Land Use Shapes the Rhizosphere Microbiome and Metabolome of Naturally Growing Barbarea vulgaris

<b>Background:</b> Land use change fundamentally alters soil microbial communities and biochemical processes, yet the integrated effects on rhizosphere microbiome-metabolome networks remained poorly understood. <b>Objective:</b> This study investigated land uses as forest, grassland and intermediary edge shape the rhizosphere biochemical networks of naturally grown <i>Barbarea vulgaris</i>. <b>Methods:</b> Rhizosphere soils of <i>Barbarea vulgaris</i> were analysed for microbial community structure abundance, and metabolomic profile applying phospholipid fatty acid (PLFA) profiling and mass spectrometric untargeted metabolomics (GC-MS/MS and MALDI-TOF/TOF MS). These were coupled with co-inertia analysis to assess microbiome-metabolome interactions. <b>Results:</b> Microbial community analysis revealed significant effects of land use on bacterial community structure (G+/G-, <i>p</i> < 0.001). Untargeted metabolomics identified 248 metabolites, of which 161 were mapped to KEGG pathways. Amino acids and derivatives (21.1%) followed by organic acids (16.8%) were the most representative among identified metabolites. Pathway enrichment analysis revealed coordinated reprogramming of central carbon and nitrogen metabolism across land use gradients, particularly in the amino acid metabolism, TCA cycle, and glycolysis/gluconeogenesis pathways. Microbiome-metabolome coupling analysis revealed distinct correlation patterns between microbial phenotypes and metabolite classes, with forest environments showing the strongest biochemical network integration (RV = 0.91). Edge habitats presented intermediate signatures, supporting their role as transitional zones with unique biochemical properties. <b>Conclusions:</b> The environmental context fundamentally shapes rhizosphere biochemical network organization through coordinated shifts in bacterial community structure and metabolic pathway activity. These habitat-specific metabolic signatures suggest that land use change triggers adaptive biochemical responses that may influence plant performance and ecosystem functioning across environmental gradients.