Resumo. Eugenia involucrata DC. (Myrtaceae) is a native forest species to several Brazilian states which has several features of interest in the sectors of forestry, fruit faming, environment and medicine. Considering the difficulty of propagation by seeds, which are recalcitrant and lose viability rapidly after harvest, this study evaluated the influence of the use of apical and nodal segments cultured in different nutrient media on the in vitro establishment and development of this species. Apical and nodal segments collected from three- year- old plants kept in the greenhouse were inoculated in the nutrient media MS, ½ MS and WPM. First, 1 ¿M of NAA and 5 ¿M of TDZ were added to the culture media. After that, the explants were transferred to flasks containing the respective fresh nutrient media in absence of growth regulators, but with addition of 1 g L-1 activated charcoal, and then cultured for 30 additional days. Medium ½ MS is most suitable for the in vitro establishment and development of apical segments while media ½ MS and WPM are appropriate for nodal segments. Medium ½ MS enabled the best results of rooting in explants, along with WPM; the in vitro establishment and rooting were made possible by the use of apical and nodal segments of E. involucrata DC. Nutritional media ½ MS and WPM are most suitable for in vitro culture of nodal segments of Eugenia involucrata DC; however, medium ½ MS is recommended due to economical and practical features. For apical segments of this species, the use of culture medium ½ MS is more appropriate than media MS and WPM.