Recent rDNA sequencing of 25 isolates from a previous study, during which limonene-utilizing yeasts were isolated from monoterpene-rich environments by using 1,4-disubstituted cyclohexanes as sole carbon sources, led to the identification of four hitherto unknown Rhodotorula species. Analyses of the 26S rDNA D1/D2 region as well as the internal transcribed spacer (ITS) domain indicated that two isolates (CBS 8499T and CBS 10736) were identical and were closely related to Rhodotorula cycloclastica, a previously described limonene-utilizing yeast. These novel isolates differed from known yeast species and could be distinguished from R. cycloclastica by standard physiological tests. The other three isolates represent three novel Rhodotorula species, closely related to Sporobolomyces magnisporus. These three species could also be distinguished from other Rhodotorula species by standard physiological tests. Based on these results, we suggest that the new isolates represent novel species, for which the names Rhodotorula eucalyptica sp. nov. (type strain CBS 8499T = NRRL Y-48408T), Rhodotorula pini sp. nov. (type strain CBS 10735T = NRRL Y-48410T), Rhodotorula bloemfonteinensis sp. nov. (type strain CBS 8598T = NRRL Y-48407T) and Rhodotorula orientis sp. nov. (type strain CBS 8594T = NRRL Y-48719T) are proposed. R. eucalyptica and R. pini can also utilize limonene.