Plastidial fatty acid signaling modulates salicylic acid- and jasmonic acid-mediated defense pathways in the Arabidopsis ssi2 mutant.

A mutation in the Arabidopsis gene ssi2/fab2, which encodes stearoyl-acyl carrier protein desaturase (S-ACP-DES), results in the reduction of oleic acid (18:1) levels in the mutant plants and also leads to the constitutive activation of NPR1-dependent and -independent defense responses. By contrast, ssi2 plants are compromised in the induction of the jasmonic acid (JA)-responsive gene PDF1.2 and in resistance to the necrotrophic pathogen Botrytis cinerea. Although S-ACP-DES catalyzes the initial desaturation step required for JA biosynthesis, a mutation in ssi2 does not alter the levels of the JA precursor linolenic acid (18:3), the perception of JA or ethylene, or the induced endogenous levels of JA. This finding led us to postulate that the S-ACP-DES-derived fatty acid (FA) 18:1 or its derivative is required for the activation of certain JA-mediated responses and the repression of the salicylic acid (SA) signaling pathway. Here, we report that alteration of the prokaryotic FA signaling pathway in plastids, leading to increased levels of 18:1, is required for the rescue of ssi2-triggered phenotypes. 18:1 levels in ssi2 plants were increased by performing epistatic analyses between ssi2 and several mutants in FA pathways that cause an increase in the levels of 18:1 in specific compartments of the cell. A loss-of-function mutation in the soluble chloroplastic enzyme glycerol-3-phosphate acyltransferase (ACT1) completely reverses SA- and JA-mediated phenotypes in ssi2. In contrast to the act1 mutation, a loss-of-function mutation in the endoplasmic reticulum-localized omega6 oleate desaturase (FAD2) does not alter SA- or JA-related phenotypes of ssi2. However, a mutation in the plastidial membrane-localized omega6 desaturase (FAD6) mediates a partial rescue of ssi2-mediated phenotypes. Although ssi2 fad6 plants are rescued in their morphological phenotypes, including larger size, absence of visible lesions, and straight leaves, these plants continue to exhibit microscopic cell death and express the PR-1 gene constitutively. In addition, these plants are unable to induce the expression of PDF1.2 in response to the exogenous application of JA. Because the act1 mutation rescues all of these phenotypes in ssi2 fad6 act1 triple-mutant plants, act1-mediated reversion may be mediated largely by an increase in the free 18:1 content within the chloroplasts. The reversion of JA responsiveness in ssi2 act1 plants is abolished in the ssi2 act1 coi1 triple-mutant background, suggesting that both JA- and act1-generated signals are required for the expression of the JA-inducible PDF1.2 gene. Our conclusion that FA signaling in plastids plays an essential role in the regulation of SSI2-mediated defense signaling is further substantiated by the fact that overexpression of the N-terminal-deleted SSI2, which lacks the putative plastid-localizing transit peptide, is unable to rescue ssi2-triggered phenotypes, as opposed to overexpression of the full-length protein.