Induction of high affinity nitrate and ammonium uptake systems in wheat

Abstract

The ability of 7 day old wheat seedlings to take up nitrate or ammonium from hydroponic solution was measured. Seedlings were grown under fully aerated hydroponic conditions. The growth solution consisted of either 0.5 mM CaSO₄ alone or in combination with high nitrate (5 mM NO₃ ^‐ ), high ammonium (2 mM NH₄ ⁺) or modified 1/10 Hoaglands solution with nitrate N only (14 mM) or ammonium N only (2 mM). After washing the roots for one hour in CaSO₄, nitrate or ammonium uptake was measured with an ion selective electrode. Plants grown in high nitrate were unable to take up nitrate from a 0.1 mM external solution. Those grown in CaSO₄ were able to take up nitrate at the same external concentration (flux = 10.2 +/‐ 3.0 μmol nitrate/g dry wtlbh). The same result was seen for plants grown in high ammonium vs those grown in CaSO₄ (flux = 21.0 +/‐ 10.0 μmol/g dry wtlbh). Similar results were obtained when modified Hoagland's solution was substituted for the high N solutions. These data indicate that wheat roots possess both high and low affinity nitrate and ammonium uptake systems. The data further indicate that, for a given ion, the high and low affinity systems do not operate simultaneously under high N conditions. The high affinity system is switched off in the range of 1 mM for both ionic forms of N. Developmental studies show that the expression of the high affinity trait is reversible and may be induced (repressed) by conditioning for 24 h in low (high) N media. Plants grown in high N solutions showed efflux of the ion under assay conditions. Neither ion interfered with the induction/repression of the high affinity trait for the other under the conditions used in this study.     

Ammonium uptake capacity and response of cytosolic glutamine synthetase 1;2 to ammonium supply are key factors for the adaptation of ammonium nutrition in Arabidopsis thaliana

Plant requires nitrogen for the growth, and it use nitrate and ammonium from the environment. Plant suffers from the toxicity when excess ammonium is supplied as a sole nitrogen, although it could be a good nitrogen source for plant growth. We hypothesized that the different responses of ecotypes to ammonium nutrient could partly account for the adaptation of Arabidopsis to an ammonium environment. The purpose of this study is to understand the different responses of ecotypes in ammonium environment. The growth of Arabidopsis thaliana ecotypes, Columbia was compared to those of Arabidopsis thaliana ecotypes, Landsberg erecta in ammonium nutrient. The ratio of shoot dry weight to root dry weight was compared to evaluate the adaptation of two ecotypes. The shoot:root ratio of Landsberg was significantly higher than that of Columbia. T-DNA insertion in cytosolic glutamine synthetase 1;2, one of the essential ammonium assimilatory enzymes, led a decrease of shoot:root ratio. We also measured the isotope-labeled ammonium uptake and the expression levels of ammonium transporter genes, and also the expression of ammonium assimilatory genes, glutamine synthetase genes and glutamate synthase genes, in roots after ammonium re-supply using real-time polymerase chain reaction analysis. We found that (1) ammonium uptake of Landsberg erecta was higher than that of Columbia, when ammonium was supplied at higher concentration, and (2) cytosolic glutamine synthetase 1;2 was highly increased by ammonium supply in the root of Landsberg erecta. The present study suggested the importance of these two factors for adaptation of Arabidopsis to an ammonium-rich environment.     

Natural variation of Arabidopsis thaliana root architecture in response to nitrate availability

Root system architecture (RSA) plays a crucial role in nutrient and water uptake in plants. RSA can be modulated to improve nitrogen use efficiency (NUE) of crops, decreasing the need for fertilization. In this study, we analyzed 12 root parameters in 48 Arabidopsis accessions grown in media with low or high nitrogen (N). Low N repressed six root traits and enhanced six others. A principal component analysis showed that six components contributed to 90.08% of N availability. The primary component contributed to 35.64% of the variation and was associated with LR characteristics. The 48 Arabidopsis accessions were divided into five response groups. Stw-0 had the highest biomass production and a high N concentration in both roots and shoots, independent of growth conditions. H55 had numerous LRs at low N. Our results provide N-responsive parameters suitable for mapping of quantitative trait loci related with root morphology.     

