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1.
The intact nodules attached to the upper part of soybean roots were exposed to 15N2 and the incorporation of 15N into various soluble nitrogen constituents was investigated. Results indicated that ammonia, a primary product of N2 fixation, was located in more than two compartments. Ammonia reduced from N2 gas seemed to be incorporated firstly into glutamine especially amido-group nitrogen. Newly fixed nitrogen was secondly incorporated into glutamic acid and alanine in this sequence. These results suggested that fixed ammonia was assimilated by glutamine synthetase/glutamate synthase pathway. Turn-over rate of allantoin plus allantoic acid and serine was relatively high, although apparently these compounds were not primary products of newly fixed ammonia. 15N content of allantoin was always higher than that of allantoic acid. 15N incorporation to aspartic acid and asparagine was relatively slow, especially in early period. In bacteroid fraction there is much amount of ammonia comparing with other compounds, while allantoin and asparagine were presented exclusively in cytosol. 15N was incorporated into nitrate within a few minutes especially in bacteroids.  相似文献   

2.
The assimilation of ammonium and nitrate nitrogen into amino acids of mature sunflower leaves and their transport to the other plant parts were investigated using nitrogen-15 as a tracer. In the leaf, to which 15N-labelled ammonium was vacuum-infiltrated, the 15N content of glutamine was always the highest of the amino acids tested and that of alanine was higher than that of glutamic acid and aspartic acid at 15 min after the infiltration. On the other hand in the leaf to which 15N-labelled nitrate was vacuum-infiltrated, the 15N content of glutamic acid and aspartic acid was superior to that of glutamine. The incorporation of 15N into serine was not active in the case of either 15N-labelled ammonium or nitrate. In the internodes above and below the treated leaf, through which photosynthates were transported into other parts, the 15N content of γ-aminobutyric acid and glutamine was markedly high when both nitrogen sources were supplied. There were no differences in the labelling patterns of amino acids between the upper and lower internodes. From these results it may be concluded that glutamine, glutamic acid, and aspartic acid play an important role in the assimilation of ammonium and nitrate nitrogen in leaves and that nitrogen is transported mainly in the forms of γ-aminobutyric acid and glutamine from the leaves to the other plant parts,  相似文献   

3.
The kinetics of 15NH4 + assimilation into free amino acids and total reduced nitrogen were monitored in both roots and shoots of two week old tomato seedlings supplied with 5 mM 99% (15NH4)2SO4 via the aerated root medium in hydroponic culture, in the presence and absence of a 2 h pre‐incubation with 1 mM methionine sulfoximine (MSX). The labeling kinetics of amino acids in roots of tomato plants in the presence of MSX show that continued assimilation of 15NH4 + can occur when the GS/GOGAT cycle is inhibited. In the presence of MSX, three amino acids [glutamate, alanine, and y‐amino butyrate (GABA)] of the root tissue continue to become labeled with 15N under conditions where labeling of the amino‐N moiety of glutamine is completely inhibited. This indicates primary ammonia assimilation via GDH, or alternatively, assimilation of ammonia into alanine via alanine dehydrogenase. Free ammonia accumulates rapidly in both shoots and roots of tomato in response to MSX. The labeled ammonia accumulated in the roots in the presence of MSX must be largely derived from the medium whereas in shoots this ammonia appears to be derived from catabolism of unlabeled amino acids and proteins. The pools of glutamine, glutamate and alanine after 24 h exposure to 15NH4 + were, on the average, 5‐ to 10‐fold lower in the MSX‐treated than in the control (‐MSX) shoots and roots. In contrast, the pools of valine, leucine, isoleucine, proline, threonine, phenylalanine, lysine, and tyrosine increased 5‐ to 10‐fold above the control values in the shoots of MSX‐treated plants, and 2‐ to 4‐fold above control values in the roots of MSX‐treated tomato plants after 24 h. The latter amino acids all exhibited low isotope abundance, and presumably were derived from protein turnover.  相似文献   

4.
14C-labelled sodium bicarbonate and 15N-labelled ammonium sulfate were simultaneously vacuum-infiltrated into detached sunflower leaves, and the incorporation of 14C and 15N into free amino acids was chased during 60-min period in the light and in the dark.

