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Plant Protein Secretion pp 257–266Cite as

Using Microscopy Tools to Visualize Autophagosomal Structures in Plant Cells

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1662))

Abstract

Macroautophagy (hereafter as autophagy), is a metabolic process for sequestration of cytoplasmic cargos into a double membrane structure named as autophagosome. In plants, autophagy is required for nutrition mobilization/recycling and clearance of protein aggregates or damaged organelles during starvation or other unfavorable conditions, as well as for plant immunity during pathogen infection. Multiple experimental approaches have been developed to elucidate the autophagic activity. To facilitate further investigations on the potential involvement of autophagy in protein secretion process in plant cells, here we describe detailed protocols to measure the autophagic activity in model plant Arabidopsis. Using the autophagosome marker ATG8 and a novel autophagic regulator SH3P2 as examples, we illustrate the major cell biology tools and methods using microscopy to analyze the autophagosomal structures in plant cells, including BTH-induced autophagic response, transient expression and colocalization analysis, as well as immuno-EM labeling.

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Acknowledgments

This work was supported by grants from the Research Grants Council of Hong Kong (CUHK465112, 466313, 14130716, 14102417, CUHK2/CRF/11G, C4011-14R, C4012-16E, and AoE/M-05/12), Germany/Hong Kong Joint Research Scheme, CUHK Research Committee Direct Grant, NSFC (31670179, 31270226 and 31470294), CAS-Croucher Joint Lab Scheme, and Shenzhen Peacock Project (KQTD201101).

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Authors and Affiliations

  1. State Key Laboratory of Agrobiotechnology, Centre for Cell & Developmental Biology, School of Life Sciences, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China

    Weili Lin & Xiaohong Zhuang

Authors
  1. Weili Lin
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  2. Xiaohong Zhuang
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Correspondence to Xiaohong Zhuang .

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Editors and Affiliations

  1. School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong, China

    Liwen Jiang

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Lin, W., Zhuang, X. (2017). Using Microscopy Tools to Visualize Autophagosomal Structures in Plant Cells. In: Jiang, L. (eds) Plant Protein Secretion. Methods in Molecular Biology, vol 1662. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7262-3_23

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  • DOI: https://doi.org/10.1007/978-1-4939-7262-3_23

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7261-6

  • Online ISBN: 978-1-4939-7262-3

  • eBook Packages: Springer Protocols

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