Phospholipase A2 Is Required for PIN-FORMED Protein Trafficking to the Plasma Membrane in the Arabidopsis Root

Phospholipase A(2) (PLA(2)), which hydrolyzes a fatty acyl chain of membrane phospholipids, has been implicated in several biological processes in plants. However, its role in intracellular trafficking in plants has yet to be studied. Here, using pharmacological and genetic approaches, the root hair bioassay system, and PIN-FORMED (PIN) auxin efflux transporters as molecular markers, we demonstrate that plant PLA(2)s are required for PIN protein trafficking to the plasma membrane (PM) in the Arabidopsis thaliana root. PLA(2)alpha, a PLA(2) isoform, colocalized with the Golgi marker. Impairments of PLA(2) function by PLA(2)alpha mutation, PLA(2)-RNA interference (RNAi), or PLA(2) inhibitor treatments significantly disrupted the PM localization of PINs, causing internal PIN compartments to form. Conversely, supplementation with lysophosphatidylethanolamine (the PLA(2) hydrolytic product) restored the PM localization of PINs in the plA(2)alpha mutant and the ONO-RS-082-treated seedling. Suppression of PLA(2) activity by the inhibitor promoted accumulation of trans-Golgi network vesicles. Root hair-specific PIN overexpression (PINox) lines grew very short root hairs, most likely due to reduced auxin levels in root hair cells, but PLA(2) inhibitor treatments, PLA(2)alpha mutation, or PLA(2)-RNAi restored the root hair growth of PINox lines by disrupting the PM localization of PINs, thus reducing auxin efflux. These results suggest that PLA(2), likely acting in Golgi-related compartments, modulates the trafficking of PIN proteins.