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Micropropagation of Eucalyptus grandis  ×  E. urophylla AEC 224 clone

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Abstract

Genetic transformation systems require protocols that allow regenerating transgenic plants from transformed tissues. This study aimed to establish a protocol for indirect organogenesis in leaf explants of a Eucalyptus grandis  ×  E. urophylla AEC 224 clone. During callogenesis stage, several concentrations of NAA and then NAA or 2,4-D combined with TDZ were tested in JADS culture medium for 30 days, followed by subculture of the explants in the regeneration medium, containing 5.0 µM BA and 0.5 µM NAA for another 30 days. In these media, the explant oxidation rate was high (95 %). Thus, in order to reduce oxidation, different culture media were compared: WPM, MS, JADS and modified QL, followed by explant transfer onto regeneration medium. The highest percentage of regeneration and the lowest oxidation rate were achieved on WPM medium. Then, NAA and 2,4-D were tested in combination with TDZ and also TDZ and BA combined with NAA in WPM medium. The most efficient culture media in terms of shoot regeneration were WPM supplemented with 0.25 µM TDZ and 0.1 µM NAA during 30 days for callus induction and then with 5.0 µM BA and 0.5 µM NAA for another 30 days. This protocol yielded a regeneration rate of 43 %, with a low oxidation of tissues. A rooting experiment was conducted using half strength MS medium and comparing three concentrations of IBA (2.46, 4.90 and 7.35 µM). The highest rooting percentage (35 %) was obtained on medium containing 2.46 µM IBA. Once the shoots were rooted, acclimatization in a greenhouse was not challenging and plant survival reached 100 %.

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Abbreviations

2,4-D:

2,4-dichlorphenoxyacetic acid

BA:

6-benzyladenine

IAA:

Indole-acetic acid

IBA:

Indole-3-butyric acid

JADS:

Culture medium of Correia et al. 1995

LD:

Culture medium of Lainé and David, 1994

MS:

Culture medium of Murashige and Skoog, 1962

NAA:

Naphthalene acetic acid

QLm:

Modified culture medium of Quoirin and Lepoivre, 1977

SIM:

Shoot Induction Medium

TDZ:

Thidiazuron

WPM:

Woody plant medium of Lloyd and McCown, 1980

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Acknowledgments

The authors thank Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Capes) for PhD scholarship and Eileen Bagyary for editing the manuscript.

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Authors and Affiliations

  1. Department of Plant Science, Federal University of Parana, 1540, Rua dos Funcionários, Curitiba, Parana, Brazil

    Cassiana de Oliveira

  2. Embrapa-Forest, 111, Estrada da Ribeira, Colombo, Parana, Brazil

    Juliana Degenhardt-Goldbach & Luziane Franciscon

  3. Department of Technology, Federal University of Parana, 210, avenida Cel, Francisco Heráclito dos Santos, Curitiba, Parana, Brazil

    Gisela Manuela de França Bettencourt

  4. Department of Botany, Federal University of Parana, 210, avenida Cel, Francisco Heráclito dos Santos, Curitiba, Parana, Brazil

    Erika Amano & Marguerite Quoirin

Authors
  1. Cassiana de Oliveira
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  2. Juliana Degenhardt-Goldbach
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  3. Gisela Manuela de França Bettencourt
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  4. Erika Amano
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  5. Luziane Franciscon
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  6. Marguerite Quoirin
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Corresponding author

Correspondence to Cassiana de Oliveira.

Additional information

Project: This work was financially supported by CAPES and EmpresaBrasileira de PesquisaAgropecuária (Embrapa, Brazil).

The online version is available at http://www.springerlink.com

Corresponding editor: Hu Yanbo

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de Oliveira, C., Degenhardt-Goldbach, J., de França Bettencourt, G.M. et al. Micropropagation of Eucalyptus grandis  ×  E. urophylla AEC 224 clone. J. For. Res. 28, 29–39 (2017). https://doi.org/10.1007/s11676-016-0282-6

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  • DOI: https://doi.org/10.1007/s11676-016-0282-6

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