Glutamate metabolism and amino acid translocation were investigated in the control and cadmium stressed shoots of tomato (Solanum lycopersicum - 63/5 F1), using (15N) ammonium and (15N) glutamate tracers. Regardless of organ type, [15N] ammonium assimilation occurred via glutamine synthetase (EC 6.3.1.2), in the control and stressed plants, and it did not depend on glutamate dehydrogenase (EC 1.4.1.2). The [15N] ammonium and ammonium accumulation patterns support the role of glutamate dehydrogenase in the deamination of [15N] glutamate to provide 2-oxoglutarate and [15N] ammonium. In the presence of cadmium, excess [15N] ammonium was incorporated into asparagine, which served as an additional detoxification molecule. In the presence both of methionine sulfoximine and cadmium, glutamate, alanine, and γ-amino butyrate of leaf tissue continued to become labelled with 15N. The labelling kinetics of amino acids in leaves of tomato plants in the presence of cadmium show that continued assimilation of [15N] ammonium can occur when the glutamine synthetase-glutamate synthase cycle is inhibited. The data provided evidence that the glutamine synthetase pathway and glutamate dehydrogenase play distinct roles in the source-sink nitrogen cycle of tomato leaves under cadmium stress conditions.
ammonium; glutamate dehydrogenase; methionine sulfoximine; 15N-NH4+
