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This work evaluated the influence of seal type and substrate type on pre-acclimatization, and symptomatological aspects relating to acclimatization of ipê-roxo. For the pre-acclimatization, ipê-roxo plants obtained through an in vitro zygotic embryo culture were aseptically inoculated in test tubes with different types of seal (cotton plug, plastic cap + parafilm and plastic lid) and different substrates (agar, vermiculite and Plantmax®) supplemented with a WPM culture medium with 1 gL-1 of activated charcoal, 10 mgL-1 of citric acid and 30 gL-1 of sucrose added. For the acclimatization, in vitro ipê-roxo plants were transplanted into 56-cm3 plastic tubes containing vermiculite and Plantmax® at a 2:1 ratio which had been previously autoclaved at 121oC and 1 atm for 20 minutes. Once transplanted, the plants were irrigated with different concentrations of the MS medium (25%, 50%, 75%, 100% and 150%), using distilled water as control. After 30 days, the effects of seal type and substrate type on pre-acclimatization were examined. Cotton can be recommended where Plantmax® or vermiculite is used as substrate. In the acclimatization it was noted that where plants were irrigated with distilled water and concentrations < 50% they showed symptoms of nutritional deficiency. Similar symptoms were observed when using MS medium at a concentration equal to or greater than 100%. The 75% MS concentration was found to be the most effective at maintaining the nutritional vigor of ipê-roxo seedlings in the process of acclimatization.