Recent studies have shown that the cosmopolitan diatom Skeletonema costatum sensu lato is composed of several morphologically and genetically distinct species. To assess whether the separate species have a cosmopolitan distribution, we analysed 184 strains from marine and estuarine sites worldwide. We identified the strains using light and electron microscopy, and we sequenced the hyper-variable region of nuclear LSU rDNA. All recently described species were genetically distinct, and all but two were morphologically distinct. Variability was found for the only ultrastructural character used to distinguish Skeletonema dohrnii and S. marinoi, which cannot be identified based on morphology alone. Furthermore, multiple genetically distinct taxa, which may represent cryptic species, were found within the S. menzelii and S. tropicum clades. We found that all currently recognized species of Skeletonema are widespread, however, gaps seem to occur in their geographical ranges. For example, some species are found in both the northern and southern temperate latitudes whereas other species appear to have only subtropical to tropical ranges. Skeletonema pseudocostatum and S. grethae seem to have more restricted geographical ranges because the former was not found along American coasts and the latter was encountered only in US waters. A taxonomic update is provided for Skeletonema strains currently available in several culture collections, which could aid reinterpretation of results obtained in comparative studies using these strains.