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During an investigation of fungal diversity on fruit trees in Guizhou Province, 23 <i>Cladosporium</i> strains were isolated from various locations in Guizhou Province. Culture characteristics, morphology and molecular phylogenetic analysis of three genetic markers, namely, the internal transcribed spacer regions (ITS) of the rDNA, partial fragments of actin (<i>act</i>), and the translation elongation factor 1-α (<i>tef</i>1-ɑ) loci were used to characterize these isolates. Seven new <i>Cladosporium</i> species and new host records for five other species were introduced, with detailed descriptions and illustrations. This study showed that there is a rich diversity of <i>Cladosporium</i> spp. in fruit trees in Guizhou Province.