Ascogenous yeasts were isolated from a decaying mangosteen fruit (Garcinia mangostana L., Clusiaceae). Based on colony morphology and RAPD analysis, the strains were grouped into four groups (G-I, G-II, G-III and G-IV). Phenotypic properties and analysis of the D1/D2 domain sequence of the 26S rRNA gene identified representative members of groups G-II, G-III and G-IV as Pichia manshurica (=Pichia galeiformis), Pichia fermentans and Pichia anomala, respectively. Representatives of group G-I, YS110T and YS111, showed similar phenotypic traits, 99.9 % similarity in the sequence of the D1/D2 domain of the 26S rRNA gene and ITS1–5.8S rRNA gene–ITS2 sequence and 92 % DNA–DNA relatedness. Hence, YS110T and YS111 are regarded as members of the same species. Based on D1/D2 domain and ITS region sequences, the nearest phylogenetic neighbours of YS110T and YS111 were identified as Pichia membranifaciens and P. manshurica. However, YS110T differs from these two nearest phylogenetic neighbours by >3 % in the D1/D2 domain sequence and by >18 % with respect to the ITS region sequence. In addition, YS110T and YS111 differ from P. membranifaciens NRRL Y-2026T and P. manshurica NRRL Y-27978T with respect to a number of phenotypic traits. The strains show 52–57 % DNA–DNA relatedness with P. membranifaciens NRRL Y-2026T and P. manshurica NRRL Y-27978T. Strains YS110T and YS111 are proposed as two strains of a novel species, for which the name Pichia garciniae sp. nov. is proposed. The type strain is YS110T (=NRRL Y-48422T =CBS 10758T).