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Antagonistic yeasts suppress plant pathogenic fungi by various mechanisms, but their biocontrol efficacy also depends on the ability to compete and persist in the environment. The goal of the work presented here was to quantify the composition of synthetic yeast communities in order to determine the competitiveness of different species and identify promising candidates for plant protection. For this purpose, colony counting of distinct species and matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS; MALDI biotyping) were used to distinguish different yeast species and to quantify the composition of a synthetic community of six yeasts ( Aureobasidium pullulans ,  Candida subhashii ,  Cyberlindnera sargentensis ,  Hanseniaspora  sp.,  Metschnikowia pulcherrima  and  Pichia kluyveri ) over time, on apples and in soil, and in different growth media. These studies revealed important characteristics that predispose the different species for particular applications. For example, the competitiveness and antagonistic activity of  C. subhashii  was strongly increased in the presence of  N ‐acetylglucosamin as the sole carbon source,  M. pulcherrima  and  A. pullulans  were the strongest competitors on apple, and  C. sargentensis  competed the best in soil microcosms. Based on these laboratory studies,  M. pulcherrima  and  A. pullulans  are promising candidates for biocontrol applications against fungal phyllosphere diseases, while  C. sargentensis  may hold potential for use against soilborne fungal pathogens. These results document the potential of MALDI‐TOF MS for the quantitative analysis of synthetic yeast communities and highlight the value of studying microorganisms with relevant functions in moderately complex, synthetic communities and natural substrates rather than as individual isolates.

Characterization of antagonistic yeasts for biocontrol applications on apples or in soil by quantitative analyses of synthetic yeast communities