Nitrogen and phosphorus requirements for raising mycorrhizal seedlings of Leucaena leucocephala in containers

A greenhouse study was undertaken to determine the nitrogen and phosphorus fertilization requirements for raising mycorrhizal seedlings in soil in containers. Seedlings of Leucaena leucocephala were grown for 40 days in dibble tubes containing fumigated or nonfumigated soil uninoculated or inoculated with Glomus aggregatum. The soil was fertilized with NH₄NO₃ solution to obtain 25–200 mg N kg^-1 soil, and with a KH₂PO₄ solution to establish target soil solution P concentrations of 0.015–0.08 mg P l^-1. At the end of 40 days, seedlings were transplanted into pots containing 5-kg portions of fumigated soil. Posttransplant vesicular arbuscular mycorrhizal fungal (VAMF) effectiveness, measured as pinnule P content, plant height, shoot dry weight and tissue N and P concentrations, was significantly increased by pretransplant VAMF colonization in both soils. The best posttransplant mycorrhizal colonization and mycorrhizal growth responses were observed if the nonfumigated pretransplant soil was amended with 50 mg N kg^-1 soil and 0.04 mg P l^-1 or if the fumigated pretransplant soil was amended with 100 mg N kg^-1 soil and 0.04 mg P 1^-1. There was no relationship between NP ratios of nutrients added to the pretransplant soil medium and shoot NP ratios observed after transplanting. Shoot NP ratio was also not correlated with root colonization level.Contribution from the Hawaii Institute of Tropical Agriculture and Human Resources Journal Series No. 4025     

Emission of gaseous nitrogen oxides from an extensively managed grassland in NE Bavaria, Germany.

We analysed the stable isotope composition of emitted N₂O in a one-year field experiment (June 1998 to April 1999) in unfertilized controls, and after adding nitrogen by applying slurry or mineral N (calcium ammonium nitrate). Emitted N₂O was analysed every 2–4 weeks, with additional daily sampling for 10 days after each fertilizer application. In supplementary soil incubations, the isotopic composition of N₂O was measured under defined conditions, favouring either denitrification or nitrification. Soil incubated for 48 h under conditions favouring nitrification emitted very little N₂O (0.024 mol g_dw ^–1) and still produced N₂O from denitrification. Under denitrifying incubation conditions, much more N₂O was formed (0.91 mol g_dw ^–1 after 48 h). The isotope ratios of N₂O emitted from denitrification stabilized at ¹⁵N = –40.8 ± 5.7 and ¹⁸O = 2.7 ± 6.3. In the field experiment, the N₂O isotope data showed no clear seasonal trends or treatment effects. Annual means weighted by time and emission rate were ¹⁵N = –8.6 and ¹⁸O = 34.7 after slurry application, ¹⁵N = –4.6 and ¹⁸O = 24.0 after mineral fertilizer application and ¹⁵N = –6.4 and ¹⁸O = 35.6 in the control plots, respectively. So, in all treatments the emitted N₂O was ¹⁵N-depleted compared to ambient air N₂O (¹⁵N = 11.4 ± 11.6, ¹⁸O = 36.9 ± 10.7). Isotope analyses of the emitted N₂O under field conditions per se allowed no unequivocal identification of the main N₂O producing process. However, additional data on soil conditions and from laboratory experiments point to denitrification as the predominant N₂O source. We concluded (1) that the isotope ratios of N₂O emitted from the field soil were not only influenced by the source processes, but also by microbial reduction of N₂O to N₂ and (2) that N₂O emission rates had to exceed 3.4 mol N₂O m^–2 h^–1 to obtain reliable N₂O isotope data.     

