Abstract
The plant cell wall comprises various types of macromolecules whose abundance and spatial distribution change dynamically and are crucial for plant architecture. High-resolution live cell imaging of plant cell wall components is, therefore, a powerful tool for plant cell biology and plant developmental biology. To acquire suitable data, the experimental setup for staining and imaging of non-fixed samples must be straightforward and avoid creating stress-induced artifacts. We present a detailed sample preparation and live image acquisition protocol for fluorescence visualization of cell wall components using commercially available probes and stains.
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References
Houston K, Tucker MR, Chowdhury J, Shirley N, Little A (2016) The plant cell wall: a complex and dynamic structure as revealed by the responses of genes under stress conditions. Front Plant Sci 7:984
Altartouri B, Geitmann A (2015) Understanding plant cell morphogenesis requires real-time monitoring of cell wall polymers. Curr Opin Plant Biol 23:76–82
Baskin TI (2005) Anisotropic expansion of the plant cell wall. Annu Rev Cell Dev Biol 21:203–222
Cosgrove DJ (2016) Plant cell wall extensibility: connecting plant cell growth with cell wall structure, mechanics, and the action of wall-modifying enzymes. J Exp Bot 67(2):463–476
Echevin E, Le Gloanec C, Skowrońska N, Routier-Kierzkowska A-L, Burian A, Kierzkowski D (2019) Growth and biomechanics of shoot organs. J Exp Bot 70(14):3573–3585
Somerville C, Bauer S, Brininstool G, Facette M, Hamann T, Milne J et al (2004) Toward a systems approach to understanding plant cell walls. Science 306(5705):2206–2211
Zhang T, Tang H, Vavylonis D, Cosgrove DJ (2019) Disentangling loosening from softening: insights into primary cell wall structure. Plant J 100(6):1101–1117
Bidhendi AJ, Chebli Y, Geitmann A (2020) Fluorescence visualization of cellulose and pectin in the primary plant cell wall. J Microsc 278(3):164–181
Mravec J, Kračun SK, Rydahl MG, Westereng B, Miart F, Clausen MH et al (2014) Tracking developmentally regulated post-synthetic processing of homogalacturonan and chitin using reciprocal oligosaccharide probes. Development 141(24):4841–4850
Rivero L, Scholl R, Holomuzki N, Crist D, Grotewold E, Brkljacic J (2014) Handling Arabidopsis plants: growth, preservation of seeds, transformation, and genetic crosses. In: Sanchez-Serrano J, Salinas J (eds) Arabidopsis protocols. Methods in molecular biology (methods and protocols), vol 1062. Humana Press, Totowa, pp 3–25
Acknowledgment
This study was supported by a Discovery grant from the Natural Sciences and Engineering Research Council of Canada (NSERC) and by the Canada Research Chairs Program. Sample preparation and image acquisition were performed at the McGill University ECP3-Multi-Scale Imaging Facility, Sainte-Anne-de-Bellevue, Québec, Canada. We thank Dr. Jozef Mravec from the University of Copenhagen for providing us with COS488.
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Chebli, Y., Geitmann, A. (2023). Live Fluorescence Visualization of Cellulose and Pectin in Plant Cell Walls. In: Pellicciari, C., Biggiogera, M., Malatesta, M. (eds) Histochemistry of Single Molecules. Methods in Molecular Biology, vol 2566. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2675-7_22
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DOI: https://doi.org/10.1007/978-1-0716-2675-7_22
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