The present study evaluated the seasonal changes of the soil microbial population by selected media in an organic farming system (OFS) with rye rotation cropping management compared to those in a conventional farming system (CFS) with chemical fertilizers, pesticide and herbicide from May 2009 to October 2010 in an upland field. With the exception of fungi, populations of aerobic bacteria, Gram-negative bacteria, and Bacillus spp. were higher in the OFS soil during soybean-growing stages. In addition, populations of aerobic bacteria, Gram-negative bacteria, and Bacillus spp. in the OFS soil were nearly two times more than those in the CFS soil on reproductive growth stages. Our findings suggested that Bacillus spp. should be considered as responsible factor for microbial population differentiation observed between the OFS and the CFS in upland fields.