Leaf senescence is a developmentally programmed cell death process that constitutes the final step of leaf development, and it can be regulated by multiple environmental cues and endogenous signals. The mitogen-activated protein kinase (MAPK) cascades play diverse roles in intracellular and extracellular signaling in plants. Roles of the MAPK signaling module in leaf senescence are unknown. Here, a MAPK cascade involving MKK9-MPK6 is shown to play an important role in regulating leaf senescence in Arabidopsis (Arabidopsis thaliana). Both MKK9 and MPK6 possess kinase activities, with MPK6 an immediate target of MKK9, as revealed by in vitro, in vivo, and in planta assays. The constitutive and inducible overexpression of MKK9 causes premature senescence in leaves and in whole Arabidopsis plants. The premature senescence phenotype is suppressed when MKK9 is overexpressed in the mpk6 null background. When either MKK9 or MPK6 is knocked out, leaf senescence is delayed.