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Journal of Experimental Botany, Vol. 54, No. 381, pp. 203-211, January 2, 2003
© 2003 Oxford University Press

Effect of pmt gene overexpression on tropane alkaloid production in transformed root cultures of Datura metel and Hyoscyamus muticus

Received 5 March 2002; Accepted 14 August 2002

Elisabet Moyano1, Katja Jouhikainen6,2,3, Päivi Tammela2, Javier Palazón4, Rosa M. Cusidó4, M. Teresa Piñol4, Teemu H. Teeri7,3 and Kirsi-Marja Oksman-Caldentey8,5

1 Departament de Ciències Experimentals i de la Salut, Universitat Pompeu Fabra, Avda. Dr Aiguader 80, E-08003 Barcelona, Spain
2 Division of Pharmacognosy, Department of Pharmacy, PO Box 56, FIN-00014 University of Helsinki, Finland
3 Institute of Biotechnology, PO Box 56, FIN-00014 University of Helsinki, Finland
4 Sección de Fisiología Vegetal, Facultad de Farmacia, Universidad de Barcelona, Avda. Diagonal 643, E-08028 Barcelona, Spain
5 VTT Biotechnology, PO Box 1500, FIN-02044 VTT (Espoo), Finland

6 Deceased.
7 Present address: Department of Applied Biology, PO Box 27, FIN-00014 University of Helsinki, Finland.
8 To whom correspondence should be addressed. Fax: +358 9 4552103. E-mail: kirsi-marja.oksman{at}vtt.fi

In order to increase the production of the pharmaceuticals hyoscyamine and scopolamine in hairy root cultures, a binary vector system was developed to introduce the T-DNA of the Ri plasmid together with the tobacco pmt gene under the control of CaMV 35S promoter, into the genome of Datura metel and Hyoscyamus muticus. This gene codes for putres cine:SAM N-methyltransferase (PMT; EC. 2.1.1.53), which catalyses the first committed step in the tropane alkaloid pathway. Hairy root cultures overexpressing the pmt gene aged faster and accumulated higher amounts of tropane alkaloids than control hairy roots. Both hyoscyamine and scopolamine production were improved in hairy root cultures of D. metel, whereas in H. muticus only hyoscyamine contents were increased by pmt gene overexpression. These roots have a high capacity to synthesize hyoscyamine, but their ability to convert it into scopolamine is very limited. The results indicate that the same biosynthetic pathway in two related plant species can be differently regulated, and overexpression of a given gene does not necessarily lead to a similar accumulation pattern of secondary metabolites.

Key words: Datura metel, hairy root cultures, hyoscyamine, Hyoscyamus muticus, 35S-pmt gene, scopolamine, tropane alkaloids.


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