丛枝菌根根外菌丝对铵态氮和硝态氮吸收能力的比较

采用空气隔板分室法并结合15N标记技术,以玉米为宿主植物并接种Glomus mosseae和Glomus intraradices,比较了这两种真菌根外菌丝对铵态氮和硝态氮吸收传递能力的差异。结果表明,丛枝菌根根外菌丝吸收传递氮的能力因菌种和氮素形态而异。两种真菌根外菌丝吸收传递NH4+-N能力均高于NO3--N;G. intraradices根外菌丝吸收传递氮的能力高于G. mosseae,这可能与两种真菌根外菌丝生长量有关。     

海绵铁缓解污水厌氧氨氧化反应器中硝酸盐积累的效果

该文旨在通过向厌氧氨氧化反应器中投加海绵铁来减轻厌氧SBR(sequencingbatchreactoractivatedsludge process)反应器中的硝酸盐积累,试验研究了海绵铁与硝酸盐和亚硝酸盐在静态条件下的反应。在静态条件下,部分硝酸盐和亚硝酸盐被海绵铁还原成了氨。对比动态试验表明投加海绵铁可以将SBR出水硝酸盐质量浓度控制在25～30 mg/L左右。相同条件下不投加海绵铁出水硝酸盐质量浓度不断累积,直至超过55 mg/L。这可能是由于铁将硝酸盐还原为亚硝酸盐并与厌氧氨氧化进行了耦合。采用高通量测序发现投加海绵铁的反应器中厌氧氨氧化菌在微生物群落中所占的比例(22.55%)约为不投加反应器(8.85%)的3倍,表明投加海绵铁有利于反应器中厌氧氨氧化(ANAMMOX)菌的生长和厌氧氨氧化反应器的启动。     

中国某些水稻土中硝态氮向铵态氮的还原研究

Three paddy soils were examined for their capacities of dissimilatory reduction of nitrate to ammonium (DRNA). 15N-labelled KNO3 was added at the rate of 100 mg N/kg. Either glucose or rice straw powder was incorporated at the rate of 1.0 or 2.0 mg C/kg respectively. Three treatments were designed to keep the soil saturated with water: (1) a 2-cm water layer on soil surface (with beaker mouth open); (2) a 2-cm water layer and a 1-cm liquid paraffin layer (with beaker mouth open); and (3) water saturated under an O2-free Ar atmosphere. The soils were incubated at 28℃ for 5 days. There was almost no 15N-labelled NH4+-N detected in Treatment 1. However, there was 1.4 to 3.4 mg N/kg 15N-labelled NH4+-N in Treatment 2, and 2.1 to 13.8 mg N/kg in Treatment 3. Glucose was more effective than straw powder in ammonium production. Because there was sufficient amount of non-labelled NH4+-N in the original soils, 15N-labelled NH4+-N produced as such should be the result of dissimilatory reduction. Studies on microbial population showed that there were plenty of bacteria responsible for DRNA process (DRNA bacteria) in the soils examined, indicating that number of DRNA bacteria was not a limiting factor for ammonium production. However, DRNA bacteria were inferior in number to denitrifiers. The DRNA process in soil suspension started after 5 days of incubation. Glycerol and sodium succinate, though both are readily available carbon sources to organisms, did not facilitate DRNA process. DRNA occurred only when glucose was available and at a C:NO3--N ratio >12. Both availability and quality of the carbon sources affected DRNA.     

