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The analysis of primary and secondary nitrogen metabolism in plants by nuclear magnetic resonance (NMR) spectroscopy is comprehensively reviewed. NMR is a versatile analytical tool, and the combined use of (1)H, (2)H, (13)C, (14)N and (15)N NMR allows detailed investigation of the acquisition, assimilation and metabolism of nitrogen. The analysis of tissue extracts can be complemented by the in vivo NMR analysis of functioning tissues and cell suspensions, and by the application of solid state NMR techniques. Moreover stable isotope labelling with (2)H-, (13)C- and (15)N-labelled precursors provides direct insight into specific pathways, with the option of both time-course and steady state analysis increasing the potential value of the approach. The scope of the NMR method, and its contribution to studies of plant nitrogen metabolism, are illustrated with a wide range of examples. These include studies of the GS/GOGAT pathway of ammonium assimilation, investigations of the metabolism of glutamate, glycine and other amino acids, and applications to tropane alkaloid metabolism. The continuing development of the NMR technique, together with potential applications in the emerging fields of metabolomics and metabolic flux analysis, leads to the conclusion that NMR will play an increasingly valuable role in the analysis of plant nitrogen metabolism.

Chamaegigas intrepidus Dinter is a poikilohydric aquatic plant that lives in rock pools on granite outcrops in central Namibia. The pools are filled with water only intermittently during the wet season, and the plants may pass through up to 20 rehydration/dehydration cycles during the summer rains. The potential nitrogen sources for the rehydrated plants are ammonium, which is only present at 10-20 µM, amino acids, particularly glycine, and urea, which is generally present at 20-30 µM. We show that urea can be utilised by plants in the field through the presence of urease in the sediments of the rock pools. Urease activity is higher in non-submerged than in submerged sediments, and it can survive 6 months of complete dryness at temperatures up to 60°C. Experiments with [ 14 C]urea under laboratory conditions show that the roots of C. intrepidus are unable to take up urea; while 15 N-nuclear magnetic resonance experiments show that [ 15 N]urea is only metabolised to labelled glutamine and glutamate after ammonium has been released by the action of urease. Thus urease plays a vital role in allowing urea to be utilised as a major N source in this nutrient-limited aquatic ecosystem.

The distribution of inorganic phosphate (Pi) between the cytoplasm and the vacuole of Humulus lupulus L. cells grown in suspension culture at different exogenous Pi levels was examined by 31-P nuclear magnetic resonance. In growing cells excess Pi accumulated in the vacuoles and the inhibitory effect of high exogenous Pi was not associated with a change in the cytoplasmic Pi level or with a change in the cytoplasmic pH.