Cell wall strengthening by phenylpropanoid dehydrodimers during the plant hypersensitive cell death
Infection of Arabidopsis with avirulent Pseudomonas syringae and exposure to nitrogen dioxide (NO2) both trigger hypersensitive cell death (HCD) that is characterized by the emission of bright blue-green (BG) autofluorescence under UV illumination. The aim of our current work was to identify the BG fluorescent molecules and scrutinize their biosynthesis and functions during the HCD. Compared to wild-type (WT) plants, the phenylpropanoid-deficient mutant fah1 developed normal HCD except for the absence of BG fluorescence. Ultrahigh resolution metabolomics combined with mass difference network analysis revealed that WT but not fah1 plants rapidly accumulate dehydrodimers of sinapic acid, sinapoylmalate, 5-OH-ferulic acid, and 5-OH-feruloylmalate during the HCD. FAH1-dependent BG fluorescence appeared exclusively within dying cells of the upper epidermis as detected by microscopy. Saponification released dehydrodimers from extracted cell wall material. Collectively, our data suggest that HCD induction leads to the formation of free BG fluorescent dehydrodimers from monomeric sinapates and 5-hydroxyferulates. Reactive oxygen species from de-regulated photosynthesis likely contribute to the radical-radical coupling. The formed dehydrodimers move from upper epidermis cells into the apoplast where they esterify and thereby cross-link cell wall polymers. Both, free as well as wall-bound phenylpropanoid dehydrodimers are defense-related compounds in Arabidopsis. We propose that other plants also employ dehydrodimers of highly abundant phenylpropanoids for rapid defense against pathogen attack.