Ultrastructural studies of 2,4-D (2,4 dichlorophenoxyacetic acid) induced coconut calli and of untreated controls enabled us to characterize early events in cellular reorganization leading to embryogenic cell individualization and subsequent development into proembryos. Embryogenic cells were characterized by special features that chie¯y aected the nucleus, cytoplasm and cell wall: deep invaginations of the nuclear envelope, proliferation of dictyosomes, with emission of Golgi vesicles, directly related to an increase in cell wall thickness. Modi®cation of the cell wall structure was studied and particular attention was paid to the cytolocalization of b-1,4-glucans, and of callose and pectin epitopes, using gold-conjugated probes. The ®rst changes (detected 7±14 d after 2,4-D increase) involved the closure of plasmodesmata, breaking of symplastic continuity, and callose deposition. The acquisition of embryogenic competence was linked to the appearance of an outer layer of ®brillar material containing pectin epitope (mainly un-methyl-esteri®ed), fully coating the embryogenic cells (21 d after the induction treatment). Some of the ultrastructural changes observed during the reprogramming of somatic cells towards embryogenesis can be likened to those accompanying the maturation of female gamete cells in many plant species. The possible signi®cance of these observations is discussed.
Immature inflorescences of coconut belonging to three different genotypes were cultured on a solid medium supplemented with activated charcoal (2%) and a range of 2,4-dichlorophenoxyacetic acid (2,4-D) concentrations (from 1.5 to 3.5 × 10(-4)M). Globular white callus formed from immature floral meristems, depending on inflorescence age and 2,4-D concentration. Acquisition of embryogenic competence is described histologically. Somatic embryos presented a functional bipolar organization with a completely differentiated shoot meristem which is reported here for the first time in coconut tissue culture. Embryo maturation allowed reliable plant regeneration of this in vitro recalcitrant species. Details are given of exogenous hormonal requirements for the acquisition of embryogenic competence and embryo maturation.
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