The effect of mixed cropping on disease suppressiveness of soils was tested for two cropping systems, Brussels sprouts-barley and triticale-white clover. Disease suppressiveness of field soils was evaluated in bioassays for the soilborne pathogens Rhizoctonia solani, Fusarium oxysporum f. sp. lini, and Gaeumannomyces graminis var. tritici. For both cropping systems, mixed cropping did not enhance disease suppressiveness of the soils. In some cases, soil cropped to barley alone was significantly more suppressive to F. oxysporum f. sp. lini than soils cropped to Brussels sprouts or the mixture of Brussels sprouts and barley. Analyses of the diversity of the indigenous bacterial and fungal microflora by denaturing gradient gel electrophoresis of amplified 16S- and 18S-rDNA fragments, respectively, revealed, in most cases, no significant differences between mixed and mono-cropped soils. In conclusion, in this study, mixed cropping of soils with Brussels sprouts and barley or with triticale and white clover did not enhance microbial diversity or disease suppressiveness of soils to three different soilborne plant pathogens.