Enhancement of scopolamine production in Hyoscyamus muticus L. hairy root cultures by genetic engineering

In order to test the possibility of enhancing the production of pharmaceutically valuable scopolamine in transgenic cultures, the 35S-h6h transgene that codes for the enzyme hyoscyamine-6 beta-hydroxylase (EC 1.14.11.11) was introduced into Hyoscyamus muticus L. strain Cairo (Egyptian henbane). This...

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Bibliographic Details
Published in:Planta 1999-06, Vol.208 (4), p.545-551
Main Authors: Jouhikainen, K, Lindgren, L, Jokelainen, T, Hiltunen, R, Teeri, T.H, Oksman-Caldentey, K.M
Format: Article
Language:English
Subjects:
Agrobacterium
Agronomy. Soil science and plant productions
Alkaloids
Amines
Biological and medical sciences
biosynthesis
Biotechnology
callus
chemical constituents of plants
Cultured cells
Fundamental and applied biological sciences. Psychology
gene expression
genes
genetic engineering
Genetic engineering applications
genetic variation
Genetics and breeding of economic plants
Hyoscyamus muticus
oxygenases
Plant breeding: fundamental aspects and methodology
Plant cells
plant morphology
Plant roots
Plants
roots
tissue culture
transformation
Transgenes
Transgenic plants
tropane alkaloids
Tropanes
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Summary:In order to test the possibility of enhancing the production of pharmaceutically valuable scopolamine in transgenic cultures, the 35S-h6h transgene that codes for the enzyme hyoscyamine-6 beta-hydroxylase (EC 1.14.11.11) was introduced into Hyoscyamus muticus L. strain Cairo (Egyptian henbane). This plant was chosen for its capability to produce very high amounts of tropane alkaloids (up to 6% of the dry weight in the leaves of mature plant). To our knowledge, this is the first time such a large population of transgenic cultures has been studied at the morphological, chemical and genetic levels. A great variation was observed in the tropane alkaloid production among the 43 positive transformants. The best clone (KB7) produced 17 mg/l scopolamine, which is over 100 times more than the control clones. However, conversion of hyoscyamine to scopolamine was still incomplete. The expression of h6h was found to be proportional to the scopolamine production, and was the main reason behind the variation in the scopolamine/hyoscyamine ratio in the hairy-root clones. These results indicate that H. muticus strain Cairo has a potential for even more enhanced scopolamine production with more efficient gene-expression systems.
ISSN:0032-0935
1432-2048
DOI:10.1007/s004250050592