PNNL

Abstract

Membrane lipids serve as substrates for the generation of numerous signaling lipids when plants are exposed to environmental stresses, and jasmonic acid, an oxidized product of 18-carbon unsaturated fatty acids (e.g., linolenic acid), has been recognized as the essential signal in wound-induced gene expression. Yet, the contribution of individual membrane lipids in linolenic acid generation is ill-defined. In this work we performed spatial lipidomic experiments to track lipid changes that occur locally at the sight of leaf injury to better understand the potential origin of linoleic acid from individual membrane lipids. The central veins of tomato leaflets were crushed using surgical forceps, leaves were cryosectioned and analyzed by two orthogonal MALDI MS imaging platforms for insight into lipid spatial distribution. Significant changes in lipid composition are only observed 30 min after wounding, while after 60 min lipidome homeostasis has been re-established. PCs exhibit inconsistent pattern of spatial behavior in individual plants. Among lysolipids, LPCs strongly co-localize with the injured zone of wounded leaflets, while, for example, LPG (16:1) accumulated preferentially toward the apex in the injured zone of wounded leaflets. In contrast, two other LPGs (LPG (18:3) and LPG (18:2)) are depleted in the injured zone.Our high-resolution co-localization imaging analyses suggest that linolenic acids are predominantly released from PCs with 16/18 fatty acid composition along the entire leaf, while it seems that in the apex zone PG 16:1_18:3 significantly contributes to the linoleic acid pool. These results also indicate distinct localization and/or substrate preferences of phospholipase isoforms in leaf tissue.