his study was carried out to investigate antimicrobial activities of water extracted Pleurotus ostreatus var Heuktari (PO), and their application to F. fujikuroi Nirenberg growth inhibition material. Various organic solvents (chloroform, hexane, ethyl acetate, ethanol) were tested to investigate the antimicrobial activities of Pleurotus ostreatus var Heuktari against F. fujikuroi Nirenberg. Chloroform, hexane, ethyl acetate and hot water extracts had no antimicrobial activities, but butanol extract showed 2.5% strong activities in order of F. fujikuroi Nirenberg disk diffusion test. Then we observe Antifungal activity using green house. Bed soil and mushroom extract mixed at 10%, 7.5%, 5%, 2.5%, 1% total bed soil mass. Screening of Antifungal activity was tested two periode 18day and 25day. The Antifungal activity rate of each period and extract density was 7.5% mixed bedsoil has investigated 80% Antifungal activity rate. The result indicates that the Pleurotus ostreatus var Heuktari extract using F. fujikuroi Nirenberg registance, which shows the development of organic agriculture seed disinfectant.