In this study, we examined the chemical properties and microbial community characteristics in 25 controlled horticultural soils (CHS) sampled from Gyeongnam Province by fatty acid methyl ester (FAME) method. The electrical conductivity of watermelon CHS was significantly (p < 0.05) higher than those of red pepper CHS, pumpkin CHS, and strawberry CHS. The amounts of total FAMEs, total bacteria, gram-negative bacteria, gram-positive bacteria, and fungi were significantly (p < 0.05) higher in red pepper CHS than those in strawberry CHS and pumpkin CHS. In addition, higher (p < 0.05) ratios of cy19:0 to $18:1{omega}7c$ were detected in tomato CHS than those in watermelon CHS, pumpkin CHS, and red pepper CHS. This implied that microbial communities of tomato CHS were stressed more than other species of cultivation soils. Actinomycetes community in red pepper CHS was significantly (p < 0.05) higher than those in tomato CHS, strawberry CHS, and watermelon CHS. Differences in soil microbial community composition were highly associated with cultivated crop species which might result from the management inputs such as fertilizer, herbicide, and irrigation.