This is the first report on the Zoantharia fauna (Cnidaria: Hexacorallia) of the isolated oceanic island of Ascension, southern Atlantic, where zoantharians are a dominant component of the shallow-water benthos. Specimens from two expeditions (1983, 2012) were examined regarding external morphology and molecular phylogeny using three DNA markers, which confirmed the presence of four zoantharian species: Palythoa caribaeorum; Palythoa aff. clavata; Parazoanthus swiftii; and Zoanthus sp. Two of these, Palythoa aff. clavata and Parazoanthus swiftii, were previously only known from the western Atlantic and Caribbean. Molecular examination of the zooxanthellae (=Symbiodinium spp.) of Palythoa specimens showed that they were in association with only one type, subclade C1.