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PLANT PHYSIOLOGY , Vol 105, Issue 2 483-490, Copyright © 1994 by American Society of Plant Biologists
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MOLECULAR BIOLOGY AND GENE REGULATION |
Species-Dependent Expression of the Hyoscyamine 6[beta]-Hydroxylase Gene in the Pericycle
T. Kanegae, H. Kajiya, Y. Amano, T. Hashimoto and Y. Yamada
Department of Agricultural Chemistry, Faculty of Agriculture, Kyoto University, Kyoto 606-01, Japan
The tropane alkaloid scopolamine is synthesized in the pericycle of branch
roots in certain species of the Solanaceae. The enzyme responsible for the
synthesis of scopolamine from hyoscyamine is hyoscyamine
6[beta]-hydroxylase (H6H). The gene for H6H was isolated from Hyoscyamus
niger. It has an exon/intron organization very similar to those for
ethylene-forming enzymes, suggesting a common evolutionary origin. The
827-bp 5[prime] flanking region of the H6H gene was fused to the
[beta]-glucuronidase (GUS) reporter gene and transferred to three
solanaceous species by Agrobacterium-mediated transformation systems: H.
niger and belladonna (Atropa belladonna), which have high and low levels,
respectively, of H6H mRNA in the root, and tobacco (Nicotiana tabacum),
which has no endogenous H6H gene. Histochemical analysis showed that GUS
expression occurred in the pericycle and at the root meristem of transgenic
H. niger hairy roots, but only at the root meristem of hairy roots and
plants of transgenic tobacco. In transgenic hairy roots and regenerated
plants of belladonna, the root meristem was stained with GUS activity,
except for a few transformants in which the vascular cylinder was also
stained. These studies indicate that the cell-specific expression of the
H6H gene is controlled by some genetic regulation specific to
scopolamine-producing plants.
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