In the light, the ue specific activity of aspartic acid, alanine, serine and glycine rapidly increased for 5 min and thereafter decreased. On the other hand, that of gultamic acid continued to increase slowly during the entire 60-min period. In the dark, aspartic acid most actively incorporated 14C. The difference of changes in 14C specific activity between glutamic acid and other amino acids was also observed in the dark as in the light. These results suggest that the carbon skeleton of glutamic acid is synthesized from aspartic acid, alanine, serine and glycine.

15N content of glutamine was the highest of all amino acids investigated in the light, and it was followed by glutamic acid. alanine, aspartic acid, serine and glycine, in this order. In the dark, 15N content of glutamic acid fell remarkably and was lower than that of alanine up to 5 min. From these 15N tracer experiments, it is suggested that the incorporation of ammonium into glutamic acid is strictly dependent on light and that alanine incorporates ammonium by the direct amination besides the transamination from glutamic acid.  相似文献   

5.
Assimilation of ammonium (NH4) into free amino acids and total reduced nitrogen (N) was monitored in both roots and shoots of two‐week old rice seedlings supplied with 5 mM 99% (15NH4)2SO4 in aerated hydroponic culture with or without a 2 h preincubation with 1 mM methionine sulfoximine (MSX), an inhibitor of glutamine synthetase (GS) activity. 15NH4 was not assimilated into amino acids when the GS/GOGAT (glutamate synthase) cycle was inhibited by MSX. Inhibition of glutamine synthetase (GS) activity in roots with MSX increased both the amount of NH4 and the abundance of 15N labeled NH4. In contrast, the amount of Gln and Glu, and their proportions as 15N, decreased in roots when GS activity was inhibited. This research confirms the importance of GS/GOGAT in NH4 assimilation in rice roots.

15N‐labeled studies indicate that NH4 ions incorporated by roots of rice are transformed primarily into glutamine (Gln) and glutamic acid (Glu) before being converted to other amino acids through transamination (15). The formation of amino acids such as aspartic acid (Asp) and alanine (Ala) directly from free NH4 in roots also has been reported (4,15). Translocation of free NH4 to plant shoots, based on the concentration of free NH4 in xylem exudate, has been reported in tomato (13), although NH4 in shoots primarily originates from nitrate reduction in the shoot. Photorespiration also can contribute to the accumulation of NH4 in leaves (7).

The GS/GOGAT cycle appears to be primarily responsible for the assimilation of exogenously supplied NH4 and NH4 derived from nitrate reduction in leaves, as well as NH4 derived from photorespiration (2,3,6,8). Genetic evidence cited to support this conclusion includes the lethal effect of photorespiratory conditions on plant mutants deficient in chloroplast‐localized GS and GOGAT activities (2,3,9), and the rapid accumulation of free NH4 in GS‐deficient mutants under photorespiratory conditions (2,3,5).

The present study was initiated to quantify the in vivo amino acid synthesis in rice roots and shoots by analysis of 15N labeling, and should provide a more complete understanding of this important system for NH4 utilization.  相似文献   

6.
15N‐labeled ammonia was rapidly assimilated by Azolla caroliniana and incorporated into plant material even though sustained growth of the fern‐algae symbiosis cannot be maintained with ammonia as nitrogen source. During ammonia uptake, the nitrogenous pool of the fern rapidly increases and contains large amounts of free ammonia and glutamine. N2 fixation activity of the algal symbiont declines during assimilation of ammonia, but it is restored to a high level upon transfer of plants to nitrogen‐free media, as the pool ammonia content decreases. During growth of the fern on N2, the algal symbiont supplies ammonia in a manner permitting sustained growth of the plant. Exogenous ammonia, therefore, appears to interrupt regulation of inorganic nitrogen metabolism of the plant‐algal symbiosis.  相似文献   

