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Background: The plasma membrane (PM) proteins function in a highly dynamic state, including protein trafficking and protein homeostasis, to regulate various biological processes. The dwell time and colocalization of PM proteins are considered to be two important dynamic features determining endocytosis and protein interactions, respectively. Dwell-time and colocalization detected using traditional fluorescence microscope techniques are often misestimated due to bulk measurement. In particular, analyzing these two features of PM proteins at the single-molecule level with spatiotemporal continuity in plant cells remains greatly challenging. Results: We developed a single molecular (SM) kymograph method, which is based on variable angle-total internal reflection fluorescence microscopy (VA-TIRFM) observation and single-particle (co-)tracking (SPT) analysis, to accurately analyze the dwell time and colocalization of PM proteins in a spatial and temporal manner. Furthermore, we selected two PM proteins with distinct dynamic behaviors, including AtRGS1 (Arabidopsis regulator of G protein signaling 1) and AtREM1.3 (Arabidopsis remorin 1.3), to analyze their dwell time and colocalization upon jasmonate (JA) treatment by SM kymography. First, we established new 3D (2D+t) images to view all trajectories of the interest protein by rotating these images, and then we chose the appropriate point without changing the trajectory for further analysis. Upon JA treatment, the path lines of AtRGS1-YFP appeared curved and short, while the horizontal lines of mCherry-AtREM1.3 demonstrated limited changes, indicating that JA might initiate the endocytosis of AtRGS1. Analysis of transgenic seedlings coexpressing AtRGS1-YFP/mCherry-AtREM1.3 revealed that JA induces a change in the trajectory of AtRGS1-YFP, which then merges into the kymography line of mCherry-AtREM1.3, implying that JA increases the colocalization degree between AtRGS1 and AtREM1.3 on the PM. These results illustrate that different types of PM proteins exhibit specific dynamic features in line with their corresponding functions. Conclusions: The SM-kymograph method provides new insight into quantitively analyzing the dwell time and correlation degree of PM proteins at the single-molecule level in living plant cells. [ABSTRACT FROM AUTHOR]

Dear Editor, The phytohormone jasmonate (JA), including jasmonic acid and its oxylipin derivatives, is essential for plant resistance against various stresses[1],[2]. Here, we identify I Jasmonate-Induced Dioxygenase1 i ( I JID1 i ) that encodes a 2OGD modifying OPDA and reducing the conversion of OPDA into jasmonic acid and JA-Ile. Interestingly, the amounts of OPDA, jasmonic acid, and JA-Ile in loss-of-function mutants I jid1-1 i and I jid1-5 i were similar to those of WT (Supplementary Figs. [Extracted from the article]

Plants and microbial pathogens often engage in a fierce war that determines their survival. Host plants have evolved sophisticated regulatory mechanisms to fine-tune defense responses to counter attacks from pathogens, while pathogens often hijack the lipid-derived phytohormone jasmonate to cause hormonal signaling imbalances for efficient infection. This review focuses on the jasmonate-based warfare between host plants and pathogenic intruders, and further discusses approaches to uncouple plant growth and defense tradeoffs in crop breeding. [ABSTRACT FROM AUTHOR]

Jasmonate (JA) is one of the key phytohormones essential for plant development and defense processes. The core JA biosynthetic and signaling pathways have been well-characterized. Notably, post-translational modifications (PTMs), which affect the protein structures and functions, have emerged as critical mechanisms to modulate JA output at different spatiotemporal levels. Disruption of PTMs in JA biosynthesis and signaling would cause the dysfunction of vital biological processes. Here, we give an overview of the PTMs that have been identified in JA biosynthetic and signaling pathways, and provide insights into the mechanisms by which PTMs define JA responses. [ABSTRACT FROM AUTHOR]

Phytohormones integrate external environmental and developmental signals with internal cellular responses for plant survival and multiplication in changing surroundings. Jasmonate (JA), which might originate from prokaryotes and benefit plant terrestrial adaptation, is a vital phytohormone that regulates diverse developmental processes and defense responses against various environmental stresses. In this review, we first provide an overview of ligand–receptor binding techniques used for the characterization of phytohormone–receptor interactions, then introduce the identification of the receptor COI1 and active JA molecules, and finally summarize recent advances on the regulation of JA perception and its evolution. Mechanisms of hormone perceptionis a topic of extraordinary significance, and perception of the hormone jasmonate (JA) is essential for plant defense and growth. This review highlights the methodology of ligand–receptor binding techniques and the interaction between bioactive JA and its receptor COI1. Recent advances on the regulation of JA perception and its evolution are also discussed. [ABSTRACT FROM AUTHOR]