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Isolation, culture and division of olive (Olea europaea L.) protoplasts


Protoplasts from Olea europaea L. have been compared in terms of their yield, viability, cell division and callus differentiation. Viable protoplasts were isolated from in vitro cultured leaves and cotyledons by an overnight incubation in an enzyme solution containing 1–1.5% driselase and 0.5M sucrose. This method allowed high yield of purified protoplasts, which floated and formed a dark green band at the meniscus, after centrifugation. Purified protoplasts were diluted to 3×104 protoplasts·ml−1 in culture medium. After cell wall regeneration, protoplasts gradually increased their volumes under appropriate conditions. The first divisions occurred during the second week in culture. Division efficiency ranged from 5.2 to 9.8% after 20 days in culture. Two weeks later visible microcolonies developed only from cotyledon protoplasts. After 6 weeks in culture, the microcalli were transferred to a solidified culture medium with 0.6% agarose, which induced active callus growth.

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olive proliferation medium, Rugini 1984


OM for the germination of olive embryos


for root induction


for protoplasts


for callus


Bourgin and Nitsch medium 1967


indol-3-butyric acid


naphthalene acetic acid


dichlorophenoxyacetic acid.


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Correspondence to L. A. Cañas.

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Communicated by A. M. Boudet

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Cañas, L.A., Wyssmann, A.M. & Benbadis, M.C. Isolation, culture and division of olive (Olea europaea L.) protoplasts. Plant Cell Reports 6, 369–371 (1987).

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  • Sucrose
  • Cell Wall
  • Agarose
  • Cell Division
  • Solidify Culture Medium