Nitrogen Transfer Within and Between Plants Through Common Mycorrhizal Networks (CMNs)

Mycorrhizae play a critical role in nutrient capture from soils. Arbuscular mycorrhizae (AM) and ectomycorrhizae (EM) are the most important mycorrhizae in agricultural and natural ecosystems. AM and EM fungi use inorganic NH₄ ⁺ and NO₃ ^?, and most EM fungi are capable of using organic nitrogen. The heavier stable isotope ¹⁵N is discriminated against during biogeochemical and biochemical processes. Differences in ¹⁵N (atom%) or δ¹⁵N (‰) provide nitrogen movement information in an experimental system. A range of 20 to 50% of one-way N-transfer has been observed from legumes to nonlegumes. Mycorrhizal fungal mycelia can extend from one plant's roots to another plant's roots to form common mycorrhizal networks (CMNs). Individual species, genera, even families of plants can be interconnected by CMNs. They are capable of facilitating nutrient uptake and flux. Nutrients such as carbon, nitrogen and phosphorus and other elements may then move via either AM or EM networks from plant to plant. Both ¹⁵N labeling and ¹⁵N natural abundance techniques have been employed to trace N movement between plants interconnected by AM or EM networks. Fine mesh (25～45 μm) has been used to separate root systems and allow only hyphal penetration and linkages but no root contact between plants. In many studies, nitrogen from N₂-fixing mycorrhizal plants transferred to non-N₂–fixing mycorrhizal plants (one-way N-transfer). In a few studies, N is also transferred from non-N₂–fixing mycorrhizal plants to N₂-fixing mycorrhizal plants (two-way N-transfer). There is controversy about whether N-transfer is direct through CMNs, or indirect through the soil. The lack of convincing data underlines the need for creative, careful experimental manipulations. Nitrogen is crucial to productivity in most terrestrial ecosystems, and there are potential benefits of management in soil-plant systems to enhance N-transfer. Thus, two-way N-transfer warrants further investigation with many species and under field conditions.     

Variation in 15N enrichment of crimson clover labeled under field conditions

Plant material labeled with ¹⁵N is often used to determine recovery of N from green manure crops by subsequent crops. In this study, ¹⁵N enriched crimson clover (Trifolium incarnatum L.) was grown at a field site where it was to be utilized in a subsequent experiment. A foliar spray of (NH₄)₂SO₄ (99 atom % excess ¹⁵N) was applied to a 1.2 m × 8.8 m plot of crimson clover at a rate of 10 kg N ha^–1 in early March 1990, immediately prior to the period of rapid vegetative growth. Clover shoots harvested in April contained 1.72 atom % excess ¹⁵N. Total N concentration of enriched clover was similar to that in adjacent untreated clover. Clover shoots contained 20% of the applied ¹⁵N, and an additional 27% was recovered from the surface soil horizon (0 to 15 cm). A gradient was observed across the plot, with clover enrichment increasing from 1.3 to 2.2 atom % excess ¹⁵N. Recovery of applied ¹⁵N in soil was highest in the subplots with lowest clover enrichment. Variability in ¹⁵N enrichment was also observed among plant parts: leaves from the basal half of shoots had 2.2 atom % excess ¹⁵N; while leaves from the terminal half of shoots, terminal stems, and basal stems had between 1.1 and 1.4 atom % excess ¹⁵N.Abbreviation %Ndf source the percentage of the N atoms in a sample derived from a labeled source     

Transfer of nitrogen from damaged roots of white clover (Trifolium repens L.) to closely associated roots of intact perennial ryegrass (Lolium perenne L)

An experiment is described in which the magnitude of N transferred from damaged white clover roots to perennial ryegrass was determined, using ¹⁵N labelling of the grass plant. There was no effect on the growth and N-fixation of the clover plants after removing part of the root system. The ¹⁵N data suggested that N had been acquired by all grass plants, even in plants grown alone with no further N supplied after labelling. However, after quantifying the mobile and stored N pools of the grass plants it was evident that significant transfer of N from clover to grass only took place from damaged clover roots. Dilution of the atom% ¹⁵N in the roots of the grass plants grown alone, and in association with undamaged clover roots, was explained by remobilisation of N within the plant.     

