Assimilation and transport of nitrogen in rice I. 15N-labelled ammonium nitrogen

The assimilation and transport of ¹⁵N-labelled ammonium nitrogenin rice plants (Oryza sativa L.) was studied. Plants assimilatedlarge amounts of nitrogen from labelled ammonium into theiramides and amino acids, particularly in the roots and stem,at the end of a 4-day ¹⁵N feeding and 10 days later in the upperleaves, especially in the blades. Although the incorporationof ¹⁵N into all the nitrogen fractions of the newly emergedpanicle was evident, it was particularly pronounced in the amidesand amino acids of the soluble fractions. The upper leaves hada greater ¹⁵N incorporation in their organic N-fractions thandid the lower ones. Amides and amino acids are considered tobe the main forms of nitrogen transported to the shoot fromthe ammonium assimilated in the roots. The transport of theorganic forms of nitrogen was possibly greater to the upperleaves than to the lower ones. The nitrite fraction had more ¹⁵N than did the nitrate fractionin all parts of the plant, particularly in the upper leaf blades.It appeared that some of the ammonia might have been oxidizedto nitrite, then to nitrate in some parts of the plant; probablyin the upper leaves. The synthesis of protein and nucleic acid occurred rapidly inthe upper leaves, especially in the blades, also in the rootsas evidenced by the considerable incorporation of ¹⁵N in theinsoluble fractions of these parts. The variation in ¹⁵N-distribution,during the 10 days, in the different plant parts suggests thatthe nitrogen incorporated during protein synthesis in the rootsand tillers was remobilized and transported to the upper partsof the shoot. A concept for the transport of organic nitrogenouscompounds from the roots to shoot through the phloem and xylemof the rice plant stem is discussed. (Received May 11, 1974; )     

Uptake of fertilizer nitrogen and soil nitrogen by rice using15N-labelled nitrogen fertilizer

Data from five field experiments using labelled nitrogen fertilizer were used to determine the relative effects of soil nitrogen and fertilizer nitrogen on rice yield. Yield of grain was closely correlated with total aboveground nitrogen uptake (soil+fertilizer), less closely correlated with soil nitrogen uptake and not significantly correlated with fertilizer nitrogen uptake. When yield increase rather than yield was correlated with fertilizer nitrogen uptake, the correlation coefficient was statistically significant.Contribution from the Laboratory for Flooded Soils and Sediments, Agronomy Dept., Louisiana Agri. Exper. Sta., Louisiana State Univ., Baton Rouge, LA 70803, and Univ. of Florida, Agricultural Research and Education Center, Sanford, FL 32771.     

A kinetic study of the assimilation of 15N-labelled nitrate in rice seedlings

A ¹⁵N kinetic-analysis of the assimilation of nitrate nitrogenin the roots of rice seedlings indicated that (1) nitrate wasrapidly reduced to ammonia in the roots, where it was incorporatedinto glutamine and glutamic acids; (2) the pattern of nitrateassimilation into amino acids was very similar to that of ammoniumassimilation; and (3) the pattern of nitrogen incorporationinto protein was also similar to that of the incorporation withNH₄-feeding. In the shoots, alanine, serine, glutamic acid, -amino butyricacid and aspartic acid were relatively strongly labelled with¹⁵N as compared with the other amino acids. A different mechanismof nitrogen assimilation seems to operate in between the photosyntheticand non-photosynthetic organs of the plants. (Received August 19, 1974; )     

Availability of nitrogen from 15N-labelled Azolla pinnata and urea to flooded rice

In view of the recently generated interest in Azolla and the high cost of N fertilizers, this field study was aimed at measuring the availability of Azolla-N applied in two split application in comparison to urea-N. Azolla was cultivated and labelled with ¹⁵N isotope in the field. A total of about 60 kg N ha^-1 was applied as Azolla, urea or Azolla and urea in combination, in two equal splits at transplanting and at maximum tillering, i.e. 30 days after transplanting (30 DAT).The recovery by the crop of Azolla-N applied at 30 DAT was significantly higher than that applied at transplanting, viz. 30.2% and 20.2%, respectively. The recoveries of urea-N applied at the same stages were similarly low, viz. 22.5% at transplanting and 38.6% at 30 DAT. Total recoveries of fertilizer N at the time of harvest were 26.8% from Azolla, 30.7% from urea applied in the same two splits and 49.1% from urea applied in locally recommended three splits. Recoveries of labelled Azolla-N in succeeding rice crop were twice higher than those of labelled urea-N. The recoveries ranged from 1.9 to 2.1% from urea-N and 4.0 to 4.9% from Azolla-N. There were no differences in residual ¹⁵N recovery in the succeeding crop between Azolla and urea either applied at transplanting or at 30 DAT.     

