Resumo. This study aimed to evaluate the usefulness of an air layering technique for vegetative rescue of annatto (Bixa orellana L.) genotypes and obtainment of propagules for vegetative propagation. Ten 12 year-old annatto stock plants, whose genotypes are derived from artificial crossing between "Hairy-green capsule" X "Smooth-red capsule" were used in this study. Air layering adventitious roots were induced in branches (1-2 cm diameter) using as substrate a mixture of vermiculite and moss. The effects of (1) the type of girdling (total or partial), 1 cm in length, (2) IBA (indole-3-butyric acid at 0 and 4.92 mM) solution in paper filter and (3) the covering with either transparent or dark plastic films and tencel were evaluated. The technique of air layering provided rooted branches for the 10 genotypes evaluated, with efficiency ranging from 20 to 100%, and a genotype effect was observed on rooting frequency. Under greenhouse conditions, the survival of layers derived from totally girdled stems treated with IBA and protected with transparent plastic was 100%. Established layers provided vigorous branching from which shoot apexes or nodal segment explants were obtained for in vitro establishment and propagation of this species.