The effect of successive subcultures on the microshoots production from organogenic lines of cigotic material of Eucalyptus globulus was studied. Nodal segments were subcultured every 50 days on proliferation medium (MS + 4.4 μM BAP + 0.05 μM ANA) and the microshoots per explant were counted. The results indicated a decrease in the quantity of microshoots from the 3rd to the 6th subculture (1.58 to 0.18 microshoots per explant), accompanied by a high density of mass proliferation. This effect was reverted by maintaining the mass proliferation in a free hormone MS medium for 30 days and later transferring it to a shoots proliferation medium. The proliferation index increased from 0.18, in the 6th subculture, and up to 1.03 shoots per explant in the 7th subculture. The results indicated that maintaining the explants in cytokinins and auxins presence during successive subcultures induces a decrease in the caulogenic competition and consequently, a smaller microshoots production. Transferring the explants to free hormones medium helps to recover the lost or diminished competition, allowing an increase in the microshoots production with exogenous cytokinins and auxins.