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Plant Chromatin Dynamics pp 443–454Cite as

Cell-Type Specific Chromatin Analysis in Whole-Mount Plant Tissues by Immunostaining

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1675))

Abstract

Chromatin organization in eukaryotes is highly dynamic, playing fundamental roles in regulating diverse nuclear processes including DNA replication, transcription, and repair. Thus, the analysis of chromatin organization is of great importance for the elucidation of chromatin-mediated biological processes. Immunostaining coupled with imaging is one of the most powerful tools for chromatin analysis at the cellular level. However, in plants, it is sometimes technically challenging to apply this method due to the inaccessibility of certain cell types and/or poor penetration of the reagents into plant tissues and cells. To circumvent these limitations, we developed a highly efficient protocol enabling the analysis of chromatin modifications and nuclear organization at the single-cell level with high resolution in whole-mount plant tissues. The main procedure consists of five steps: (1) tissue fixation; (2) dissection and embedding; (3) tissue processing; (4) antibody incubation; and (5) imaging. This protocol has been simplified for the processing of multiple samples without the need for laborious tissue sectioning. Additionally, it preserves cellular morphology and chromatin organization, allowing comparative analyses of chromatin organization between different cell types or developmental stages. This protocol was successfully used for various tissues of different plant species, including Arabidopsis thaliana, Oryza sativa (rice), and Zea mays (maize). Importantly, this method is very useful to analyze poorly accessible tissues, such as female meiocytes, gametophytes, and embryos.

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Acknowledgments

This work was supported by the University of Zürich, the Swiss National Foundation and, in part, DuPont-Pioneer. We are thankful to Christof Eichenberger, Valeria Gagliardini, Arturo Bolanos, Matthias Philipp, Anja Frey, and Peter Kopf for general lab support, and to Karl Huwiler and Christian Frey for plant care.

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Authors and Affiliations

  1. Department of Plant and Microbial Biology, Zürich-Basel Plant Science Center, University of Zürich, Zollikerstrasse 107, 8008, Zürich, Switzerland

    Wenjing She, Célia Baroux & Ueli Grossniklaus

Authors
  1. Wenjing She
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  2. Célia Baroux
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  3. Ueli Grossniklaus
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Corresponding author

Correspondence to Wenjing She .

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Editors and Affiliations

  1. Department of Molecular Biology, Wageningen University & Research, Wageningen, The Netherlands

    Marian Bemer

  2. Department of Plant and Microbial Biology, Zürich-Basel Plant Science Center, University of Zürich, Zürich, Switzerland

    Célia Baroux

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She, W., Baroux, C., Grossniklaus, U. (2018). Cell-Type Specific Chromatin Analysis in Whole-Mount Plant Tissues by Immunostaining. In: Bemer, M., Baroux, C. (eds) Plant Chromatin Dynamics. Methods in Molecular Biology, vol 1675. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7318-7_25

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  • DOI: https://doi.org/10.1007/978-1-4939-7318-7_25

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7317-0

  • Online ISBN: 978-1-4939-7318-7

  • eBook Packages: Springer Protocols

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