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In vitro propagation and production of cardiotonic glycosides in shoot cultures of Digitalis purpurea L. by elicitation and precursor feeding

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Abstract

Digitalis purpurea L. (Scrophulariaceae; Foxglove) is a source of cardiotonic glycosides such as digitoxin and digoxin which are commercially applied in the treatment to strengthen cardiac diffusion and to regulate heart rhythm. This investigation deals with in vitro propagation and elicited production of cardiotonic glycosides digitoxin and digoxin in shoot cultures of D. purpurea L. In vitro germinated seedlings were used as a primary source of explants. Multiple shoot formation was achieved for three explant types (nodal, internodal, and leaf) cultured on Murashige and Skoog (MS) medium with several treatments of cytokinins (6-benzyladenine—BA; kinetin—Kin; and thidiazuron—TDZ) and auxins (indole-3-acetic acid—IAA; α-naphthaleneacetic acid—NAA; and 2,4-dichlorophenoxy acetic acid—2,4-D). Maximum multiple shoots (12.7 ± 0.6) were produced from nodal explants on MS + 7.5 μM BA. Shoots were rooted in vitro on MS containing 15 μM IAA. Rooted plantlets were successfully acclimatized. To further maintain the multiple shoot induction, mother tissue was cut into four equal parts and repeatedly sub-cultured on fresh shoot induction liquid medium after each harvest. On adaptation of this strategy, an average of 18 shoots per explant could be produced. This strategy was applied for the production of biomass and glycosides digitoxin and digoxin in shoot cultures on MS medium supplemented with 7.5 μM BA and several treatments with plant growth regulators, incubation period, abiotic (salicylic acid, mannitol, sorbitol, PEG-6000, NaCl, and KCl), biotic (Aspergillus niger, Helminthosporium sp., Alternaria sp., chitin, and yeast extract) elicitors, and precursors (progesterone, cholesterol, and squalene). The treatment of KCl, mycelial mass of Helminthosporium sp., and progesterone were highly effective for the production of cardenolides. In the presence of progesterone (200 to 300 mg/l), digitoxin and digoxin accumulation was enhanced by 9.1- and 11.9-folds respectively.

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Acknowledgments

Financial support from UGC, SAP-DRS III, DST-FIST and DST-PURSE program, Government of India to the Department of Botany, University of Pune, Pune is gratefully acknowledged. The author Patil JG is thankful to the UGC for the junior research fellowship under meritorious student program.

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  1. Department of Botany, University of Pune, Pune, 411 007, Maharashtra, India

    Jitendra Gopichand Patil, Mahendra Laxman Ahire, Kirti Manik Nitnaware, Sayantan Panda & Tukaram Dayaram Nikam

  2. Herbal Research and Development Institute, Mandal, Gopeshwar, 246 401, Chamoli, Uttarakhand, India

    Vijay P. Bhatt

  3. Department of Genetics, Osmania University, Hyderabad, 500 007, India

    Polavarapu B. Kavi Kishor

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  1. Jitendra Gopichand Patil
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  2. Mahendra Laxman Ahire
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Patil, J.G., Ahire, M.L., Nitnaware, K.M. et al. In vitro propagation and production of cardiotonic glycosides in shoot cultures of Digitalis purpurea L. by elicitation and precursor feeding. Appl Microbiol Biotechnol 97, 2379–2393 (2013). https://doi.org/10.1007/s00253-012-4489-y

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