Secretion of Phospholipase Dd Functions as a Regulatory Mechanism in Plant Innate Immunity

Jingjing Xing, Xiaojuan Li, Xiaohua Wang, Xueqin Lv, Li Wang, Liang Zhang, Yingfang Zhu, Qianhua Shen, Frantisek Balu ska, Jozef Samaj,  and Jinxing Lin*

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) PLDd 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 PLDd density in the plasma membrane after chitin treatment; PLDd also underwent rapid exocytosis. Fluorescence resonance energy transfer with fluorescence lifetime imaging microscopy showed that the interaction between PLDd and the microdomain marker AtREMORIN1.3 (AtREM1.3) increased in response to chitin, indicating that exocytosis facilitates rapid, efficient sorting of PLDd into microdomains upon pathogen stimulus. We further unveiled a trade-off between brefeldin A (BFA)每resistant and 每sensitive pathways in secretion of PLDd under diverse conditions. Upon pathogen attack, PLDd 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 PLDd 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 PLDd to papillae is modulated by exocytosis, thus triggering PA-mediated signaling in plant innate immunity.