7.
Nitrogen Assimilation in Roots and the Transport of Nitrogen Compounds in the Bleeding Sap of Roots in relation to Manganese Nutrition. The assimilation of nitrogen in the roots of 27 days old pumpkin plants was examined in relation to manganese nutrition. The transport of nitrogen compounds in the xylem was determined in roots and in the bleeding sap of roots using nitrate as the N-source. The maximum NO3 content in the roots was observed in the Mn treatment which resulted in the highest shoot yields (0.05 ppm Mn). The bleeding sap of this treatment was lowest in nitrate concentration, but showed the highest rate of transport of organic nitrogen compounds. In experiments with 15N in the nutrient solution the isotope was found in the roots in organic and in inorganic compounds. The composition of the fraction of free amino acids differed between roots and xylem sap. In the bleeding sap glutamine was especially dominant. In the roots the amino acid composition depended on the extent of Mn-supply. Lowest glutamine concentrations were found in the xylem sap from the treatment with maximum shoot yields. A numerical difference was found in the xylem sap between organic N (N(org)) and the amino acid nitrogen. This difference which account for more than 50 % of the organically bound nitrogen is suggested to be made up in part by low molecular weight peptides, amino sugars and other compounds. In Mn deficiency a general reduction in the intensity of nitrogen metabolism was found. With Mn toxicity the N assimilation activity was more intensive than for the low Mn supply. Simultaneously, however, the transport of organic N compounds from the root was lower.  相似文献   

8.
Abstract

Extract

Leguminous plants consist of two groups, amide-exporting and ureide-exporting plants. The former legumes export a large fraction of fixed-N in the form of amides (asparagine and glutamine), and the latter legumes in the form of ureides (allantoic acid and allantoin). Another characteristic of the nodules is the enrichment in 15N. There are two types of legumes: one characterized by the enrichment with 15N in N2-fixing nodules, in contrast to the other where the enrichment does not occur. The first investigation by Shearer et al. (1982) suggested that the nodules exporting fixed-N in the form of ureides were enriched in 15N unlike those exporting it in the form of amides. Soybeans, mungbean, and cowpea belong to the former group and groundnut, alfalfa, white clover to the latter. Although pea and faba bean were first classified into the latter group (Shearer et al. 1982), a recent investigation (Yoneyama 1988) showed that these nodules were also enriched in 15N.  相似文献   

9.
Summary We studied the effect of three successive cuttings on N uptake and fixation and N distribution in Leucaena leucocephala. Two isolines, uninoculated or inoculated with three different Rhizobium strains, were grown for 36 weeks and cut every 12 weeks. The soil was labelled with 50 ppm KNO3 enriched with 10 atom % 15N excess soon after the first cutting. Except for the atom % 15N excess in branches of K28 at the second cutting, both the L. leucocephala isolines showed similar patterns of total N, fixed N2, and N from fertilizer distribution in different parts of the plant at each cutting. The Rhizobium strain did not influence the partitioning of 15N among the different plant parts. Significant differences in 15N enrichment occurred in different parts. Live nodules of both isolines showed the lowest atom % 15N excess values (0.087), followed by leaves (0.492), branches (0.552), stems (0.591), and roots (0.857). The roots contained about 60% of the total plant N and about 70% of the total N derived from fertilizer over the successive cuttings. The total N2 fixed in the roots was about 60% of that fixed in the whole plant, while the shoots contained only 20% of the fixed N2. We conclude that N reserves in roots and nodules constitute another N source that must be taken into account when estimating fixed N2 or the N balance after pruning or cutting plants. 15N enrichment declined up to about fivefold in the reference and the N2-fixing plants over 24 weeks following the 15N application. The proportion and the amounts of N derived from fertilizer decreased, while the amount derived from N2 fixation increased with time although its proportion remained constant.  相似文献   

10.
Summary Under greenhouse and field conditions, after the harvest of maize-cowpea intercropping, soils were analysed for total, ammonium and organic N fractions and fertilizer 15N residues. Growing cowpea as the sole crop or in intercropping with maize results in increased relative amounts of the acid hydrolysable organic N fractions in soil. After sole cropping of maize 70% of the residual fertilizer N was found in the acid hydrolysable fraction while after intercropping it was 80%–92%. The fertilizer and soil N labelling with 15N in identical but alternate series provided information on the nitrogen fixed by cowpea and left in the soil as crop residues. Under field conditions the cowpea plant residues left after cropping contained 170 kg N ha–1 in sole cropping and 105 kg N ha–1 in intercropping with maize. The N assimilated by cowpea-Rhizobium symbiosis was mainly present in the acid hydrolysable forms, particularly in the -amino N fraction and ammonium N fraction.  相似文献   