Bi-directional transfer of nitrogen between alfalfa and bromegrass: Short and long term evidence

Transfer of N from legumes to associated non-legumes has been demonstrated under a wide range of conditions. Because legumes are able to derive their N requirements from N₂ fixation, legumes can serve, through the transfer of N, as a source of N for accompanying non-legumes. Studies, therefore, are often limited to the transfer of N from the legume to the non-legume. However, legumes preferentially rely on available soil N as their source of N. To determine whether N can be transferred from a non-legume to a legume, two greenhouse experiments were conducted. In the short-term N-transfer experiment, a portion of the foliage of meadow bromegrass (Bromus riparius Rhem.) or alfalfa (Medicago sativa L.) was immersed in a highly labelled ¹⁵N-solution and following a 64 h incubation, the roots and leaves of the associated alfalfa and bromegrass were analyzed for ¹⁵N. In the long-term N transfer experiment, alfalfa and bromegrass were grown in an ¹⁵N-labelled nutrient solution and transplanted in pots with unlabelled bromegrass and alfalfa plants. Plants were harvested at 50 and 79 d after transplanting and analyzed for ¹⁵N content. Whether alfalfa or bromegrass were the donor plants in the short-term experiment, roots and leaves of all neighbouring alfalfa and bromegrass plants were enriched with ¹⁵N. Similarly, when alfalfa or bromegrass was labelled in the long-term experiment, the roots and shoots of neighbouring alfalfa and bromegrass plants became enriched with ¹⁵N. These two studies conclusively show that within a short period of time, N is transferred from both the N₂-fixing legume to the associated non-legume and also from the non-legume to the N₂-fixing legume. The occurrence of a bi-directional N transfer between N₂-fixing and non-N₂-fixing plants should be taken into consideration when the intensity of N cycling and the directional flow of N in pastures and natural ecosystems are investigated.     

The nitrogen mineralization rate of legume residues in soil as influenced by their polyphenol,lignin, and nitrogen contents

A 12-week greenhouse experiment was conducted to determine the effect of the polyphenol, lignin and N contents of six legumes on their N mineralization rate in soil and to compare estimates of legume-N release by the difference and ¹⁵N-recovery methods. Mature tops of alfalfa (Medicago sativa L.), round leaf cassia (Cassia rotundifolia Pers., var. Wynn), leucaena (Leucaena leucocephala Lam., deWit), Fitzroy stylo (Stylosanthes scabra Vog., var Fitzroy), snail medic (Medicago scutellata L.), and vigna (Vigna trilobata L., var verde) were incorporated in soil at the rate of 100 mg legume N kg^-1 soil. The medic and vigna were labeled with ¹⁵N. Sorghum-sudan hybrid (Sorghum bicolor, L. Moench) was used as the test crop. A non-amended treatment was used as a control. Net N mineralization after 12 weeks ranged from 11% of added N with cassia to 47% of added N for alfalfa. With the two legumes that contained less than 20 g kg^-1 of N, stylo and cassia, there was net N immobilization for the first 6 weeks of the experiment. The legume (lignin + polyphenol):N ratio was significantly correlated with N mineralization at all sampling dates at the 0.05 level and at the 0.01 level at 6 weeks (r²=0.866). Legume N, lignin, or polyphenol concentrations or the lignin:N ratio were not significantly correlated with N mineralization at any time. The polyphenol:N ratio was only significantly correlated with N mineralization after 9 weeks (r²=0.692). The (lignin + polyphenol):N ratio appears to be a good predictor of N mineralization rates of incorporated legumes, but the method for analyzing plant polyphenol needs to be standardized. Estimates of legume-N mineralization by the difference and ¹⁵N recovery methods were significantly different at all sampling dates for both ¹⁵N-labeled legumes. After 12 weeks, estimates of legume-N mineralization averaged 20% more with the difference method than with the ¹⁵N recovery method. This finding suggests that estimates of legume N available to subsequent crops should not be based solely on results from ¹⁵N recovery experiments.     

Amino acids as a nitrogen source for tomato seedlings: The use of dual-labeled (C, N) glycine to test for direct uptake by tomato seedlings

