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Somatic embryogenesis and plant regeneration in date palm Phœnix dactylifera L., cv. Boufeggous is significantly improved by fine chopping and partial desiccation of embryogenic callus

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Abstract

Plant regeneration through somatic embryogenesis from young leaf explants (5–10 mm long) adjacent to the apex of 5–6 year old offshoots of Tunisian date palm (Phœnix dactylifera L.), cultivar Boufeggous was successfully achieved. Factors affecting embryogenic callus initiation, including plant growth regulators and explant size, were investigated. The highest induction frequencies of embryogenic calli occurred after 6–7 months on MS medium supplemented with 10 mg l−1 2,4-D and 0.3 mg l−1 activated charcoal. The subculture of these calli onto maintenance medium resulted in the formation of proembryos. Fine chopping and partial desiccation (6 and 12 h) of embryogenic calli with proembryos prior to transfer to MS medium supplemented with 1 mg l−1 ABA stimulated the rapid maturation of somatic embryos. Maturated somatic embryo yield per 0.5 g FW of embryogenic callus was 51 embryos with an average maturation time of 55 days. This was increased to 422 with finely chopped callus, and 124 and 306 embryos following 6 and 12 h desiccation treatments, respectively. The average time to maturation for these 3 treatments was 35, 43 and 38 days, respectively. Subsequent substitution of ABA in MS medium with 1 mg l−1 NAA resulted in the germination and conversion of 81% of the somatic embryos into plantlets with normal roots and shoots. The growth of regenerated somatic plants was also monitored in the field.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid

ABA:

Abscisic acid

AC:

Activated charcoal

BAP:

6-Benzylaminopurine

FW:

Fresh weight

IBA:

Indole-3-butyric acid

MS:

Murashige and Skoog medium

NAA:

α-Naphthalenacetic acid

PGRs:

Plant growth regulators

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Acknowledgments

The authors thank Dr Alain Poupet for critical reading of the manuscript. This work was financial supported by grants from the IPGRI “CRPh Degache, Projet FEM-PNUD-IPGRI, RAB 98 G31”.

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Authors and Affiliations

  1. Immunologie et Biotechnologie, Faculté des Sciences de Tunis, Laboratoire de Génétique Moléculaire, 2092, El-Manar Tunis, Tunisia

    A. Othmani, M. Marrakchi & M. Trifi

  2. Laboratoire de Culture in vitro, Centre Régional de Recherches Agronomiques Oasiennes, 2260, Degache, Tunisia

    C. Bayoudh

  3. Faculté des Sciences de Sfax, Laboratoire des Biotechnologies Végétales Appliquées à l’Amélioration des Cultures, B.P. 802, 3018, Sfax, Tunisia

    N. Drira

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  1. A. Othmani
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  2. C. Bayoudh
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  3. N. Drira
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Correspondence to A. Othmani.

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Othmani, A., Bayoudh, C., Drira, N. et al. Somatic embryogenesis and plant regeneration in date palm Phœnix dactylifera L., cv. Boufeggous is significantly improved by fine chopping and partial desiccation of embryogenic callus. Plant Cell Tiss Organ Cult 97, 71–79 (2009). https://doi.org/10.1007/s11240-009-9500-7

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