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Measuring Callose Deposition, an Indicator of Cell Wall Reinforcement, During Bacterial Infection in Arabidopsis

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Plant Pattern Recognition Receptors

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1578))

Abstract

The plant cell wall responds dynamically during interaction with various pathogens. Upon recognition of “nonself” components, plant cells deploy a variety of immune responses including cell wall fortification. Callose, a β-(1, 3)-d-glucan polymer, is a component of the material deposited at the site of infection between the plasma membrane and the preexisting cell wall that is hypothesized to serve as a physical barrier and platform for directed antimicrobial compound deposition. The defense-associated function of callose deposition is supported by its induction during pathogen-associated molecular patterns (PAMP)-triggered immunity (PTI) and its inhibition by defense suppressing virulence effectors. Thus, callose deposition is a commonly monitored read-out in plant defense. This protocol describes the use of aniline blue staining and fluorescent microscopy to measure callose deposition in bacteria-infected or elicitor-challenged Arabidopsis leaf tissues.

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Acknowledgments

This protocol is adapted from Kim and Mackey, 2008, MiMB [22]. Funding for this work was provided by the US Department of Agriculture (National Institute of Food and Agriculture, grant #2015-11870612), the Korean Rural Development Administration Next-Generation BioGreen Program (System and Synthetic Agro-Biotech Center and grant nos. PJ009088 and PJ011091), and the Ohio Agricultural Research and Development Center of the Ohio State University.

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Authors and Affiliations

  1. Department of Horticulture and Crop Science, The Ohio State University, Columbus, OH, 43210, USA

    Lin Jin & David M. Mackey

  2. Department of Molecular Genetics, The Ohio State University, Columbus, OH, 43210, USA

    David M. Mackey

Authors
  1. Lin Jin
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  2. David M. Mackey
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Corresponding author

Correspondence to David M. Mackey .

Editor information

Editors and Affiliations

  1. Department of Plant Pathology and Microbiology Institute for Plant Genomics and Biotechnology, Texas A&M University, College Station, Texas, USA

    Libo Shan

  2. Department of Biochemistry and Biophysics Institute for Plant Genomics and Biotechnology, Texas A&M University, College Station, Texas, USA

    Ping He

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Jin, L., Mackey, D.M. (2017). Measuring Callose Deposition, an Indicator of Cell Wall Reinforcement, During Bacterial Infection in Arabidopsis. In: Shan, L., He, P. (eds) Plant Pattern Recognition Receptors. Methods in Molecular Biology, vol 1578. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6859-6_16

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  • DOI: https://doi.org/10.1007/978-1-4939-6859-6_16

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6858-9

  • Online ISBN: 978-1-4939-6859-6

  • eBook Packages: Springer Protocols

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