Alternative splicing refers to the process of producing different splicing isoforms from the same pre-mRNA through different alternative splicing events, which almost participates in all stages of plant growth and development. In order to understand its role in the fruit development ofOsmanthus fragrans, transcriptome sequencing and alternative splicing analysis was carried out on three stages ofO. fragransfruit (O. fragrans“Zi Yingui”). The results showed that the proportion of skipping exon events was the highest in all three periods, followed by a retained intron, and the proportion of mutually exclusive exon events was the lowest and most of the alternative splicing events occurred in the first two periods. The results of enrichment analysis of differentially expressed genes and differentially expressed isoforms showed that alpha-Linolenic acid metabolism, flavonoid biosynthesis, carotenoid biosynthesis, photosynthesis, and photosynthetic-antenna protein pathways were significantly enriched, which may play an important role in the fruit development ofO. fragrans. The results of this study lay the foundation for further study of the development and maturation ofO. fragransfruit and further ideas for controlling fruit color and improving fruit quality and appearance.