Abstract
Early changes in the Arabidopsis thaliana membrane phosphoproteome in response to oligogalacturonides (OGs), a class of plant damage-associated molecular patterns (DAMPs), were analyzed by two complementary proteomic approaches. Differentially phosphorylated sites were determined through phosphopeptide enrichment followed by LC-MS/MS using label-free quantification; differentially phosphorylated proteins were identified by 2D-DIGE combined with phospho-specific fluorescent staining (phospho-DIGE). This large-scale phosphoproteome analysis of early OG-signaling enabled us to determine 100 regulated phosphosites using LC-MS/MS and 46 differential spots corresponding to 34 pdhosphoproteins using phospho-DIGE. Functional classification showed that the OG-responsive phosphoproteins include kinases, phosphatases and receptor-like kinases, heat shock proteins (HSPs), reactive oxygen species (ROS) scavenging enzymes, proteins related to cellular trafficking, transport, defense and signaling as well as novel candidates for a role in immunity, for which elicitor-induced phosphorylation changes have not been shown before. A comparison with previously identified elicitor-regulated phosphosites shows only a very limited overlap, uncovering the immune-related regulation of 70 phosphorylation sites and revealing novel potential players in the regulation of elicitor-dependent immunity.
Highlights
Plants have developed various mechanisms to defend themselves against biotic stresses
Seedlings In Arabidopsis, treatment with OGs induces the activation, by phosphorylation, of MPK3 and Mitogen-activated protein kinase 6 (MPK6) within few minutes, a temporal kinetics similar to that observed upon treatment with Pathogen-associated molecular patterns (PAMPs) (Galletti et al, 2011)
Two-week-old liquid-grown Arabidopsis ecotype Columbia 0 (Col-0) seedlings were treated with OGs (50 μg/ml) or water as a control
Summary
Plants have developed various mechanisms to defend themselves against biotic stresses. Oligogalacturonide-Regulated Phosphoproteome of endogenous molecular patterns that are present only when the tissue is infected or damaged (damage-associated molecular patterns or DAMPs). In these cases, the discrimination between an intact self and an altered self leads to the activation of the immune system (Benedetti et al, 2015). The discrimination between an intact self and an altered self leads to the activation of the immune system (Benedetti et al, 2015) Recognition of both PAMPs and DAMPs is mediated by the so-called pattern recognition receptors (PRRs; Boller and Felix, 2009)
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