Contribution of N2 fixing cyanobacteria to rice production: availability of nitrogen from 15N-labelled cyanobacteria and ammonium sulphate to rice

This study investigate the potential contribution of nitrogen fixation by indigenous cyanobacteria to rice production in the rice fields of Valencia (Spain). N₂-fixing cyanobacteria abundance and N₂ fixation decreased with increasing amounts of fertilizers. Grain yield increased with increasing amounts of fertilizers up to 70 kg N ha^-1. No further increase was observed with 140 kg N ha^-1. Soil N was the main source of N for rice, only 8–14% of the total N incorporated by plants derived from ¹⁵N fertilizer. Recovery of applied ¹⁵N-ammonium sulphate by the soil–plant system was lower than 50%. Losses were attributed to ammonia volatilization, since only 0.3–1% of applied N was lost by denitrification. Recovery of ¹⁵N from labeled cyanobacteria by the soil–plant system was higher than that from chemical fertilizers. Cyanobacterial N was available to rice plant even at the tillering stage, 20 days after N application. This revised version was published online in June 2006 with corrections to the Cover Date.     

Uptake of 15N-labelled alanine,ammonium and nitrate in Pinus sylvestris L. ectomycorrhiza growing in forest soil treated with nitrogen,sulphur or lime

The uptake of ¹⁵N-labelled alanine, ammonium and nitrate was studied in ectomycorrhizal morphotypes of intact Pinus sylvestris seedlings. PCR-RFLP analysis of the ITS-region of fungal rDNA was used to identify the morphotypes. Seedlings were grown in forest soil collected at an experimental site in southern Sweden. The treatments compared were a control, N fertilisation (600 kg N ha^-1 as urea), sulfur application (1200 kg S ha^-1) and lime application (6000 kg CaCO₃ ha^-1). The forest, which had been dominated by Picea abies, was clear-cut two years before the forest soil was sampled. Soil was also collected from an adjacent standing forest. The aim of the present study was to detect changes in the ectomycorrhizal communities in forest soils and relate these changes to the functional parameter of uptake of nitrogen from organic (alanine and protein) and inorganic (ammonium and nitrate) sources.Liming resulted in the detection of a morphotype not found in other samples, and one morphotype was only found in samples from the standing forest (the fungi in these two morphotypes could not be identified). All mycorrhizal root tips showed a higher ¹⁵N concentration after exposure to different nitrogen forms than non-mycorrhizal long roots. Uptake of¹⁵ N from a labelled solution of alanine or ammonium was higher (about tenfold) than uptake from a ¹⁵N-labelled solution of nitrate. Uptake of ammonium and alanine varied between 0.2 and 0.5 mg N g^-1 h^-1 and between 0.1 and 0.33 mg N g^-1 h^-1, respectively, among the different morphotypes.In seedlings grown in the control soil and in soil from standing forest, alanine and ammonium were taken up to a similar degree from a supply solution by all morphotypes, whereas ammonium uptake was higher than alanine uptake in seedlings grown in lime-treated soil (about twofold) and, to a lesser extent, in the nitrogen- and sulfur-treated soils. The higher ammonium uptake by morphotypes from the limed soil was confirmed in pure culture studies. In cases where ammonium was used as the N source, an isolate of the S. variegatus morphotype collected in the limed soil produced more biomass compared with isolates of S. variegatus collected in nitrogen- or sulphur-treated soil. One isolate of a silvery white morphotype produced about equal amounts of biomass on alanine and ammonium, whereas all S. variegatus isolated performed better with ammonium as their N source. Based on the results it is hypothesised that liming can induce a shift in the ectomycorrhizal community, favouring individuals that mainly utilise inorganic nitrogen over those that primarily utilise organic nitrogen.     