11.
The natural 15N abundances (δ15N values) were measured for nitrate and free and bound amino acids from the leaves of field-grown spinach (Spinacia oleracea L.) and komatsuna (Brassica campestris L.), as well as ureides and free and bound amino acids in the leaves and roots of hydroponically grown soybean (Glycine max L.) totally depending on dinitrogen. Nitrate from the spinach and komatsuna leaves and ureides from leaves and roots of soybean showed higher δ15N values than the total tissue N and N in free or bound amino acid fractions. The δ15N values of individual free and bound amino acids, determined by GC/C/MS using their acetylpropyl derivatives, were similar in leaf tissues except for proline but varied in soybean root tissues. The order of 15N enrichment was similar in the four samples: aspartic acid > glutamic acid > threonine, proline, valine > glycine + alanine +serine, γ-amino butyric acid, and phenylalanine.  相似文献   

12.
The quantitative analysis of the initial transport of fixed isotope 15-nitrogen (15N) in intact nodulated soybean plants (Glycine max [L.] Merr. cv. Williams) was investigated at the vegetative stage (36 days after planting, DAP) and pod-filling stage (91 DAP) by the 15N pulse-chase experiment. The nodulated roots were exposed to N2 gas labeled with a stable isotope 15N for 1 h, followed by 0, 1, 3 and 7 h of exposure with normal air. Plant roots and shoots were separated into three sections (basal, middle and distal parts) with the same length of the main stem or primary root. Approximately 80 and 92% of fixed N was distributed in the basal part of the nodulated roots at the vegetative and pod-filling stages by the end of 1 h of 15N2 exposure, respectively. In addition, about 90% of fixed 15N was retained in the nodules and 10% was exported to root and shoot after 1 h of 15N2 exposure at 91 DAP. The percentage distribution of 15N in the nodules at the pod-filling stage decreased from 90% to 7% during the 7 h of the chase period, and increased in the roots (14%), stems (54%), leaves (12%), pods (10%) and seeds (4%). The 15N distribution was negligible in the distal root segment, suggesting that N fixation activity was negligible and recycling fixed N from the shoot to the roots was very low in the initially short time of the experiment.  相似文献   

13.
Stable isotope composition of carbon and nitrogen in the sediment and pore water of a eutrophic freshwater lake was studied. Based on changes in the δ11C and δ15N values of dissolved components and sediment fraction. possible processes involved in the decomposition of sedimentary organic matter are outlined. The relative importance of acetate fermentation and CO2 reduction was estimated using known mathematical models, and ammonia assimilation by methanogenic bacteria is hypothesised to be the main process governing the isotope fractionation of dissolved nitrogen in pore water.  相似文献   

14.
Summary The dynamics of basally applied 15N-labeled ammonium sulfate in inorganic and organic soil fractions of five wetland rice soils of the Philippines was studied in a greenhouse experiment. Soil and plant samples were collected and analyzed for 15N at various growth stages. Exchangeable NH4 + depletion continued after 40 days after transplanting (DAT) and corresponded with increased nitrogen uptake by rice plants. Part of the applied fertilizer was fixed by 2:1 clay minerals, especially in Maligaya silty clay loam, which contained beidellite as the dominant clay mineral. After the initial fixation, nonexchangeable 15N was released from 20 DAT in Maligaya silty clay loam, but fixation delayed fertilizer N uptake from the soil. Part of the applied N was immobilized into the organic fraction. In Guadalupe clay and Maligaya silty clay loam, immobilization increased with time while the three other soils showed significant release of fertilizer N from the organic fraction during crop growth. Most of the immobilized fertilizer N was recovered in the nondistillable acid soluble (alpha-amino acid + hydrolyzable unknown-N) fraction at crop maturity. Between 61% and 66% of applied N was recovered from the plant in four soils while 52% of fertilizer N was recovered from the plant in Maligaya silty loam. Only 20% – 30% of the total N uptake at maturity was derived from fertilizer N. Nmin (mineral N) content of the soil before transplanting significantly correlated with N uptake. Twenty-two to 34% of applied N was unaccounted for possibly due to denitrification and ammonia volatilization.  相似文献   

