Ethylene, an important hormone in plant growth, development and response to environmental stimuli, is rapidly induced by mechanical injury or wounding. Although induction of ACS (1-aminocyclopropane-1-carboxylic acid synthase) gene expression has been associated with this process, the detailed regulatory mechanism is unclear. Here, we report that the wounding-induced ethylene production is modulated by both mitogen-activated protein kinase (MAPK) pathway and calcium-dependent protein kinase (CPK) pathway. Study using acs mutants demonstrated that four ACS isoforms, including ACS2, ACS6, ACS7 and ACS8, contribute to ethylene production in response to wounding. Loss-of-function analysis defines the role of MPK3 and MPK6, and their upstream MKK4 and MKK5, in wounding-induced ethylene production. They play an important role in the wounding-induced up-regulation of all four ACS genes expression. Independent of MAPK pathway, CPK5 and CPK6 are also involved in the wounding-induced ethylene production by regulating the expression of ACS2, ACS6 and ACS8 genes. Taken together, we demonstrate that two independent kinase signalling pathways, MPK3/MPK6 cascade and CPK5/CPK6, are involved in the wounding-induced ethylene biosynthesis via differential regulation of ACS genes at transcriptional level.