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The influence of salicylic acid elicitation of shoots, callus, and cell suspension cultures on production of naphtodianthrones and phenylpropanoids in Hypericum perforatum L.

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Abstract

Hypericum perforatum is a well known medicinal plant. The main pharmacological properties are due to the presence of naphtodianthrones such as hypericin and pseudohypericin. Unfortunately the levels of these compounds vary under different environmental conditions. Elicitation of in vitro cultures is a useful approach to enhance and extend production of desirable products. Therefore, the effects of salicylic acid were characterized on different explants of H. perforatum L. (cells, calli and shoots) cultured in vitro. It appears at first that salicylic acid did not affect growth and development of these explants. In addition, the production of both hypericin and pseudohypericin has doubled in elicited cell suspension cultures but not in the two other cultures. Furthermore, phenylpropanoids that are among the most frequently observed metabolites affected upon treatment of in vitro culture material with elicitors, were produced and the enzymatic activities of phenylalanine ammonia lyase and of chalcone isomerase were stimulated upon elicitation. These effects were dependant of the type of in vitro culture, the concentration of salicylic acid and the duration post-elicitation. The H. perforatum cells were globally more sensitive to salicylic acid elicitation when maintained in an undifferentiated state and particularly in cell suspension cultures. In the absence of glands considered as the sites of naphtodianthrones biosynthesis, cells and calli were capable of producing these compounds. This implies that salicylic acid could act at biosynthesis level but not for the accumulation of both hypericin and pseudohypericin. Consequently, the regulation of this process is more complex than cited in the literature involving the responsibility of only Hyp-1 gene, encoding a hypericin biosynthetic enzyme, cloned and characterized from H. perforatum.

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Abbreviations

BA:

Benzyladenine

CHI:

Chalcone isomerase

2,4-D:

2,4-Dichlorophenoxyacetic acid

HPLC:

High-performance liquid chromatography

JA:

Jasmonic acid

NAA:

α-Naphtaleneacetic acid

PAL:

Phenylalanine ammonia lyase

pKS:

Polyketide synthase

SA:

Salicylic acid

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Acknowledgements

This work was supported by the Ministère des Affaires Etrangères – France (Programme COCOP: Réseau d’Enseignement regional Postgraduate en Biologie, grant no. DSUR-NGE-4B1-505). We are very grateful to Professor C. Jay-Allemand from University of Montpellier (France) for helpful advice for secondary metabolites analyses. We thank Dr Jan Kosuth and Eva Cellarova from University in Kosice (Slovakia) for their help in the RTPCR analyses. We acknowledge to Professor F. Brignolas for his help in statistical analyses and Mrs Alison Price for carefully reading of the manuscript. We thank Dr S. Ounnar and G. Moreau for their technical help.

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Authors and Affiliations

  1. Laboratoire de Biologie des Ligneux et des Grandes Cultures, UPRES EA 1207, Faculté des Sciences, Université d’Orléans, rue de Chartres, B.P. 6759, 45067, Orléans Cedex 2, France

    Sonia Gadzovska, Stéphane Maury, Alain Delaunay, Daniel Hagège & Claude Joseph

  2. INRA, USC 1328, Arbres et réponses aux Contraintes Hydriques et Environnementales (ARCHE), 45067, Orléans Cedex 2, France

    Stéphane Maury, Alain Delaunay & Claude Joseph

  3. Faculty of Natural Sciences and Mathematics, Institute of Biology, P.O. Box 162, 1000, Skopje, Macedonia

    Sonia Gadzovska & Mirko Spasenoski

  4. Centre R et D Nestlé-Tours, 101 avenue Gustave Eiffel, B.P. 49716, 37097, Tours Cedex 2, France

    Didier Courtois

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  1. Sonia Gadzovska
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  2. Stéphane Maury
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Correspondence to Daniel Hagège.

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Gadzovska, S., Maury, S., Delaunay, A. et al. The influence of salicylic acid elicitation of shoots, callus, and cell suspension cultures on production of naphtodianthrones and phenylpropanoids in Hypericum perforatum L.. Plant Cell Tiss Organ Cult 113, 25–39 (2013). https://doi.org/10.1007/s11240-012-0248-0

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