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An important tool for the conservation of biomes is the application of technology to support biodiversity maintenance and recovery. Thus, plant tissue culture could be used as strategic tool to support the production of woody species for reforestation purposes. The use of fruit trees is particularly important because they attract seed dispersing animals that could help environment recovery. Although black pitanga (Eugenia sulcata Spreng ex Mart.) is difficult to find in natural habitat, it presents potential relevance for initiatives aimed at the reforestation of the Atlantic Forest because it is highly appreciated by wild birds. However, the low seed production and germination rate in natural environments impairs the maintenance of the genetic diversity. In this way, in vitro cultivation is an alternative to produce seedlings of arboreal species. The objective of the present work was to evaluate the potential use of in vitro culture for the germination of black pitanga seeds from the field to produce viable seedlings used in initiatives to recover Atlantic Forest areas. Seeds of two E. sulcata donor plants were cultured in vitro in the Wood Plant Medium (WPM) and Murashige Skoog medium (MS) with and without activated charcoal. During in vitro cultivation, development parameters (germination, leaf emission, root emission) and contamination were evaluated. Plants obtained were successfully acclimatized. The results demonstrated that the in vitro cultivation of E. sulcata seeds is a viable alternative to produce seedlings for reintroduction under in vivo conditions. However, the genetic background of donor plants could interfere in seed germination and plant development. These results are a probable reflect of the natural genetic diversity present in seeds.