This study employed Illumina high-throughput sequencing technology to investigate the effects of Bacillus velezensis GF-46 treatment on miRNA expression in maize B73. A total of 37 differentially expressed miRNAs were identified, including conserved families (such as miR164, miR171, miR166, etc.) and novel miRNAs (such as 8_36419, 3_16586, etc.). KEGG pathway enrichment analysis revealed 135 significantly modulated metabolic pathways (Q<0.05), with pronounced representation in: (i) phytohormone signaling (auxin and jasmonic acid signaling pathways), (ii) phenylpropanoid biosynthesis (PAL, 4CL enzymatic nodes), (iii) plant-pathogen recognition systems (RLK-mediated signaling pathways). Notably, miRNA-8_36419 exhibited superior regulatory capacity, targeting approximately 50% of annotated genes within these key pathways. These findings clarify the mechanisms underlying GF-46-mediated growth promotion and systemic resistance in maize, and deepen the understanding of microbiomeregulated transcriptional networks in monocot crops.