The low acclimatization efficiency of pineapple seedlings obtained by in vitro culture has been one of the main limitations to massively scale micropropagation protocols. Various factors may be affecting the successful establishment of in vitro plants to ex vitro conditions, related to the plant and/or the environment and the management of seedlings. The objective of this study was to establish an efficient micropropagation and acclimatization system for the MD2 variety, with a high in vitro multiplication rate and a high greenhouse and field survival of pineapple in vitro plants. In the multiplication phase, the axillary buds were isolated and placed on a semisolid medium and three induction treatments with different concentrations of BAP (1.0, 3.0, 5.0 mg/L) and ANA (2.0 mg/L), were evaluated. After the third subculture, multiple shoots were transferred to a temporary immersion system (BIOMINT). During acclimatization and nursery phases, six treatments with different substrate compositions wereevaluated. The induction treatment IT3 (BAP 5.0 mg/L and ANA 2.0 mg/L) was selected for a significantly high multiplication rate. AT4 treatment (Soil + Coconut fiber + perlite 1:1:1) showed the highest survival rate (95%) and allowed the obtaining of well-developed plants. This system constitutes a valuable technology to introduce in vitro plants to the pineapple production scheme on a commercial scale
