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In vitro establishment of shoot meristems of Ilex paraguariensis and identification of endophytic bacteria

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Abstract

Yerba mate (Ilex paraguariensis A. St.-Hil.) is a species of great economic, social and environmental importance for the southern regions of Brazil, Uruguay and Argentina. Currently the most diverse products are obtained from mate leaves, including mate tea. The objective of this study was to establish shoot meristem cultures (meristematic dome and a few primordia) of elite clones and identify the endophytic bacteria present in the explants. We tested the effect of clones (F1, F2, A03 and A07), culture media (MS, 1/2MS, 1/4MS and WPM), cytokinins (kinetin, BA and 2iP), activated charcoal (1, 2 and 3 g L−1), and disinfecting agent (sodium hypochlorite and mercuric chloride) on in vitro establishment. F1 and F2 clones were the most responsive for shoot meristem in vitro culture. WPM medium supplemented with 8.8 µM 2iP, 0.2 µM NAA and 3 g L−1 activated charcoal was the most suitable for the in vitro establishment of the F1 clone. No phytotoxic effect of the disinfectant was observed and some meristems sprouted. The isolated endophytic bacterium was identified for the first time in yerba mate as Agrobacterium larrymoorei. To conclude, we were able to establish in vitro culture of yerba mate using meristems as explants but the tissues were not free of endophytic microorganisms which could interfere with explant response.

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Acknowledgements

The authors thank EMBRAPA Forestry for providing plant material and laboratory facilities, the Laboratories of Biochemistry and of Structural Botany, Federal University of Parana, Brazil and CAPES for providing a Grant to J. C. Tomasi.

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Authors and Affiliations

  1. Department of Crop Production and Protection, Federal University of Parana, 1540 Rua dos Funcionários, Curitiba, PR, 80035-060, Brazil

    Jéssica de Cássia Tomasi & Renata Lucia Grunennvaldt

  2. Brazilian Agricultural Research Corporation (EMBRAPA) Forestry, Colombo, PR, Brazil

    Juliana Degenhardt-Goldbach & Luziane Franciscon

  3. Department of Basic Pathology, Federal University of Parana, Curitiba, Brazil

    Germana Dávila dos Santos & Vania Aparecida Vicente

  4. Department of Botany, Federal University of Parana, Curitiba, Brazil

    Cleusa Bona & Marguerite Quoirin

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  1. Jéssica de Cássia Tomasi
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  2. Juliana Degenhardt-Goldbach
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  3. Renata Lucia Grunennvaldt
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  4. Germana Dávila dos Santos
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  5. Vania Aparecida Vicente
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  8. Marguerite Quoirin
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Correspondence to Jéssica de Cássia Tomasi.

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Project funding: Brazilian Agricultural Research Corporation (EMBRAPA) Forestry.

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Corresponding editor: Zhu Hong.

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de Cássia Tomasi, J., Degenhardt-Goldbach, J., Lucia Grunennvaldt, R. et al. In vitro establishment of shoot meristems of Ilex paraguariensis and identification of endophytic bacteria. J. For. Res. 30, 1765–1777 (2019). https://doi.org/10.1007/s11676-018-0763-x

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