Date palm is one of the most economically powerful fruit tree grown in Egypt. Although Egypt tops the list of date's product in the world. Egyptian date palm industry suffers from many constraints. Low-quality varieties and limited conventional propagation methods are the main obstacles. Extension of date palm plantations in various areas of Egypt and replanting trees to compensate loss due to diseases or human factors are reduced expected to lack adequate planting materials. The knowledge of the interplay between some fundamental medium parameters allows not only the optimization of the micropropagation system but also gives models to investigate and rationalize the process of induction and development of somatic embryogenesis itself. In vitro growth and development of produced embryos and plantlets has been improved by using of full MS salt strength. MS medium modification of some nutrients concentrations, i.e. NH4NO3(1237.5 mg/l) which recorded (13.20 and 9.60 embryos/explant for Medjool and Khalas respectively) at the end of three subculture. KNO3 which gave a higher score of mature somatic embryos (14.40 and 10.00 embryo/culture for Medjool and Khalas respectively) was obtained with 1425 mg/l KNO3 after three subcultures as compared to those obtained from any other treatment combination. The morphological response of somatic embryos production is controlled by some internal factors that fall under the influence of the genetic make-up of the plant and specialized for each genotype that is responsible for the production of somatic embryos (Medjool and Khalas cultivars). The genetic make-up is a decisive factor during somatic embryos production. There are differences between the two cultivars named, Medjool and Khalas. The produced somatic embryos at germination stage were transferred from the two experiments namely ammonium nitrate and potassium nitrate to multiplication and rooting stages for more in vitro growth for 12 weeks at three re-cultures. Individual shoots were cultured on modified MS basal medium in addition to IBA (1.0 mg/l), sucrose (30 g/l) and solidified with phyto-agar (8.0 g/l) for more in vitro growth and development. In vitro plantlets were transferred to acclimatization stage in plastic pots 5*18 cm diameter containing peatmoss, perlite and washed sand at equal volume. In vitro date palm plantlets produced from rooting stage grow well in the greenhouse during acclimatization stage without morphological abnormality. The growth and development of Medjool cultivar were better than Khalas during in vitro culture and ex vitro acclimatization.
Background: Green synthesis of nanoparticles has provided a cost-effective, environmentally friendly method, as well as raised safe strategies for the synthesis of nanomaterials. In this study, we investigated the synthesis of selenium nanoparticles from the leaf aqueous extract of Moringa oleifera (MO-SeNPs) and studied the effect of MO-SeNPs and Moringa extract with various concentrations on growth, active constituents, and antioxidant activity (scavenging DPPH) of Populus alba callus. MS medium supplemented with 2, 4-D at 2 mg/l + 0.5 mg/l BA took the maximum callus induction percentage (100%), increasing the callus fresh weight, dry weight, and degree of callus formation compared with 2.0 mg/l 2,4-D + 0.5 mg/l Kin. Methods: The highest values of callus fresh and dry weight were recorded on MS containing 20% MO-SeNPs. Regarding the active constituents, the GC-MS chromatogram revealed the presence of 23 major compounds identified in the chloroform extract. The phytoconstituents with maximum peak area in callus extract are Tetradecane, Hexadecen-1-ol,trans-9-, 5-Octadecene, (Benzene, 1-pentyloctyl), (Benzene, 1-butylnonyl), Cycloeicosane, and 10-Heneicosene in all treatments. Moringa extract and MO-SeNPs stimulate the increase of compounds in the callus compared to control. In this regard, MO-SeNPs 5% led to a higher increase in the area percent of active substances compared to the treatment of Moringa extract and control. Results: Concerning the antioxidant activity, the results indicated that Moringa extract and MO-SeNPs had a positive effect on scavenging DPPH compared with the control. Conclusion : The highest percentages of scavenging DPPH were recorded when Populus alba L. callus was treated with MO-SeNPs 20%, 10%, and Moringa extract 20%, percentages reached 84.65, 75.35, and 71.94 %, respectively.
Abstract:Date palm, like all other crops, is very sensitive to the injury by many insect pests, which may lead to the death of the affected plant and causes decrease in yield. In the present study, an efficient Agrobacterium for genetic transformation was successfully achieved for well known date palm (Phoenix dactylifera L.) cv. Medjool and Khalas using callus as explant. Embryogenic callus were recorded 100% mortality when cultured on MS medium containing 100 mg/L kanamycin with different cultivars, thus it was chosen for the selection of transformed explants. Embryogenic callus of Medjool and Khalas were incubated with Agrobacterium tumefaciens strain LBA 4404 for 0.5, 1, 2, 4 and 24 h on LB medium. After the incubation periods, embryogenic callus was transferred to MS medium with 0.1 mg/L NAA, 0.05 mg/L BA, 250 mg/L carbenicillin and 100 mg/L kanamycin for detection of transgenic embryogenic callus. Polymerase chain reaction (PCR) was used for the rapid screening of Cry3Aa gene. For screening, total genomic DNA was isolated from transformants. Using primer specific to Cry3Aa gene (forward and reverse), a PCR product with a size of about 2,000 bp was amplified when all nucleic acid from the transformants were utilized as templates. PCR analysis confirmed the appearance of the transgene of 2,000 bp in one individual plantlet. Presence and integration of foreign Cry3Aa gene in regenerated kanamycin resistant embryogenic callus was also confirmed by Southern blot hybridization. It was found that one transgenic embryogenic callus for both Medjool and Khalas showed a single copy of gene integration. These results signify the successful transfer of Cry3Aa gene into date palm plant.
Immature female inflorescence plays a significant role in date palm micropropagation because inflorescences are available with no practical limit as the source of explants. Moreover, using floral buds for propagation helps in the conservation of date palm biodiversity and the enhancement of socioeconomically valuable landraces. With the goal of avoiding undesirable genetic variability, the optimal combinations and concentrations of plant growth regulators and other medium compositions were investigated to achieve direct organogenesis and multiplication from the immature female inflorescence of date palm (Phoenix dactylifera L.) cultivar Amri. For the initiation stage, the best response was achieved using Murashige and Skoog (MS) medium containing 1.0 mg L−1 zeatin and 1.0 mg L−1 thidiazuron (TDZ) after 16 wk of culturing. For the multiplication stage, the best culture medium contained 0.5 mg L−1 TDZ solidified with GelriteTM, without activated charcoal for four subcultures, and then supplementing 30 mg L−1 glutathione to this medium composition for two additional subcultures. Plantlets were multiplied and grown for 12 wk on elongation medium and then transferred to the rooting stage in two steps. Compared with other treatments, foliar spraying and watering with 30 g L−1 sorbitol and 40 g L−1 salicylic acid twice a week yielded the best results in terms of survival percentage (95%), leaf width (2.9 cm), and growth vigor (4.4 lateral branch). This was the best combination of plant growth regulators and other medium compositions for micropropagation of date palm (Phoenix dactylifera L.) cultivar Amri without the need for callus formation to avoid undesirable genetic variability.
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