15N natural abundance studies in Australian commercial sugarcane

The measurement of natural ¹⁵N abundance is a well-established technique for the identification and quantification of biological N₂ fixation in plants. Associative N₂ fixing bacteria have been isolated from sugarcane and reported to contribute potentially significant amounts of N to plant growth and development. It has not been established whether Australian commercial sugarcane receives significant input from biological N₂ fixation, even though high populations of N₂ fixing bacteria have been isolated from Australian commercial sugarcane fields and plants. In this study, ¹⁵N measurements were used as a primary measure to identify whether Australian commercial sugarcane was obtaining significant inputs of N via biological N₂ fixation. Quantification of N input, via biological N₂ fixation, was not possible since suitable non-N₂ fixing reference plants were not present in commercial cane fields. The survey of Australian commercially grown sugarcane crops showed the majority had positive leaf ¹⁵N values (73% >3.00, 63% of which were >5.00), which was not indicative of biological N₂ fixation being the major source of N for these crops. However, a small number of sites had low or negative leaf ¹⁵N values. These crops had received high N fertiliser applications in the weeks prior to sampling. Two possible pathways that could result in low ¹⁵N values for sugarcane leaves (other than N₂ fixation) are proposed; high external N concentrations and foliar uptake of volatilised NH₃. The leaf ¹⁵N value of sugarcane grown in aerated solution culture was shown to decrease by approximately 5 with increasing external N concentration (0.5–8.0 mM), with both NO₃ ^– and NH₄ ⁺ nitrogen forms. Foliar uptake of atmospheric NH₃ has been shown to result in depleted leaf ¹⁵N values in many plant species. Acid traps collected atmospheric N with negative ¹⁵N value (–24.45±0.90) from above a field recently surface fertilised with urea. The ¹⁵N of leaves of sugarcane plants either growing directly in the soil or isolated from soil in pots dropped by 3.00 in the same field after the fertiliser application. Both the high concentration of external N in the root zone (following the application of N-fertilisers) and/or subsequent foliar uptake of volatilised NH₃ could have caused the depleted leaf ¹⁵N values measured in the sugarcane crops at these sites.     

Failure of an iterative curve-fitting procedure to successfully estimate two organic N pools

Fitting a double negative exponential function to N mineralization data can be used to characterize two organic nitrogen pools; an easily decomposable (N_dpm) and a resistant one (N_rpm). The relevance of those two calculated N mineralization pools was investigated by adding easily decomposable organic material to soils. Soil amended with crop residues of sugar-beet or bean was mixed with an equal amount of coarse sand, incubated at 35 °C and leached at specific time-intervals. Upon leaching, NH₄ ⁺ and NO₃ ^- were measured in the extracts. A double negative exponential function was fitted to the data and two organic N pools were defined. Fitting a double negative exponential function to N mineralization data to characterize an active and resistant organic N pool was sometimes impossible; the N mineralization data did not always resemble a negative exponential function. Additionally, the size of the two pools calculated were not constant with time and were often meaningless; the N_rpm pool was greater than the soil organic N content, the size of the N_rpm pool was smaller than the N_dpm pool or one of the N pools was negative. Relevant values for both N_rpm and N_dpm which were consistent with incubation time were only obtained when excessive amounts of organic material, normally not dealt with in the field, were applied.     

δ15N as an indicator of N2-fixation by cyanobacterial mats in tropical marshes

Cyanobacterial mats (CBM) are important components of wetland ecosystems in limestone-based regions of the Caribbean. During two sampling periods (July 1999 and January 2000) we measured N₂-fixation in samples from 23 different marshes simultaneously with measurements of relevant environmental factors. Samples were evaluated for abundance of five groups of cyanobacteria: (1) Leptolyngbya, (2) Oscillatoria, (3) Chroococcales, (4) Nostoc-& Stigonematales, and (5) dead sheaths. Differences in nitrogen fixation, expressed as nitrogenase activity in nmol C₂H₄ cm^–2 h^–1, were best explained by the proportion of heterocyst-forming cyanobacteria. The samples were analyzed for the natural abundance of ¹⁵N. ¹⁵N values ranged from –1.99 to 11.44 and were strongly negatively correlated with N₂-fixation. With all data included, ¹⁵N was also strongly correlated with nitrates in water. With the samples from Little Belize (high nitrate content marshes) excluded, the effect of nitrate became insignificant. N₂-fixation predicted from ¹⁵N measured on an independent data set from September 2000 was moderately accurate (r² = 0.68, 0.52 and 0.54 for predictions based on July 1999, January 2000 and combined data sets, respectively). When individual sample sets were divided into two groups with ¹⁵N < 2 and ¹⁵N > 2, the two groups were always highly significantly different in terms of their N₂-fixation. The presented evidence suggests that ¹⁵N can be used as a reliable indicator of N₂-fixation by CBM.     

