This work was carried out in Tissue Culture Laboratory in the Vegetable and Floriculture Dept. Fac. of Agric.. Mansoura Univ. from 2002 to 2005 years to find out an ideal method of propagation through tissue culture technique for Eucalyptus gomphocephaia trees. In this concern. shoot tips (terminal part) and basal part (one node from the base of the stem) each explant part was about 1 — 1.5 cm prepared from E. gomphocephala trees. After sterilization, the expla nts were initiated on culture medium i.e. Murashige and Skoog (MS) sUpplemented with 1.0 mglL BA (6-benzyl adenine) and 0.1 mglL IBA (lndol-3-butyric acid) for establishment stage. The newly formed shoots were transferred to the same medium supplemented with BA at the concentration of (0. 0.5, 1, 2 and 4 mglL) and with or without activated charcoal (2 glL) through proliferation stage to study the effect of the explant type, activated charcoal and the plant growth regulators on proliferation of shoots and some vegetative charcaters (shoot number and length and number of leaflets). Highest number of formed shoots were regarded when culturing the basal explant part on MS without activated charcoal, supplemented with 0.2 mglL BA. While. the tallest shoots were resulted from culturing the shoot tip on MS media with 2 glL activated charcoal. Microshoots were rooted in the same full and half strength media supplemented with IBA at the concentration of (0. 0.5. 1. 2 and 4 mglL). Full strength MS medium supplemented with 2 mglL IBA was superior and had the greatest for the rooting growth measurements (mumber of rootslplantlet and average root length). At acclimatization the plantlets were transplanted to (300 ml) plastic pots containing autoclaved transplanting media (sand: peatmoss: vermiculite: perlite) mixed by volume (1:1:1:1) and maintained in greenhouse for four weeks to investigate their effect on survival % per plant during acclimatization stage on media mixed. Produced the highest survival percentage of Eucalypts was 81.25% in case of using (1 : 1 :2:1) by volume. respectively. After transfer of plantlets from in vitro cultures to the greenhouse or field to study the effect of acclimatization time (48 h. 7. 14 and 30 days) on leaf structure, substantial changes in leaf morphology and anatomy are observed. above all in epidermal characteristics. leaf thickness. differentiation of leaf mesophylt. and chloroplast number and structure.