<p>Advances in molecular biology and functional genomics have elucidated the critical roles of microRNAs (miRNAs) in regulating plant stress response pathways. In sugarcane, several miRNAs have been identified with diverse functions related to pathogen infection, temperature fluctuation, salinity, drought stress, and developmental processes. By regulating immune receptor levels, hormone signaling, and the synthesis of defense-related metabolites, miRNAs play a pivotal role in mediating both biotic and abiotic stress tolerance. Owing to their dual role in preventing autoimmunity and conferring resistance, they have emerged as key targets for genetic improvement. Recent identification of novel miRNAs in sugarcane through high-throughput sequencing and functional validation has expanded opportunities for biotechnological applications aimed at enhancing disease resistance in this economically important crop. This review highlights the association of specific miRNAs: such as miR398, miR169, miR156, and miR393, with responses to heat, salinity, drought, and pathogen attack. Their varied regulatory effects on sugarcane biology extend to developmental timing, nutrient acquisition, and the biosynthesis of secondary metabolites. Furthermore, advanced tools including artificial miRNA technology, CRISPR/Cas9-mediated gene editing, degradome analysis, and small RNA sequencing have deepened our understanding of miRNA-mediated regulation. Collectively, these advances establish miRNAs as powerful components of the molecular breeding toolkit. In the context of climatic uncertainty and the pursuit of sustainable agriculture, miRNAs represent crucial molecular resources for sugarcane improvement programs supported by continued investment in genomic resources, functional validation, and integrated omics approaches. This review, therefore, provides comprehensive insights into the diverse roles of miRNAs in conferring stress tolerance and promoting crop improvement.</p>