Shoot-tip explants obtained from offshoots of adult date palms are an excellent source for callus induction and subsequent somatic embryogenesis. In this protocol, the shoot-tip explants are transferred sequentially to a series of media containing gradually reduced concentrations of plant growth hormones: (a) 10 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D) and 3 mg/L 2-isopentenyl adenine (2iP), (b) 7 mg/L 2,4-D and 1 mg/L 2iP, (c) 5 mg/L 2,4-D and 1 mg/L 2iP, and (d) 3 mg/L 2,4-D and 1 mg/L 2iP. Embryogenic callus differentiates into somatic embryos upon transfer to MS medium containing 0.5 mg/L abscisic acid (ABA) and 0.1 mg/L naphthalene acetic acid (NAA). Well-matured somatic embryos germinate on a medium containing 0.1 mg/L NAA. Repeated, multiple, and secondary somatic embryos are induced to produce normal well-developed somatic embryos upon transfer to MS medium containing 0.1 mg/L NAA and 0.05 mg/L benzyladenine (BA). This protocol is potentially applicable for commercial micropropagation of date palm.
Hyperhydricity (or vitrification) is a fundamental physiological disorder in date palm micropropagation. Several factors have been ascribed as being responsible for hyperhydricity, which are related to the explant, medium, culture vessel, and environment. The optimization of inorganic nutrients in the culture medium improves in vitro growth and morphogenesis, in addition to controlling hyperhydricity. This chapter describes a protocol for controlling hyperhydricity during the embryogenic callus stage by optimizing the ratio of nitrogen salts of the Murashige and Skoog (MS) nutrient culture medium. The best results of differentiation from cured hyperhydric callus are obtained using modification at a ratio of NH/NO at 10:15 (825:1425 mg/L) of the MS culture medium to remedy hyperhydric date palm callus and achieve the recovery of normal embryogenic callus and subsequent regeneration of plantlets. Based on the results of this study, nutrient medium composition has an important role in avoiding hyperhydricity problems during date palm micropropagation.
Abstract. Date palm (Phoenix dactylifera L.), commonly grown in the hot arid zones predominantly in the Middle East and North Africa, became one of the highly important cultivated palms around the world, because of the multiple processing utilization of the edible fruit, and the various industry- uses of the whole tree parts. Moreover, there are intensive studies indicated the higher nutraceutical value of the essential biological compounds in the date palm tissues like (carotenoids, phenols, lignin, flavonoids, tannins and sterols) and their therapeutic aspects, such as antioxidants (lutein, β-carotene and vitamin A), antibacterial (syringic acid, vanillic acid and gallic acid), antifungal (tannic acid) and anti-cancer (quercetin) and anti-sterility (β-sitosterol and stigmasterol). Meanwhile, the biotechnology approach provides the production possibilities of the plants' secondary metabolites, using cell suspension cultures and the scale-up by bioreactors. Also, using the biotic and abiotic elicitors as important factors inducing bioactive compounds accumulation in plants tissue cultures. This review describes the progress in studying the in vitro production of some important secondary metabolites from the date palm tissues.
This current investigation was conducted to study the effect of different sugar type (sucrose, sorbitol or mannitol at 0.3M) with different ABA concentration (0.0, 2.0, 4.0, 6.0 and 8.0 mg/l) on the shoot tip explants of date palm Zaghlool cv. conserved at 5 or 15 C° under complete darkness for six and 12 months by observing the physiological changes and growth degree of the conserved explants during the conservation period and also their recovery when they were returned and recultured under normal growth condition in order to achieve the best minimal growth condition for in vitro conservation of shoot tip explants. All shoot tip explants conserved at 5 C° for six month or at 15 C° for 12 months on conservation medium supplemented with 0.3M sucrose or sorbitol combined with different concentration of ABA able to survive.Yet, there are few information about conservation of date palm shoot tip explants or embryonic callus by in vitro slow growth storage so this approach needs more studies to keep continuously viable sterilized and contaminated free new source from mother plants for labor commercial production.
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