Abstract: Plants inhabit dynamic environments and have evolved a variety of complex mechanisms to cope with stress under extreme conditions. Alternative splicing (AS), a post-transcriptional regulatory mechanism prevalent in eukaryotes, expands transcriptomic and proteomic diversity and plays a pivotal role in plant resilience. Exposure to high or low temperature, drought, and pathogen challenge triggers AS-mediated regulatory pathways that enable stress mitigation while maintaining normal growth and development. This review consolidates recent advances in elucidating how AS regulates plant stress responses, underscoring its central role in regulatory networks and the transformative impact of high-throughput sequencing technologies on this field. It also examines the theoretical and applied significance of AS in plant breeding, providing a framework for advancing research on stress resistance and for developing cultivars with enhanced resilience.