Abstract: In order to clarify the mechanism of MADS-box transcription factor gene family member in flowering and floral organ development in Macadamia integrifolia,a MiMADS-box gene was cloned from the leaves of M. integrifolia“GUIRE1”cultivar by PCR technique and analyzed its structure and function by Bioinformatics method. The cloned cDNA sequence of MiMADS-box gene was 1 179 bp in length,729 bp open reading frame（ORF）,encoded 242 amino acids. MiMADS-box,an unstable hydrophilic protein that had no transmembrane structure,no signal peptide and localized in the nucleus,had MADS superfamily and K-box superfamily conserved domains belonging to the MIKCc-type family. Phylogenetic analysis clustered MiMADS-box and AGL7/AP1,AGL10/CAL1,AGL8/FUL,AGL79 into FUL-AP1（SQUA）subfamily. The transcriptome data were used to analyze the expression patterns of MiMADS-box gene in branch,flower and leave of M. integrifolia“GUIRE1”and“695”cultivar. Results indicated that MiMADS-box had the lowest expression in the flower of“GUIRE1”,but the expression level of“695”was the highest. The green fluorescent protein fusion vector pGREEN-MiMADS-box-ORF-GFP was constructed by homologous recombination. It was speculated that MiMADS-box was closely related to flowering and floral organ development. The study provided a theoretical and technical guide for clarification the role MiMADS-box in flowering regulation mechanism of M. integrifolia.