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Gibberellin (GAs) plays an important regulatory role in the development and growth of pineapple (<i>Ananas comosus</i> (L.) Merr.). Bioinformatics was used to confirm the differential expression of GA2 gibberellin oxidase gene <i>AcGA2oxs</i> in the pineapple genome, which laid the foundation for exploring its role in pineapple. In this study, 42 <i>GA2ox</i> genes <i>(AcGA2oxs)</i> were identified in the pineapple genome, named from <i>AcGA2ox1</i> to <i>AcGA2ox42</i>, and divided into four groups according to phylogenetic analysis. We also analyzed the gene structure, conserved motifs and chromosome localization of AcGA2oxs. AcGA2oxs within the same group had similar gene structure and motifs composition. Collinear analysis and cis-element analysis provided the basis for understanding the evolution and function of <i>GA2ox</i> genes in pineapple. In addition, we selected different tissue parts to analyze the expression profile of <i>AcGA2oxs</i>, and the results show that 41 genes were expressed, except for <i>AcGA2ox18</i>. <i>AcGA2ox18</i> may not be expressed in these sites or may be pseudogenes. qRT-PCR (real-time fluorescence quantitative PCR) was used to detect the relative expression levels of the <i>GA2ox</i> gene family under different concentrations of GA3 treatment, and it was found that <i>AcGA2ox</i> gene expression was upregulated in different degrees under GA3 treatment. These results provide useful information for further study on the evolution and function of the <i>GA2ox</i> family in pineapple.