Nuclear Magnetic Resonance

This second volume in Modern Methods of Plant Analysis, New Series concentrates on nuclear magnetic resonance (NMR) spectroscopy, a powerful technique that is now being used for plant studies in a major way. This is the first time that a series of chapters by leading experts on the application of NMR to plant cells has been assembled in a single volume. The methods are described in detail so that even beginners can apply them directly in the laboratory. Topics include general metabolism in higher plants and fungi, intercellular pH, energy status, DNA structure analysis, multiple quantum two-dimensional NMR spectroscopy, chloroplast orientation, rubber analysis, and the use of NMR to determine pollen viability. This volume should provide not only an excellent practical guide to the possibilities of NMR application to the plant sciences, but also give impetus to its future use.

Contenu
13C-NMR in Metabolic Studies.- 1 Introduction.- 2 The Design and Interpretation of Labelling Experiments.- 3 Radioisotope Labelling.- 4 13C NMR Spectroscopy.- 5 Biosynthetic Methodology.- 6 13C Enrichment Studies.- 7 13C, 15N Doubly Labelled Precursors.- 8 Isotope-Induced Shifts in 13C NMR.- 9 Conclusion.- References.- Determination of the Energy Status of Plant Cells by 31P-Nuclear Magnetic Resonance Spectroscopy.- 1 Introduction.- 2 Principles.- 3 Experimental Considerations.- 4 Quantitation of Metabolite Concentrations of vivo by NMR.- 5 Measurement of Nucleoside Di- and Triphosphate Levels by 31P-NMR.- 6 Estimation of the Extent of Magnesium Ion Binding to Nucleoside Triphosphates in vivo by 31P-NMR.- 7 Measurement of the Gibbs' Free Energy for Hydrolysis of ATP.- 8 Relationship of in vivo 31P-NMR Measurements to What Is Seen by Other Methods.- 9 Conclusions.- References.- The Use of NMR Spectroscopy to Follow Deuterium in Studies of Fungal Metabolism.- 1 Introduction.- 2 Deuterium NMR Spectroscopy.- 3 Detection of Deuterium Indirectly in the 13C-NMR Spectrum.- 4 Conclusions.- References.- Proton NMR Studies on DNA Structure.- 1 Introduction.- 2 Theoretical Background.- 3 Examples.- 4 Summary.- References.- NMR Methods for Determination of Intracellular pH.- 1 Introduction.- 2 Some Principles.- 3 Determinants of Chemical Shift.- 4 Measurement of Chemical Shifts.- 5 Estimating Intracellular pH from Chemical Shifts: Titration Curves.- 6 Other Factors Contributing to Potential Errors in Intracellular pH Measurements Using NMR.- 7 Observation of pH Heterogeneity in Cell Populations and Tissues by NMR.- 8 NMR Signals That Have Been Used as Intracellular pH Indicators.- 9 Assignment of Resonances.- 10 Can the pH in Compartments Other Than Cytoplasm and Vacuole beMeasured in Vivo by NMR Methods?.- 11 Conclusions.- References.- Orientation of Chloroplasts in Leaves by 1H NMR Spectroscopy.- 1 Introduction.- 2 Experimental Technique.- 3 Data Analysis.- 4 Examples of Interpreted Spectra.- 5 Future Directions.- References.- 13C-NMR Determination of Rubber Content in Guayule Bushes.- 1 Introduction.- 2 Experimental.- 3 Results and Discussion.- 4 Conclusions.- Appendix A 13C-NMR Method.- References.- Nuclear Magnetic Resonance and Pollen Quality.- 1 What is the Pollen Grain?.- 2 1H-NMR Water Content.- 3 31P, 13C-NMR and Membrane State.- 4 Conclusions.- References.