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Cardiotonic glycosides from biomass of Digitalis purpurea L. cultured in temporary immersion systems

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Abstract

Cardiotonic glycosides are extracted mostly from leaves of Digitalis plants. Commercial production of bioactive secondary metabolites by traditional agriculture is an inefficient process and can be affected by climatic and soil conditions. Strategies, based on in vitro culture methods, have been extensively studied to improve the production of specific plant derived chemicals. The aim of the present research was to obtain biomass of D. purpurea using the temporary immersion system (TIS) and to determine the content of cardiotonic glycosides (digitoxin, digoxin and lanatoside C) as secondary metabolites of commercial value for the pharmaceutical industry. Shoots were cultured in 1,000 ml TIS during 28 days. The effect of four immersion frequencies (once every 2, 4, 6, and 12 h) was studied. Biomass accumulation was influenced by immersion frequency. The maximum biomass accumulation (values in respect of fresh and dry weight) was obtained with immersions every 4 h (six immersions per day). HPLC analysis revealed the presence of digoxin and digitoxin for all immersion frequencies. No lanatoside C was detected in the biomass cultured in TIS. Digoxin concentrations varied depending on the frequencies tested. In contrast, the digitoxin content showed no dependency on the immersion frequency. Net production of digoxin and digitoxin per TIS were found to be higher with immersions every 4 h. The best net production of digitoxin and digoxin per TIS were 167.6 and 119.9 μg, respectively. The development of organ culture based on temporary immersion system can be a reliable method for the steady production of biomass for cardiotonic glycosides production, which is reported for the first time for Digitalis genus in this communication.

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Abbreviations

6-BAP:

6-Benzylaminopurine

DW:

Dry weight

FW:

Fresh weight

IAA:

Indole-3-acetic acid

TIS:

Temporary immersion system

MS:

Murashige and Skoog medium

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Acknowledgments

The authors wish to thank the support of the EU through the ALFA Network CARIBIOTEC (project AML/B7-311/97/0666/II-0201), the German Ministry for Education and Research (BMBF) and the Cuban Ministry of Science, Technology and Environment (CITMA). Also to Emeritis Prof. Dr. ir. Oswald Van Cleemput from the Faculty of Bioscience Engineering at Gent University for the revision of the manuscript.

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Authors and Affiliations

  1. Instituto de Biotecnología de las Plantas, Universidad Central “Marta Abreu” de Las Villas, Carretera a Camajuaní km 5,5, 54830, Santa Clara, Villa Clara, Cuba

    Naivy Pérez-Alonso, Alina Capote-Perez & Elio Jiménez

  2. BioPlanta GmbH, Deutscher Platz 5, 04103, Leipzig, Germany

    Dirk Wilken & André Gerth

  3. Institute of Non-Classic Chemistry, Leipzig University (INC), Permoserstraße 15, 04318, Leipzig, Germany

    Annett Jähn & Horst-Michael Nitzsche

  4. Saxon Institute for Applied Biotechnology, Leipzig University, Permoserstraße 15, 04318, Leipzig, Germany

    Gerhard Kerns

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  1. Naivy Pérez-Alonso
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  2. Dirk Wilken
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  3. André Gerth
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  7. Alina Capote-Perez
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  8. Elio Jiménez
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Correspondence to Naivy Pérez-Alonso.

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Pérez-Alonso, N., Wilken, D., Gerth, A. et al. Cardiotonic glycosides from biomass of Digitalis purpurea L. cultured in temporary immersion systems. Plant Cell Tiss Organ Cult 99, 151–156 (2009). https://doi.org/10.1007/s11240-009-9587-x

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  • DOI: https://doi.org/10.1007/s11240-009-9587-x

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