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In vitro culture of Hypericum rumeliacum Boiss. and production of phenolics and flavonoids

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Abstract

The Balkan endemic species, Hypericum rumeliacum, Guttiferae was introduced in vitro for the first time with the aim to study the type of morphogenetic response to plant growth regulators and ability to produce phenolics and flavonoid compounds. The morphoregulatory effect of 2,4-dichlorophenoxyacetic acid (2,4-D), 1-naphtaleneacetic acid (NAA), 6-benzyladenine (BA) and combination of BA with NAA in Murashige–Skoog's basal medium on leaf lamina, internode stem segment, stem node and root cuttings was studied. Histological analysis of the structures regenerated from the primary explants proved the presence of both, embryoids and meristemoids. The node explants cultivated on BA-supplemented medium were the most favourable for regeneration through meristemoids. Therefore a double-stage culture approach, allowing an effective multiplication of large quantities of plant shoots in vitro along with maintenance of the biosynthetic capacity of the culture was developed. It comprised one subculture of three-nodal stem explants derived from the stock shoot cultures on MS medium supplemented with 0.2 mg/l BA followed by subculture of the induced multiple shoots on cytokinin-free MS medium. Determination of the total phenolics and flavonoids showed that the decrease of the levels of these secondary metabolites is transitional, as the exclusionof BA from the medium resulted in an increase of their total content.

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Acknowledgments

This work was supported by the grant of bilateral Slovak - Bulgarian project under contract BG/SK-209/08 and mobility grant of the National Scholarship Programme of the Slovak Republic, grant SU - №83/2008 of the Sofia University “St. Kliment Ohridski”, Bulgaria. This work was also supported by the Slovak Research and Development Agency under contracts No VVCE-0001-07, 0321-07 and SK-BUL 01306 and by Scientific Grant Agency of the Slovak Republic 1/0049/08. We are thankful to Assoc Prof Petka D. Yurukova – Grancharova PhD for the helpful discussions. The authors wish to thank Mrs. Zdenka Lacková for technical assistance.

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Authors and Affiliations

  1. Sofia University, St. Kliment Ohridski, Sofia, Bulgaria

    Kalina Danova & Veneta Kapchina-Toteva

  2. Pavol Jozef Šafárik University, Košice, Slovakia

    Eva Čellárová, Anna Macková & Zuzana Daxnerová

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  1. Kalina Danova
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  2. Eva Čellárová
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  3. Anna Macková
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  4. Zuzana Daxnerová
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  5. Veneta Kapchina-Toteva
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Correspondence to Kalina Danova.

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Editor: H. M. Schumacher

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Danova, K., Čellárová, E., Macková, A. et al. In vitro culture of Hypericum rumeliacum Boiss. and production of phenolics and flavonoids. In Vitro Cell.Dev.Biol.-Plant 46, 422–429 (2010). https://doi.org/10.1007/s11627-010-9299-2

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