The taxonomic positions of four novel actinomycetes isolated from soil samples, designated KT2142T, PM2084T, K236Tand A4038T, were established by using a polyphasic approach. The organisms had chemical and morphological features that were consistent with their classification in the genusPseudonocardia. Whole-cell hydrolysates of the four strains containedmeso-diaminopimelic acid and arabinose and galactose as the diagnostic sugars (cell-wall type IV). Their predominant menaquinone was found to be MK-8(H4). The major fatty acid was iso-C16 : 0. 16S rRNA gene sequence data supported the classification of the isolates in the genusPseudonocardiaand showed that they formed four distinct branches within the genus. DNA–DNA relatedness studies between the isolates and their phylogenetic neighbours showed that they belonged to distinct genomic species. The four isolates were readily distinguished from one another and from the type strains of species classified in the genusPseudonocardiabased on a combination of phenotypic and genotypic properties. In conclusion, it is proposed that the four isolates be classified in four novel species of the genusPseudonocardia, for which the namesPseudonocardia cypriacasp. nov. (type strain KT2142T = KCTC 29067T = DSM 45511T = NRRL B-24882T),Pseudonocardia hierapolitanasp. nov. (type strain PM2084T = KCTC 29068T = DSM 45671T = NRRL B-24879T),Pseudonocardia salamisensissp. nov. (type strain K236T = KCTC 29100T = DSM 45717T) andPseudonocardia kujensissp. nov. (type strain A4038T = KCTC 29062T = DSM 45670T = NRRL B-24890T) are proposed.