In this study, the different stages of somatic embryogenesis (SE) of the African oil palm (Elaeis guineensis Jacq.) were characterized biochemically. The total soluble sugars, starch, total free amino acids, and total proteins were extracted, identified and quantified at various stages of embryogenesis: zygotic embryos (initial explants), primary calluses, embryogenic calluses, calluses with pro-embryos, globular embryos, differentiated somatic embryos, and regenerated plants. It was found that at the onset of induction of SE, the level of soluble sugars in the tissues of the explants fell by half. During this period, the total soluble sugars present in the cultures consisted basically of glucose and fructose. In the process of regeneration and maturation, the concentrations of soluble sugars gradually increased, reaching the highest values in the last two stages of development. At this stage, the disaccharide sucrose accounts for more than 80 % of the composition of total soluble sugars in the explants. Compared to starch, we found that the concentrations thereof in developing tissues are inversely proportional to that of soluble sugars virtually throughout embryogenic development. As for free amino acids, we found that after 30 days of induction until formation of the embryogenic calluses, there is an accentuated synthesis of total free amino acids in the explant tissue. In this stage, there was a significant increase in the levels of alanine and serine in the tissues. However, after the formation of the embryogenic calluses, the levels of total free amino acids present in the cultures become stabile and remain constant until the end of cultivation. Similar results were found for total protein, which also showed a significant increase at the onset of induction, undergoing slight changes during the remainder of the cultivation.
420 days of cultivation, the propagules formed somatic embryos with no connection to source tissues, initially composed (globular stage) of a very characteristic ground meristem and protoderm. After 480 days of cultivation, as the cultures matured (torpedo stage), procambial strands, a structural characteristic also observed in mature zygotic embryos, were reported. The results provide an in-depth understanding of somatic embryogenesis of immature leaves of oil palm. Further, current analysis develops morphological markers at different stages of development obtained during the process.
In species of the Arecaceae, initial conversion of somatic embryos to non-rooted plantlets (those with only shoot development and no roots) is common. A consistent methodology for improving rooting and survival of oil palm plantlets derived from somatic embryos was developed. Two experiments were carried out. In the first, non-rooted shoots regenerated via somatic embryogenesis were inoculated in nutrient medium supplemented with 0, 53.7, or 107.4 μM indole-3-butyric acid (IBA) or 1-naphthaleneacetic acid (NAA). In the second, the optimum treatment as determined in the first experiment was evaluated in three culture systems: semi-solid medium (SS), stationary liquid (SL), and doublephase system (DP), with and without the addition of activated charcoal. Next, the plantlets were acclimatized in a greenhouse, where the influence of the presence or absence of roots, the number of roots, the length of the main root, and the height of the shoot on survival were assessed. Supplementation with 53.7 μM IBA and the DP system without activated charcoal provided the highest percentage of rooted plantlets (92.9%) and number of roots (6.3) and improved the length of the aerial parts (11.0 cm) and the survival of the plantlets during the acclimatization (82.1%). The best survival of plantlets was obtained when they were acclimatized with at least one root and with a height greater than 7.5 cm.
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