Abstract
Accumulation of callose (β-1,3 glucans) at the plasmodesmata (PD) neck region dynamically regulates symplastic intercellular transport. Here we describe a 2–3-day immuno-labelling protocol to determine callose levels in the cell wall region at PD. The method relies on exposure of internal cell walls by hand-sectioning of the sample and digestion of the cell wall with enzymes in order to improve antibody penetration to deep tissue layers. By using this protocol, combined with high-resolution confocal imaging, we successfully detected PD-associated callose in Arabidopsis root apical meristem, vascular tissue, and developing lateral root primordia.
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Acknowledgments
We would like to thank Grant Calder and Prof. Peter Shaw for critical reading and editing of the manuscript and Andrew Davis for help with production of the video. Financial support for this research was provided by the UK Biotechnology and Biological Sciences Research Council (BBSRC).
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Pendle, A., Benitez-Alfonso, Y. (2015). Immunofluorescence Detection of Callose Deposition Around Plasmodesmata Sites. In: Heinlein, M. (eds) Plasmodesmata. Methods in Molecular Biology, vol 1217. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1523-1_6
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DOI: https://doi.org/10.1007/978-1-4939-1523-1_6
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Online ISBN: 978-1-4939-1523-1
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