Plant phospholipase Ds (PLDs), essential regulators of phospholipid signaling, function in multiple signal transduction cascades; however, the mechanisms regulating PLDs in response to pathogens remain unclear. Here, we found that Arabidopsis (Arabidopsis thaliana) PLDδ accumulated in cells at the entry sites of the barley powdery mildew fungus, Blumeria graminis f. sp hordei Using fluorescence recovery after photobleaching and single-molecule analysis, we observed higher PLDδ density in the plasma membrane after chitin treatment; PLDδ also underwent rapid exocytosis. Fluorescence resonance energy transfer with fluorescence lifetime imaging microscopy showed that the interaction between PLDδ and the microdomain marker AtREMORIN1.3 (AtREM1.3) increased in response to chitin, indicating that exocytosis facilitates rapid, efficient sorting of PLDδ into microdomains upon pathogen stimulus. We further unveiled a trade-off between brefeldin A (BFA)-resistant and -sensitive pathways in secretion of PLDδ under diverse conditions. Upon pathogen attack, PLDδ secretion involved syntaxin-associated VAMP721/722-mediated exocytosis sensitive to BFA. Analysis of phosphatidic acid (PA), hydrogen peroxide, and jasmonic acid (JA) levels and expression of related genes indicated that the relocalization of PLDδ is crucial for its activation to produce PA and initiate reactive oxygen species and JA signaling pathways. Together, our findings revealed that the translocation of PLDδ to papillae is modulated by exocytosis, thus triggering PA-mediated signaling in plant innate immunity.plantcell;31/12/3015/FX1F1fx1.

Beijing Advanced Innovation Center for Tree Breeding by Molecular Design and College of Biological Sciences and Biotechnology Beijing Forestry University Beijing 100083 China

Centre of the Region Hana for Biotechnological and Agricultural Research Faculty of Science Palacky University Olomouc 78301 Czech Republic

Institute of Botany Chinese Academy of Sciences Beijing 100093 China

Institute of Cellular and Molecular Botany Rheinische Friedrich Wilhelms University Bonn Department of Plant Cell Biology Bonn D 53115 Germany

Key Laboratory of Plant Stress Biology School of Life Sciences Henan University Kaifeng 457004 China

State Key Laboratory of Plant Cell and Chromosome Engineering Centre for Molecular Agrobiology Institute of Genetics and Developmental Biology Chinese Academy of Sciences Beijing100101 China

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Plant Cell. 2019 Dec;31(12):2819-2820. doi: 10.1105/tpc.19.00822. PubMed

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Spatial proteomics of vesicular trafficking: coupling mass spectrometry and imaging approaches in membrane biology

DIACYLGLYCEROL KINASE 5 participates in flagellin-induced signaling in Arabidopsis

Imaging plant cells and organs with light-sheet and super-resolution microscopy