Based on the transcriptome sequencing of flower development in Lycium barbarum, it is suggested that the Squamosa promoter binding proteinlike (SPL) transcription factor may play an important role in flower development of L. barbarum. In this study, we used RACE method to clone the LbSPL6 gene from a characteristic plant resource in Ningxia——L. barbarum. Then we studied the gene function through bioinformatics and gene expression analysis. The results showed that： (1) the open reading frame of LbSPL6 gene was 1 524 bp in length, which encoded 507 amino acids, and a molecular weight of 55.34 kD; Sequence analysis showed that the LbSPL6 protein contained three conserved motifs; The amino acid sequence of LbSPL6 protein was similar to that of Solanaceae homologous protein. (2) qRTPCR analysis results showed that the LbSPL6 gene was expressed in the floral organs of L. barbarum. Moreover, the expression of LbSPL6 was higher in the tetrad stage and single nucleus pollen stage of anther development. Subcellular localization confirmed the nuclear localization of LbSPL6 protein. These results further elucidated the function of LbSPL6 transcription factor in the development of L. barbarum flowers.