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31P NMR for the study of P metabolism and translocation in arbuscular mycorrhizal fungi
Authors:Nanna Rasmussen  David C Lloyd  R George Ratcliffe  Poul Erik Hansen  Iver Jakobsen
Institution:1. Plant Biology and Biogeochemistry Department, Ris? National Laboratory, PBK-330, P.O. Box 49, DK-4000, Roskilde, Denmark
2. Department of Plant Sciences, University of Oxford, South Parks Road, Oxford, OX1 3RB, UK
3. Department of Life Sciences and Chemistry, Roskilde University, DK-4000, Roskilde, Denmark
4. Plant Biology and Biogeochemistry Department, Ris? National Laboratory, PBK-330, P.O. Box 49, DK-4000, Roskilde, Denmark
Abstract:31P nuclear magnetic resonance (NMR) spectroscopy was used to study phosphate (P) metabolism in mycorrhizal and nonmycorrhizal roots of cucumber (Cucumis sativus L) and in external mycelium of the arbuscular mycorrhizal (AM) fungus Glomus intraradices Schenck & Smith. The in vivo NMR method allows biological systems to be studied non-invasively and non-destructively. 31P NMR experiments provide information about cytoplasmic and vacuolar pH, based on the pH-dependent chemical shifts of the signals arising from the inorganic P (Pi) located in the two compartments. Similarly, the resonances arising from α, β and γ phosphates of nucleoside triphosphates (NTP) and nucleoside diphosphates (NDP) supply knowledge about the metabolic activity and the energetic status of the tissue. In addition, the kinetic behaviour of P uptake and storage can be determined with this method. The 31P NMR spectra of excised AM fungi and mycorrhizal roots contained signals from polyphosphate (PolyP), which were absent in the spectra of nonmycorrhizal roots. This demonstrated that the Pi taken up by the fungus was transformed into PolyP with a short chain length. The spectra of excised AM fungi revealed only a small signal from the cytoplasmic Pi, suggesting a low cytoplasmic volume in this AM fungus. This revised version was published online in June 2006 with corrections to the Cover Date.
Keywords:Arbuscular mycorrhizal fungi            Cucumis sativus                      Glomus intraradices            NMR spectroscopy  phosphorus transport  polyphosphate
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