The removal of nitrate reductase phosphorylation enhances tolerance to ammonium nitrogen deficiency in rice

Nitrate reductase(NR) is a key enzyme for nitrogen assimilation in plants,and its activity is regulated by posttranslational phosphorylation.To investigate the effects of dephosphorylation of the NIA1 protein on the growth and the physiological and biochemical characteristics of rice under different forms of nitrogen supplies,the phenotypes,nitrogen metabolism and reactive oxygen metabolism were measured in NIA1 phosphorylation site-directed mutant lines(S532 D and S532 A),an Os Nia1 over-expression line(OE) and Kitaake(wild type,WT).Compared with WT and OE,S532 D and S532 A have stronger nitrogen assimilation capacities.When ammonium nitrate served as the nitrogen source,the plant heights,dry weights of shoots and chlorophyll(Chl) contents of S532 D and S532 A were lower than those of the WT and OE,whereas hydrogen peroxide(H_(2)O_(2)),malondialdehyde(MDA) and nitrite contents were higher.When potassium nitrate served as the nitrogen source,the plant heights,dry weights of shoots and Chl contents of S532 D and S532 A were higher than those of the WT and OE,there were no significant differences in the contents of H_(2)O_(2) and MDA in the leaves of the test materials,and the difference in nitrite contents among different lines decreased.When ammonium sulfate served as the nitrogen source,there were no significant differences in the physiological indexes of the test materials,except NR activity.Compared with ammonium nitrate and ammonium sulfate,the content of NH_(4)^(+)-N in the leaves of each plant was lower when potassium nitrate was used as the nitrogen source.The q PCR results showed that Os GS and Os NGS1 were negatively regulated by downstream metabolites,and Os Nrt2.2 was induced by nitrate.In summary,when ammonium nitrate served as the nitrogen source,the weak growth of NIA1 phosphorylation site-directed mutant lines was due to the toxicity caused by the excessive accumulation of nitrite.When potassium nitrate served as the nitrogen source,the assimilation rates of nitrate,nitrite and ammonium salt were accelerated in NIA1 phosphorylation site-directed mutant lines,which could provide more nitrogen nutrition and improve the tolerance of rice to ammonium nitrogen deficiency.These results could provide a possible method to improve the efficiency of nitrogen utilization in rice under low-nitrogen conditions.

Nodule study in Albizia chinensis in relation to nitrogen metabolism,morphology and biomass

Morphology,biomass,nitrate reductase(NR)and nitrogenase activity in Albizia chinensis(Osb.)Merr.nodules were assessed on monthly and seasonal basis for 1 year.Average NR and nitrogenase activity was higher during the rainy season,reaching a maximum in August.Thereafter,activity decreased through autumn and reached a minimum value during winter.Fresh and dry biomass of nodules increased gradually from summer to the rainy season and then started decreasing with the onset of winter as nodules began to senesce.Among four developmental stages of the nodules that correspond to their increasing age,NR and nitrogenase activity remained low in stage 1 nodules and peaked in stage 2.The activity of both enzymes further decreased with increasing age(stage 2 to stage 4).Morphological features such as shape,diameter and color varied considerably among the developmental stages.Stage 1 nodules were cream-colored,oval to heartshaped with smallest average diameter whereas at stage 2,they became bilobed to tetralobed.On the other hand,stage 3 nodules had the largest average diameter and were multilobed in structure.Stage 4 nodules that correspond to the senescing stage were dark brown to black,multilobed,flattened and hollow due to degeneration of nodular tissue.