Nitrogen mineralization in relation to C:N ratio and decomposability of organic materials

A desk study was made on N mineralization of various organic materials. Data were obtained from the literature containing information on mineralized organic N, C/N of the substrate and some index for decomposability. Per class of substrate decomposability, linear regression lines between N mineralization and substrate C/N were established, from which apparent initial age and humification coefficient were calculated to indicate decomposability. For quantitative grading, a resistance index was formulated comprising the concentrations of lignin and polyphenols. The results confirmed that the fraction of mineralized organic N is linearly related to substrate C/N for equally decomposable materials. The nature of the organic materials and the processes to which they had been subjected were reflected in the ranking by decomposability.     

Formation of nucleoside 5′-polyphosphates from nucleotides and trimetaphosphate

Summary When solutions of nucleoside 5-phosphates and trimetaphosphate are dried out at room temperature, nucleoside 5-polyphosphates are formed. The Mg⁺⁺ ion shows a superior catalytic function in this reaction when compared with other divalent metal ions. Starting with nucleoside 5-phosphates, Mg⁺⁺ and trimetaphosphate, the predominant products in the nucleoside 5-polyphosphate series p_nN are p₄N, p₇N and P₁₀N. Nucleoside 5-diphosphates yield p₅N and p₈N, nucleoside 5-triphosphates give p₆N and p₉N. The prebiological relevance of these reactions is discussed.Abbreviations P_n (n = 1,2,3,) linear polyphosphate containing n phosphate residues - P₃! trimetaphosphate - A adenosine - U uridine - dA 2-dexyadenosine - T thymidine - P_nN nucleoside 5-polyphosphate containing n phosphate residues, e.g. with N = A and n = 4 - p₄A adenosine 5-tetraphosphate     

Mineralization of labeled N from cowpea [Vigna unguiculata (L.) Walp.] plant parts at two growth stages in sandy soil

Cowpea [Vigna unguiculata (L). Walp.] has great potential as green manure due to its rapid N accumulation and efficient N₂ fixation. The objective of this study was to measure the rate of N mineralization from cowpea plant parts harvested at onset of flowering (5 weeks) and mid pod-fill (7 weeks) under near optimum conditions. Cowpeas were grown in a greenhouse and supplied with ¹⁵NH₄ ¹⁵NO₃ to isotopically label tissue. Cowpea leaves, stems, and roots were incorporated into a sandy soil (Psammentic Paleustalf) and net N mineralized was measured several times during a 10 week incubation. The amount of N accumulated in 7-week old cowpeas was more than double that in 5-week old cowpeas. The portion of N mineralized after 10 weeks was 24% for 5-week old cowpeas and 27% for 7-week old cowpeas. The rate of N mineralization from leaves and stems increased with plant age, but decreased for roots. The amount of N mineralized from 7-week old cowpeas was more than double (235%) that from 5-week old cowpeas due to greater N accumulation and a more rapid rate of N mineralization of the more mature cowpeas. The greatest amount of N was released from leaves, which amounted to 74 and 65% of total N mineralization from 5- and 7-week old cowpeas, respectively. The percentage of N mineralized by 10 weeks was linearly related to the tissue N concentration of the plant parts and to their C/N ratio. These relationships allow a quick estimation of the amount of N that would mineralize from cowpea residues incorporated into soil based on their N concentration or C/N ratio.     

Method of measuring invertase activity in soils

Summary Invertase (-D-fructofuranoside fructohydrolase, EC [Enzyme Commission] 3.2.1.26) is the enzyme that catalyzes the hydrolysis of sucrose and yields glucose and fructose. The activity of this enzyme was monitored by systematically developing a sensitive and rapid method to detect reducing sugars with the precision of 1.4 to 6.1% C.V. The method involves the colorimetric determination of reducing sugars which react with 3,5-dinitrosalicylic acid when soil is incubated with buffered sucrose solution and toluene at 37°C for 24 h. The detection limit for the method described is 100 g of reducing sugar per ml of soil extract. The color intensity remained constant up to 24 h. Comparative studies showed that the method described was in good agreement to other invertase assay procedures reported in the literature.Studies on the stability and distribution of invertase in soils by using the method described showed that air-drying of field-moist soil samples resulted in decreased activity ranging from 15.3 to 23.7% (avg.=19.8%). Statistical analyses indicated that invertase activity was significantly correlated with total N (r=0.78^***) and organic C (r=0.70^***) in the topsoil of 19 diverse samples. There was no significant correlation between invertase activity and soil pH, cation exchange capacity, percentage of clay and percentage of sand. The activity of this enzyme was concentrated in surface soils and decreased with profile depth. Regression analyses showed that invertase activity was significantly correlated with organic carbon content of three soil profiles examined.     

