FAB1/PIKfyve is a key enzyme that catalyzes phosphatidylinositol 3-phosphate (PtdIns3P) to form phosphatidylinositol 3,5-bisphosphate (PtdIns (3,5) P₂). Its product PtdIns (3,5) P₂ plays an important role in the development of eukaryotic cells. In order to figure out the function of PtdIns (3,5) P₂ in rice reproductive development, this study combined bioinformatics and genetics methods to identify the rice FAB1/PIKfyve genes, analyzed the physical and chemical properties, gene structures, conserved domains, phylogenetic tree, cis-acting elements, tissue expression profiles and used CRISPR/Cas9 gene editing technology to obtain osfab1b mutants. Bioinformatics analysis results showed that nine FAB1 gene family members were identified from the whole genome of Oryza Sativa. Gene structure analysis indicated the differences existed in the gene structure of FAB1 family, the number of exons are 8-12. Conserve domain analysis showed that OsFAB1A and OsFAB1B contained the N-terminal FYVE domain, and the remaining members contained the Cpn60_TCP1 domain and PIPKc kinase domain. Phylogenetic analysis indicated that the functions of FAB1 family were highly conserved in mono- and dicotyledonous plants. Prediction of cis-element in the upstream regulatory region of FAB1 genes revealed a variety of growth-related, light-responsive, and hormone and stress-responsive cis-elements. Tissue expression profiles showed that most of FAB1 genes were global-expressed, and the high expression of FAB1C sub-cluster genes in lemma and palea suggested that they might be involved in floral organ development. Finally, the osfab1b mutants were obtained through the CRISPR/Cas9 system, and the pollen vitality of the osfab1b mutants showed no significantly abnormality, implied that FAB1 family had functional redundancy in the reproductive development of rice. The present results provide a theoretical reference for biological function studies of the phosphatidylinositol regulatory network in rice.