The chemical composition of propolis from Aegean Uşak (Balcıdamı and Kaşbelen), Afyon ( Emirdağ and Dinar), Manisa (Salihli and Kula), Denizli (Merkez and Çivril), Muğla (Milas and Merkez), İzmir (Kemalpaşa and Menemen), Aydın (Söke and Kuşadası) and Kütahya (Hisarcık and Tavşanlı) was studied in order to determine the major compounds by using GC-MS. In this study, 8 % ethanol extract of propolis prepared by mixing 920 ml of 70 % ethanol and 80 g of propolis was used. Chemical analysis of propolis extracts indicated that the propolis samples had high concentrations of the aromatic acids, esters and other derivatives which are responsible for the antibacterial, antifungal, antiviral, anti-inflammatory and anti-cancer properties of propolis such as benzyl cinnamate, methyl cinnamate, caffeic acid, cinnamyl cinnamate and cinnamoylglcine besides the most common compounds as fatty acid, terpenoids, esters, alcohols hydrocarbons and aromatic acids. Also, in this research the antifungal effects of 7 concentrations (6.25, 12.5, 25, 50 ppm) of propolis ethanol extract (PEE) against Fusarium oxysporum was investigated in vitro conditions. Propolis was mixed alone or in combination with potato dextrose agar (PDA) medium at various concentrations. The results indicated that the mycelial growth of the tested fungi decreased with each increase in PEE concentrations. Propolis extract collected from Muğla province showed 77.81% antifungal effect against Fusarium oxysporum at the highest concentration (50 ppm). The lowest antifungal effect (64.52%) against the pathogen was detected in the propolis extract collected from Denizli province.