Transformation of15N-labelled urea and its modified forms in tropical wetland rice culture

Summary The fate of 100 kg N ha^–1 applied as¹⁵N-urea and its modified forms was followed in 4 successive field-grown wetland rice crops in a vertisol. The first wet season crop recovered about 27 to 36.6% of the applied N depending upon the N source. In subsequent seasons the average uptake was very small and it gradually decreased from 1.4 to 0.5 kg N ha^–1 although about 18 to 20, 12 to 17 and 14 to 18 kg ha^–1 residual fertilizer N was available in the root zone after harvest of first, second and third crops, respectively. The average uptake of the residual fertilizer N was only 7.6% in the second crop and it decreased to 4.5% in the third and to 3.2% in the fourth crop although all these crops were adequately fertilized with unlabelled urea. The basal application of neem coated urea was more effective in controlling the leaching loss of labelled NH₄+NO₃–N than split application of uncoated urea. In the first 3 seasons in which¹⁵N was detectable, the loss of fertilizer N through leaching as NH₄+NO₃–N amounted to 0.5 kg ha^–1 from neem-coated urea, 1.5 kg from split urea and 4.1 kg from coal tar-coated urea. At the end of 4 crops, most of the labelled fertilizer N (about 69% on average) was located in the upper 0–20 cm soil layer showing very little movement beyond this depth. In the profile sampled upto 60 cm depth, totally about 13.8 kg labelled fertilizer N ha^–1 from neem-coated urea, 12.7 kg from coal-tar coated urea, and 11.8 kg from split urea were recovered. The average recovery of labelled urea-N in crops and soil during the entire experimental period ranged between 42 and 51%. After correcting for leaching losses, the remaining 47 to 56% appeared to have been lost through ammonia volatilization and denitrification.     

^15N标记稻草中N,C在羊体内的转化和利用

应用~(13)N标记稻草饲喂3只山羊,以探明羊对稻草N、C化合物的消化、吸收、排泄和转化规律。结果表明,已宰杀的2只羊消化、吸收、转化为羊机体的~(15)N占试验日粮中~(15)N富集总量的38.54和23.78％,平均为31.16％。3只羊从粪尿中排泄的~(15)N各占饲料中~(15)N的34.78、33.88和33.18％,平均为33.95±0.80％,已屠宰的2只羊对饲料~(15)N总回收率为73.32和56.96％,损失率为26.68和43.04％。饲料~(15)N的回收利用率与饲料中氨基酸的消化率(％)相吻合。1、2、3号羊对饲料碳水化合物的消化率分别为76.40、68.66和65.19％。其中饲喂2、3号羊的饲料中都含稻草50％左右,羊对碳水化合物的平均消化率为66.93％。     

A kinetic study of the assimilation of 15N-labelled ammonium in rice seedling roots

The pathway of ammonium nitrogen assimilation, its incorporationinto amino acids and synthesis of protein was studied with theaid of nitrogen-15. The analysis of ¹⁵N involves the use ofoptical emission spectrometry. Kinetic analysis of nitrogen assimilation by the roots indicatesthat glutamine and glutamic acid were the primary products ofammonium assimilation. Possibly some of the amino acids, suchas aspartic acid and alanine received their amino nitrogen directlyfrom free ammonia in the roots. Amino groups were transformedinto other amino acids from these primary products, especiallyfrom glutamic acid through transamination. (Received April 1, 1974; )     

Biochar application to rice with 15N-labelled fertilizers,enhanced leaf nitrogen concentration and assimilation by improving morpho-physiological traits and soil quality

Leaf nitrogen (N) concentration plays an important role in biochemical and physiological functions, and N availability directly influences rice yield. However, excessive N fertilization is considered to be a root cause of environmental issues and low nitrogen use efficiency. Therefore, the selection of appropriate nutrient management practices and organic amendments is key to maximizing nitrogen uptake and maintaining high and sustainable rice production. Here, we evaluated the effects of different ¹⁵N-labelled nitrogen sources (urea, ammonium nitrate, and ammonium sulfate at 315 kg ha⁻¹) with or without biochar (30 t ha⁻¹) on paddy soil properties, root growth, leaf gas exchange, N metabolism enzymes, and N uptake in the early and late seasons of 2019. We found significant differences among N fertilizer sources applied with or without biochar (P < 0.05). Across the seasons, the combination of biochar with N fertilizers significantly increased soil organic carbon by 51.21% and nitrogen availability by 27.51% compared with N fertilizers alone. Correlation analysis showed that rice root morphological traits were strongly related to soil chemical properties, and higher root growth was measured in the biochar treatments. Similarly, net leaf photosynthetic rate averaged 9.34% higher, chlorophyll (Chl) a concentration 12.91% higher, and Chl b concentration 10.05% higher in the biochar treatments than in the biochar-free treatments across the seasons. Notably, leaf ¹⁵N concentration was 23.19% higher in the biochar treatments in both seasons. These results illustrated higher activities of N metabolism enzymes such as NR, GS, and GOGAT by an average 23.44%, 11.26% and 18.16% in the biochar treatments across the seasons, respectively. The addition of biochar with synthetic N fertilizers is an ecological nutrient management strategy that can increase N uptake and assimilation by ameliorating soil properties and improving the morpho-physiological factors of rice.     

