Intron retention dominates alternative splicing reprogramming during anaerobic stress in Chlamydomonas reinhardtii.

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conclusionFull-length transcriptomic analysis indicates that alternative splicing, particularly intron retention, is a prominent component of the anaerobic stress response in Chlamydomonas reinhardtii and may contribute to metabolic adaptation through transcript isoform remodeling. Alternative splicing (AS) is a pivotal biological process that enhances transcriptomic plasticity in eukaryotes, especially under environmental stress. In this study, we integrated PacBio Iso-Seq and Illumina RNA-seq technologies to characterize the full-length transcriptome of Chlamydomonas reinhardtii under dark anaerobic conditions. Anaerobic treatment was associated with extensive remodeling of transcript structures, with intron retention emerging as the predominant splicing type. Transcript-structure changes were concentrated in genes related to carbon metabolism, pyruvate conversion, and fatty acid biosynthesis. Several key genes showed clear condition-dependent transcript isoform changes, including putative isoform switching events, which were further supported by qRT-PCR analysis. These findings support the view that AS-associated transcript-structure remodeling contributes to the anaerobic response of Chlamydomonas reinhardtii and provide a high-quality transcriptomic resource for future studies of stress adaptation and gene regulation in green algae.