A preliminary study was conduct to determine the effect of  iso-osomatic water deficit on three fig (Ficus carica L.) cultivars using mannitol as a casual agent for drought stress.   The shoots of Fig  cultivars (i.e., Black Mission, Brown Turkey and Brunswick) were sub-cultured on MS medium supplemented with 3 mg L-1 BAP and 0.5 mg L-1 2iP  at different concentrations of mannitol (0.0, 50, 100, 150, 200, 250, and 300 mM L-1) for 35 days under in vitro culture conditions. Number of newly formed shoots, shoot length, leaves number per shoot,  necrosis %, fresh weight, dry weight, chlorophyll content, relative water content were determined at regular intervals. The Obtained results showed that all growth parameters were reduced by increasing the mannitol concentration except for the percentage of necrosis %  which was increased with a significant difference among all treatments. Black Mission was found to resist Mannitol up to concentration of 300 mM L-1, while Brown Turkey and Brunswick cultivars was found to resist mannitol up to the concentration of 250 mM L-1only , where both of this cultivars was not survived under 300 mM L-1. Genomic DNA of the  Ficus carica L cvs. Black Mission, Brown Turkey and Brunswick were extracted and were used in performing Randomly Amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) markers. Three arbitrary oligonucleotide primers showed that the number of amplified fragments different from one cultivars to another and this  indicating that the fig cultivars are not always identical in their DNA ability to be amplified, which may due to unsimilarity in their genetic structure and common selection history. The use of RAPD molecular markers will be more effectiveness than ISSR to distinguish between the different fig cultivars.