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This study aimed at evaluating the effect of genotypes of Eucalyptus dunnii on in vitro establishment and also the influence of genotypes grown in different concentrations of 6-Benzylaminopurine (BAP) on in vitro multiplication. Explants were obtained from 10 parent plants selected in the field as a function of their superior phenotype characteristics. For in vitro establishment, 10 genotypes were evaluated, while in vitro multiplication consisted of 30 treatments which corresponded to combinations of six genotypes found to succeed in the in vitro establishment and five BAP concentrations (0, 0.25, 0.50, 0.75, and 1.0 mg L-1). Different behaviors were observed regarding the genotypes as to in vitro establishment rates, in which genotypes 3, 6 and 7 had establishment rates of over 70%, against 40% to 6.6% for the other genotypes. Genotypes 1, 5, 8 and 9 were later discarded due to the reduced number of explants successfully established. Factors such as microbial contamination and phenol oxidation posed a threat to in vitro establishment. The genotypes of Eucalyptus dunnii interacted differently with BAP in in vitro multiplication, noting that the concentration 0.50 mg L-1 positively influenced bud formation on the explants in most genotypes. Hyperhydricity was relatively low when 0.50 mg L-1 BAP was used and thus does not pose a threat to in vitro multiplication of nodal segments of Eucalyptus dunnii.