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Micropropagation of Eucalyptus benthamii to form a clonal micro-garden

  • Micropropagation
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Abstract

Eucalyptus benthamii is an important component of forestry plantations in cold regions, but it is difficult to obtain clonal plants of this species, especially by low rooting. In this study, we developed a method for cloning selected genotypes of E. benthamii using a micropropagation technique, enabling the formation of a clonal micro-garden. Nodal segments from sprouts of mini-stumps in the clonal mini-garden were used as explants. After in vitro establishment of the explants, we tested two selected clones (BP101 and BP118), three culture media (Wood Plant Medium (WPM), Correia and colleagues JADS medium, and Murashige and Skoog medium), and two plant growth regulators (6-benzylaminopurine (BAP) and α-naphthaleneacetic acid (NAA)) for the multiplication of adventitious buds. Additionally, combinations of two other plant growth regulators (BAP and gibberellic acid (GA3)) were tested for the elongation of shoots. The in vitro and ex vitro rooting of micro-plantlets prior to acclimatization were compared. The in vitro bud multiplication of E. benthamii depended on the clone, culture medium, and concentration of plant growth regulators. The best results were obtained with WPM supplemented with 0.5 mg L−1 BAP and 0.05 mg L−1 NAA. The elongation of shoots depended on the clone and plant growth regulator, and the best results were obtained with nutrient medium free of GA3 and BAP. Histological analysis showed that both in vitro and ex vitro rooting were successful, resulting in normal development of adventitious roots showing a vascular connection with the vascular cambium. The new protocol is efficient for micro-plantlet production of E. benthamii and can be used for the formation of a clonal micro-garden for other Eucalyptus or tree species.

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Acknowledgments

We thank FAPESP (São Paulo Research Foundation, Brazil) and CAPES (Coordination for the Improvement of Higher Level, Brazil) for their financial support. We also thank Planflora for providing selected genotypes of E. benthamii. Finally, we are grateful for the technical support provided by José Roberto Romanini.

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Authors and Affiliations

  1. Department of Forest Sciences, “Luiz de Queiroz” College of Agriculture, University of São Paulo (ESALQ/USP), Avenida Pádua Dias, 11, POB 9, 13418-900, Piracicaba, SP, Brazil

    Gilvano Ebling Brondani, Heron Wilhelmus de Wit Ondas, Francisco José Benedini Baccarin & Antonio Natal Gonçalves

  2. Department of Biological Sciences, “Luiz de Queiroz” College of Agriculture, University of São Paulo (ESALQ/USP), Avenida Pádua Dias, 11, POB 9, 13418-900, Piracicaba, SP, Brazil

    Marcilio de Almeida

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  1. Gilvano Ebling Brondani
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  2. Heron Wilhelmus de Wit Ondas
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  3. Francisco José Benedini Baccarin
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  4. Antonio Natal Gonçalves
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  5. Marcilio de Almeida
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Correspondence to Gilvano Ebling Brondani.

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Editor: J. Forster

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Brondani, G.E., de Wit Ondas, H.W., Baccarin, F.J.B. et al. Micropropagation of Eucalyptus benthamii to form a clonal micro-garden. In Vitro Cell.Dev.Biol.-Plant 48, 478–487 (2012). https://doi.org/10.1007/s11627-012-9449-9

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