The success of a biological control programme depends on the isolation and selection of antagonists. There is an enormous diversity of culturable microbial species in the soil, rhizosphere, phylloplane, spermosphere and carposphere, which can be used in the isolation and selection of antagonists. The structures of fungal plant pathogens concerned with survival and infection may also be sources of antagonists. Although non-culturable microorganisms and microbiome-based strategies have great potential for development as commercial products in disease control, more knowledge is needed to understand the mechanisms involved in interactions between plants and complex microbial communities. Methods of isolation and selection of the most commercially exploited groups of antagonists and their advantages and disadvantages are discussed in this chapter as well as those of non-traditional antagonists. Finally, possible strategies for engineering the soil and host microbiome to actively promote plant protection against pathogens are discussed.