Ficus spp. are used as medicine and are used to reducing the risk of cancer and heart disease. The phytochemical screening of the Ficus palmata plant extracts showed the presence of alkaloids, tannins, flavonoids, terpenoids and cardiac glycosides. This investigation was undertaken by using tissue culture technique on Ficus palmata & Ficus carica subsp rupestris. Study the effect of gamma rays doses of 0.0, 20, 30 and 40 Gy on callus production and the secondary plant constituents. The highest callus fresh weightand callus induction were achieved by callus derived from (leaves & stem) explants, which were cultured on (MS) medium supplemented with different concentrations of 2,4-D at 2.0 (mg/l) and Kin at 0.2 (mg/l). Polymerase chain reaction (PCR) amplification using eight ISSR primers showed polymorphic patterns with different response to gamma radiation doses. found genetic differences at the molecular level using ISSR with gamma doses compared to control. The total number of polymorphic bands was 239 bands (99.58% polymorphism). Found that there was genetic stability between the mother plants and product tissue culture for plants (Ficus palmata & Ficus carica subsp rupestris). The chemical composition of Ficus palmata was studied by means of GC-MS analysis under the effect of gamma radiation. The best doses of gamma were 30 & 40 Gy, as they increased the secondary compounds of Ficus palmata.