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JXB Advance Access originally published online on January 10, 2005
Journal of Experimental Botany 2005 56(412):645-652; doi:10.1093/jxb/eri067
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Journal of Experimental Botany, Vol. 56, No. 412, © Society for Experimental Biology 2005; all rights reserved

RESEARCH PAPER

Overexpression of tropinone reductases alters alkaloid composition in Atropa belladonna root cultures

Ute Richter, Grit Rothe, Anne-Katrin Fabian, Bettina Rahfeld and Birgit Dräger*

Institute of Pharmaceutical Biology, Martin-Luther-University Halle-Wittenberg, Hoher Weg 8, D-06120 Halle/Saale, Germany

* To whom correspondence should be addressed. Fax: +49 345 55 27021. E-mail: draeger{at}pharmazie.uni-halle.de

The medicinally applied tropane alkaloids hyoscyamine and scopolamine are produced in Atropa belladonna L. and in a small number of other Solanaceae. Calystegines are nortropane alkaloids that derive from a branching point in the tropane alkaloid biosynthetic pathway. In A. belladonna root cultures, calystegine molar concentration is 2-fold higher than that of hyoscyamine and scopolamine. In this study, two tropinone reductases forming a branching point in the tropane alkaloid biosynthesis were overexpressed in A. belladonna. Root culture lines with strong overexpression of the transcripts contained more enzyme activity of the respective reductase and enhanced enzyme products, tropine or pseudotropine. High pseudotropine led to an increased accumulation of calystegines in the roots. Strong expression of the tropine-forming reductase was accompanied by 3-fold more hyoscyamine and 5-fold more scopolamine compared with control roots, and calystegine levels were decreased by 30–90% of control. In some of the transformed root cultures, an increase of total tropane alkaloids was observed. Thus, transformation with cDNA of tropinone reductases successfully altered the ratio of tropine-derived alkaloids versus pseudotropine-derived alkaloids.

Key words: Atropa belladonna, calystegine, gene transformation, hyoscyamine, overexpression, scopolamine, tropane alkaloids, tropinone reductase


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