ABSTRACT
The role of the δ-ornithine amino transferase (OAT) pathway in proline synthesis is still controversial and was assessed in leaves of cashew plants subjected to salinity. The activities of enzymes and the concentrations of metabolites involved in proline synthesis were examined in parallel with the capacity of exogenous ornithine and glutamate to induce proline accumulation. Proline accumulation was best correlated with OAT activity, which increased 4-fold and was paralleled by NADH oxidation coupled to the activities of OAT and Δ(1)-pyrroline-5-carboxylate reductase (P5CR), demonstrating the potential of proline synthesis via OAT/P5C. Overall, the activities of GS, GOGAT and aminating GDH remained practically unchanged under salinity. The activity of P5CR did not respond to NaCl whereas Δ(1)-pyrroline-5-carboxylate dehydrogenase was sharply repressed by salinity. We suggest that if the export of P5C from the mitochondria to the cytosol is possible, its subsequent conversion to proline by P5CR may be important. In a time-course experiment, proline accumulation was associated with disturbances in amino acid metabolism as indicated by large increases in the concentrations of ammonia, free amino acids, glutamine, arginine and ornithine. Conversely, glutamate concentrations increased moderately and only within the first 24h. Exogenous feeding of ornithine as a precursor was very effective in inducing proline accumulation in intact plants and leaf discs, in which proline concentrations were several times higher than glutamate-fed or salt-treated plants. Our data suggest that proline accumulation might be a consequence of salt-induced increase in N recycling, resulting in increased levels of ornithine and other metabolites involved with proline synthesis and OAT activity. Under these metabolic circumstances the OAT pathway might contribute significantly to proline accumulation in salt-stressed cashew leaves.
Subject(s)
Anacardium/metabolism , Nitrogen/metabolism , Ornithine-Oxo-Acid Transaminase/metabolism , Ornithine/metabolism , Proline/metabolism , Salt Tolerance/physiology , Stress, Physiological/physiology , Enzyme Assays , Glutamate Dehydrogenase/metabolism , Glutamate Synthase (NADH)/metabolism , Glutamate-Ammonia Ligase/metabolism , Glutamates/administration & dosage , Glutamates/metabolism , NAD/metabolism , Ornithine/administration & dosage , Plant Leaves/metabolism , Proline/biosynthesis , Pyrroline Carboxylate Reductases/metabolism , Salinity , delta-1-Pyrroline-5-Carboxylate ReductaseABSTRACT
In the present study, the effect of acid stress on ammonium assimilation in Bradyrhizobium sp. SEMIA 6144 (Arachis hypogaea L.) microsymbiont was analyzed. The bacterial growth rate was decreased by 50%, and a significant increase in intracellular glutamate concentration was detected when the strain grew at acid pH (5.5). Assays of the enzymes involved in glutamate synthesis showed increased activities of glutamine synthetase (GS) and glutamate synthase (NADPH-GOGAT) under acid stress condition. This would support the contention that the GS/NADPH-GOGAT pathway contributes to the increase of glutamate synthesis as a compatible solute in response to acid stress.