Abstract
Plasmodesmata are nanopores in the plant cell wall that allow direct cell-to-cell communication. They are key for plant growth, development, and defense. However, studying these pores is challenging due to their small size, with diameters of 30–50 nm and lengths that match cell wall thickness. One particular challenge is measuring how much cell-to-cell trafficking is facilitated by the plasmodesmata in a tissue or between particular cells. Here, we present an approach for studying plasmodesmata-mediated trafficking in the model plant Arabidopsis thaliana by using an easy-to-build and affordable low-pressure particle bombardment apparatus. Using low-pressure particle bombardment at around 60 psi, we are able to transform individual cells in the leaf epidermis and study by confocal fluorescence microscopy the subsequent cell-to-cell trafficking of the diffusible molecule green fluorescent protein (GFP). The technique and equipment could be used by any plant biologist to measure intercellular trafficking through plasmodesmata under varying growth conditions including exposure to different stresses, light conditions, chemical treatments, or in various mutant backgrounds.
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Acknowledgments
We thank George Butler, an undergraduate researcher in the lab, for excellent technical assistance. The Staff of the Biological Service Facility at the University of Tennessee, Knoxville, are acknowledged for assembling the bombardment apparatus and providing technical details for this article. Research reported in this publication was supported by the National Institute of General Medical Sciences of the National Institutes of Health under Award Number F31GM131671. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Additional support was from the National Science Foundation through grant MCB 1846245.
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Fernandez, J.C., Burch-Smith, T.M. (2022). Investigating Plasmodesmata Function in Arabidopsis Thaliana Using a Low-Pressure Bombardment System and GFP Movement Assay. In: Benitez-Alfonso, Y., Heinlein, M. (eds) Plasmodesmata. Methods in Molecular Biology, vol 2457. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2132-5_18
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DOI: https://doi.org/10.1007/978-1-0716-2132-5_18
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