15.
Abstract

Effect of the treatment of soybean nodules with nitrate on the permeability of the outer membrane of bacteroids of Bradyrhizobium japonicurn strain 138NR was examined using the hydrophobic fluorescent probe N-phenyl-1-naphthylamine (NPN). The incorporation of NPN into the outer and inner membranes of the bacteroid cells isolated from nodules of soybean (Glycine max L. Merro cv. Tamahomare) was followed by the measurement of fluorescence at 30°C. The NPN-uptake activity of the bacteroids was expressed by two parameters, the initial rate of NPN-uptake, k, and the final fluorescence intensity, F max. Acetylene reduction activity (ARA) of nodules, k, and F max decreased exponentially during three weeks following the 20 d period after planting (DAP), resulting in high correlations between logarithmic transforms of k-values or F max per bacteroid and those of ARA per bacteroid. Application of 20 mM KNO3 to the nodulated roots at 24 DAP inhibited ARA of the roots by 85% of the control after 4 d of treatment. There were concomitant decreases in k and F max by 75% and 65% of the control, respectively. These results suggest that treatment of soybean nodules with nitrate results in some changes in the permeability of the outer membrane of bacteroids with a concomitant decline of N2-fixing activity of bacteroids.  相似文献   

16.
Urine patches are significant hot‐spots of C and N transformations. To investigate the effects of urine composition on C and N turnover and gaseous emissions from a Danish pasture soil, a field plot study was carried out in September 2001. Cattle urine was amended with two levels of 13C‐ and 15N‐labeled urea, corresponding to 5.58 and 9.54 g urea‐N l–1, to reflect two levels of protein intake. Urine was then added to a sandy‐loam pasture soil equivalent to a rate of 23.3 or 39.8 g urea‐N m–2. Pools and isotopic labeling of nitrous oxide (N2O) and CO2 emissions, extractable urea, ammonium (NH4+), and nitrate (NO3), and plant uptake were monitored during a 14 d period, while ammonia (NH3) losses were estimated in separate plots amended with unlabeled urine. Ammonia volatilization was estimated to account for 14% and 12% of the urea‐N applied in the low (UL) and high (UH) urea treatment, respectively. The recovery of urea‐derived N as NH4+ increased during the first several days, but isotopic dilution was significant, possibly as a result of stress‐induced microbial metabolism. After a 2 d lag phase, nitrification proceeded at similar rates in UL and UH despite a significant difference in NH4+ availability. Nitrous oxide fluxes were low, but generally increased during the 14 d period, as did the proportion derived from urea‐N. On day 14, the contribution from urea was 23% (UL) and 13% (UH treatment), respectively. Cumulative total losses of N2O during the 14 d period corresponded to 0.021% (UL) and 0.015% (UH) of applied urea‐N. Nitrification was probably the source of N2O. Emission of urea‐derived C as CO2 was only detectable within the first 24 h. Urea‐derived C and N in above‐ground plant material was only significant at the first sampling, indicating that uptake of urine‐C and N via the leaves was small. Urine composition did not influence the potential for N2O emissions from urine patches under the experimental conditions, but the importance of site conditions and season should be investigated further.  相似文献   

17.
本研究利用同位素15N尿素,采用尿素熔融工艺分别制备了普通尿素(U)、 海藻酸增值尿素(AU)、 腐植酸增值尿素(HAU)和谷氨酸增值尿素(GU)试验产品,运用土柱栽培试验研究等氮量条件下(N 0.1 g/kg 干土,以030 cm土层干土重量计算)3种增值尿素产品对小麦产量、 氮肥利用率和肥料氮在土壤剖面中分布的影响。主要结果, 1)与U相比,AU、 HAU和GU处理均可显著提高小麦籽粒产量,增加幅度分别为7.12%、 13.63%和3.65%; 2)小麦吸收的氮素中有44.8%~48.0%来自肥料氮,AU、 HAU、 GU处理的小麦地上部吸收的肥料氮量均显著高于U处理,分别高出4.83%、 7.41% 和 3.12%; 3)虽然所有处理土壤残留的肥料氮均主要集中在050 cm土层中,U处理在5090 cm层次土壤肥料氮累积量显著高于AU和HAU处理; 4)AU、 HAU和GU处理的小麦氮肥表观利用率分别较U处理显著提高6.38、 15.63、 3.08个百分点,HAU和AU处理的15N利用率分别较普通尿素高出3.70和2.41个百分点,AU、 HAU和GU处理的肥料氮的损失率分别比普通尿素显著降低7.64、 9.52和2.19个百分点。  相似文献   

