A polyphasic study was undertaken to establish the taxonomic position of a rhizosphere isolate that had been assigned provisionally to the genus Kitasatospora. The organism, isolate F18-98(T), was found to have chemical and morphological properties that were consistent with its classification as a Kitasatospora strain. Direct 16S rDNA sequence data confirmed the taxonomic position of the strain, following the generation of phylogenetic trees by using three treeing algorithms. The organism formed a 16S rDNA subclade with Kitasatospora azatica and Streptomyces kifunensis, but was distinguished readily from the latter by using a combination of biochemical and physiological properties. Genotypic and phenotypic data show that strain F18-98(T) should be classified in the genus Kitasatospora as a novel species, for which the name Kitasatospora putterlickiae sp. nov. is proposed; the type strain has been deposited in culture collections as strain F18-98(T) (=DSM 44665(T)=NCIMB 13932(T)). It is also proposed that Streptomyces kifunensis should be transferred to the genus Kitasatospora as Kitasatospora kifunensis comb. nov. An emended description of Streptomyces aureofaciens Duggar 1948 is also given.