This experiment investigates the characteristics of microorganisms isolated from a medium during cultivation process and reveals the relationship between these microorganisms and the growth of Agaricus bisporus. The domestically grown strains of Agaricus bisporus displayed a higher inhibition growth rate against microorganisms isolated from straw, chicken manure, and medium than imported strains. As for inhibition of mycelial growth among mushroom cultivars of the microorganisms separated by each fermentation step from the mushroom medium, the domestic cultivar, ‘Saedo,’ grew more vigorously among other cultivars. As the fermentation progressed, it was confirmed that inhibitation of microorganisms against Agaricus bisporus was weakened. A total of 21 strains of microorganisms that promote mushroom growth were isolated in the 4th turning process, and the microorganisms isolated from the mushroom medium affect the growth and as yield of the mushroom through secretory substances.