Adventitious shoot formation and plant regeneration from Pinus pinaster Sol. ex Aiton

  • Filomena Calixto
  • M. Salomé Pais


Pinus pinaster plants were regenerated from cotyledons excised from in vitro germinated seeds and axenically cultured on induction medium (GMD). 6-Benzyladenine (2.2 µM) induced the highest frequency of direct bud formation from cotyledons. An average of 13.1 ± 2.1 elongated shoots per cotyledon was obtained. Germination time influenced shoot induction, and the organogenic potential decreased with explant age. Cotyledons remained for 21 d on induction medium, and in order to promote adventitious shoot elongation, they were transferred to Gupta and Durzan’s DCR medium without growth regulators, containing 0.5% (wt/vol) activated charcoal and 3% (wt/vol) sucrose. Rooting was achieved by application of an indole-3-butyric acid, (396.6 µM) pulse (24 h at 4° C), followed by transfer to a sterile mixture of peat plus perlite (1:1 vol/vol). Ninety-eight to 100% of the regenerated plants were successfully acclimatized. All plants have survived after transfer to the field.

Key words

cotyledon micropropagation organogenesis Pinus pinaster rhizogenesis 


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Copyright information

© Society for In Vitro Biology 1997

Authors and Affiliations

  • Filomena Calixto
    • 1
  • M. Salomé Pais
    • 1
  1. 1.Centro de Biotecnologia VegetalFaculdade de Ciências de LisboaLisboaPortugal

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