Short-term ammonium supply stimulates glutamate dehydrogenase activity and alternative pathway respiration in roots of pea plants

Abstract

The changes of C and N metabolism after switching Pisum sativum L. plants from nitrate to ammonium were studied. Pea plants were grown for three weeks in nutrient solution containing 0.5 mmol/L nitrate, and then randomly divided into five sets for five different nitrogen treatments: control (0.5 mmol/L nitrate) and four ammonium concentrations (0.5, 1, 2.5 and 5 mmol/L). After 72 hours, activities of enzymes related to C and N metabolism were measured. Ammonium content in roots increased with ammonium concentration showing saturation from a concentration of 2.5 mmol/L. Increasing external ammonium concentration also increased free amino acid content (mainly glutamine and asparagine), whereas starch content decreased and neither organic acid or soluble carbohydrates changed. Glutamine synthetase (GS; EC 6.3.1.2) activity decreased and root glutamate dehydrogenase (GDH; EC 1.4.1.2) activity increased with ammonium regardless of the concentration used. Root respiration rate increased with ammonium, due mainly to an increase of the alternative pathway. These results could be consistent with the assumption of a possible role for GDH in ammonium detoxification. Our results show a close relationship between GDH activity and respiration rate through alternate pathways in order to ensure the supply of C skeletons for ammonium assimilation, whereas surplus NADH is oxidised directly via the non-phosphorylating route.