Resumo. The present study aimed to evaluate the behavior of Eucalyptus grandis x Eucalyptus globulus and Eucalyptus urophylla x Eucalyptus globulus clones and the effect of different in vitro insertions (at 30, 90 and 150 days after pruning the donor plants) in the in vitro establishment phase. The mini-stump plants, explant providers for the in vitro introductions, were cultivated under an indoor semi-hydroponic clonal mini-hedging. Nodal segment explants of 21 Eucalyptus urophylla x Eucalyptus globulus clones and eight Eucalyptus grandis x Eucalyptus globulus clones were harvested, disinfected and inoculated in an MS culture medium, supplemented with 0.5 mg L-1 of BAP and 0.1 mg L-1 of NAA. After 30 days in culture, evaluations were made and it was concluded that there were variations among the clones, as well as in vitro introductions, for the studied variables, where the in vitro establishment was possible for most clones.