Tomato growth responses to foliar application of ammonium sulfate in hydroponic culture

This study was done to evaluate the growth responses of tomato plants (Lycopersicon esculentum Mill.) to foliar application of ammonium sulfate in different concentrations of 0, 50, 100, or 200 mM once per week, as well as 50 mM in every other day under greenhouse and hydroponic culture system. The results showed that foliar application of ammonium restricted the plant height, yield, and vitamin C content of fruits, but it increased the chlorophyll content in treated plants. The highest and the lowest yield were obtained from zero (control) and 200 mM treated plants, respectively. Fruit hardness was not affected by treatments, while blossom end rot disorder was increased by higher concentrations of ammonium sulfate. In general, the result indicated that tomato plant is rather sensitive to foliar application of ammonium sulfate particularly to higher concentrations and application numbers.     

Constraining the conditions conducive to dissimilatory nitrate reduction to ammonium in temperate arable soils

Here we offer the first assessment of conditions conducive to dissimilatory nitrate reduction to ammonium (DNRA) in temperate arable soils, through an examination of the potential for this process to occur in a range of soils of contrasting characteristics. NH₄¹⁵NO₃ (6.2 g N m⁻², 25 atom % excess ¹⁵N) was applied, and recovery of ¹⁵N in the pool taken as indicative of occurrence of DNRA. Up to 5% of applied ¹⁵N was recovered in the pool 2 d after addition of N, glucose (44.6 g C m⁻²) and l-cysteine (7.7 g m⁻², 0.9 g N m⁻², 2.3 g C m⁻²). concentrations were positively correlated with soil pH, ratio, bulk density, sand content and concentration, but negatively correlated with soil C and organic N content. Our results demonstrate the potential for DNRA to contribute to N cycling in temperate arable soils, but its detection and significance is likely to depend on the provision of a low molecular weight C source.     

太湖湖积物及黄海潮间海积物中氮的反硝化及硝态氮还原成铵的研究

Denitrification and nitrate reduction to ammonium in Taihu Lake and Yellow Sea inter-tidal marine sediments were studied.The sediment samples were made slurry containing 150g dry matter per liter.Various of glucose-C to nitrate-N.Acetylene inhibition technique was applied to measure denitrification in the slurres,All samples were incubated anaerobically under argon atmosphere,Data showed that Taihu Lake sediment produced more N2O than marine sediment,Denitrification potential was higher in Taihu Lake sediment than in marine one,Glucose added increase denitrification activity but not the denitrification potential of the sediments.Dissimilatory nitrate reduction to ammonium seemed to occur in marine sediment,but not in freshwater one.When the marine sediment was treated with 25mmol L^-1 glucose,its denitrification potentail,as indicated by maximum N2O production by acetylene blockage,was lower than that treated with no or 2.5mmol L^-1 glucose.Acetylene was suspected to have inhibitory effect on dissimilatory nitrate reduction to ammonium.     

不同氮素形态比例条件下接种AMF对玉米氮同化关键酶的影响

以珍珠岩为基质,通过供应3种不同的NH4+∶NO3ˉ比例营养液,研究了接种丛枝菌根真菌对玉米氮同化关键酶活性的影响。结果看出,与不接种的玉米植株相比,接种Glomus intraradices和Glomus mosseae分别在NH4+∶NO3ˉ=3∶1和NH4+∶NO3ˉ=1∶3形态下提高了植物叶片的硝酸还原酶活性;接种AMF对叶片谷氨酰胺合成酶活性（GS）影响不大,但在NH4+∶NO3ˉ=3∶1形态下接种3种AMF处理均显著提高了根系GS活性,相对提高了铵态氮在地下部的同化比例。在铵态氮比例较高时,接种AMF的促生效应较好,且AMF提高根系GS活性作用较大。表明丛枝菌根真菌在促进宿主植物对铵态氮的利用作用较大。     

Evaluation of denitrification losses by the acetylene inhibition technique in a permanent ryegrass field (Lolium perenne L.) fertilized with animal slurry or ammonium nitrate

