Molecular Basis and Regulation of Ammonium Transporter in Rice

Rice grows in flooded paddy fields and takes up ammonium as the preferred nitrogen （N） source. Ammonium uptake is facilitated by a family of integral membrane proteins known as ammonium transporters found in all domains of life. However, the molecular mechanism and functional characteristics of the ammonium transporters （AMT） in rice have not been determined in detail yet. In this review, we report a genome-wide search for AMT genes in rice, resulting in the increase of the number of potential AMT proteins to at least 12, including members of both the alpha and beta sub-groups. Analysis of the predicted protein sequences for the 12 OsAMT proteins identified many conserved phosphorylation sites in both the alpha and beta group members, which could potentially play a role in controlling the activity of the transporters. Present knowledge of the expression of rice AMT genes is also summarized in detail. Future studies should focus on the structural and functional characteristics of OsAMT proteins to provide insight into the mechanism of ammonium uptake and its regulation in rice. Such research could improve utilization and decrease wastage of N fertilizer in rice cultivation.

Ammonium-dependent regulation of ammonium transporter ZmAMT1s expression conferred by glutamine levels in roots of maize

In maize,two root epidermis-expressed ammonium transporters ZmAMT1;1a and ZmAMT1;3 play major roles in highaffinity ammonium uptake.However,the transcriptional regulation of ZmAMT1s in roots for ensuring optimal ammonium acquisition remains largely unknown.Here,using a split root system we showed that ZmAMT1;1a and ZmAMT1;3transcript levels were induced by localized ammonium supply to nitrogen-deficient roots.This enhanced expression of Zm AMT1s correlated with increases in ^(15)NH_(4)^(+)influx rates and tissue glutamine concentrations in roots.When ammonium was supplied together with methionine sulfoximine,an inhibitor of glutamine synthase,ammonium-induced expression of ZmAMT1s disappeared,suggesting that glutamine rather than ammonium itself regulated ZmAMT1s expression.When glutamine was supplied to nitrogen-deficient roots,expression levels of ZmAMT1s were enhanced,and negative feedback regulation could subsequently occur by supply of glutamine at a high level.Thus,our results indicated an ammonium-dependent regulation of ZmAMT1s at transcript levels,and a dual role of glutamine was suggested in the regulation of ammonium uptake in maize roots.

The OsAMT1.1 gene functions in ammonium uptake and ammoniumepotassium homeostasis over low and high ammonium concentration ranges

Rice （Oryza sativa） grown in paddy fields is an ammonium （NH4^＋）-preferring crop; however, its AMT-type NH4^＋ transporters that mediate root N acquisition have not been well characterized yet. In this study, we analyzed the expression pattern and physiological function of the OsAMT1.1 gene of the AMT1 subfamily in rice. OsAMT1.1 is located in the plasma membrane and is mainly expressed in the root epidermis, stele and mesophyll cells. Disruption of the OsAMTI.1 gene decreased the uptake of NH4^＋, and the growth of roots and shoots under both low NH4^＋ and high NH4^＋ conditions. OsAMT1.1 contributed to the short-term （5 min） ^15NH4^＋ influx rate by approximately one-quarter, irrespective of the NH4^＋ concentration. Knockout of OsAMTI.I significantly decreased the total N transport from roots to shoots under low NH4^＋ conditions. Moreover, compared with the wild type, the osamt1.1 mutant showed an increase in the potassium （K） absorption rate under high NH4^＋ conditions and a decrease under low NH4^＋ conditions. The mutants contained a significantly high concentration of K in both the roots and shoots at a limited K （0.1 mmol/L） supply when NH4^＋ was replete. Taken together, the results indicated that OsAMT1.1 significantly contributes to the NH4^＋ uptake under both low and high NH4^＋ conditions and plays an important role in N-K homeostasis in rice.

A Critical Role of AMT2;1 in Root-To-Shoot Translocation of Ammonium in Arabidopsis

Ammonium uptake in plant roots is mediated by AMT/MEP/Rh-type ammonium transporters. Out of five AMTs being expressed in Arabidopsis roots, four AMT1-type transporters contribute to ammonium uptake, whereas no physiological function has so far been assigned to the only homolog belonging to the MEP subfamily, AMT2;1. Based on the observation that under ammonium supply, the transcript levels of AMT2;1 increased and its promoter activity shifted preferentially to the pericycle, we assessed the contribution of AMT2;1 to xylem loading. When exposed to ^15N-labeled ammonium, amt2;1 mutant lines translocated less tracer to the shoots and contained less ammonium in the xylem sap. Moreover, in an amtl;1 amtl;2 amtl ;3 amt2;1 quadruple mutant （qko）, co-expression of AMT2;1 with either AMT1;2 or AMT1;3 significantly enhanced ^15N translocation to shoots, indicating a cooperative action between AMT2;1 and AMT1 transporters. Under N deficiency, proAMT2;1-GFP lines showed enhanced promoter activity predominantly in cortical root cells, which coincided with elevated ammonium influx conferred by AMT2;1 at millimolar sub- strate concentrations. Our results indicate that in addition to contributing moderately to root uptake in the low-affinity range, AMT2;1 functions mainly in root-to-shoot translocation of ammonium, depending on its Cell-type-specific expression in response to the plant nutritional status and to local ammonium gradients.

Migration of ammonium nitrogen in ion-absorbed rare earth soils during and post in situ mining: a column study and numerical simulation analysis

Ion-absorbed rare earth mines,leached in situ,retain a large amount of ammonium nitrogen(NH4–N)that continuously releases into the surrounding environments.However,quantitative descriptions and predictions of the transport of NH4–N across mining area with hill slopes are not fully established.Here,laboratory column experiments were designed with an inclined slope(a sand box)to examine the spatial temporal transport of NH4–N in soils collected from the ionic rare earth elements(REE)mining area.An HYDRUS-2D model simulation of the experimental data over time showed that soils had a strong adsorption capacity toward NH4–N.Chemical non-equilibrium model(CNEM)could well simulate the transport of NH4–N through the soil-packed columns.The simulation of the transport-adsorption processes at three flow rates of leaching agents revealed that low flow rate enabled a longer residence time and an increased NH4-N adsorption,but reduced the extraction efficiency for REE.During the subsequent rainwater washing process,the presence of slope resulted in the leaching of NH4–N on the surface of the slope,while the leaching of NH4–N deep inside the column was inhibited.Furthermore,the high-intensity rainfall significantly increased the leaching,highlighting the importance of considering the impact of extreme weather conditions during the leaching process.Overall,our study advances the understanding of the transport of NH4–N in mining area with hills,the impact of flow rates of leaching agents and precipitation intensities,and presents as a feasible modeling method to evaluate the environmental risks of NH4–N pollution during and post REE in situ mining activities.

Unimolecular artificial transmembrane channel with terminal dihydrogen phosphate groups showing transport selectivity for ammonium

A new artificial transmembrane channel molecule bearing dihydrogen phosphate groups has been synthesized.The terminal dihydrogen phosphate groups enable the channel to be highly negatively charged at both ends of the channel structures.The artificial channel could incorporate into the lipid bilayer efficiently under low concentration.The channel displays high NH4+/K+selectivity due to the electrostatic interaction and hydrogen bonding between NH4+and the terminal dihydrogen phosphate groups.