Guard Cell Microfilament Analyzer Facilitates the Analysis of the Organization and Dynamics of Actin Filaments in Arabidopsis Guard Cells
Abstract
:1. Introduction
2. Results
2.1. Guard Cell Microfilament Analyzer (GCMA) Allows Semiautomatic Measurements of Angles of Individual Actin Filaments Formed with the Stomatal Pore Edge in the Guard Cell
2.2. The Amount of Longitudinally Oriented Actin Filaments in Guard Cells Gradually Increases with the Closing of Stomata during Diurnal Cycle
2.3. Actin Nucleation Occurs at both Dorsal and Ventral Sides of Guard Cells, and the Majority of Actin Filaments Elongate Radially and Longitudinally in Guard Cells of Open and Closed Stomata, Respectively
2.4. Longitudinally Oriented Actin Filaments Elongate Substantially Faster than Actin Filaments in Other Orientations in Guard Cells of Closed Stomata
2.5. Longitudinally Oriented Actin Filaments Tend to be Destroyed by Severing and Depolymerization in Guard Cells of Open Stomata whereas They Tend to be Retained in Guard Cells of Closed Stomata
2.6. Actin Filaments in Different Orientations Have Different Actin Bundling Frequency in Guard Cells of Open and Closed Stomata
3. Discussion
3.1. GCMA is a Useful Tool to Evaluate the Orientations of Individual Guard Cell Actin Filaments
3.2. The Majority of De Novo Nucleated Actin Filaments Grow Radially and Longitudinally in Open and Closed Stomata, Respectively
3.3. Radially Oriented Actin Cables in Open Stomata whereas Longitudinally Oriented Actin Cables in Closed Stomata Tend to be Preserved
4. Materials and Methods
4.1. Plant Materials and Growth Conditions
4.2. Stomatal Bioassay and Visualization of Actin Filaments by Confocal Laser Scanning Microscopy (CLSM)
4.3. Measurement of Angles of Actin Filaments in Guard Cells
4.4. Visualization of the Dynamics of Actin Filaments in Guard Cells by Total Internal Reflection Fluorescence Microscopy (TIRFM)
4.5. Quantitative Analyses of the Dynamics of Actin Filaments in Guard Cells
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
GCMA | Guard Cell Microfilaments Analyzer |
MATLAB | Matrix Laboratory |
R | Radial |
O | Oblique |
L | Longitudinal |
D-V R | Dorsal-to-Ventral Radial |
V-D R | Ventral-to-Dorsal Radial |
CLSM | Confocal Laser Scanning Microscopy |
TIRFM | Total Internal Reflection Fluorescence Microscopy |
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Li, X.; Diao, M.; Zhang, Y.; Chen, G.; Huang, S.; Chen, N. Guard Cell Microfilament Analyzer Facilitates the Analysis of the Organization and Dynamics of Actin Filaments in Arabidopsis Guard Cells. Int. J. Mol. Sci. 2019, 20, 2753. https://doi.org/10.3390/ijms20112753
Li X, Diao M, Zhang Y, Chen G, Huang S, Chen N. Guard Cell Microfilament Analyzer Facilitates the Analysis of the Organization and Dynamics of Actin Filaments in Arabidopsis Guard Cells. International Journal of Molecular Sciences. 2019; 20(11):2753. https://doi.org/10.3390/ijms20112753
Chicago/Turabian StyleLi, Xin, Min Diao, Yanan Zhang, Guanlin Chen, Shanjin Huang, and Naizhi Chen. 2019. "Guard Cell Microfilament Analyzer Facilitates the Analysis of the Organization and Dynamics of Actin Filaments in Arabidopsis Guard Cells" International Journal of Molecular Sciences 20, no. 11: 2753. https://doi.org/10.3390/ijms20112753