•Phospholipase D (PLD) hydrolyzes phospholipids to produce phosphatidic acid (PA) and a head group, and is involved in the response to various environmental stresses, including salinity. Here, we determined the roles of PLD and PA in the mediation of salt (NaCl)-stress signaling through the regulation of mitogen-activated protein kinase (MAPK or MPK) in Arabidopsis thaliana.•NaCl-induced changes in the content and composition of PA were quantitatively profiled by electrospray ionization–tandem mass spectrometry (ESI-MS/MS). A specific PA species (a MAPK 16:0–18:2 PA), which was increased in abundance by exposure to NaCl, bound to a MPK6, according to filter binding and ELISA. The effect of... More
•Phospholipase D (PLD) hydrolyzes phospholipids to produce phosphatidic acid (PA) and a head group, and is involved in the response to various environmental stresses, including salinity. Here, we determined the roles of PLD and PA in the mediation of salt (NaCl)-stress signaling through the regulation of mitogen-activated protein kinase (MAPK or MPK) in Arabidopsis thaliana.•NaCl-induced changes in the content and composition of PA were quantitatively profiled by electrospray ionization–tandem mass spectrometry (ESI-MS/MS). A specific PA species (a MAPK 16:0–18:2 PA), which was increased in abundance by exposure to NaCl, bound to a MPK6, according to filter binding and ELISA. The effect of PA on MPK6 activity was tested using in-gel analysis.•16:0–18:2 PA stimulated the activity of MPK6 immunoprecipitated from Arabidopsis leaf extracts. Treatment with NaCl induced a transient activation of MPK6 in wild-type plant, but the activation was abolished in the pld1 plant mutant. A plasma membrane Na+/H+ antiporter (SOS1) was identified as a downstream target of MPK6. MPK6 phosphorylated the C-terminal fragment of SOS1. The MPK6 phosphorylation of SOS1 was stimulated by treatment with NaCl, as well as directly by PA.•These results suggest that PA plays a critical role in coupling MAPK cascades in response to salt stress.
