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JXB Advance Access published online on July 28, 2003
Journal of Experimental Botany, doi:10.1093/jxb/erg227
© 2003 by Oxford University Press
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Received February 14, 2003; accepted June 3, 2003
© 2003 Society for Experimental Biology

RESEARCH PAPER

Alkaloids in plants and root cultures of Atropa belladonna overexpressing putrescine N-methyltransferase

Grit Rothe 1, Akira Hachiya 2, Yasuyuki Yamada 2, Takashi Hashimoto 2, and Birgit Dräger 1*

1 Institute of Pharmaceutical Biology, Martin-Luther-University Halle-Wittenberg, Hoher Weg 8, D-06120 Halle/Saale, Germany
2 Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama-cho, Ikoma, Nara 630-0192, Japan

* To whom correspondence should be addressed. E-mail: draeger{at}pharmazie.uni-halle.de.


  Abstract

Putrescine N-methyltransferase (PMT) is the first alkaloid-specific enzyme for nicotine and tropane alkaloid formation. The pmt gene from Nicotiana tabacum was fused to the CaMV 35S promoter and integrated into the Atropa belladonna genome. Transgenic plants and derived root cultures were analysed for gene expression and for levels of alkaloids and their precursors. Scopolamine, hyoscyamine, tropine, pseudotropine, tropinone, and calystegines were found unaltered or somewhat decreased in pmt-overexpressing lines compared to controls. When root cultures were treated with 5% sucrose, calystegine levels were elevated in control roots, but were not affected in pmt-overexpressing roots. 1 µM auxin reduced calystegine levels in control roots, while in pmt-overexpressing roots all alkaloids remained unaltered. Expression level of pmt alone is apparently not limiting for tropane alkaloid formation in A. belladonna.

Key words: Atropa belladonna, calystegine, hyoscyamine, overexpression, putrescine N-methyltransferase, scopolamine, tropane alkaloids.


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