Direct uptake of organic nitrogen (ON) compounds, rather than inorganic N, by plant roots has been hypothesized to constitute a significant pathway for plant nutrition. The aim of this study was to test whether tomatoes (Solanum lycopersicum cv. Huying932) can take up ON directly from the soil by using ¹⁵NH₄Cl, K¹⁵NO₃, 1, 2-¹³C₂¹⁵N-glycine labeling techniques. The ¹³C and ¹⁵N in the plants increased significantly indicating that a portion of the glycine-N was taken up in the form of intact amino acids by the tomatoes within 48 h after injection into the soil. Regression analysis of excess ¹³C against excess ¹⁵N showed that approximately 21% of the supplied glycine-N was taken up intact by the tomatoes. Atom% excesses of ¹⁵N and ¹³C in the roots were higher than in any shoots. Results also indicated rapid turnover of amino acids (e.g., glycine) by soil microorganisms, and the poor competitive ability of tomatoes in absorbing amino acids from the soil solution. This implies that tomatoes can take up ON in an intact form from the soil despite the rapid turnover of organic N usually found under such conditions. Given the influence of climatic change and N pollution, further studies investigating the functional ecological implications of ON in horticultural ecosystems are warranted.     

Effects of timing and supply of nitrogen on nitrogen remobilization from vegetative organs and redistribution to developing seeds of sunflower

A glasshouse study was made of the distribution of ¹⁵N among vegetative organs of sunflower and its later remobilization and redistribution to seeds, as influenced by the developmental stage at which ¹⁵N was provided, and by the N status of the plants. Plants of Hysun 30 sunflower were grown in sand culture and provided with K¹⁵NO₃ for a 3-day period at: (a) 3 days before the end of floret initiation; (b) 3 days before anthesis; (c) the start of anthesis; (d) full anthesis; and (e) 8 days after full anthesis. The plants were grown on a range of N supply rates, from severely deficient to more than adequate for maximum growth. Nitrogen-15 was distributed to all parts of the plant at the end of the ¹⁵N uptake periods. With the exception of the most N-stressed plants, subsequent remobilization of ¹⁵N from roots, stems and leaves occurred irrespective of the time the ¹⁵N was taken up. However, the percentage redistribution to seeds of ¹⁵N taken up at the end of floret initiation was less than for ¹⁵N taken up at anthesis. Remobilization of ¹⁵N from leaves and roots was higher (70%) for ¹⁵N taken up during and after anthesis than for ¹⁵N taken up at the end of floret initiation (45%), except for plants grown on the lowest N supply. By contrast, remobilization of ¹⁵N from the stem was lower for ¹⁵N taken up after full anthesis (40%) than before or during anthesis (>70%). The proportion of ¹⁵N remobilized from the top third of the stem was less than that from the bottom third, and decreased with increasing plant N status. Nitrogen-15 taken up over the 3-day supply periods during anthesis contributed from 2 to 11% of the total seed N at maturity; the contribution to seeds was greatest for plants grown on the highest N supply. Nitrogen taken up just before and during anthesis contributed most of the N accumulated in mature seeds of plants grown on an adequate N supply, but N taken up between the end of floret initiation and just before anthesis, or after full anthesis seemed to make an equally important contribution to mature seeds as N taken up during anthesis for plants grown on a very low N supply. It was concluded that the development of florets and seeds of sunflower is supported by N taken up by the plant between the end of floret initiation and anthesis, and by N redistributed from vegetative organs. Unless soil N is so low as to impair early growth, split applications of N fertilizer would be best made just before the end of floret initiation (‘star stage’) and just before anthesis.     

Soil aggregate sequestration of cover crop root and shoot-derived nitrogen

Cover crop roots and shoots release carbon (C) and nitrogen (N) compounds in situ during their decomposition. Depending upon the season, these C and N compounds may be sequestered, the C may be respired or the N may be leached below the root zone. A field study was established to identify the contributions of cover crop root and shoot N to different regions within aggregates in the A_p horizon of a Kalamazoo loam soil. Fall-planted rye plants (Secale cerealeL.) were labeled the next May with foliar applications of solutions containing 99% atom (¹⁵NH₄)₂SO₄. Isotopic enrichment of soil aggregates ranging from 2.0 to 4.0, 4.0–6.3 and 6.3–9.5 mm across was determined following plant residue applications. Concentric layers of aggregates were removed from each aggregate by newly designed meso soil aggregate erosion (SAE) chambers. Non-uniform distributions of total N and recently derived rye N in soil macroaggregates, across time, suggested that the formations and functions of macroaggregates are very dynamics processes and soil aggregates influence where N is deposited. Early in the season, more ¹⁵N migrated to the interior regions of the smallest aggregates, 2–4 mm across, but it was limited to only surfaces and transitional regions of the larger aggregates, 6.3–9.3 mm across. Exterior layers of aggregates between 6.0 and 9.5 mm retained 1.6% of the N_{derived from roots} in July 1999, which was three times more than their interior regions. This was slightly greater than the % N_{derived from shoot.} One month later, as the maize root absorption of N increased rapidly, % N_{derived from roots} and % N_{derived from shoot} were nearly equal in exterior layers and interior regions of soil aggregates. This equilibrium distribution may have been from either greater diffusion of N within the aggregates and/or maize root removal form aggregate exteriors. Results supported that most of roots grew preferentially around surfaces of soil aggregates rather than through aggregates. Cover crop roots contributed as much N as cover crop shoots to the total soil N pool. Subsequent crops use N from the most easily accessible zones of soil structure, which are surfaces of larger soil aggregates. Therefore maintaining active plant roots and aggregated soil structure in the soil enhances N sequestration and maximize soil N availability. These studies suggest that the rapid and perhaps bulk flow of soil N solutions may bypass many of the central regions of soil aggregates, resulting in greater leaching losses.     

