We describe the leaf anatomy of 42 of the 60 species of the genus Banisteriopsis, one of the largest and most diverse genera of Malpighiaceae in the Cerrado domain. Our aim was to identify anatomical leaf characters useful for the taxonomy and systematics of this morphological diverse genus. Expanded leaves from herbarium specimens were rehydrated and processed according to usual anatomical methods for analysis under light microscopy. All characters were compared using the Unweighted Pair Group Method with Arithmetic Mean algorithm and expressed in dendrograms. The most useful leaf anatomical characters were petiole contour, presence/absence of vascular bundles in the petiole, the position of the vascular bundles in the petiole, shape of the midrib, presence/absence of sheath extension, mesophyll type, and both the presence/absence and shape of leaf glands. Our study indicates that the morphological groups proposed in the literature for Banisteriopsis are natural groups.