Alkaloid production by a Cinchona officinalis 'Ledgeriana' hairy root culture containing constitutive expression constructs of tryptophan decarboxylase and strictosidine synthase cDNAs from Catharanthus roseus |
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Authors: | A Geerlings D Hallard A Martinez Caballero I Lopes Cardoso R van der Heijden R Verpoorte |
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Institution: | (1) Leiden/Amsterdam Center for Drug Research, Division of Pharmacognosy, Leiden University, Gorlaeus Laboratories, P.O. Box 9502, 2300 RA Leiden, The Netherlands e-mail: verpoort@lacdr.leidenuniv.nl Fax: +31–71–5274511, NL |
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Abstract: | Cinchona officinalis 'Ledgeriana', former called Cinchona ledgeriana, hairy roots were initiated containing constitutive-expression constructs of cDNAs encoding the enzymes tryptophan decarboxylase
(TDC) and strictosidine synthase (STR) from Catharanthus roseus, two key enzymes in terpenoid indole and quinoline alkaloid biosynthesis. The successful integration of these genes and the
reporter gene gus-int was demonstrated using Southern blotting and the polymerase chain reaction. The products of TDC and STR, tryptamine and
strictosidine, were found in high amounts, 1200 and 1950 μg g–1 dry weight, respectively. Quinine and quinidine levels were found to rise up to 500 and 1000 μg g–1 dry weight, respectively. The results show that genetic engineering with multiple genes is well possible in hairy roots of
C. officinalis. However, 1 year after analyzing the hairy roots for the first time, they had completely lost their capacity to accumulate
alkaloids.
Received: 15 October 1997 / Accepted after revision: 21 March 1999 |
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Keywords: | Quinoline alkaloids Genetic engineering Catharanthus roseus Cinchona officinalis 'Ledgeriana' Terpenoid indole alkaloids |
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