Despite intensive morphological and chemical studies on the Myrtales, the circumscription of the order remains poorly defined. To test the monophyly of Myrtales sensu Dahlgren and Thorne (Annals of the Missouri Botanical Garden 71: 633‐694, 1984), determine the relationships of some controversial families, and identify the most likely sister group of Myrtales, we conducted parsimony analyses on 80 rbcL sequences representing 36 taxa from families traditionally included in Myrtales and 44 taxa from other Rosidae. The consensus tree resulting from these analyses supports the monophyly of Myrtales and is substantially congruent with the circumscription of the order proposed by Dahlgren and Thorne (Annals of the Missouri Botanical Garden 71: 633‐694, 1984), with one notable exception: in the rbcL tree Vochysiaceae are placed in Myrtales. A reanalysis of morphological attributes of Vochysiaceae revealed that the inclusion of the family in Myrtales is also supported by the combined occurrence of two typical myrtalean features of the wood: vestured pits and bicollateral vascular bundles. Furthermore, our analyses excluded Thymelaeaceae, Lecythidaceae, Haloragaceae, and Gunneraceae from Myrtales, suggesting that the association of these families with Myrtales, as previously proposed by other authors, may not reflect common ancestry. Finally, our analyses support a sister group relationship between the order Myrtales and a clade formed by an expanded Malvales, Sapindales, and an expanded Capparales.