Fate of 15N-labelled nitrogen fertilizer applied to kiwifruit (Actinidia deliciosa) vines

The fate of ¹⁵N-labelled ammonium fertilizer applied once to six-year-old field-grown kiwifruit (Actinidia deliciosa Hayward) vines was measured over three years. The three main treatments were nitrogen (N) applied singularly at 100 or 200 kg N ha^–1 in early spring (two weeks before bud burst) or split with 100 kg N ha^–1 (unlabelled) in early spring and 100 kg N ha^–1 (¹⁵N-labelled) ten weeks later. All N treatments were applied to vines with a history of either 50 or 200 kg N ha^–1 yr^–1. For three years after ¹⁵n application, components of the vines and soil (0–600 mm depth) were sampled at harvest in late autumn and the N and ¹⁵N contents determined.By the first harvest, all plant uptake of ¹⁵N had occurred and this represented 48–53% of the ¹⁵N applied. There was no significant effect of current N fertilizer treatment or of N history on ¹⁵N recovery by vines. Removal of ¹⁵N in harvested fruit was small at 5–6% in the first year and 8% over 3 years. After 2–3 years, most plant ¹⁵N occurred in the roots and this component declined only slowly over time. In contrast, there was a large temporal decline in ¹⁵N in above-ground plant components due to the annual removal in leaf fall and pruning. An associated experiment showed that when ¹⁵N-labelled prunings and leaves were mulched and returned to the soil, only about 9% was recovered by plants within 2 years. Almost all remaining mulched material had been immobilised into the soil organic N.In all treatments, about 20% of the added ¹⁵N remained in soil at the first harvest. This was almost entirely in organic fractions (<0.4% in inorganic N) and mostly in the surface 150-mm layer. The ¹⁵N content in soil changed little over time (from 20 to 17% between the first and third harvests respectively) and indicated that most of the N had been immobilised into stable humus forms.     

The fate of 15N-labelled nitrate additions to a northern hardwood forest in eastern Maine,USA

We followed the movements of ¹⁵N-labelled nitrate additions into biomass and soil pools of experimental plots (15×15 m each) in a mid-successional beech-maple-birch-spruce forest in order to identify sinks for nitrate inputs to a forest ecosystem. Replicate plots (n=3) were spray-irrigated with either 28 or 56 kg N ha^–1 year^–1 using ¹⁵N-labelled nitric acid solutions (¹⁵N = 344 ) during four successive growing seasons (April–October). The ¹⁵N contents of foliage, bolewood, forests floor and mineral soil (0–5 cm) increased during the course of treatments. Mass balance calculations showed that one-fourth to one-third of the nitrate applied to forest plots was assimilated into and retained by above ground plant tissues and surface soil horizons at both rates of nitrate application. Plant and microbial assimilation were of approximately equal importance in retaining nitrate additions to this forest. Nitrate use among tree species varied, however, with red spruce showing lower rates of nitrate assimilation into foliage and bolewood than American beech and other deciduous species.     

Incorporation of nitrate nitrogen in rice seedlings transferred to anaerobic conditions

Summary Incorporation of¹⁵NO₃- into amino acids was studied in 3-day-old aerobic rice seedlings (with coleoptile and root) subjected for 24h to anaerobic conditions. The incorporation of¹⁵N into glutamate, glutamine and alanine accounted for 89% and 84% of total incorporation in coleoptile and root, respectively. These findings indicate that, after the primary incorporation of¹⁵N into glutamate and glutamine, the main fate of nitrate nitrogen in rice seedlings subjected to anoxia is alanine.     

