A molecular mechanosensor for real-time visualization of appressorium membrane tension in Magnaporthe oryzae

Ryder, Lauren S., Lopez, Sergio G., Michels, Lucile, Eseola, Alice B., Sprakel, Joris, Ma, Weibin and Talbot, Nicholas J. ORCID: https://orcid.org/0000-0001-6434-7757 (2023) A molecular mechanosensor for real-time visualization of appressorium membrane tension in Magnaporthe oryzae. Nature Microbiology, 8. pp. 1508-1519. ISSN 2058-5276

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Abstract

The rice blast fungus Magnaporthe oryzae uses a pressurized infection cell called an appressorium to drive a rigid penetration peg through the leaf cuticle. The vast internal pressure of an appressorium is very challenging to investigate, leaving our understanding of the cellular mechanics of plant infection incomplete. Here, using fluorescence lifetime imaging of a membrane-targeting molecular mechanoprobe, we quantify changes in membrane tension in M. oryzae. We show that extreme pressure in the appressorium leads to large-scale spatial heterogeneities in membrane mechanics, much greater than those observed in any cell type previously. By contrast, non-pathogenic melanin-deficient mutants, exhibit low spatially homogeneous membrane tension. The sensor kinase ∆sln1 mutant displays significantly higher membrane tension during inflation of the appressorium, providing evidence that Sln1 controls turgor throughout plant infection. This non-invasive, live cell imaging technique therefore provides new insight into the enormous invasive forces deployed by pathogenic fungi to invade their hosts, offering the potential for new disease intervention strategies.

Item Type: Article
Additional Information: Data availability statement: The M. oryzae genome database used in this study was http://fungi.ensembl.org/Magnaporthe_oryzae/Info/Index. All M. oryzae strains used in this study are freely available upon request from the corresponding authors. Detailed protocols for synthesis of both the N+-BDP rotor probe and the chemical polarity probe NR125 are available on request from J.S. Source data are provided with this paper. Code availability: The Python script used to produce the M. oryzae FLIM movie has been publicly deposited in GitHub at https://github.com/SergioGabrielLopez/movie_script. Funding Information: This project was funded by the BBSRC grant BB/V016342/1 and by the Gatsby Charitable Foundation. We thank C. Faulkner for providing us with C. higginsianum strain IMI 349061.
Uncontrolled Keywords: microbiology,immunology,applied microbiology and biotechnology,genetics,microbiology (medical),cell biology ,/dk/atira/pure/subjectarea/asjc/2400/2404
Faculty \ School: Faculty of Science > The Sainsbury Laboratory
UEA Research Groups: Faculty of Medicine and Health Sciences > Research Centres > Norwich Institute for Healthy Aging
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Depositing User: LivePure Connector
Date Deposited: 04 Nov 2024 16:30
Last Modified: 10 Nov 2024 07:30
URI: https://ueaeprints.uea.ac.uk/id/eprint/97464
DOI: 10.1038/s41564-023-01430-x

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