The objective of this study was to evaluate the effect of sucrose and substrates on the acclimatization of micropropagated Luehea divaricata plants. First, the effect of sucrose on in vitro growth and development of nodal segments in Woody Plant Medium reduced to its half concentration was tested. After that, these shoots were then acclimatized and the combined effect of sucrose, as previously described, plus the commercial substrates vermiculite, MecPlant® or H-decker® were tested. In the in vitro culture, root and leaf formation were superior (44%, 8.44 leaves) with 20 g L-1 sucrose, and then the presence of sucrose continued to be more efficient in the acclimatization with the MecPlant® or H-decker® commercial substrates. Sucrose at 20 g L-1 is more efficient for in vitro leaf and root formation, and the MecPlant® or H-decker® substrates may be used in acclimatization.