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An optimised protocol for plant regeneration from embryogenic suspension cultures of date palm, Phoenix dactylifera L., cv. Deglet Nour

  • Cell Biology and Morphogenesis
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Abstract

An improved protocol is described for the large-scale micropropagation of an elite date palm (Phoenix dactylifera L.) cultivar, Deglet Nour. Clonal plants were regenerated from somatic embryos derived from highly proliferating suspension cultures. Friable embryogenic calli were initiated from both leaf and inflorescence explants. Suspension cultures consisting of pro-embryonic masses were established from calli showing a high competency for somatic embryogenesis. The subculture of suspensions in liquid medium enriched with low amounts of plant growth regulators (1 mg l-1 2,4-dichlorophenoxyacetic acid with 300 mg l-1 charcoal) resulted in the differentiation of large numbers of somatic embryos. The productivity of the cultures increased 20-fold (from 10 to 200 embryos per month per 100 mg fresh weight of embryogenic callus) when embryogenic suspensions were used instead of standard cultures on solid media. The overall production of somatic embryos reached 10,000 units per litre per month. Partial desiccation of the mature somatic embryos, corresponding to a decrease in water content from 90% to 75%, significantly improved germination rates (from 25% to 80%). The cutting back of the cotyledonary leaf was also found to stimulate embryo germination. Flow cytometric analysis showed that the micropropagation protocol followed here did not affect the ploidy level of somatic embryo-derived plantlets.

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Abbreviations

BAP :

6-Benzylaminopurine

DAPI :

4′, 6-Diamidino−2-phenylindole

2,4-D :

2,4-Dichlorophenoxyacetic acid

NAA :

α-Naphthaleneacetic acid

PEMs :

Pro-embryonic masses

PGR :

Plant growth regulator

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Acknowledgements

We are grateful to Dr. B.S. Ahloowalia, Plant Breeding and Genetics Section, Joint FAO/IAEA Division, International Atomic Energy Agency, Vienna, Austria, for his help with the manuscript, comments and encouragement. This investigation was supported by the International Atomic Energy Agency, under TC Project RAF/5/035.

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Authors and Affiliations

  1. Laboratoire des Biotechnologies Végétales, Faculté des Sciences de Sfax, 802, 3018, Sfax, Tunisia

    L. Fki, R. Masmoudi & N. Drira

  2. UMR 1098, Programme Palmier, TA80-PSIII, CIRAD-CP, 34398 , Montpellier Cedex 5, France

    A. Rival

Authors
  1. L. Fki
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  2. R. Masmoudi
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  3. N. Drira
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  4. A. Rival
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Correspondence to A. Rival.

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Communicated by P. Debergh

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Fki, L., Masmoudi, R., Drira, N. et al. An optimised protocol for plant regeneration from embryogenic suspension cultures of date palm, Phoenix dactylifera L., cv. Deglet Nour. Plant Cell Rep 21, 517–524 (2003). https://doi.org/10.1007/s00299-002-0558-5

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  • DOI: https://doi.org/10.1007/s00299-002-0558-5

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