Carbon distribution and nitrogen partitioning in a soil-plant system with barley (Hordeum vulgare L.), ryegrass (Lolium perenne) and rape (Brassica napus L.) grown in a14CO2-atmosphere

To examine the influence of plant-microorganism interactions on soil-N transformations (e.g. net mineralization, net immobilization) a pot experiment was conducted in a¹⁴C-labelled atmosphere by using different (two annuals, one perennial) plants species. It was assumed that variation in below-ground, microorganism-available C would influence N transformations in soil. Plant species were fertilized (low rate) with¹⁵N-labelled nitrogen and grown, during days 13 and 62 after germination, in a growth chamber with a¹⁴C-labelled atmosphere. Nitrification was inhibited by using nitrapyrin (N-Serve). During the chamber period, shoots were harvested, and associated roots and soil were collected on two sampling occasionm, e.g. after 4 and 7 weeks in the growth chamber.The distribution of net (%) assimilated¹⁴C was significantly affected by both plant and time factors, and there was a significant plant × time interaction. There were significant differences between plants in all plant-soil compartments examined as well as in the degree of the plant × time interaction.Differences in the¹⁴C distribution between plants were due to both interspecific and developmental variation. In general, when comparing¹⁵N and¹⁴C quantities between species, many of the differences found between plants can be explained by the differences determined in the weight of shoot or root parts. Despite the fact that amounts of C released were greater in ryegrass than in the other plant-treatments no unequivocal evidence was found to show that the effects of plant-microorganism interactions on soil-N mineralization were greater under ryegrass. Possible mechanisms accounting for the partitioning of N found among plant biomass, soil biomass and soil residues are discussed.     

Uptake of carbon and nitrogen derived from carbon-13 and nitrogen-15 dual-labeled maize residue compost applied to radish, komatsuna, and chingensai for three consecutive croppings

A pot experiment was conducted to determine the effects of the application of ¹³C (1.256 atom%) and ¹⁵N (1.098 atom%) dual-labeled maize residue compost (MRC) on the nitrogen and carbon uptake by radish, komatsuna, and chingensai as compared with the effect of inorganic fertilizer (IF). The vegetables were grown over three consecutive growing seasons over 4 months; compost was applied at the rate of 24 g kg^–1 soil. Nonlabeled nitrogen fertilizer was applied to the compost treatments in the second and third crops to compare the effects of blends of compost with N fertilizer to fertilizer alone. The N uptake and yield of vegetables were significantly higher with the recommended inorganic N treatment. The vegetables took up significantly (P < 0.05) lower amounts of N from MRC than from IFs during the three cultivations. The values of the N uptake derived by fertilizer application to the plant exhibited significant differences among different vegetables. Nitrogen recovered by komatsuna and chingensai from MRC was 7.3 (6.6%), 2.7 (1.8%), and 2.3, (1.7%) in the first, second, and third crops, respectively. Radish, komatsuna, and chingensai recovered significant amounts of C from MRC in the first and second crops, with negligible C recovery in the third crop. The initial loss of fertilizer C in soil at the first crop indicates that the microbial decomposition decoupled substantial amounts of ¹³C/¹⁵N-labeled compounds early in plant development, thus giving the microorganisms a preemptive competitive advantage in the acquisition of easily available ¹³C/¹⁵N-labeled substrates. It is concluded that a combination of compost and inorganic N did not supply sufficient plant-available N to increase vegetables yields or N uptake over those of fertilizer alone. The data suggested that higher productivity of vegetables might be achieved after the accumulation of a certain amount of residual compost N.     

