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Establishment of adventitious root culture of Stevia rebaudiana Bertoni in a roller bottle system

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Abstract

Cultures of adventitious roots of Stevia rebaudiana (Bert.) Bertoni were performed in a roller bottle system for the production of both primary and secondary metabolites. Adventitious roots were induced from 1-cm-long root tip explants derived from in vitro regenerated plantlets on solid Murashige and Skoog (MS 1962) media supplemented with 10.7 μM of α-naphthaleneacetic acid. These cultures were successfully maintained in the same medium for 6 months with regular subcultures after 4 weeks. Thereafter, the roots were cut into 1.0- to 1.5-cm-long segments and transferred to the roller bottle system containing a fresh root tissue culture on liquid MS medium supplemented with 10.7 μM NAA. The apparatus consisted of a flask rolling system adjusted to 4g, and 3° of flask inclination. The roots were allowed to grow in the absence of light for adaptation and adventitious root formation. The best conditions for cultivation were investigated, considering culture volume (25 ml), culture period (4 weeks), salt concentrations in the nutrient medium (33%) and optimal initial inoculum (0.2 g) of S. rebaudiana roots. These results could give important information on how to improve the development of adventitious roots of S. rebaudiana for the production of primary and secondary metabolites.

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Abbreviations

ESI-MS:

Electrospray ionization mass spectrometric analysis

FW:

Fresh weight

GI:

Growth index

HPLC:

High-performance liquid chromatography

MS:

Murashige and Skoog media

NAA:

α-Naphthaleneacetic acid

RBS:

Roller bottle system

Rf:

Retention factor

SGs:

Diterpenoid steviol-glycosides

TLC:

Thin layer chromatography

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Acknowledgements

The authors are grateful to SteviaFarma Industrial S/A, the Brazilian funding agencies National Council of Scientific Development (CNPq), Brazilian Commission for Personnel Improvement (CAPES) and Araucária Foundation for financial support.

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Authors and Affiliations

  1. Departamento de Farmácia, Universidade Estadual de Maringá, Avenida Colombo, 5790, Maringá, Paraná, 87020-900, Brazil

    Rafael Valim Reis, Ana Paula Patrão Luis Borges, Talita Perez Cantuaria Chierrito, Arildo José Braz de Oliveira & Regina Aparecida Correia Gonçalves

  2. Departamento de Agronomia, Universidade Estadual de Maringá, Avenida Colombo, 5790, Maringá, Paraná, 87020-900, Brazil

    Eliezer Rodrigues de Souto

  3. Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Paraná, Curitiba, Paraná, Brazil

    Lauro Mera de Souza & Marcello Iacomini

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  1. Rafael Valim Reis
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  2. Ana Paula Patrão Luis Borges
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  3. Talita Perez Cantuaria Chierrito
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  4. Eliezer Rodrigues de Souto
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  7. Arildo José Braz de Oliveira
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  8. Regina Aparecida Correia Gonçalves
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Correspondence to Regina Aparecida Correia Gonçalves.

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Reis, R.V., Borges, A.P.P.L., Chierrito, T.P.C. et al. Establishment of adventitious root culture of Stevia rebaudiana Bertoni in a roller bottle system. Plant Cell Tiss Organ Cult 106, 329–335 (2011). https://doi.org/10.1007/s11240-011-9925-7

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  • DOI: https://doi.org/10.1007/s11240-011-9925-7

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