Abstract:SPL (squamosa promoterbinding proteinlike) is a kind of gene family unique to plants. It plays an important role in the process of flower development. The putative OfSPLs were explored and identified from Osmanthus fragrans transcriptome data and their physicochemical properties, phylogeny, gene structure and expression pattern were analyzed. The key genes involved in the process of flower development were obtained by homologous sequence alignment qRTPCR, subcellular localization and yeast selfactivation. We further provided genetic resources for the mechanism of action and function of the corresponding transcription factors of O. fragrans in flower development. The results showed that: (1) 29 putative OfSPL were confirmed, they are distributed uniformly on 15 chromosomes with SBP domains. (2) According to the phylogenetic tree with Arabidopsis thaliana, they were divided into 8 subfamilies, of which have relatively conservative gene structures. (3) RNASeq data analysis showed thatOfSPL9/10/17/19/21/27/28 had higher FPKM values and had certain specificity in flower tissues. qRTPCR analysis showed that the OfSPL10/21 expression level generally exhibited an upward trend followed by a downward trend. (4) The subcellular localization analysis showed that OfSPL10/21 proteins of O. fragrans localized in the nucleus, and OfSPL10/21 had no selfactivating activity and toxicity. The study found that OfSPL10/21 may participate in the synthesis and regulation of color and aroma during the flowering of O. fragrans.