In a field experiment, the effect of animal slurry, (with and without the nitrification inhibitor dicyandiamide on total denitrification losses estimated by the C₂H₂ inhibition technique was measured over 2 years (1989–1990). During this period, four different plots (each with four replicates) were fertilized six times with 150 kg N ha^-1 in the form of cattle-pig slurry or NH₄NO₃. Soil samples (0–20 cm) were analysed at regular intervals for NH _inf4 ^sup+ and NO _inf3 ^sup– concentrations. The soil water content was determined gravimetrically. During the first year (1989) total denitrification losses from unfertilized, mineral-fertilized, and animal slurry-amended plots (with or without dicyandiamide) were estimated as 0.2, 3.1, 0.7, and 0.6 kg N ha^-1, respectively. During the second year (1990) the denitrification losses were 0.4, 1.3, 0.7, and 0.7 kg N ha^-1, respectively. There was a clear relationship between the NO _inf3 ^sup– concentration or soil water content and the denitrification rate. The results are siteund experiment-specific and cannot be generalized so far.     

黄瓜幼苗干物质积累、膨压及光合速率对铵态氮和硝态氮的响应

采用溶液培养方法,研究了氮素形态及水平对黄瓜干物质积累、膨压及光合速率的影响。结果表明,同一氮素水平时,NO3--N处理黄瓜干物质重量、叶片面积、叶片含水量、水势、膨压等均高于NH4+-N处理。低水平NH4+-N(5mmol/L)对黄瓜光合速率影响不大,甚至高于同水平NO3--N处理的,叶片面积稍有减小;高水平NH4+-N不仅显著抑制黄瓜干物质的积累,叶片面积仅是同水平NO3--N处理的12.7%,最小光合速率（Pn）为CO2 1.6mol/（m2·s）,仅为同水平NO3--N处理的10%左右。NH4+-N处理引起黄瓜叶片面积的减小与叶片膨压降低相关性较大,相关系数为0.98;膨压与叶片水势直接相关,而渗透势对此影响不大。高水平NO3--N对黄瓜产生了渗透胁迫,抑制黄瓜生长,表现在渗透调节物质的大量积累。不同黄瓜品种对氮素形态及水平响应差异较大,所有处理中新泰密刺长势均好于吉尼罗。低水平的NH4+-N对黄瓜干物质的积累影响不大,高水平则明显抑制其生长。说明高浓度铵态氮对植物产生了毒害,而高浓度的硝态氮则是渗透胁迫。     

N2O产生法测定土壤无机态氮15N丰度

用化学方法分别将土壤中微量的铵、硝酸盐和亚硝酸盐转化为N2O气体,然后用带自动预浓缩装置的同位素比值质谱仪测定N2O中的15N丰度.N2O中的15N丰度测量值完全符合铵、硝酸盐和亚硝酸盐的15N参考值.方法快速、简单和准确,不受空气氮的污染.特别是方法的检测限很低,每批次样品中只需含5～ 20μg N.它将有助于土壤氮素的矿化作用、硝化作用和反硝化作用的研究.     

养猪场处理尾水灌溉对太湖地区水稻土硝酸根异化还原为铵的影响

以太湖地区的湖白土和乌栅土为研究对象,通过室内15N 示踪实验研究了低浓度废水灌溉对土壤异化硝酸根还原为铵（DNRA）的影响。乌栅土DNRA速率和相对潜势分别为0.68 ~ 0.79 mg N kg-1干土 天-1和34.61 ~ 44.45%;湖白土DNRA速率和相对潜势分别为1.14 ~ 1.41 mg N kg-1干土 天-1和54.24 ~ 106.70%。低浓度废水对2种土壤的DNRA速率均影响不大;低浓度废水对湖白土DNRA相对潜势影响不大而明显降低了乌栅土DNRA相对潜势。相关分析表明,土壤DNRA相对潜势与培养开始（r = 0.836,p < 0.05,n = 6）和结束（r = 0.936,p < 0.01,n = 6）时的土壤溶解有机碳 / 硝态氮（DOC / NO3-N）均显著正相关,而与培养始末土壤Eh和DOC含量的相关性不显著。以上研究结果表明,太湖地区乌栅土具有较高的DNRA潜势,实践上有可能通过调控DNRA过程实现保持土壤氮素而减少农田氮损失的目的;尾水灌溉主要通过改变土壤DOC / NO3-N而影响DNRA对NO3-异化还原的贡献且其影响因土壤类型而异。     

