Soil microbes are widely well known to play an important role for sustainable agriculture in terms of crop healthy cultivation and environmental conservation. In this context, the distributional characteristics of soil microbes according to cropping systems were investigated under rice (R)-rice (R), rice (R)-barley (B)-rice (R), and soybean (S)-barley (B)-soybean (S) cropping condition to get basic informations for sustainable agriculture, where barley was grown for winter, in mid-mountainous loam paddy located at the altitude of 285 m above sea level in Sangju area from 2014 to 2015. Estimating from microbial communities by fatty acid methyl ester (FAME) method, a total biomass of bacteria, actinomycetes, and fungi in R-B-R plot was 37% and 40% higher than that in S-B-S and R-R plots, respectively (p < 0.05). In especial, bacteria and fungi were more in R-B-R plot than those in any other ones. B. japonicum, AMF, and mesophilic Bacillus sp. were also greater in S-B-S plot than those. In the community distribution, however, bacteria and actinomycetes showed comparatively high values in S-B-S plot relative to either R-R or R-B-R plot including rice, in which fungi outstanding. In the correlation between microbial biomass and soil properties changed by the cropping, bacteria was positively correlated with C:N ratio; actinomycetes with exchangeable Ca; fungi with available P 2 O 5 (p < 0.05). While these microbes showed negative response to water stable aggregates of soil.