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Flow Cytometry and Sorting in Arabidopsis

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Arabidopsis Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2200))

Abstract

Flow cytometry and sorting represents a valuable and mature experimental platform for the analysis of cellular populations. Applications involving higher plants started to emerge around 40 years ago and are now widely employed both to provide unique information regarding basic and applied questions in the biosciences and to advance agricultural productivity in practical ways. Further development of this platform is being actively pursued, and this promises additional progress in our understanding of the interactions of cells within complex tissues and organs. Higher plants offer unique challenges in terms of flow cytometric analysis, first since their organs and tissues are, almost without exception, three-dimensional assemblies of different cell types held together by tough cell walls, and, second, because individual plant cells are generally larger than those of mammals.

This chapter, which updates work last reviewed in 2014 [Galbraith DW (2014) Flow cytometry and sorting in Arabidopsis. In: Sanchez Serrano JJ, Salinas J (eds) Arabidopsis Protocols, 3rd ed. Methods in molecular biology, vol 1062. Humana Press, Totowa, pp 509–537], describes the application of techniques of flow cytometry and sorting to the model plant species Arabidopsis thaliana, in particular emphasizing (a) fluorescence labeling in vivo of specific cell types and of subcellular components, (b) analysis using both conventional cytometers and spectral analyzers, (c) fluorescence-activated sorting of protoplasts and nuclei, and (d) transcriptome analyses using sorted protoplasts and nuclei, focusing on population analyses at the level of single protoplasts and nuclei. Since this is an update, details of new experimental methods are emphasized.

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Acknowledgements

Part of the development of the methods described in this chapter involved support to DG from the NSF Plant Genome Research program, and the National Institutes of Health. DG also acknowledges on-going support from the USDA through the University of Arizona College of Agriculture and Life Sciences. We also acknowledge support from the Chinese National Key Research and Development Program (2OI6YFDOl0l006), the National Natural Science Foundation of China (31770300), the Program for Innovative Research Teams in Science and Technology at a University of Henan Province (18IRTSTHNO23), and the 111 Project (D16014) of China. We finally thank Lisa Villalobos-Menuey (Beckman-Coulter), Laurie Appling (SONY), and Matt Alexander and Alex Rodriguez (BioRad), for valuable assistance in generating the data of Figs. 2, 3, and 4.

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Authors and Affiliations

  1. University of Arizona, School of Plant Sciences and Bio5 Institute, Tucson, AZ, USA

    David W. Galbraith

  2. Henan University, Institute of Plant Stress Biology, School of Life Sciences, Kaifeng, China

    David W. Galbraith & Guiling Sun

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  1. David W. Galbraith
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  2. Guiling Sun
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Correspondence to David W. Galbraith .

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  1. Centro Nacional de Biotecnología, CSIC Centro Nacional de Biotecnología, Madrid, Spain

    Jose J. Sanchez-Serrano

  2. Centro de Investigaciones Biologicas, CSIC Madrid Centro de Investigaciones Biologicas, Madrid, Spain

    Julio Salinas

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Galbraith, D.W., Sun, G. (2021). Flow Cytometry and Sorting in Arabidopsis. In: Sanchez-Serrano, J.J., Salinas, J. (eds) Arabidopsis Protocols . Methods in Molecular Biology, vol 2200. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0880-7_12

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