Cladosporium rot (Cladosporium spp.) of grapevine (Vitis vinifera) is a common disease in Chile, particularly in Cabernet Sauvignon and other red wine grape cultivars. It is favored by delayed harvest to obtain the phenolic maturity necessary for high-quality red wine. This study expands on previous investigations of the specific causal agents, the histopathological host:pathogen relationship, and the population dynamics of Cladosporium spp. during the seasonal development of grape clusters. Over 100 isolates were obtained and identified as C. cladosporioides and C. herbarum, confirming previous results. The identity of a subset of isolates was confirmed by molecular analysis. Isolates of both C. cladosporioides and C. herbarum from grapevines were pathogenic on inoculated table grapes and wine grapes. These pathogens were reisolated, fulfilling Koch's postulates. Berry injuries and total soluble solids content above 15% were necessary for Cladosporium spp. to infect wine grapes. The mycelia of C. cladosporioides and C. herbarum grew between 0 and 30°C, but no growth was obtained at 35°C in vitro. The histological studies showed that Cladosporium spp. superficially colonize mature V. vinifera berries, invading the epidermis but scarcely penetrating the hypodermis. The Cladosporium populations obtained on apparently healthy berries of V. vinifera cvs. Cabernet Sauvignon and Chardonnay were significantly larger (P = 0.05) than the populations obtained under similar conditions on berries of V. champini cv. Ramsey and hybrids Kober 5BB and Couderc 1613. Considering the importance of Cladosporium rot in Chile compared with other grape production areas, the development of control strategies is needed to prevent high disease severity, which affects both yield and wine quality.