Quantifying below-ground nitrogen of legumes

Quantifying below-ground nitrogen (N) of legumes is fundamental to understanding their effects on soil mineral N fertility and on the N economies of following or companion crops in legume-based rotations. Methodologies based on ¹⁵N shoot-labelling with subsequent measurement of ¹⁵N in recovered plant parts (shoots and roots) and in the root-zone soil have proved promising. We report four glasshouse experiments with objectives to develop appropriate protocols for in situ ¹⁵N labelling of the four legumes, fababean (Vicia faba), chickpea (Cicer arietinum), mungbean (Vigna radiata) and pigeonpea (Cajanus cajan). Treatments included ¹⁵N-urea concentration (0.1–2.0% w/w), feeding technique (leaf-flap and petiole), leaflet/petiole position (top and bottom of shoot) and frequency of feeding (one and two occasions). ¹⁵N-labelling via the leaf-flap was best for fababean, mungbean and pigeonpea, whilst petiole feeding was best for chickpea, in all cases at the lower-stem nodes 3 or 4 using 0.2 mL volumes of 0.5% urea (98 atom% ¹⁵N excess). Fed leaflets and petioles were removed within 2 weeks of labelling. Uneven ¹⁵N enrichment of the nodulated roots because of effects of the less-enriched nodules meant that root derived N in soil would be overestimated if recovered roots were more heavily nodulated than unrecovered roots. One possible solution would be to assume crown nodulation of the plants. Thus, recovered roots would be nodulated; root-derived N remaining in soil may be without nodules. The ratios of nodulated root to unnodulated root enrichments could then be used as an adjustment in the calculations, i.e. in the case of fababean and chickpea, by dividing calculated root-derived N in soil by 1.12 (fababean) and 1.56 (chickpea).     

Influence of light intensity on the distribution of carbon and consequent effects on mineralization of soil nitrogen in a barley (Hordeum vulgare L.)-soil system

A pot experiment was conducted in a ¹⁴C-labelled atmosphere to study the influence of living plants on organic-N mineralization. The soil organic matter had been labelled, by means of a 200-days incubation, with ¹⁵N. The influence of the carbon input from the roots on the formation of microbial biomass was evaluated by using two different light intensities (I). Mineralization of ¹⁵N-labelled soil N was examined by following its fate in both the soil biomass and the plants. Less dry matter accumulated in shoots and roots at the lower light intensity. Furthermore, in all the plant-soil compartments examined, with the exception of rhizosphere respiration, the proportion of net assimilated ¹⁴C was lower in the low-I treatment than in the high-I treatment. The lower rates of ¹⁴C and ¹⁵N incorporation into the soil biomass were associated with less root-derived ¹⁴C. During the chamber period (¹⁴CO₂-atmosphere), mineralized amounts of ¹⁵N (measured as plant uptake of ¹⁵N) were small and represented about 6.8 to 7.8% of the initial amount of organic ¹⁵N in the soil. Amounts of unlabelled N found in the plants, as a percentage of total soil N, were 2.5 to 3.3%. The low availability of labelled N to microorganisms was the result of its stabilization during the 210 days of soil incubation. Differences in carbon supply resulted in different rates of N mineralization which is consistent with the hypothesis that roots induce N mineralization. N mineralization was higher in the high-I treatment. On the other hand, the rate of mineralization of unlabelled stable soil N was lower than labelled soil ¹⁵N which was stabilized. The amounts of ¹⁵N mineralized in planted soil during the chamber period (43 days) which were comparable with those mineralized in unplanted soil incubated for 210 days, also suggested that living plants increased the turnover rate of soil organic matter.     

Carbon and nitrogen in the root-zone of barley (Hordeum vulgare L.) supplied with nitrogen fertilizer at two rates

