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Somatic embryogenesis and plantlet regeneration from immature anthers of Opuntia ficus-indica

Bouamama, B., Ben Salem, A., Zoghlami, N., Zemni, H., Mliki, A., Ghorbel, A.
Journal of horticultural science & biotechnology 2011 v.86 no.4 pp. 313-318
2,4-D, Opuntia ficus-indica, activated carbon, anthers, biotechnology, callus, cladodes, cultivars, culture media, greenhouses, horticulture, photoperiod, plantlets, protocols, roots, scanning electron microscopy, somatic embryogenesis, somatic embryos, thidiazuron
An in vitro protocol has been developed for callus induction, somatic embryogenesis, and plant regeneration in the Barbary fig (Opuntia ficus-indica) cultivars ‘Gialla’ and ‘Moore’. Immature anthers cultivated in the dark on induction medium which consisted of Chée and Pool medium containing 2.0 mg l–1 2,4-dichlorophenoxyacetic acid (2,4-D) and 2.5 mg l–1 thidiazuron (TDZ), produced primary nodular and greenish calli within 6 – 8 weeks. Periodic transfer of nodular calli on induction medium in the dark induced the formation of compact embryogenic masses after 4 – 5 months of cultivation, with induction rates of 58.97 ± 1.10% and 61.15 ± 1.16% for ‘Moore’ and ‘Gialla’, respectively. Various distinct stages were observed during the development of somatic embryos on induction medium, including pro-embryogenic, globular, heart-, torpedo-, and cotyledonary-shaped somatic embryos, as well as secondary somatic embryos. Further development of somatic embryos was accomplished using half-strength Murashige and Skoog medium containing 1% (w/v) activated charcoal under a 16-h photoperiod. Bipolarity of the somatic embryos was confirmed by environmental scanning electron microscopy. Regenerated plantlets with well-developed cladodes and roots were transferred to a greenhouse and acclimatised successfully