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Clearing of Vascular Tissue in Arabidopsis thaliana for Reporter Analysis of Gene Expression

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Xylem

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2722))

Abstract

To study the gene regulatory mechanisms modulating development is essential to visualize gene expression patterns at cellular resolution. However, this kind of analysis has been limited as a consequence of the plant tissues’ opacity. In the last years, ClearSee has been increasingly used to obtain high-quality imaging of plant tissue anatomy combined with the visualization of gene expression patterns. ClearSee is established as a major tissue clearing technique due to its simplicity and versatility.

In this chapter, we outline an easy-to-follow ClearSee protocol to analyze gene expression of reporters using either β-glucuronidase (GUS) or fluorescent protein (FP) tags, compatible with different dyes to stain cell walls. We detail materials, equipment, solutions, and procedures to easily implement ClearSee for the study of vascular development in Arabidopsis thaliana, but the protocol can be easily adapted to a variety of plant tissues in a wide range of plant species.

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Acknowledgments

Work in JB’s group is funded by a grant from the Spanish Ministry of Science PID2021-1274610B-I00. JB is sponsored by a Ramon y Cajal contract RYC2019-026537-I. Research in ASM's group is funded by a grant from the Valencian Government (CISEJI/2022/28, Plan GenT).

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Authors and Affiliations

  1. Instituto de Biología Molecular y Celular de Plantas, (CSIC-Universitat Politècnica de València), Valencia, Spain

    Antonio Serrano-Mislata & Javier Brumós

Authors
  1. Antonio Serrano-Mislata
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  2. Javier Brumós
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Corresponding authors

Correspondence to Antonio Serrano-Mislata or Javier Brumós .

Editor information

Editors and Affiliations

  1. UPV-CSIC, Universitat Politècnica de València, Valencia, Spain

    Javier Agusti

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Serrano-Mislata, A., Brumós, J. (2024). Clearing of Vascular Tissue in Arabidopsis thaliana for Reporter Analysis of Gene Expression. In: Agusti, J. (eds) Xylem. Methods in Molecular Biology, vol 2722. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3477-6_15

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  • DOI: https://doi.org/10.1007/978-1-0716-3477-6_15

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3476-9

  • Online ISBN: 978-1-0716-3477-6

  • eBook Packages: Springer Protocols

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