Mineralization of15N-labelled legume residues in soils with different nitrogen contents and its uptake by Rhodes grass

Summary Soil was collected from pots that had grown 1,3 or 6 soybean (Glycine max) or Siratro (Macroptillium atropurpureum) crops that had received organic residue returns from each crop.¹⁵N-labelled residues were added to half the pots in the experiment and the other half left unamended. Half of each group was then sown to Rhodes grass (Chloris gayana) which was grown, under glasshouse conditions, for 12 weeks.Ten grams of organic matter residues were added to each pot (1.5 kg soil) and the pots subjected to two wetting and drying cycles. At the end of the second wet cycle, soil mineral N values ranged from 6 to 64 ppm in unamended soils and from 19 to 177 ppm in amended soils. These levels generally declined over a 12 week period both in the presence and absence of sown grass.Nitrogen uptake by the grass increased with the number of previous cycles and was higher in Siratro than soybean soils. Recovery of¹⁵N by plant growth from the incorporated soybean residues was little effected by previous crop history and averaged 15.4%. On the other hand, Siratro recoveries were 13.7, 42.4 and 55.5% from soils that had grown 1, 3 and 6 previous Siratro crops, respectively.The addition of organic residues stimulated the release of native organic N (positive priming effect) on all soils.These results show that the turnover rate of nitrogen from organic residues can be high and the net result of these additions depends on the nature of the organic residues and the soil system to which they are added. These data emphasise the need to consider the rate of nutrient turnover from organic sources rather than concentrate on the nature and size of the resident nutrient pools.     

Assimilation of ammonium ions and reutilization of nitrogen in rice (Oryza sativa L.)

A major source of inorganic nitrogen for rice plants grown in paddy soil is ammonium ions. The ammonium ions are actively taken up by the roots via ammonium transporters and subsequently assimilated into the amide residue of glutamine (Gln) by the reaction of glutamine synthetase (GS) in the roots. The Gln is converted into glutamate (Glu), which is a central amino acid for the synthesis of a number of amino acids, by the reaction of glutamate synthase (GOGAT). Although a small gene family for both GS and GOGAT is present in rice, ammonium-dependent and cell type-specific expression suggest that cytosolic GS1;2 and plastidic NADH-GOGAT1 are responsible for the primary assimilation of ammonium ions in the roots. In the plant top, approximately 80% of the total nitrogen in the panicle is remobilized through the phloem from senescing organs. Since the major form of nitrogen in the phloem sap is Gln, GS in the senescing organs and GOGAT in developing organs are important for nitrogen remobilization and reutilization, respectively. Recent work with a knock-out mutant of rice clearly showed that GS1;1 is responsible for this process. Overexpression studies together with age- and cell type-specific expression strongly suggest that NADH-GOGAT1 is important for the reutilization of transported Gln in developing organs. The overall process of nitrogen utilization within the plant is discussed.     

Gene structure and expression of the high-affinity nitrate transport system in rice roots

Rice has a preference for uptake of ammonium over nitrate and can use ammonium-N efficiently. Consequently, transporters mediating ammonium uptake have been extensively studied, but nitrate transporters have been largely ignored. Recently,some reports have shown that rice also has high capacity to acquire nitrate from growth medium, so understanding the nitrate transport system in rice roots is very important for improving N use efficiency in rice. The present study identified four putative NRT2 and two putative NAR2 genes that encode components of the high-affinity nitrate transport system (HATS) in the rice (Oryza sativa L. subsp, japonica cv. Nipponbare) genome. OsNRT2.1 and OsNRT2.2 share an identical coding region sequence, and their deduced proteins are closely related to those from monocotyledonous plants. The two NAR2 proteins are closely related to those from mono-cotyledonous plants as well. However, OsNRT2.3 and OsNRT2.4 are more closely related to Arabidopsis NRT2 proteins. Relative quantitative reverse tranecdption-polymerase chain reaction analysis showed that all of the six genes were rapidly upregulated and then downregulated in the roots of N-starved rice plants after they were re-supplied with 0.2 mM nitrate, but the response to nitrate differed among gene members.The results from phylogenetic tree, gene structure and expression analysis implied the divergent roles for the individual members of the rice NRT2 and NAR2 families. High-affinity nitrate influx rates associated with nitrate induction in rice roots were investigated and were found to be regulated by external pH. Compared with the nitrate influx rates at pH 6.5, alkaline pH (pH 8.0) inhibited nitrate Influx, and acidic pH (pH 5.0) enhanced the nitrate influx In I h nitrate induced roots, but did not significantly affect that in 4 to 8 h nitrate induced roots.