Abstract

Root exudates influence soil microbial community assembly and function, yet plant species-specific effects on microbially-driven metabolic transformations remain insufficiently characterized under controlled conditions. Here, we present a multi-omic dataset of exudate-microbe interactions across six plant species, including tallgrass prairie species (Helianthus pauciflorus, Carex brevior, Eragrostis spectabilis, Astragalus canadensis, and Panicum virgatum) and a model grass (Brachypodium distachyon), generated using an in vitro incubation experiment. Sterile root exudates were used to amend a soil-mimicking medium inoculated with a native tall-grass prairie soil microbiome. The dataset includes full-length 16S rRNA gene sequencing for microbial community profiling, untargeted LC-MS/MS metabolomics for exometabolomic profiling, and optical density measurements of microbial growth. We describe the axenic plant growth protocols, experimental design, data acquisition, processing workflows, and technical validation. This dataset provides a resource for investigating microbially mediated exometabolite transformations across diverse plant species to help understand rhizosphere processes, microbiome assembly, and ecosystem function.