The genetic stability of date palm shoots regenerated from leaves explant

Rasha N. Arafa, Sayed A.A. Elsayh, Marwa M. Abdalgaleel, Salwa El-Habashy, Emadeldin A.H. Ahmed, Eman H. Afifi, Alaa N. Draz, Rabab W. El Aramany, Abdallah Sharkh, Mohamed T. Abd El-Kader, Ahmed A.M. Baraka, Dosoky Hoda A.A.
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Abstract

The purpose of this research was to develop a micro-propagation method for the date palm Zaghlol cv. using juvenile leaves. To produce the necessary results, different plant growth regulator combinations were used. The leaves were grown on MS medium supplemented with PVP to prevent the explants from browning. The results showed that adding PVP at a concentration of 1.0 g/l considerably reduced browning. On the induction medium, callus formation occurred during the fourth week of culture; however, callus formation (87.5%) was more prevalent on the ¾ MS medium containing with 10.0 mg/l NAA, 1.0 mg/l BA and 2.0 mg/l 2ip. The greatest development of embryogenic callus (94.50%) occurred on a ¾ MS medium supplemented with 5.0 mg/l NAA and 2.0 mg/l BA. The largest fresh callus weight (3.53 g) was produced by this treatment after four months in culture. On MS medium supplemented with 2.0 TDZ, 1.5 BA and 0.5 NAA, which was regarded as the optimum medium for increasing the number of embryos to 32.10 embryos/culture, the best results (65.67%) were obtained. Further investigation into the stimulation and development of somatic embryogenesis involved using MS basal medium supplemented with BA at 0.5 mg/l, kin at 0.1 mg/l and NAA at 0.05 mg/l; this treatment formed the most leaves (20.11 leaf/cluster). The cluster of shoots grown on MS basal medium supplemented with BA at 2.0 mg/l and NAA at 0.5 mg/l had the highest leaf number (34.25 leaf/cluster) and leaves length values after three sub-cultures (4.75 cm). The DNA-based fingerprinting technology ISSR was used to confirm the genetic stability of this protocol. The mother tree and tissue culture-derived shoots evaluated exhibited no differences in the ISSR banding patterns. The micro-propagation method could be used to produce genetically stable date palm plants.
枣椰树叶片外植体再生嫩枝的遗传稳定性
本研究的目的是为枣椰树Zaghlol cv开发一种微繁方法。使用幼叶。为了得到必要的结果,使用了不同的植物生长调节剂组合。为了防止外植体褐变,将叶片生长在添加PVP的MS培养基上。结果表明,添加浓度为1.0 g/l的PVP显著降低了褐变。在诱导培养基上,愈伤组织在培养第4周形成;而在添加10.0 mg/l NAA、1.0 mg/l BA和2.0 mg/l 2ip的培养基中,愈伤组织形成率为87.5%。在添加5.0 mg/l NAA和2.0 mg/l BA的培养基中,胚性愈伤组织发育率最高(94.50%)。培养4个月后,该处理产生的愈伤组织鲜重最大,为3.53 g。在MS培养基中添加2.0 TDZ、1.5 BA和0.5 NAA是将胚数增加到32.10个胚/次的最佳培养基,其效果最好(65.67%)。进一步研究了在MS基础培养基中添加0.5 mg/l BA、0.1 mg/l kin和0.05 mg/l NAA对体细胞胚胎发生的促进作用;该处理形成的叶片最多(20.11片/簇)。在添加2.0 mg/l BA和0.5 mg/l NAA的MS基生培养基上生长的芽簇经3次传代后叶数最高(34.25片/簇),叶长值最高(4.75 cm)。采用基于dna的指纹识别技术ISSR验证了该方案的遗传稳定性。母树和组织培养衍生的枝条在ISSR条带模式上没有差异。该方法可用于生产遗传稳定的枣椰树。
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