The study examined how altering ethylene biosynthesis (ACO1) or perception (etr1.1) in a hybrid poplar (P. tremula × P. tremuloides T89) influences the assembly of root and shoot fungal and bacterial communities, using amplicon sequencing and confocal microscopy. Ethylene modulation had limited impact on the sterile plant metabolome but triggered distinct primary and secondary metabolic changes in microbe‑colonized plants, correlating with reduced fungal colonisation of shoots and increased root fungal colonisation, while arbuscular mycorrhizal fungi and bacterial communities were largely unchanged.