Soils from rural zones with high natural value (HNV) agriculture systems are an important source of beneficial microbial species that can be useful for various biotechnological purposes, such as transfer of suppressiveness against plant pathogens from suppressive to disease-inducing soils by using inoculation with antagonistic selected strains. The main goal of the paper was to present the results of the research carried out on strains isolated from soil microbial populations in HNV agriculture system (Mureș county, Romania) responsible for specific suppressiveness against soil-borne phytopathogens. The dual culture method was used for assessing the mechanisms involved in antagonism against a plant pathogenic strain from genus Fusarium. The global microbial activity measured as soil respiration was intense. Total counts of bacteria and fungi estimated by dilution plate were also high. The community of heterotrophic aerobic bacteria included 13 species. Associations of fluorescent pseudomonads and actinomycetes were dominant and presented antagonistic activity against Fusarium. Twenty fungal species presented cellulolytic capability evidenced by growth on culture media with cellulose as sole source of carbon. Over cellulolytic capacity, the selected isolate of Trichoderma viride presented antagonistic activity against pathogenic Fusarium strain. Both biochemical mechanism and hyperparasitism were evidenced as involved in its antifungal activity.