Interpretation of sulfur cycling in two catchments in the Black Forest (Germany) using stable sulfur and oxygen isotope data

The isotopic composition of SO ₄ ^2- in bulk precipitation, canopy throughfall, seepage water at three different soil depths, stream water, and groundwater was monitored in two forested catchments in the Black Forest (Germany) between November 1989 and February 1992. Isotope measurements on aqueous sulfate were complemented by ³⁴S-analyses on SO₂ in the air, total sulfur and inorganic sulfate in the soil, and bedrock sulfur, in order to identify sources and biogeochemical processes affecting S cycling in catchments with base poor, siliceous bedrock. Stable S isotope data indicated that atmospheric deposition and not mineral weathering is the major source of S in both catchments since ³⁴S-values for sulfate in the soil, in seepage water, and in stream water were generally found to be similar to the mean ³⁴S-values of precipitation SO ₄ ^2- (+2.1. However, ¹⁸O-values of seepage water SO ₄ ^2- at 30 cm and especially at 80 cm depth were depleted by several per mil with respect to those of the atmospheric deposition (+7.5 to +13.5. This indicates that in both catchments a considerable proportion of the seepage water SO ₄ ^2- is derived from mineralization of carbon-bonded soil S and must therefore have cycled through the organic soil S pool. ³⁴S-values for different S compounds in the solid soil were found to differ markedly depending on S fraction and soil depth. Since atmospheric S deposition with rather constant ³⁴S-values was identified as the dominant S source in both catchments, this is interpreted as a result ofin situ isotope fractionation rather than admixture of isotopically different S. The differences between the ³⁴S-values of seepage water and soil sulfate and those of organic soil S compounds are consistent with a model in which SO ₄ ^2- uptake by vegetation and soil microorganisms favours³⁴SO ₄ ^2- slightly, whereas during mineralization of organic soil S to aqueous SOSO ₄ ^2- ,³²S reacts preferentially. However, the data provide evidence for negligible isotope fractionation during physico-chemical S transformations such as adsorption/desorption in aerated forest soils.     

Evidence suggesting energy-dependent formaldehyde transport in an RuMP-type methylotroph (T15)

Formaldehyde accumulation ratios ([¹⁴CH₂O]_i/[¹⁴CH₂O]_o) as high as 12-fold were measured in anaerobic, CH₃OH-energized, whole cell suspensions of the ribulose monophosphate (RuMP)-type methylotrophic strain T15. Uptake kinetics were extremely rapid, enabling the attainment of equilibrium in only 10–30 s. Transport appears to be energy-dependent and associated with the protonmotive force (pmf). Anaerobic incubation with 5 M carbonyl p-(trifluoromethoxy)-phenylhydrazone (FCCP) led to 70%–90% reduction of the accumulation ratio. Though not as pronounced, diminished uptake was also observed in the presence of 140 M nigericin, 161 M valinomycin and 90 mM KSCN, commensurate with their effects on pmf. Accumulation of CH₂O as a function of external pH followed a trend more similar to that of pmf than either pH or . Preventing energization by incubation with 100 M N,N-dicyclohexylcarbodiimide (DCCD) led to nearly 80% inhibition of CH₂O transport. Over short time periods it was possible to chase accumulated ¹⁴CH₂O from previously loaded cells by collapsing pmf; however, this technique also indicated that significant ¹⁴CH₂O incorporation began to occur within 3 min.Abbreviations FCCP Carbonyl cyanide p-(trifluoromethyoxy)-phenylhydrazone - DCCD N,N-dicyclohexylcarbodiimide - RuMP ribulose monophosphate - TPP⁺ tetra[U-¹⁴C]phenylphosphonium - pmf protonmotive force     

