The WRKY gene family is one of the largest and unique gene families in plants, and it is involved in various aspects of plant growth and development. But there have been no reports on WRKY12 in loquat. In this study, the EjWRKY12 gene was cloned from loquat buds and leaves using homologous cloning, and its physical and chemical properties, protein structure, evolutionary relationship, and cis-acting elements in the promoter region were analyzed using bioinformatics. The expression pattern of the gene was also analyzed in combination with transcriptome data. The results showed that the EjWRKY12 gene in loquat is 711 bp in length and encodes 236 amino acids. The homologous genes obtained by comparison are mostly related to flowering, suggesting that EjWRKY12 may be involved in loquat flowering. Promoter analysis indicated that it may be regulated by light, MeJA, gibberellin, and drought. EjWRKY12 protein was localized in the nucleus. After exogenous GA3 treatment, there was no significant difference in the expression level and trend of EjWRKY12 in leaves between the treatment group and the control group. However, in the apical bud, the expression level of EjWRKY12 in the control group was the lowest during the critical period of flowering. After GA3 treatment, the expression level of the gene increased continuously with the prolongation of treatment time, indicating that it may be a transcription factor that inhibits flowering and promotes vegetative growth. This study first cloned the EjWRKY12 gene and has guiding significance for further elucidating the molecular mechanism of loquat flowering regulation.