Bio-Protocol (Aug 2018)

In vivo and in vitro 31P-NMR Study of the Phosphate Transport and Polyphosphate Metabolism in Hebeloma cylindrosporum in Response to Plant Roots Signals

  • Christine Le Guerneve,
  • Adeline Becquer,
  • Margarita Torres-Aquino,
  • Laurie Amenc,
  • Carlos Trives-Segura,
  • Siobhan Staunton,
  • Claude Plassard,
  • Hervé Quiquampoix

DOI
https://doi.org/10.21769/BioProtoc.2973
Journal volume & issue
Vol. 8, no. 16

Abstract

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We used in vivo and in vitro phosphorus-31 nuclear magnetic resonance (31P-NMR) spectroscopy to follow the change in transport, compartmentation and metabolism of phosphate in the ectomycorrhizal fungus Hebeloma cylindrosporum in response to root signals originating from host (Pinus pinaster) or non-host (Zea mays) plants. A device was developed for the in vivo studies allowing the circulation of a continuously oxygenated mineral solution in an NMR tube containing the mycelia. The in vitro studies were performed on fungal material after several consecutive treatment steps (freezing in liquid nitrogen; crushing with perchloric acid; elimination of perchloric acid; freeze-drying; dissolution in an appropriate liquid medium).