The taxonomic position of a group of mesophilic actinomycetes isolated from arid Australian soils was determined using a polyphasic approach. The organisms shared chemical and morphological markers typical of members of the genus Amycolatopsis. They had identical 16S rRNA gene sequences and formed a distinct phyletic line in the Amycolatopsis mediterranei clade, being most closely related to A. mediterranei. In addition, they shared a range of phenotypic properties that distinguished them from representatives of all of the species classified in this clade. The combined genotypic and phenotypic data indicate that the strains merit species status within the genus Amycolatopsis. The name proposed for the novel species is Amycolatopsis australiensis sp. nov.; the type strain is GY048T (=DSM 44671T=NCIMB 14142T).