As with cuttings, the microcuttings produced from in vitro generated shoots could be affected by topophysis. Very little research has been done about such in vitro effect. This paper reports the influence of the topophysis on the regeneration response (shoot multiplication, microcutting rooting and plantlet acclimation) of explants on the in vitro culture of juvenile N. obliqua trees. The results showed that the main responsible of the physiological variability arising from all the in vitro propagation stages is topophysis. This influence was found in the first culture and can be extended to the next subculture (multiplication stage). Growth of the obtained plants was also affected (rooting of microcuttings and plantlet acclimation). The multiplication rate was lower in apical portion (1:3.66) than basal portion (1:5.8) while rooting was 48.6% and 27.0% respectively. By the way, length of acclimated explants was 52.4 mm in the apical rooted explants and 28.7 mm in the basal ones at the 30th day. In order to reduce such variability, it is necessary to take into account the initial position of the explants during the establishment and multiplication stages of micropropagation of forest trees.