Below-ground carbon (C) production and nitrogen (N) flows in the root-zone of barley supplied with high or low amounts of N-fertilizer were investigated. Interest was focused on the effect of the level of N-fertilizer on the production of root-derived C and on gross immobilization (i) and gross mineralization (m) rates. The plants were grown for 46 days in a sandy loam soil. Principles of pool dilution and changes in ¹⁵N pool abundances were used in conjunction with mathematical modelling to calculate the flows of N. N was applied at a high or a low rate, as (¹⁵NH₄)₂SO₄ solution (17.11 atom% ¹⁵N excess), before sowing. Nitrification was inhibited by using nitrapyrin (N-Serve). Pots were sampled four or five times during the experimental period, i.e. 0, 22, 30, 38 and 46 days after germination. On the three last sampling occasions, samples were also collected from pots in a growth chamber with ¹⁴C-labelled atmosphere.The release of ¹⁴C, measured as the proportion of the total ¹⁴C translocated below ground, was higher in the high-N treatment, but the differences between treatments were small. Our results were not conclusive in demonstrating that high-N levels stimulate the decomposition and microbial utilization of root-released materials. However, the internal circulation of soil-N, calculated N fluxes (m), which were in accordance with C mineralization rates and amounts of unlabelled N found in the plants (PU), suggested that the decomposition of native soil organic matter was hampered in the high-N treatment. Apparently, towards the end of the experimental period, microorganisms in the low-N treatment used C from soil organic matter to a greater extent than C they used from root released material, presumably because lower amounts of mineral N were available to microorganisms in the low-N treatment. Immobilization of N appeared to be soil driven (organisms decomposing soil organic matter account for the N demand) at low-N and root-driven (organisms decomposing roots and root-derived C account for the N demand) at high-N.Abbreviations AU Ammonium N-unlabelled - AL Ammonium N-labelled - AT Ammonium N-labelled and unlabelled (total) - NU Nitrate N-unlabelled - OU Organic N-unlabelled - OL Organic N-labelled - OT Organic N-total - PU Plant N-unlabelled (shoots and roots) - PL Plant N-labelled (shoots and roots) - PT Plant N-total (shoots and roots) - SL Sink or source of N-labelled - S Source or sink of N, mainly to and from the outer part of the cylinder - SU Sink or source of N-unlabelled - m Mineralization rate - i Immobilization rate - ua Uptake of ammonium - un Uptake of nitrate - la Loss of ammonium.     

Regulation of nitrogen partitioning in field-grown almond trees: Effects of fruit load and foliar nitrogen applications

Two treatments were employed to influence the amount of amino nitrogen (N) transport in phloem. In walnut trees (Juglans regia L.), developing fruit significantly reduced the efflux of foliar-applied ¹⁵N-enriched urea from treated spurs over a 33-day period in comparison with similarly-treated defruited spurs. Those data suggest that local aboveground demand for N influences vascular transport of amino N. In another experiment, a 1% urea solution was applied foliarly to 5-year old `Mission' almond trees [Prunus dulcis (Mill.) D. A. Webb] to increase the concentration of amino N in the phloem. The effect of foliar N treatments on a) the transport and distribution of labelled urea N within the trees over the experimental period and b) the uptake of soil-applied labelled N were determined by replicated whole tree excavation, fractionation into various tree components and mass spectrometric analyses of the ¹⁴N/¹⁵N ratios. Concentrations and composition of amino acids in the phloem and xylem saps of control trees and trees receiving foliar-applied urea were also determined. In foliar urea-treated trees, the amino acid concentrations increased significantly in leaf and bark phloem exudate, within 24 and 96 h, respectively. Foliar-applied urea N was translocated to the roots of almond trees over the experimental period and decreased soil N uptake. The results of these experiments are consistent with the hypothesis that aboveground N demand affects the amount of amino N cycling between shoots and roots, and may be involved in the regulation of soil N uptake. This revised version was published online in June 2006 with corrections to the Cover Date.     

Short-term effects of urea amended with the urease inhibitor N-(n-butyl) thiophosphoric triamide on perennial ryegrass

