis*; D001483:Base Sequence; D017931:DNA Primers; D018744:DNA, Plant; D018517:Plant Roots / Q000737:chemistry* / Q000254:growth & development; D010946:Plants, Medicinal; D010947:Plants, Toxic; D012601:Scopolamine / Q000096:biosynthesis*; D001617:beta-Glucosidase / Q000235:genetics*
D017931:DNA Primers; D018744:DNA, Plant; D001285:Atropine; D012601:Scopolamine; C071257:cytokinin-beta-glucosidase; D001617:beta-Glucosidase