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  • Author or Editor: M.A. El-Hamady x
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Yousef I. Dlaigen, A. E. Said and M.A. El-Hamady

Several experiments were conducted in this investigation with the objective of determining the chemical components and the physical state of an optimal medium for the growth and elongation of excised date palm, cv Sukkari, roots. The chemical tests carried out included: Comparison of (MS)-salts with “White's”-salts mixture and different concentrations of (MS)-salts and its chelated iron; sugars; Modified White's Organics; inositol; adenine sulfate; growth regulators; and some antioxidants. The physical tests, on the other hand, included comparison of the growth and elongation of cultured roots in a liquid or on solidified nutrient media. The effects of various pH values were also tested. Roots were cultured in basal nutrient media composed of: (MS)-salts mixture, and (in mg·liter–1): NaH2PO4·H2O, 170; sucrose, 30,000; inositol, 200; Modified White's Organics; adenine sulfate, 120; activated charcoal, 1500; (2,4-D), 1; kinetin, 2. pH was adjusted at 5.7 ± 01. (MS)-salts mixture was found to be superior to “White's”-salts. No significant difference was observed between (1/2MS) and full-strength (MS)-salts. However, twice the concentration was found to be inhibitory. The normal concentration of (MS)-Fe was found to be optimum for root growth and elongation. The optimal concentration most suitable for the growth and elongation of excised date palm roots has been determined for each of: sugars; Modified White's Organics; inositol; and adenine sulfate. The only growth regulator that needs to be added to the nutrient medium is 2,4-D at 0.1–1.0 mg·liter–1. The study showed the importance of the inclusion of activated charcoal to the nutrient medium. The growth and elongation of roots were both stimulated at all concentrations tested. (PVP), on the other hand, was inhibitory at all concentrations tested. Shaken liquid media was recommended for better root growth and elongation at pH 7.0–8.0. Incidentally, the medium developed was found to support the growth and elongation of roots excised from two other cultivars, namely `Khudri' and `Khaias'.

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Yousef I. Dlaigan, A.E. Said and M.A. El-Hamady

The objective of this investigation was to study the effects of some antioxidants on the growth and elongation of in vitro-cultured date palm roots. Several trials were conducted to determine the antioxidant and the concentration of antioxidants used in date palm tissue culture. Three types of antioxidants were tested—activated charcoal, polyvinylpyrrolidone (PVP), and sodium metabisulfite (NA2S2O5)—in various concentrations, singly, or in combinations. The medium used for root culture consisted of MS salts mixture, 1/2 modified White's organics, 60% sucrose, 0.004% inositol, 240 mg adenine sulfate/liter, 1.0 mg 2,4-D/liter, and 2.0 mg kinetin/liter. pH was adjusted at 5.7 ± 0.1. Concentrations of activated charcoal tested (in gliter–1) were: 0.0, 1, 2, 3, 4, 6, 8, 10, and those of PVP (in gliter–1) were: 0.0, 5, 10, 20, 40 (with or without 4 g activated charcoal/liter). NA2S2O5 concentrations (in mM) tested were: 0.0, 0.2, 0.4, 0.8, 1.6 (with 4 g activated charcoal/liter). The addition of activated charcoal to the culture medium of date palm roots is of vital importance. Four grams per liter gave the best growth and elongation, and there was no significant difference between it and 10 gliter–1. No growth or elongation occurred in the control. PVP, however, significantly inhibited growth and elongation of cultured roots at all concentrations tested. NA2S2O5, however, stimulated growth and elongation at 0.2 mM. Higher concentrations inhibited growth and elongation.

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Yousef I. Dlaigan, A.E. Said and M.A. El-Hamady

The effects of the physical state of nutrient media on the growth and elongation of excised date palm roots were investigated. Roots were cultured in a nutrient medium containing MS salts mixture, 1/2 modified White's organics, and (in mgliter–1): NaH2PO4 H2O, 170; sucrose, 60,000; inositol, 40; adenine sulfate, 240; activated charcoal, 4000; 2,4-D, 1; kinetin, 2. pH was adjusted at 5.7 ± 0.1. Both agar and Gelrite were singly used as solidifying agents. Liquid media were either stationary or rotated on gyratory shakers at 70 to 80 rpm. The effects of incubation of cultured roots under light or dark conditions were also studied. Media pH and its effects on growth and elongation of cultured roots were tested at various ranges (3, 4, 5, 6, 7, 8, 9, 10). Trials were also made to determine the passage length for transfer and subculture of cultured roots to a newly prepared medium. Liquid media highly supported better growth and elongation of cultured roots compared to solidified media. There was no significant difference in growth or elongation between agar or Gelrite media. Shaking liquid medium resulted in significantly better growth and elongation compared to stationary medium. No difference was observed between dark- and light-incubated cultured roots. Cultured roots grew and elongated better at pH 7.0–8.0. No growth or elongation occurred at pH 9.0. Roots continued to grow and elongate even after 12 weeks in culture. Therefore, 10 to 12 weeks after culture was determined to be the optimum passage length for date palm root culture.

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Yousef I. Dlaigan, A.E. Said and M.A. El-Hamady

Several trials were conducted with the objective of obtaining an explant for the establishment of date palm root culture in vitro. These trials included disinfecting and germinating seeds of three cultivars on several autoclaved culture media, the influence of incubation temperature on different germination parameters, and the quality of roots before excision and after culture in nutrient media. Three culture media were used: distilled water only; minimal organics that consisted of MS salts, 3% sucrose, modified White's organics, 0.01% inositol, and 0.15% activated charcoal; and 1/2 MS salts mixture, 3% sucrose, and 1/2 modified White's organics. All three media were solidified with 0.7% agar. The seeds were incubated at 25 or 35C for germination. The study revealed the difficulty of seed disinfection. We immersed seeds in 20% to 40% Clorox, with two to four drops of Tween-20, for 30 to 60 minutes and then rinsed them four to five times in deionized distilled water before culturing. The minimal organics medium supported optimal growth of excised roots, and incubation at 35C significantly improved germination. The use of 10-mm-long root tips as explants for culture initiation gave the best growth and elongation. In addition, the growth and elongation of excised root tips increased significantly as the distance from it to the apex of the cotyledonary sheath increased.