The current study investigated the short-term physiological implications of plant nitrogen uptake of urea amended with the urease inhibitor N-(n-butyl) thiophosphoric triamide (nBTPT) under both greenhouse and field conditions. ¹⁵N labelled urea amended with 0.0, 0.01, 0.1 and 0.5% nBTPT (w/w) was surface applied at a rate equivalent to 100 kg N ha^–1 to perennial ryegrass in a greenhouse pot experiment. Root, shoot and soil fractions were destructively harvested 0.75, 1.75, 4, 7 and 10 days after fertilizer application. Urease activity was determined in each fraction together with ¹⁵N recovery and a range of chemical analyses. The effect of nBTPT amended urea on leaf tip scorch was evaluated together with the effect of the inhibitor applied on its own on plant urease activity.nBTPT-amended urea dramatically reduced shoot urease activity for the first few days after application compared to unamended urea. The higher the nBTPT concentration the longer the time required for shoot activity to return to that in the unamended treatment. At the highest inhibitor concentration of 0.5% shoot urease activity had returned to that of unamended urea by 10 days. Root urease activity was unaffected by nBTPT in the presence of urea but was affected by nBTPT in the absence of urea.Transient leaf tip scorch was observed approximately 7–15 days after nBTPT + urea application and was greatest with high concentrations of nBTPT and high urea-N application rates. New developing leaves showed no visual sign of tip necrosis.Urea hydrolysis of unamended urea was rapid with only 1.3% urea-N remaining in the soil after 1.75 days. N uptake and metabolism by ryegrass was rapid with ¹⁵N recovery from unamended urea, in the plant (shoot + root) being 33% after 1.75 days. Most of the ¹⁵N in the soil following the urea+0.5% nBTPT application was still as urea after 1.75 days, yet ¹⁵N plant recovery at this time was 25% (root+shoot). This together with other evidence, suggests that if urea hydrolysis in soil is delayed by nBTPT then urea can be taken up by ryegrass as the intact molecule, albeit at a significantly slower initial rate of uptake than NH₄ ⁺-N. Protein and water soluble carbohydrate content of the plant were not significantly affected by amending urea with nBTPT however, there was a significant effect on the composition of amino acids in the roots and shoots, suggesting a difference in metabolism.Although nBTPT-amended urea affected plant urease activity and caused some leaf-tip scorch the effects were transient and short-lived. The previously reported benefit of nBTPT in reducing NH₃ volatilization of urea would appear to far outweigh any of the observed short-term effects, as dry-matter production of ryegrass is increased.     

Adaptation of methods for evaluating N2 fixation in food legumes and legume cover crops

Methods for partitioning the nitrogen assimilated by nodulated legumes, between nitrogen derived from soil sources and from N₂ fixation, are described as applied in peninsular Malaysia. The analysis of nitrogenous components translocated from the roots to the shoots of nodulated plants in the xylem sap is outlined, with some precautions to be observed for applications in the tropics. Some examples of the use of the technique in surverying apparent N₂ fixation by tropical legumes, in studying interrow cropping in plantation systems and in assessing effects of experimental treatments on N₂ fixation by food legumes, are described. Techniques for assesing N₂ fixation by means of¹⁵N abundance have been used to show that applications of nitrogenous fertilizers commonly used in Malaysia for soybeans depress N₂ fixation, that similar results are obtained with natural abundance and¹⁵N-enrichment methods and that, in at least two locations in Malaysia, differences between the natural abundance of¹⁵N in plant-available soil nitrogen and in atmospheric N₂ are great enough to permit application to measurement of N₂ fixation by leguminous crops.     

The role of VAM fungi in nitrogen dynamics in maize-bean intercrops

Nitrogen (N) transfer from N-fixing legumes via vesicular-arbuscular mycorrhizal (VAM) fungi to associated non-fixing plants has been demonstrated in greenhouse experiments. To date, this transfer has been shown only where mineral N is applied shortly before harvest, and hence is readily available. We have yet to demonstrate VAM-mediated N transfer where soil-N is limiting, a condition under which most traditional legume-nonlegume intercrops are grown.In this study, ¹⁵N-enriched soil (with 0.28%N) was used to distinguish between the uptake of soil- and atmospherically-derived N in maize grown with beans in the presence or absence of VAM fungi. VAM infection did not result in transfer of fixed N or soil N from bean to maize, despite a VAM-stimulated increase in N fixation in bean. In fact, beans were more competitive for soil N when mycorrhizal. N content in beans increased by 75% with a concomitant 22% decrease in mg N per maize plant. The competitive effect may have resulted from a VAM-mediated shift in carbon allocation in beans (but not maize) from shoots to roots.     

