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Characterizing the Secretion Systems of Magnaporthe oryzae

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Magnaporthe oryzae

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2356))

Abstract

Pharmacological approaches have made a tremendous impact on the field of microbial secretion systems. This protocol describes the inhibition of Golgi-dependent secretion in Magnaporthe oryzae though brefeldin A (BFA) treatment. State-of-the-art live-cell imaging allows tracking secreted proteins in their secretion pathways. Here we applied this protocol for defining the secretion systems of two fluorescently labeled effectors, Bas4 (apoplastic) and Pwl2 (cytoplasmic). Secretion of Bas4 is clearly inhibited by brefeldin A (BFA), indicating its Golgi-dependent secretion pathway. By contrast, secretion of Pwl2 is BFA insensitive and follows a nonconventional secretion pathway that is Snare and Exocyst dependent. The protocol is suitable to other plant-microbial systems and in vitro secreted microbial proteins.

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Acknowledgments

This work was supported by the Louisiana State University AgCenter HATCH and the NIFA-USDA grant (#2017-67013-26525).

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Authors and Affiliations

  1. Department of Plant Pathology and Crop Physiology, Louisiana State University Agricultural Center, Baton Rouge, LA, USA

    Ely Oliveira-Garcia

  2. Department of Plant Pathology, Kansas State University, Manhattan, KS, USA

    Barbara Valent

Authors
  1. Ely Oliveira-Garcia
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  2. Barbara Valent
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Corresponding author

Correspondence to Ely Oliveira-Garcia .

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Editors and Affiliations

  1. Institute of Biotechnology and Drug Research gGmbH (IBWF), Mainz, Rheinland-Pfalz, Germany

    Stefan Jacob

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Oliveira-Garcia, E., Valent, B. (2021). Characterizing the Secretion Systems of Magnaporthe oryzae . In: Jacob, S. (eds) Magnaporthe oryzae. Methods in Molecular Biology, vol 2356. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1613-0_5

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  • DOI: https://doi.org/10.1007/978-1-0716-1613-0_5

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1612-3

  • Online ISBN: 978-1-0716-1613-0

  • eBook Packages: Springer Protocols

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