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Comparison of plant-based expression platforms for the heterologous production of geraniol

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Abstract

We compared the ability of different plant-based expression platforms to produce geraniol, a key metabolite in the monoterpenoid branch of the terpenoid indole alkaloid biosynthesis pathway. A geraniol synthase gene isolated from Valeriana officinalis (VoGES) was stably expressed in different tobacco systems. Intact plants were grown in vitro and in the greenhouse and were used to generate cell suspension and hairy root cultures. VoGES was also transiently expressed in N. benthamiana. The highest geraniol content was produced by intact transgenic plants grown in vitro (48 μg/g fresh weight, fw), followed by the transient expression system (27 μg/g fw), transgenic plants under hydroponic conditions in the greenhouse and cell suspension cultures (16 μg/g fw), and finally hairy root cultures (9 μg/g fw). Differences in biomass production and the duration of cultivation resulted in a spectrum of geraniol productivities. Cell suspension cultures achieved a geraniol production rate of 1.8 μg/g fresh biomass per day, whereas transient expression produced 5.9 μg/g fresh biomass per day (if cultivation prior to agroinfiltration is ignored) or 0.5 μg/g fresh biomass per day (if cultivation prior to agroinfiltration is included). The superior productivity, strict process control and simple handling procedures available for transgenic cell suspension cultures suggest that cells are the most promising system for further optimization and ultimately for the scaled-up production of geraniol.

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Acknowledgments

This research was funded by the European Union Seventh Framework Programme SmartCell (Rational design of plant systems for sustainable generation of value-added industrial products, Grant agreement No. 222,716) and the EU COST Action FA1006 PlantEngine. VTT would like to acknowledge the technical assistance of Airi Hyrkäs, Siv Matomaa, Jaana Rikkinen, Mari Lämsä, Annika Majanen and Tuuli Teikari. We thank Dr. Richard Twyman for critical reading of the manuscript. None of the authors declare a conflict of interest.

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Authors and Affiliations

  1. Department Plant Biotechnology, Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Forckenbeckstraße 6, 52074, Aachen, Germany

    Nikolay Vasilev, Christian Schmitz, Rainer Fischer & Stefan Schillberg

  2. Laboratory of Plant Physiology, Wageningen UR, P.O. Box 658, 6700 AR, Wageningen, The Netherlands

    Lemeng Dong, Sander van der Krol & Harro Bouwmeester

  3. VTT Technical Research Centre of Finland, P.O. Box 1000, 02044 VTT, Espoo, Finland

    Anneli Ritala, Suvi T. Häkkinen & Kirsi-Marja Oksman-Caldentey

  4. Institute of Biotechnology, Biochemical Engineering and Cell Cultivation Technique, Zurich University of Applied Sciences, Campus Grüental, Wädenswil, Switzerland

    Nicole Imseng & Regine Eibl

  5. Institute for Molecular Biotechnology, RWTH Aachen University, Worringer Weg 1, 52074, Aachen, Germany

    Rainer Fischer

  6. Institute for Phytopathology and Applied Zoology, Justus-Liebig University Giessen, Heinrich-Buff-Ring 26-32, 35392, Giessen, Germany

    Stefan Schillberg

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  1. Nikolay Vasilev
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Correspondence to Nikolay Vasilev.

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Vasilev, N., Schmitz, C., Dong, L. et al. Comparison of plant-based expression platforms for the heterologous production of geraniol. Plant Cell Tiss Organ Cult 117, 373–380 (2014). https://doi.org/10.1007/s11240-014-0446-z

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  • DOI: https://doi.org/10.1007/s11240-014-0446-z

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