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Pollen and Pollen Tube Biology pp 293–306Cite as

Measuring Exocytosis Rate in Arabidopsis Pollen Tubes Using Corrected Fluorescence Recovery After Photoconversion (cFRAPc) Technique

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2160))

Abstract

Exocytosis is a fundamental process essential for many cellular functions by targeting signal peptides, proteins, and cell wall components to the plasma membrane (PM) or extracellular matrix. Conventional methods, such as FRAP, often underestimate the exocytosis rate of a specific molecule, because retrieval of the molecules from the PM by endocytosis can impact the measurement. To overcome this issue, we have previously established a novel method, corrected fluorescence recovery after photoconversion (cFRAPc), which allows us to accurately measure the exocytosis rate by monitoring both exocytosis-dependent and exocytosis-independent events. In this chapter, we provide detailed procedures for the cFRAPc method to measure the exocytosis rate of Arabidopsis receptor-like kinase PRK1 in pollen tubes. This method should be widely applicable to various cell types.

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Acknowledgments

This work is supported by the U.S. National Institute of General Medical Sciences (GM100130).

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Authors and Affiliations

  1. FAFU-UCR Joint Center for Horticultural Biology and Metabolomics, Haixia Institute of Science and Technology, Fujian Agriculture and Forestry University, Fuzhou, Fujian, China

    Jingzhe Guo

  2. Center for Plant Cell Biology, Department of Botany and Plant Sciences, Institute for Integrative Genome Biology, University of California, Riverside, CA, USA

    Jingzhe Guo & Zhenbiao Yang

Authors
  1. Jingzhe Guo
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  2. Zhenbiao Yang
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Corresponding author

Correspondence to Zhenbiao Yang .

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Editors and Affiliations

  1. Department of Plant Science, McGill University, Montreal, QC, Canada

    Anja Geitmann

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Guo, J., Yang, Z. (2020). Measuring Exocytosis Rate in Arabidopsis Pollen Tubes Using Corrected Fluorescence Recovery After Photoconversion (cFRAPc) Technique. In: Geitmann, A. (eds) Pollen and Pollen Tube Biology. Methods in Molecular Biology, vol 2160. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0672-8_21

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  • DOI: https://doi.org/10.1007/978-1-0716-0672-8_21

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0671-1

  • Online ISBN: 978-1-0716-0672-8

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