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Six strains of a novel yeast species were isolated from tree bark collected in the Atlantic Forest and the Amazon Rainforest in Brazil. Analyses of the sequences of D1/D2 domains of the large subunit rRNA gene showed that the strains belong to a species in the genus Zygotorulaspora. The species differed by 5.54 % sequence divergence (25 substitutions and five indels out of 542 bp) in the D1/D2 sequences from Zygotorulaspora mrakii, its closest relative. The ITS sequence of the type strain of the novel species differs by 27–69 nucleotide substitutions/indels from the other Zygotorulaspora species. The novel species is able to grow on trehalose, maltose, l-sorbose, inulin and at 37 °C, which are negative in Z. mrakii. The name Zygotorulaspora cariocana sp. nov. is proposed. The holotype of Z. cariocana sp. nov. is CBS 16118T. The MycoBank number is MB 833702.