Formation of nucleoside 5′-polyphosphates under potentially prebiological conditions

Summary Aqueous solutions of linear inorganic polyphosphates incubated in presence of Mg ions depolymerize to give trimetaphosphate. The presence of a nucleoside 5-phosphate has little influence upon the reaction. Drying the products obtained by incubating a linear polyphosphate with Mg ions in the presence of a nucleoside 5-phosphate yields nucleoside 5-polyphos-phates. The prebiological relevance of the reactions is discussed.Abbreviations P_n(n=1,2,3,) linear polyphosphate containing n phosphate residues - P₃! trimetaphosphate - A adenosine - p_nN nucleoside 5-polyphosphate containing n phosphate residues, e.g. with N = A, n = 4 - p₄N adenosine 5-tetraphosphate - _P ^* _lp_mA p_nA, (n = 1 + m); adenosine 5-polyphosphate containing n phosphate units with³³p-label on terminal 1 phosphate groups     

Measurement of one-bond 15N-13C′ dipolar couplings in medium sized proteins

A simple and accurate method is described for measurement of ¹ J _CN splittings in isotopically enriched proteins. The method is of the quantitative J correlation type, and the ¹ J _CN splitting is derived from the relative intensity in two 3D TROSY-HNCO spectra with ¹ J _CN dephasing intervals of 1/(2¹ J _CN) (reference intensity) and 1/¹ J _CN (residual intensity). If the two spectra are recorded under identical conditions and with the same number of scans, the random error in the ¹ J _CN value extracted in this manner is inversely related to the signal-to-noise (S/N) in the reference spectrum. A S/N of 30:1 in the reference spectrum yields random errors of less than 0.2 Hz in the extracted ¹ J _CN value. Dipolar couplings obtained from the difference in ¹ J _CN splitting in the isotropic and liquid crystalline phase for the C-terminal domain of calmodulin are in excellent agreement with its 1.68-Å crystal structure, but agree considerably less with the 2.2-Å structure.     

Nitrogen dynamics in alpine ecosystems of the northern Caucasus

Net N mineralization, nitrification, microbial biomass N and ¹⁵N natural abundance were studied in a toposequence of representative soils and plant communities in the alpine zone of the northern Caucasus. The toposequence was represented by (1) low-productive alpine lichen heath (ALH) of wind-exposed ridge and upper slope; (2) more productive Festuca varia grassland (FG) of middle slope; (3) most productive Geranium gymnocaulon/Hedusarum caucasicummeadow (GHM) of lower slope; (4) low-productive snowbed community (SBC) of the slope bottom. N availability, net N mineralization and nitrification were higher in soils of alpine grassland and meadow of the middle part of the toposequence compared with soils of lichen heath and snowbed community of extreme habitats in the alpine zone. There was no correlation between intensities of N transformation processes and favorable (low soil acidity, low C/N ratio, long vegetation period, relatively high temperature, absence of hydromorphic features) and unfavorable (opposite) factors, indicating that the intensity of N mineralization and nitrification in the alpine soils is controlled by a complex combination of these factors. Potential net N mineralization and nitrification in alpine soils determined in the short-term laboratory incubation were considerably higher than those determined in the long-term field incubation. The differences of potential nitrification between soils of various plant communities did not correspond to the field determined pattern indicating the importance of on-site climatic conditions for control of nitrification in high mountains. The result of comparison of N transformation potentials in incubated and native soils indicated that nitrification potential was significantly increased after long-term soil incubation. It means that net nitrification determined in the field was probably overestimated, especially in the meadow soils. A soil translocation experiment indicated that low temperature was an important factor limiting net N mineralization and nitrification in alpine soils: net N mineralization and especially nitrification increased when alpine soils were translocated into the subalpine zone and mean annual temperature increased by about 3°C. Additional N input increased N availability (NH₄ ⁺-N) and potential nitrification in soils of the lower part of the toposequense (GHM and SBC), and potential net N mineralization in two soils of extreme habitats (ALH and SBC). A positive correlation was found between soil ¹⁵N and net N mineralization and nitrification; the relative ¹⁵N enrichment was characteristic of grassland and meadow ecosystems. ¹⁵N of total soil N pool increased during the field mineralization experiment; there was a positive tendency between the change in ¹⁵N and net N mineralization and nitrification, however the relationship was not significant. Foliar ¹⁵N of dominant plant species varied widely within community, however, a tendency of higher foliar ¹⁵N for species growing on the soils with higher net N mineralization, nitrification and ¹⁵N was observed.     

