Laboratory experiments were carried out to evaluate the effect of three rhizobial strains (two of Rhizobium leguminosarum "127k80c and SU157" and one of Bradyrhizobium japonicum USDA110spc4) characterized previously as Plant Growth Promoting Rhizobacteria (PGPR) to affect the tested date palm micro propagation stages using tissue culture techniques. They were positive for the qualitative assessment of IAA and siderophore production. Different concentrations of each (1, 10, 50 and 100 ml/L) with around 6x109 viable cells were used in this evaluation to investigate the enhancement of callus formation, shoot and root number and length of this cultivar in comparison with the control growth regulator media containing synthesized phytohormones (2,4-D and 2ip). The survival percentage was 100% for all treatments. The results confirmed that the best callus formation value was achieved with the control medium (2,4-D) followed by the addition of MS+10 ml/L of 127k80c supernatant strain which recorded 77% and 66%, respectively. Also, all the rhizobial supernatants stimulated the shoot formation and numbers of date palm c.v. zaghloul. On the other hand, there was no significant value on shoot length between the different treatments. Rhizobial supernatants of B. japonicum USDA110spc4 and R. leguminozarum 127k8oc did stimulate strongly the root formation of this cultivar. Therefore, addition of rhizobial supernatants can be considered an effective tool, so that more studies and special attention should be paid to the possibility of using the tested rhizobial strains instead of synthesized phytohormones to reduce chemical substances and for its sheep costs, safe and ease handling.