18.
To evaluate the use of foliar application of N fertilizer and the occurrence of leaf injury in tomato plants (Lycopersicon esculentum Mill., cv. Momotaro), the effects of the form and concentration of N and solution pH on the leaf injury were studied in the first experiment (Expt. 1). The effects of solution pH and leaf surface on the absorption, translocation, and assimilation of urea were compared with those of nitrate and ammonium in the second experiment (Expt. 2). In Expt. 1, no leaf injury was observed regardless of N sources applied at the N concentration of 1.0 g L-1. Compared with nitrate or ammonium, the index of leaf injury was the lowest in the leaf to which urea had been applied (hereafter referred to as “urea-applied leaf”), when the N level increased from 2.0 to 10.0 g L-1. Leaf injury was not affected by the solution pH in the case of urea, but it increased in the case of ammonium and decreased when nitrate was applied with increasing solution pH. In Expt. 2, the absorption of nitrate and ammonium by a leaf within 4 d was 34% and 74% of that of urea, respectively. N absorption at the lower leaf surface was much greater than that at the upper leaf surface for each N source. No apparent effect of solution pH on the absorption of urea was detected. With increasing solution pH, however, the absorption of nitrate decreased. The absorption of ammonium was the greatest at solution pH 7.5. Total-15N translocation from applied leaf to other plant parts within 4 d was the largest in the urea-applied plants. Effects of solution pH and leaf surface on 15N distribution were not appreciable. 15N assimilation was the quickest in the urea-applied plants. Two days after application, 15N assimilation in the whole plant was up to 76.9% in the urea-applied plants, but only 33.7% and 43.0% in the nitrate- and ammonium-applied plants, respectively. Urea was an appropriate foliar N source due to the low ability to injure foliage because of the rapid absorption and translocation, fast assimilation, and the wide and suitable range of solution pH. Foliar application of N to the lower leaf surface was recommended.  相似文献   

19.
K. OH  T. KATO  H. L. XU 《土壤圈》2008,18(2):222-226
An experiment was carried out to study the transport process of nitrogen (N) assimilation from tea roots by monitoring the dynamic composition of N compounds in xylem sap after 15^N-NO3 and 15^N-NH4 were fed to the root of tea plants (Camellia sinensis L.). Results showed that the main amino acids were glutamine, theanine, axginine, asparic acid and glutamic acid, which accounted for 49%, 17%, 8%, 7%, and 4%, respectively, of the total amino acids in the xylem sap. After the tea plants were fed with 15^N-NO3 and 15^N-NH4 for 48 h, the amount of total amino acids in xylem sap significantly increased and those fed with 15^N-NH4 had higher increment than those with 15^N-NOa. Two hours after 15^N- NO3 and 15^N-NH4 were fed, 15N abundance in glutamine, asparagine, glutamic acid, alanine, and arginine were detected and increased quickly over time. This indicated that it took less than 2 h for NO3-N and NH4-N to be absorbed by tea roots, incorporated into the above amino acids and transported to the xylem sap. Rapid increase in 15^N-NO3 in the xylem sap of tea plants fed with 15^N-NO3 indicated that nitrate could be directly transported to the xylem sap. Glutamine, theanine, and alanine were the main amino acids transported in xylem sap of tea plants fed with both 15^N-NO3 and 15^N-NH4.  相似文献   

20.
In previous studies, it has been shown that nitrate supply may favor adventitious rooting in the rooting recalcitrant Eucalyptus globulus. Herein, the impact of various N sources on adventitious rooting and root branching in microcuttings of E. globulus was investigated. The positive effect of nitrate on adventitious root development was confirmed and extended to root branching. Urea yielded a rooting response comparable to that observed in presence of nitrate. Urease activity was observed, displaying two peaks: one at the root induction and another at the root formation step. The use of glutamic acid, glutamine or asparagine promoted higher root number, but yielded shorter roots. Rooted microcuttings derived from all nitrogen (N) sources were successfully acclimated to ex vitro conditions. The manipulation of N sources in adventitious rooting media can be a tool for improving new root density, length and branching in this species.  相似文献   

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