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Alkamide production from hairy root cultures of Echinacea

  • Metabolic Engineering/Secondary Metabolism
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Abstract

Hairy root cultures of Echinacea, one of the most important medicinal plants in the US, represent a valuable alternative to field cultivation for the production of bioactive secondary metabolites. In this study, the three most economically important species of Echinacea (Echinacea purpurea, Echinacea pallida, and Echinacea angustifolia) were readily transformed with two strains of Agrobacterium that produce the hairy root phenotype. Transformed roots of all three species exhibited consistent accelerated growth and increased levels of alkamide production. Optimization of the culture of Echinacea hairy roots was implemented to enhance both growth and alkamide production concomitantly. The use of half-strength Gamborg’s B5 medium supplemented with 3.0% sucrose was twice as effective in maintaining hairy root production than any other media tested. The addition of indolebutyric acid increased the growth rate of roots by as much as 14-fold. Alkamide production increased severalfold in response to the addition of the elicitor, jasmonic acid, but did not respond to the addition of indolebutyric acid. Induced accumulation of the important bioactive compounds, alkamides 2 and 8, was observed both in transformed roots and in response to jasmonic acid treatments. The results of this study demonstrate the efficacy of hairy root cultures of Echinacea for the in vitro production of alkamides and establish guidelines for optimum yield.

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Acknowledgements

This publication was made possible by grant no. 9 P50 AT004155-06 from the National Center for Complementary and Alternative Medicine (NCCAM). Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the NCCAM, or the National Institutes of Health. We thank Dr. Mark P. Widrlechner for providing the seeds for our experiments and Dr. Jackie Shanks for helpful suggestions on liquid cultures. Thanks to Dr. Eve Wurtele and Lankun Wu for their helpful technical assistance on the biochemical analyses. We also thank Dr. Susan Gibson, University of Minnesota, Dept. of Plant Biology, Dr. Seung-Beom Hong, Rice University, Dept. of Biochemistry and Cell Biology and Dr. Robert Thornburg, Iowa State University, Dept. of Biochemistry, Biophysics, and Molecular Biology, for providing the Agrobacterium strains.

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Authors and Affiliations

  1. Department of Horticulture, Iowa State University, Ames, IA, 50011-1100, USA

    Fredy R. Romero, Kathleen Delate & David J. Hannapel

  2. Interdepartmental Plant Biology Major, Iowa State University, Ames, IA, 50011-1100, USA

    Fredy R. Romero & David J. Hannapel

  3. Department of Agronomy, Iowa State University, Ames, IA, 50011-1100, USA

    Kathleen Delate

  4. Department of Chemistry, Iowa State University, Ames, IA, 50011-3111, USA

    George A. Kraus

  5. Department of Food Science and Human Nutrition, Iowa State University, Ames, IA, 50011-1061, USA

    Avery K. Solco & Patricia A. Murphy

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  1. Fredy R. Romero
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  2. Kathleen Delate
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Correspondence to David J. Hannapel.

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Romero, F.R., Delate, K., Kraus, G.A. et al. Alkamide production from hairy root cultures of Echinacea . In Vitro Cell.Dev.Biol.-Plant 45, 599–609 (2009). https://doi.org/10.1007/s11627-008-9187-1

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