Abstract
Atropa baetica hairy roots, over-expressing cDNA from Hyoscyamus niger encoding the gene for hyoscyamine 6β-hydroxylase (H6H), were produced by Agrobacterium rhizogenes infection. The transgenic roots over-expressing h6h had an altered alkaloid profile in which hyoscyamine was entirely converted into scopolamine. In the best h6h clone, scopolamine accumulation increased 9-fold compared to plants, amounting to 5.6 mg g dry wt−1, some of which was released into the liquid medium. Only negligible amounts of hyoscyamine were detected. In contrast, the gus control culture contained a much higher amount of hyoscyamine than scopolamine, mimicking the situation in the plant. At the molecular level, a higher conversion of hyoscyamine into scopolamine was related to a higher level of h6h mRNA; in some instances this was 5–10-fold higher
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Acknowledgements
The authors express their gratitude to El Bosque Botanic Garden and Dr. Aparicio (Spain) for providing A. baetica seeds. Thanks are also given to Drs. Schilman & Diaz (Leiden University, The Netherlands); Drs. P. Rocha & MJ. Leech for their kind gift of the h6h sequence (John Innes Centre, Norwich, UK), and to Prof. M. Yeoman for comments on the manuscript. RZ acknowledges financial support from the European Union (project no. QLK3-CT-1999-51341), Ramón y Cajal Program and SAF-2003-04200-C02-02 Project (Spanish Ministry of Science & Technology) and Instituto Canario de Investigación del Cáncer (ICIC) which also supported together with a grant from CajaCanarias to NJ.
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This article is dedicated to the memory of Professor Antonio G. González
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Zárate, R., Jaber-Vazdekis, N.e., Medina, B. et al. Tailoring Tropane Alkaloid Accumulation in Transgenic Hairy Roots of Atropa baetica by Over-expressing the Gene Encoding Hyoscyamine 6β-hydroxylase. Biotechnol Lett 28, 1271–1277 (2006). https://doi.org/10.1007/s10529-006-9085-8
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DOI: https://doi.org/10.1007/s10529-006-9085-8