减量施氮对玉米-大豆套作系统下作物氮素吸收和利用效率的影响

为探索玉米-大豆套作系统中作物对N素吸收的差异特性,揭示减量施N对玉米-大豆套作系统的N高效利用机理。利用15N同位素示踪技术,结合小区套微区多年定位试验,研究了玉米单作(MM)、大豆单作(SS)、玉米-大豆套作(IMS)及不施N(NN)、减量施N(RN:180 kg N/hm2)、常量施N(CN:240 kg N/hm2)下玉米、大豆的生物量、吸N量、N肥利用率及土壤N素含量变化。结果表明,与MM(SS)相比,IMS下玉米茎叶及籽粒的生物量、吸N量降低,15N%丰度及15N吸收量增加,大豆籽粒及植株的生物量、吸N量及15N吸收量显著提高;IMS下玉米、大豆植株的N肥利用率、土壤N贡献率、土壤15N%丰度降低,15N回收率显著增加。施N与不施N相比,显著提高了单、套作下玉米、大豆植株的生物量、吸N量、15N丰度及15N吸收量;RN与CN相比,IMS下,RN的玉米、大豆植株总吸N量提高13.4%和12.4%,N肥利用率提高213.0%和117.5%,土壤总N含量提高12.2%和11.6%,土壤N贡献率降低12.0%和11.2%,玉米植株15N吸收量与15N回收率提高14.4%和52.5%,大豆的则降低57.1%和42.8%,单作与套作的变化规律一致。玉米-大豆套作系统中作物对N素吸收存在数量及形态差异,减量施N有利于玉米-大豆套作系统对N肥的高效吸收与利用,实现作物持续增产与土壤培肥。     

Fine Roots vs. Needles: A Comparison of 13C and 15N Dynamics in a Ponderosa Pine Forest Soil

Plant allocation patterns may affect soil C and N storage due to differences in litter quality and the depth of plant C and N inputs into the soil. We studied the dynamics of dual-labeled (¹³C/¹⁵N) Pinus ponderosa needles and fine roots placed at two soil depths (O and A horizon) in a temperate conifer forest soil during 2 y. Input of C as fine roots resulted in much more C retained in soil (70.5 ± 2.2% of applied) compared with needle C (42.9 ± 1.3% of applied) after 1.5 y. Needles showed faster mass loss, rates of soil ¹³CO₂ efflux, and more ¹⁵N immobilized into microbial biomass than did fine roots. The larger proportion of labile C compounds initially present in needles (17% more needle C was water soluble than in fine roots) likely contributed to its shorter C residence time and greater degree of transformation in the soil. A double exponential decay function best described the rate of ¹³C loss, with a smaller initial pulse of C loss from fine roots (S₁k₁) and a slower decay rate of the recalcitrant C pool for fine roots (0.03 y⁻¹) compared with (0.19 y⁻¹) for needles. Soil ¹³C respiration, representing heterotrophic respiration of litter C, was much more seasonal from the O horizon than from the A. However, offsetting seasonal patterns in ¹³C dynamics in the O horizon resulted in no net effect of soil depth on total ¹³C retention in the soil after 1.5 y for either litter. Almost 90% of applied litter N was retained in the soil after 1.5 y, independent of litter quality or soil depth. Very small amounts of ¹³C or ¹⁵N (<3% of applied) moved to the horizon above or below the placement depth (i.e., O to A or A to O). Our results suggest that plant allocation belowground to fine roots results in more C retained and less N mineralized compared with allocation aboveground to needles, primarily due to litter quality differences.     

15N abundance of surface soils,roots and mycorrhizas in profiles of European forest soils

¹⁵N natural abundances of soil total N, roots and mycorrhizas were studied in surface soil profiles in coniferous and broadleaved forests along a transect from central to northern Europe. Under conditions of N limitation in Sweden, there was an increase in ¹⁵N of soil total N of up to 9% from the uppermost horizon of the organic mor layer down to the upper 0–5 cm of the mineral soil. The ¹⁵N of roots was only slightly lower than that of soil total N in the upper organic horizon, but further down roots were up to 5% depleted under such conditions. In experimentally N-enriched forest in Sweden, i.e. in plots which have received an average of c. 100 kg N ha^–1 year^–1 for 20 years and which retain less than 50% of this added N in the stand and the soil down to 20 cm depth, and in some forests in central Europe, the increase in ¹⁵N with depth in soil total N was smaller. An increase in ¹⁵N of the surface soil was even observed on experimentally N-enriched plots, although other data suggest that the N fertilizer added was depleted in¹⁵N. In such cases roots could be enriched in¹⁵N relative to soil total N, suggesting that labelling of the surface soil is via the pathway: — available pools of N-plant N-litter N. Under N-limiting conditions roots of different species sampled from the same soil horizon showed similar ¹⁵N. By contrast, in experimentally N-enriched forest ¹⁵N of roots increased in the sequence: ericaceous dwarf shrubs15N enriched compounds in fungal material, which could contribute to explain the observed ¹⁵N profiles if fungal material is enriched, because it is a precursor of stable organic matter and recalcitrant N. This could act in addition to the previous explanation of the isotopically lighter soil surface in forests: plant uptake of ¹⁵N-depleted N and its redeposition onto the soil surface by litter-fall.