This study used strawberry variety ‘Benihoppe’ as test material. The FaSPS1 gene expression in different stages of strawberry fruit development was analyzed. The FaSPS1 gene was cloned by PCR and the plant expression vector with reporter eGFP gene was constructed. The strawberry fruit was transformed by transient transgene method. The aim of this study was to identify the transgenic plants by detecting the expression of FaSPS1 gene and observe green fluorescence. The changes of the expression of FaSPS1 gene during the ripening and development of strawberry fruits were analyzed after the overexpression and antisense expression of FaSPS1 gene. The special role of FaSPS1 gene in the ripening and development of strawberry fruits was explored to provide ideas for further understanding the molecular mechanism of strawberry fruit development and ripening regulation. The results showed that: (1) FaSPS1 gene (GenBank number AB267868.1) was successfully cloned; FaSPS1 gene overexpression vector and antisense gene expression vector with reporter gene eGFP were successfully constructed, and the transgenic strawberry plants were identified by transient transgene transformation and fluorescence and target gene expression detection. (2) Overexpression of FaSPS1 gene can promote the ripening of strawberry fruit, advance the ripening period of strawberry fruit, and increase the sucrose and fructose contents in the fruit, but antisense expression can inhibit the ripening of strawberry fruit and increase the malic acid content in the fruit. (3) After gene overexpression or antisense expression, the expression of ripeningrelated genes in strawberry fruits were regulated to varying degrees. Among them, sugar metabolism genes FaSPS2/3, FaSUT1, fruit softening genes FaEXP1, FaEXP3, FaXYL1 and hormone metabolism genes FaJAZ1, FaJAZ2, FaJAZ8, FaOPR3, FaPYL1, FaPYL8, FaPYL9 and FaNCED1 showed the most significant changes. It is speculated that FaSPS1 gene may regulate strawberry fruit ripening by affecting sugar metabolism genes, fruit softening genes and hormone metabolism genes related to fruit ripening.