Importance of heterotrophic nitrification and dissimilatory nitrate reduction to ammonium in a cropland soil: Evidences from a 15N tracing study to literature synthesis

Future climate change is predicted to influence soil moisture regime, a key factor regulating soil nitrogen (N) cycling. To elucidate how soil moisture affects gross N transformation in a cultivated black soil, a ¹⁵N tracing study was conducted at 30%, 50% and 70% water-filled pore space (WFPS). While gross mineralization rate of recalcitrant organic N (N_rec) increased from 0.56 to 2.47 mg N kg⁻¹ d⁻¹, the rate of labile organic N mineralization declined from 4.23 to 2.41 mg N kg⁻¹ d⁻¹ with a WFPS increase from 30% to 70%. Similar to total mineralization, no distinct moisture effect was found on total immobilization of ammonium, which primarily entered the N_rec pool. Nitrate (NO₃⁻) was mainly produced via autotrophic nitrification, which was significantly stimulated by increasing WFPS. Unexpectedly, heterotrophic nitrification was observed, with the highest rate of 1.06 mg N kg⁻¹ d⁻¹ at 30% WFPS, contributing 31.8% to total NO₃⁻ production, and decreased with WFPS. Dissimilatory nitrate reduction to ammonium (DNRA) increased from near zero (30% WFPS) to 0.26 mg N kg⁻¹ d⁻¹ (70% WFPS), amounting to 16.7–92.9% of NO₃⁻ consumption. A literature synthetic analysis from global multiple ecosystems showed that the rates of heterotrophic nitrification and DNRA in test soil were comparative to the forest and grassland ecosystems, and that heterotrophic nitrification was positively correlated with precipitation, soil organic carbon (SOC) and C/N, but negatively with pH and bulk density, while DNRA showed positive relationships with precipitation, clay, SOC, C/NO₃⁻ and WFPS. We suggested that low pH and bulk density and high SOC and C/N in test soil might favor heterotrophic nitrification, and that C and NO₃⁻ availability together with anaerobic condition were crucial for DNRA. Overall, our study highlights the role of moisture in regulating gross N turnover and the importance of heterotrophic nitrification for NO₃⁻ production under low moisture and DNRA for NO₃⁻ retention under high moisture in cropland.     

Preferential translocation of boron to young leaves in Arabidopsis thaliana Regulated by the BOR1 Gene

A mutant of Arabidopsis thaliana, bor1-1 (Noguchi et al. 1997: Plant Physiol., 115, 901–916) requires high levels of boron (B) for normal growth. We analyzed the B-deficiency symptoms of bor1-1 mutant plants in detail. A low B supply retarded the growth of the mutant plants more evidently in leaves than in roots. In particular, cell expansion and formation of air spaces were severely impaired by B deficiency in young rosette leaves. Such defects in growth were correlated with the reduced contents of B in leaves. These defects were not observed when a sufficient amount of B was supplied. Uptake experiments with ¹⁰B-enriched tracer B demonstrated that B taken up through roots was preferentially transported to young leaves compared to old leaves in the wild-type plants under a low B supply. Such a preferential transport to young leaves was not evident in the mutant plants. In conclusion, our data demonstrated that in A. thaliana plants B is preferentially transported to young organs under a low B supply and that this transport process is controlled at least in part by the BOR1 gene.