Endophyllum osteospermi is an autoecious, endocyclic rust fungus, which has only been recorded on Chrysanthemoides monilifera ssp. monilifera (Asteraceae, Calendulae), a perennial woody shrub. Both organisms are indigenous to South Africa. Because E. osteospermi is being considered for release in Australia as a biocontrol agent against C. monilifera ssp. monilifera, it was necessary to determine its host range and natural distribution in South Africa. To address this, natural stands of Chrysanthemoides species, as well as other South African asteraceous plants, were monitored for E. osteospermi between 1992 and 2003. A morphological and molecular comparison of specimens referable to Endophyllum was undertaken. Based on these results, E. osteospermi was recorded on C. monilifera sspp. monilifera, pisifera, rotundata, canescens, and subcanescens, C. incana, and an undescribed taxon. E. osteospermi was also recorded on Osteospermum ciliatum, O. polygaloides, and O. potbergense. Furthermore, a closely related but previously undescribed species, E. dimorphothecae sp. nov. is described on Dimorphotheca cuneata. Aecidium elytropappi is transferred to Endophyllum as E. elytropappi comb. nov., being recorded on Elytropappus rhinocerostis and Stoebe plumosa. This study shows that in South Africa E. osteospermi is restricted to a small group of related plant species in the Calenduleae. This rust is therefore considered suitable as a candidate agent for the biocontrol of C. monilifera ssp. monilifera, and pending the results of host specificity testing, would most likely be safe to introduce into Australia.