Super-Resolution Imaging of Plant Receptor-Like Kinases Uncovers Their Colocalization and Coordination with Nanometer Resolution
Abstract
Plant cell signaling often relies on the cellular organization of receptor-like kinases (RLKs) within membrane nanodomains to enhance signaling specificity and efficiency. Thus, nanometer-scale quantitative analysis of spatial organizations of RLKs could provide new understanding of mechanisms underlying plant responses to environmental stress. Here, we used stochastic optical reconstruction fluorescence microscopy (STORM) to quantify the colocalization of the flagellin-sensitive-2 (FLS2) receptor and the nanodomain marker, remorin, within Arabidopsis thaliana root hair cells. We found that recovery of FLS2 and remorin in the plasma membrane, following ligand-induced internalization by bacterial-flagellin-peptide (flg22), reached ~85% of their original membrane density after ~90 min. The pairs colocalized at the membrane at greater frequencies, compared with simulated randomly distributed pairs, except for directly after recovery, suggesting initial uncoordinated recovery followed by remorin and FLS2 pairing in the membrane. The purinergic receptor, P2K1, colocalized with remorin at similar frequencies as FLS2, while FLS2 and P2K1 colocalization occurred at significantly lower frequencies, suggesting that these RLKs mostly occupy distinct nanodomains. The chitin elicitor receptor, CERK1, colocalized with FLS2 and remorin at much lower frequencies, suggesting little coordination between CERK1 and FLS2. These findings emphasize STORM’s capacity to observe distinct nanodomains and degrees of coordination between plant cellmore »
- Publication Date:
- Research Org.:
- Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER); National Institutes of Health (NIH); National Institute of General Medical Sciences (NIGMS)
- OSTI Identifier:
- 1914371
- Alternate Identifier(s):
- OSTI ID: 1964169
- Report Number(s):
- PNNL-SA-179989
Journal ID: ISSN 2077-0375; MBSEB6; PII: membranes13020142
- Grant/Contract Number:
- SC0020346; AC05-76RL01830; 2RO1GM121445-05
- Resource Type:
- Published Article
- Journal Name:
- Membranes
- Additional Journal Information:
- Journal Name: Membranes Journal Volume: 13 Journal Issue: 2; Journal ID: ISSN 2077-0375
- Publisher:
- MDPI AG
- Country of Publication:
- Switzerland
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; fluorescence microscopy; flagellin-sensitive receptors; single molecule; chitin-elicitor receptor kinase; immune response; phytology
Citation Formats
Traeger, Jeremiah, Hu, Dehong, Yang, Mengran, Stacey, Gary, and Orr, Galya. Super-Resolution Imaging of Plant Receptor-Like Kinases Uncovers Their Colocalization and Coordination with Nanometer Resolution. Switzerland: N. p., 2023.
Web. doi:10.3390/membranes13020142.
Traeger, Jeremiah, Hu, Dehong, Yang, Mengran, Stacey, Gary, & Orr, Galya. Super-Resolution Imaging of Plant Receptor-Like Kinases Uncovers Their Colocalization and Coordination with Nanometer Resolution. Switzerland. https://doi.org/10.3390/membranes13020142
Traeger, Jeremiah, Hu, Dehong, Yang, Mengran, Stacey, Gary, and Orr, Galya. Sat .
"Super-Resolution Imaging of Plant Receptor-Like Kinases Uncovers Their Colocalization and Coordination with Nanometer Resolution". Switzerland. https://doi.org/10.3390/membranes13020142.
@article{osti_1914371,
title = {Super-Resolution Imaging of Plant Receptor-Like Kinases Uncovers Their Colocalization and Coordination with Nanometer Resolution},
author = {Traeger, Jeremiah and Hu, Dehong and Yang, Mengran and Stacey, Gary and Orr, Galya},
abstractNote = {Plant cell signaling often relies on the cellular organization of receptor-like kinases (RLKs) within membrane nanodomains to enhance signaling specificity and efficiency. Thus, nanometer-scale quantitative analysis of spatial organizations of RLKs could provide new understanding of mechanisms underlying plant responses to environmental stress. Here, we used stochastic optical reconstruction fluorescence microscopy (STORM) to quantify the colocalization of the flagellin-sensitive-2 (FLS2) receptor and the nanodomain marker, remorin, within Arabidopsis thaliana root hair cells. We found that recovery of FLS2 and remorin in the plasma membrane, following ligand-induced internalization by bacterial-flagellin-peptide (flg22), reached ~85% of their original membrane density after ~90 min. The pairs colocalized at the membrane at greater frequencies, compared with simulated randomly distributed pairs, except for directly after recovery, suggesting initial uncoordinated recovery followed by remorin and FLS2 pairing in the membrane. The purinergic receptor, P2K1, colocalized with remorin at similar frequencies as FLS2, while FLS2 and P2K1 colocalization occurred at significantly lower frequencies, suggesting that these RLKs mostly occupy distinct nanodomains. The chitin elicitor receptor, CERK1, colocalized with FLS2 and remorin at much lower frequencies, suggesting little coordination between CERK1 and FLS2. These findings emphasize STORM’s capacity to observe distinct nanodomains and degrees of coordination between plant cell receptors, and their respective immune pathways.},
doi = {10.3390/membranes13020142},
journal = {Membranes},
number = 2,
volume = 13,
place = {Switzerland},
year = {Sat Jan 21 00:00:00 EST 2023},
month = {Sat Jan 21 00:00:00 EST 2023}
}
Free Publicly Available Full Text
Publisher's Version of Record
https://doi.org/10.3390/membranes13020142
https://doi.org/10.3390/membranes13020142
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Figure 1: Representative super-resolution images of A. thaliana root hairs stimulated with 200 nM flg22 for a set length of time and then fixed, indicating localization of remorin (cyan), FLS2 (yellow), and colocalized remorin-FLS2 pairs within 100 nm (red circles). The scale bars indicate 10 µm. Note the steady dropmore »
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