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Somatic embryo production from immature seeds of carob (Ceratonia siliqua L.) : Histological evidence

Ksia E., Harzallah-skhiri F., Verdeil J.L., Gouta H., Alemanno L., Bouzid S.. 2008. Journal of Horticultural Science and Biotechnology, 83 (4) : p. 401-406.

DOI: 10.1080/14620316.2008.11512398

Immature seeds isolated from pods of carob (Ceratonia siliqua L.) were used as a source of explants for callogenesis and somatic embryogenesis. Suitable developmental Stages of immature seed explants, and in vitro culture conditions for the production of translucent callus were identified. The addition of 2,4-dichlorophenoxyacetic acid (2,4-D) to the culture medium was found to be necessary for callogenesis. Both the developmental Stage of the explant and the 2,4-D concentration had significant effects on the percentage of callus production. A high proportion (67.0 ± 1.0%) of immature seed at Stage 2 gave rise to callus when cultured on a medium containing 9.0 µM 2,4-D, which was significantly higher than all the other treatments tested. Embryogenic calli consisted of a white-translucent mass proliferating all around the explants and gave rise to somatic embryos. On day-45 of culture, transverse sections of callus showed de-differentiated cells located on the surface, which gave rise to pro-embryos. On day-70, early somatic embryos were observed as globular white structures, and others were at the heart-shaped stage. On day-90, complete somatic embryos were initiated which had attained the cotyledonary stage, characterised by a defined meristematic area and separation of a protoderm and pro-vascular bundles.

Mots-clés : ceratonia siliqua; embryon somatique; culture d'embryon; embryogénèse somatique; culture in vitro; tunisie

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