Plant Cell Reports

, Volume 18, Issue 3–4, pp 271–278 | Cite as

Agrobacterium-mediated transformation of citrange: factors affecting transformation and regeneration

  • M. Cervera
  • J. A. Pina
  • J. Juárez
  • L. Navarro
  • L. Peña

Abstract

The effects of cocultivation with Agrobacterium tumefaciens, regeneration and selection conditions on the transformation efficiency of citrange (Citrus sinensis L. Osbeck×Poncirus trifoliata L. Raf.) have been investigated. Factors such as cocultivation period, preculture of explants, use of acetosyringone or feeder plates during cocultivation, cocultivation on a medium rich in auxins, postcultivation in darkness, and different kanamycin concentrations for selection were assessed. A 3-day cocultivation on a medium rich in auxins improved transformation frequencies, since it increased the number of dividing cells competent for transformation, at the cut ends of the explants. Exposure of explants to darkness for 4 weeks on selection medium resulted in further callus development and increased the regeneration frequency of transgenic shoots. Furthermore, this treatment drastically reduced the number of regenerated escape shoots. A transformation efficiency of 41.3% was achieved using the optimized transformation procedure.

Key words Citrus Citrange Genetic transformation 

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Copyright information

© Springer-Verlag Berlin Heidelberg 1998

Authors and Affiliations

  • M. Cervera
    • 1
  • J. A. Pina
    • 1
  • J. Juárez
    • 1
  • L. Navarro
    • 1
  • L. Peña
    • 1
  1. 1.Dpto. Protección Vegetal y Biotecnología, Instituto Valenciano de Investigaciones Agrarias (IVIA), Apartado Oficial, E-46113 Moncada, Valencia, Spain e-mail: lpenya@ivia.es Fax: +34-6-1390240ES

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