Beneficial fungi are promising tools for enhancing plant growth and crop yield in stressful environments. Penicillium olsonii TLL1 (POT1) was identified as a potential biofertilizer enhancing plant growth and phosphate use efficiency especially under phosphate deficiency stress. Hence, we attempted to explore bioinformatic insights into how POT1 enhances plant growth under phosphate starvation. In our study, wild-type Arabidopsis thaliana Columbia-0 roots and shoots cultivated with POT1 under phosphate-limiting conditions were employed for comparative analyses. By integrating transcriptomic and proteomic data, we identified key molecular pathways regulated by POT1 that influenced phosphate acquisition and plant stress tolerance. Comprehensive RNA-seq analysis revealed significant upregulation of genes involved in phosphate transport, root architecture, and stress-related pathways, while proteome profiling further highlighted proteins associated with lipid remodeling, phosphate metabolism, and phytohormone signaling. Bioinformatic analyses of differentially expressed genes (DEGs) and proteins (DEPs) elucidated the complex regulatory networks at both transcriptional and translational levels, with key contributions from auxin and ethylene signaling. Our study demonstrated that POT1-treated plants exhibited enhanced root development and nutrient uptake under phosphate-deficient conditions, driven by the coordinated regulation of phosphate solubilization genes and stress-responsive proteins. Our findings underscore the potential of multi-omics approaches in unraveling the molecular mechanisms behind plant–microbe interactions, with implications for improving sustainable agricultural practices.