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Floral nectaries are essential for plant reproduction but little is known about the relationship between these secretory structures and pollination system in cacti. To test phenotypic patterns in nectaries associated with pollination syndromes and/or with its pollinators, we selected from evolutionarily related genera Cleistocactus, Denmoza, and Echinopsis, a set of species with bird-pollinated flowers and floral traits that may fit with ornithophily or with sphingophily, and other set of sphingophilous species with moths as effective pollinator. Observations were made under light microscope and scanning and transmission electron microscopes. Nectaries are located at the base of the filaments welded to the tube, forming a chamber. The nectary consists of the epidermis with distinctive features in each genus, a secretory parenchyma which may be vascularized and a non-secretory vascularized parenchyma. Anatomical variants observed in nectaries of different species are not consistent with the floral pollination syndromes neither with groups of pollinators. The basic structure of the nectar chamber is relatively conserved, a fact that may be explained by phylogenetic conservatism among the genera investigated. Our results revalue the role of anatomical traits for the systematics of Cactaceae.