Acta Physiologiae Plantarum

, Volume 34, Issue 2, pp 471–477 | Cite as

Genetic transformation of three sweet orange cultivars from explants of adult plants

  • Pâmela Fávero
  • Francisco de Assis Alves Mourão Filho
  • Liliane Cristina Libório Stipp
  • Beatriz Madalena Januzzi Mendes
Original Paper


The difficulty in adult tissue genetic transformation in woody species is still an obstacle to be overcome, including in most sweet orange cultivars of the Brazilian citrus industry. This work reports that, after in vitro culture adjustments, transgenic adventitious buds of ‘Hamlin’, ‘Pêra’, and ‘Valencia’ sweet oranges (Citrus sinensis L. Osbeck) were recovered using adult material as explant source, in genetic transformation experiments via Agrobacterium tumefaciens. The transgenic buds were identified by the GUS histochemical analysis and confirmed by PCR analysis, which indicated the presence of an amplified fragment of 817 bp corresponding to the uidA gene sequence. The efficiencies of genetic transformation for ‘Hamlin’, ‘Pêra’, and ‘Valencia’ sweet orange cultivars were 2.5, 1.4, and 3.7%, respectively. Media supplemented with auxins and cytokinins during co-culture, and media with high concentrations of cytokinins (3 mg L−1) during transgenic selection led to the transformation and, consequently, the regeneration of adequate number of adventitious buds for the three cultivars. The use of sonication during the explant disinfection was not effective to reduce endophytic contamination and reduced transformation efficiency.


Citrus sinensis Tissue culture Biotechnology Genetic improvement Agrobacterium tumefaciens 


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Copyright information

© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2011

Authors and Affiliations

  • Pâmela Fávero
    • 1
  • Francisco de Assis Alves Mourão Filho
    • 1
  • Liliane Cristina Libório Stipp
    • 1
  • Beatriz Madalena Januzzi Mendes
    • 2
  1. 1.Escola Superior de Agricultura “Luiz de Queiroz”Universidade de São PauloPiracicabaBrazil
  2. 2.Centro de Energia Nuclear na AgriculturaUniversidade de São PauloPiracicabaBrazil

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