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J Biotechnol. 2014 Apr 20;176:20-8. doi: 10.1016/j.jbiotec.2014.01.031. Epub 2014 Feb 14.

Evaluation of tobacco (Nicotiana tabacum L. cv. Petit Havana SR1) hairy roots for the production of geraniol, the first committed step in terpenoid indole alkaloid pathway.

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VTT Technical Research Centre of Finland, P.O. Box 1000, Tietotie 2, 02044-VTT Espoo, Finland. Electronic address:
Laboratory of Plant Physiology, Wageningen UR, P.O. Box 658, 6700 AR Wageningen, The Netherlands.
Zurich University of Applied Sciences, Institute of Biotechnology, Biochemical Engineering and Cell Cultivation Technique, Campus Grüental, Wädenswil, Switzerland.
VTT Technical Research Centre of Finland, P.O. Box 1000, Tietotie 2, 02044-VTT Espoo, Finland.
Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Forckenbeckstrasse 6, 52074 Aachen, Germany.


The terpenoid indole alkaloids are one of the major classes of plant-derived natural products and are well known for their many applications in the pharmaceutical, fragrance and cosmetics industries. Hairy root cultures are useful for the production of plant secondary metabolites because of their genetic and biochemical stability and their rapid growth in hormone-free media. Tobacco (Nicotiana tabacum L. cv. Petit Havana SR1) hairy roots, which do not produce geraniol naturally, were engineered to express a plastid-targeted geraniol synthase gene originally isolated from Valeriana officinalis L. (VoGES). A SPME-GC-MS screening tool was developed for the rapid evaluation of production clones. The GC-MS analysis revealed that the free geraniol content in 20 hairy root clones expressing VoGES was an average of 13.7 μg/g dry weight (DW) and a maximum of 31.3 μg/g DW. More detailed metabolic analysis revealed that geraniol derivatives were present in six major glycoside forms, namely the hexose and/or pentose conjugates of geraniol and hydroxygeraniol, resulting in total geraniol levels of up to 204.3 μg/g DW following deglycosylation. A benchtop-scale process was developed in a 20-L wave-mixed bioreactor eventually yielding hundreds of grams of biomass and milligram quantities of geraniol per cultivation bag.


Geraniol; Hairy root; Terpenoid indole alkaloid; Tobacco; Transgenic

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