MADS-box transcription factors play a crucial role in regulating plant biological processes, including fruit ripening and associated events. This study aimed to understand the mechanisms involved in durian fruit ripening and carotenoid production by conducting a genome-wide analysis of MADS-box proteins in durian (Durio zibethinus L.), an economically important fruit in Southeast Asia. A total of 119 durian MADS-box proteins were identified from the genome of the ‘Musang King’ cultivar. Phylogenetic analysis classified these proteins into types I and II, displaying a similar conserved motif composition. Notably, only 16 durian MADS-box genes exhibited fruit-specific expression patterns. Among these, DzAGL6-1 was predominantly expressed during fruit development, the stage at which carotenoid biosynthesis is activated. Transient expression of DzAGL6-1 in tomato fruit increased the transcript level of the carotenoid biosynthetic gene phytoene synthase (PSY) and β-carotene content. Furthermore, DzAGL6-1 was found to directly bind to the promoter of DzPSY, as demonstrated by a dual-luciferase assay. These findings provide insights into the role of MADS-box transcription factors in regulating carotenoid accumulation during durian fruit development.