Background and objectives: Date palm (Phoenix dactylifera L.) is propagated vegetatively via offshoots, which is limited by either the offshoots numbers produced from a superior selected plant, or the occurrence of these offshoots only during the juvenile phase of the date palm life cycle. As a result, the in vitro propagation could be considered an alternative technique for large scale propagation of date palm. Obtaining well-developed root system is considered the most important step in establishment of reliable protocol for successful production of date palm and subsequently successful acclimatization of the in vitro derived plants. The aim of this study is to obtain a well-developed root system through using different antioxidants, with detecting the similarity between the in vitro derived plants and the mother plants using RAPD, long RAPD and ISSR techniques.
Materials and methods: Individual plantlets obtained from maturation of somatic embryos in vitro of about 5-7 cm in length with 2-3 leaves were used as plant material. Plantlets were cultured on half strength MS liquid medium supplemented with 0.5 mg L-1 thiamine-HCl+2.0 mg L-1 glycine+0.1 mg L-1 biotin+40 g L-1 sucrose+ 0.1mg L-1 NAA with different concentrations from either AgNO3 or citric or ascorbic acids (0.0, 0.5, 1, 2 mg L-1). Growth development, root number and root length (cm) were evaluated at the end of the second subculture (12 weeks). Data were reported as Mean±Standard deviation (SD). Data were subjected to one way-analysis of variance (p<0.05). Results were processed by Excel (2010).
Results: Among the different antioxidants with different concentrations used, generally it was found that 2 mg L-1 of each agent gave the highest values of growth development, roots number and roots length. However, using 2 mg L-1 AgNO3 gave the best results with all parameters. Regarding the response of date palm cultivar, it was remarkable that Bartamoda showed relatively better results than Sakkoty cultivar. According to PCR reactions, the results of RAPD, long RAPD and ISSR profile of tissue culture-derived plantlets grown on a medium supplemented with 2 mg L-1 AgNO3 obviously revealed high similarity to mother plants.
Conclusion: It could be concluded that using 2 mg L-1 AgNO3 gave the best results for growth development, root numbers and length of the two cultivars but Bartamoda showed relatively better results than Sakkoty cultivar. The tissue culture-derived plantlets on this medium (2 mg L-1 AgNO3) revealed high similarity to mother plant as a result to RAPD, long RAPD and ISSR profiles.
Keywords: Egyptian date palm; RAPD; acclimatization; ascorbic and citric acids; in vitro rooting; long RAPD and ISSR; silver nitrate.