Abstract: The biodiversity of phaeohyphomycotic agents in public garden habitats located in four governorates near Cairo (Faiyoum, Ismailia, Minufia, and Qalyubia) was analyzed. A total of 120 samples of soil and plant debris were collected seasonally during the period 2006-2007 and screened for the occurrence of phaeohyphomycotic agents. Dichloran chloramphenicol peptone agar (DCPA) medium was used as a selective medium for isolation by dilution plate method. Nine fungal species belonging to 5 genera, all previously implicated as agents causing phaeohyphomycosis were identified and are as follows: Alternaria alternata, A. infectoria, A. tenuissima, Cladosporium cladosporioides, C. oxysporum, C. sphaerospermum, Cochliobolus lunatus, Phialophora sp., and Veronaea botryosa. Molecular typing of the recovered species based on ITS1-5.8S-ITS2 region sequences were carried out and the obtained sequence data confirmed the morphological identification. The diversity analysis showed that the most prevalent species was C. sphaerospermum representing (56.3%) of the total recovered isolates, followed by C. oxysporum (16.31%), C. cladosporioides (14.0%), A. alternata (5.2%), C. lunatus (2.5%), Phialophora sp. (1.9%), A. infectoria and A. tenuissima (1.6% each) and finally V. botryosa representing 0.4% of the total recovered isolates. Key words: Phaeohyphomycosis, ITS1-5.8S-ITS2, biodiversity, natural environments, Egypt.