Fusarium graminearum causes the catastrophic Fusarium head blight (FHB), affecting wheat and barley and negatively impacting yields and quality. Currently, there is no effective strategy for suppressing FHB utilizing the barley phyllosphere microbiome. This pilot study aims to establish a stable microbial community with reproducible assembly in the barley phyllosphere to investigate plant-microbe interactions that suppress pathogen proliferation leading to plant protection in barley. In this case, the microbes utilized are members of the barley spike phyllosphere core microbiome as predicted from a large amplicon sequencing study that profiled the composition of 800 diseased and non-diseased barley heads of ten genotypes collected from four misted nurseries across the US. A trial Hordeum vulgare phyllosphere Synthetic Community (Hv-PSC-Fargo) constructed from a cultured microbiome collection derived from diseased and non-diseased Barley spikes collected in Fargo, ND. A number of experiments were carried out to investigate approaches to establish the Hv-PSC-Fargo community in barley plants grown in growth chamber settings. These involved optimizing the sterilization of barley seeds to remove the background seed microbiome, cultivating strains, and determining the initial community member ratios to preserve community diversity. Using these approaches, one trial involved inoculating sterilized seeds with Hv-PSC-Fargo, heat-killed Hv-PSC-Fargo, and PBS only to assess successful recruitment to the spikes using ND Genesis barley. In another experiment, short barley genotypes were grown in inert soil substitutes (sand: vermiculite) and Promix: Osmocote on Lennard jars to explore the transition to more gnotobiotic systems for growth. In the long term, this study will improve our knowledge of some of the many variables influencing the assembly of microbial communities, allowing us to tackle the shortcomings of the inconsistent efficacy of biocontrol strains and gain a deeper understanding of key drivers of microbial community assembly in the context of disease, and its impact on disease outcomes.