MADS-box genes comprise a family of transcription factors that function in the growth and development of plants. To obtain insights into their evolution in watermelon (Citrullus lanatus), we carried out a genome-wide analysis and identified 39 MADS-box genes. These genes were classified into MIKCc (25), MIKC*(3), Mα (5), Mβ (3), and Mγ (3) clades according to their phylogenetic relationship with Arabidopsis thaliana and Cucumis sativus; moreover, these 25 genes in the MIKC clade could be classified into 13 subfamilies, and the Flowering Locus C (FLC) subfamily is absent in watermelon. Analysis of the conserved gene motifs showed similar motifs among clades. Continuing chromosomal localizations analysis indicated that MADS-box genes were distributed across 11 chromosomes in watermelon, and these genes were conditioned to be differentially expressed during plant growth and development. This research provides information that will aid further investigations into the evolution of the MADS-box gene family in plants.