Three novel Gram-stain-positive bacteria, designated IY07-20(T), IY07-56(T) and IY07-113, were isolated from soil samples from Iriomote Island, Okinawa, Japan, and their taxonomic positions were investigated by a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequence comparisons showed that the three isolates were closely related to the members of the genus Agromyces, with similarity range of 95.6-98.7%. The isolates contained l-2,4-diaminobutylic acid, d-alanine, d-glutamic acid and glycine in their peptidoglycans. The predominant menaquinone was MK-12 and the major fatty acids were anteiso-C15:0 and anteiso-C17:0. The DNA G+C contents were 70.9-72.9 mol%. The chemotaxonomic characteristics of the isolates matched those described for members of the genus Agromyces. The results of phylogenetic analysis and DNA-DNA hybridization, along with differences in phenotypic characteristics between strains IY07-20(T), IY07-56(T) and IY07-113 and the species of the genus Agromyces with validly published names, indicate that the three isolates merit classification as representatives of two novel species of the genus Agromyces, for which the names Agromyces iriomotensis sp. nov. and Agromyces subtropica sp. nov. are proposed; the type strains are IY07-20(T) ( = NBRC 106452(T) = DSM 26155(T)) and IY07-56(T) ( = NBRC 106454(T) = DSM 26153(T)), respectively.