The objective of the present study was to evaluate the effect of different in vitro culture periods of micropropagated Luehea divaricata shoots. The treatments were constituted by different in vitro culture periods (14; 21; 28; 40; 56; 70; 84; 102; 116; 130; 154, 186 or 200 days), without subculture or transfer to a new nutritive medium. The survival and establishment reached around 90% up to 84 days. After that, the mortality increased. Culture period of 56 days was the one that showed the most appropriate proportion of number of leaves and senescent leaves, simultaneously, showing nine leaves and only one senescent. Averages above 50% were observed for primary roots formation after 70 days of in vitro culture, providing the best results (60%) at 200 days. For secondary roots, the highest averages were observed after 102 days (41.5%), reaching 45.3% at 200 days. The micropropagation of Luehea divaricata is significantly affected by the in vitro culture period, which can be extended up to approximately 60 days. From that period, high mortality and leaf senescence are registered.