Stomatal closure is a common adaptation response of plants to the onset of drought condition and its regulation is controlled by transcription factors. MYB60, a transcription factor involved in the regulation of light-induced stomatal opening, has been characterized in arabidopsis and grapevine. In this work, we studied the role of MYB60 homolog SIMYB60 in tomato plants. We identified, isolated, and sequenced the SIMYB60 coding sequence, and found domains and motifs characteristic of other MYB60 proteins. We determined that SlMYB60 is mainly expressed in leaves, and its expression is repressed by abscisic acid. Next, we isolated a putative promoter region containing regulatory elements responsible for guard cell expression and other putative regulatory elements related to ABA repression and vascular tissue expression. Protein localization assays demonstrated that SlMYB60 localizes to the nucleus. Finally, SlMYB60 is able to complement the mutant phenotype of atmyb60-1 in Arabidopsis. Together, these results indicate that SlMYB60 is the homologous gene in tomato and potentially offer a molecular target to improve crops.