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The brief longevity of tree peony blossoms constrains its ornamental value and economic worth. Gibberellins (GAs) are crucial in the modulation of flower senescence, and GA 20-oxidase (GA20ox), GA 3-oxidase (GA3ox), and GA 2-oxidase (GA2ox) catalyze the synthesis and deactivation of bioactive GAs. In <i>Paeonia ostii</i>, a total of three PoGA20ox, ten PoGA3ox, and twelve PoGA2ox proteins were identified and comprehensively analyzed. The analysis of the gene structures, conserved domains, and motifs revealed structural similarities and variances among the GA20ox, GA3ox, GA2ox-A, and GA2ox-B subfamilies. The synteny analysis indicated a scarcity of collinear blocks within the <i>P. ostii</i> genome, with no tandem or whole-genome duplication/segmental duplications found in <i>PoGAoxs.</i> The investigation into the binding of transcription factors to <i>PoGAox</i> promoters and the assessments of the expression levels suggest that <i>PoGA2ox1</i> and <i>PoGA2ox8.1</i> are promising candidate genes implicated in the regulation of floral senescence. Further, Pos.gene61099 (BPC6) and Pos.gene61094 (CIL2) appear to modulate <i>PoGA2ox1</i> transcription in a positive and negative manner, respectively, while Pos.gene38359 (DDF1) and Pos.gene17639 (DREB1C) likely enhance <i>PoGA2ox8.1</i>’s expression. This study lays a foundation for an in-depth understanding of <i>PoGAox</i> functions and the development of strategies to delay flower senescence in tree peony.