Many embryogenic systems have been designed to generate somatic embryos (SEs) with the morphology, biochemistry, and vigor uniformity of zygotic embryos (ZEs). During the current investigation, several antioxidants were added to the maturation media of the developing somatic embryos of date palm. Explant material was a friable embryogenic callus that was placed in maturation media containing ABA at 0.5 mg L⁻¹, 5 g L⁻¹ polyethylene glycol, and 10 g L⁻¹ phytagel. Furthermore, α-tocopherol or reduced glutathione (GSH) were used separately at (25 and 50 mg L⁻¹). These treatments were compared to a widely used date palm combination of reduced ascorbic acid (ASC) and citric acid at 150 and 100 mg L⁻¹, respectively, and to the medium free from any antioxidants. The relative growth percentage of embryogenic callus (EC), globularization degree, differentiation%, and SEs number were significantly increased with GSH (50 mg L⁻¹). Additionally, the latter treatment significantly enhanced the conversion% of SEs and the number of secondary somatic embryos (SSEs). ASC and citric acid treatment increased leaf length, while α-tochopherol (50 mg L⁻¹) elevated the number of leaves plantlet⁻¹. GSH at 50 mg L⁻¹ catalyzed the activities of polyphenol oxidase (PPO) and peroxidase (POD) in EC and enhanced the accumulation of proteins in SEs.

The study of morpho-anatomical aspects, metabolic changes of proteins, antioxidant substances, as well as phenolic compounds in embryogenic callus (EC) and degenerative embryogenic callus (DEC) was the aim of the present investigation. Ability to form somatic embryos (SEs) was associated with the softness of the EC, which exhibited a white or creamy color and was composed of isodiametric cells containing dense cytoplasm, conspicuous nuclei and minimal vacuoles with observed mitotic activity. Furthermore, protein, reduced glutathione (GSH) and ascorbic acid (ASC) concentrations and the ratio between ASC and dehydroascorborbic acid (DHA) were increased significantly in the EC in comparison to the DEC. In addition, the phenolic extract of the EC was proved to have higher scavenging activity than the extract from the DEC. A loss of embryogenic competence in the DEC was correlated with the presence of more rigid clumps and such calli had a yellowish to brown color and no cell division could be observed in the cells of such aggregates as the cells had large vacuoles and they have very thick walls. Moreover, these morphological and anatomical observations of the DEC were accompanied by accumulations of the oxidized form of ascorbic acid (DHA), H₂O_2, total soluble phenolic compounds and overaccumulation of naringenin. Alternations in cellular metabolism can affect and regulate the morphogenesis of somatic embryos.

The survival and growth of in vitro plantlets are still problematic for the date palm; thus, the application of nanotechnology may provide date palm plantlets that develop faster with enhanced growth and biochemical parameters. Moreover, the usage of Fe-modified activated carbon (Fe-AC) prepared from date palm pits instead of activated charcoal (AC) in tissue culture media during the current investigation can be considered an innovative approach. Furthermore, the effect of zinc oxide nanoparticles (ZnO-NPs) and bulk particles (ZnO-BPs) on the rooting and growth of date palm plantlets and on some physiological indices was also investigated. X-ray diffraction, dynamic light scattering, scanning electron microscopy, and transmission electron microscopy were used to characterize Fe-AC and ZnO-NPs. As detected from the characterization, the particle size was 720 nm for Fe-AC and 60 nm for ZnO-NPs. The obtained results indicated that AC significantly enhanced plantlet growth, plantlet height, number of leaves/plantlet, root length, number of roots/plantlet, and the concentrations of IAA, chlorophyll a and chlorophyll b compared with Fe-AC. On the other hand, sugars and phenols concentrations, and POD and PAL activities were highly increased with Fe-AC. There was no significant difference in terms of plant fresh weight (FW), leaf length, leaf width, protein, amino acids, and carotenoids. ZnO nanoparticles significantly improved plant height, leaf length, leaf width, root length, and the concentrations of IAA, chlorophylls a and b, and carotenoids; in contrast, the bulk form increased roots/plantlet, amino acids, and total sugars. Supplying the MS medium with combinations of ZnO-NPs or ZnO-BPs and AC or Fe-AC significantly increased all growth and physiological parameters. Plant height, leaf length, leaf width, root length, and the concentrations of IAA, protein, and chlorophylls a and b significantly increased when ZnO-NPs were combined with AC. Amino acids, total sugars, carotenoids, and phenols concentrations and the activities of POD and PAL increased when ZnO-BPs and Fe-AC were added together.