Leaf 15N abundance of subarctic plants provides field evidence that ericoid,ectomycorrhizal and non-and arbuscular mycorrhizal species access different sources of soil nitrogen

The natural abundance of the nitrogen isotope 15, ¹⁵N, was analysed in leaves of 23 subarctic vascular plant species and two lichens from a tree-line heath at 450 m altitude and a fellfield at 1150 m altitude close to Abisko in N. Sweden, as well as in soil, rain and snow. The aim was to reveal if plant species with different types of mycorrhizal fungi also differ in their use of the various soil N sources. The dwarf shrubs and the shrubs, which in combination formed more than 65% of the total above-ground biomass at both sites, were colonized by ericoid or ectomycorrhizal fungi. Their leaf ¹⁵N was between–8.8 and–5.5 at the heath and between–6.1 and –3.3 at the fellfield. The leaf ¹⁵N of non- or arbuscular mycorrhizal species was markedly different, ranging from –4.1 to –0.4 at the heath, and from –3.4 to+2.2 at the fellfield. We conclude that ericoid and ectomycorrhizal dwarf shrubs and shrubs utilize a distinct N source, most likely a fraction of the organic N in fresh litter, and not complexed N in recalcitrant organic matter. The latter is the largest component of soil total N, which had a ¹⁵N of –0.7 at the heath and +0.5 at the fellfield. Our field-based data thus support earlier controlled-environment studies and studies on the N uptake of excised roots, which have demonstrated protease activity and amino acid uptake by ericoid and ectomycorrhizal tundra species. The leaves of ectomycorrhizal plants had slightly higher ¹⁵N (fellfield) and N concentration than leaves of the ericoids, and Betula nana, Dryas octopetala and Salix spp. also showed NO _inf3 ^sup- reductase activity. These species may depend more on soil inorganic N than the ericoids. The ¹⁵N of non- or arbuscular mycorrhizal species indicates that the ¹⁵N of inorganic N available to these plants was higher than that of average fresh litter, probably due to high microbial immobilization of inorganic N. The ¹⁵N of NH _inf4 ^sup+ -N was +12.3 in winter snow and +1.9 in summer rain. Precipitation N might be a major contributer in species with poorly developed root systems, e.g. Lycopodium selago. Our results show that coexisting plant species under severe nutrient limitation may tap several different N sources: NH _inf4 ^sup+ , NO _inf3 ^sup- and organic N from the soil, atmospheric N₂, and N in precipitation. Ericoid and ectomycorrhizal fungi are of major importance for plant N uptake in tundra ecosystems, and mycorrhizal fungi probably exert a major control on plant ¹⁵N in organic soils.     

Sources of nitrate in rivers draining sixteen watersheds in the northeastern U.S.: Isotopic constraints

The feasibility of using nitrogen and oxygenisotope ratios of nitrate (NO₃ ^–) forelucidating sources and transformations ofriverine nitrate was evaluated in a comparativestudy of 16 watersheds in the northeastern U.S.A. Stream water was sampled repeatedly at theoutlets of the watersheds between January andDecember 1999 for determining concentrations,¹⁵N values, and ¹⁸Ovalues of riverine nitrate.In conjunction with information about land useand nitrogen fluxes,¹⁵N_nitrate and¹⁸O_nitrate values providedmainly information about sources of riverinenitrate. In predominantly forested watersheds,riverine nitrate had mean concentrations ofless than 0.4 mg NO₃ ^–-N L^–1,¹⁵N_nitrate values of lessthan +5, and ¹⁸O_nitratevalues between +12 and +19. This indicatesthat riverine nitrate was almost exclusivelyderived from soil nitrification processes withpotentially minor nitrate contributions fromatmospheric deposition in some catchments. Inwatersheds with significant agricultural andurban land use, concentrations of riverinenitrate were as high as 2.6 mg NO₃ ^–-NL^–1 with ¹⁵N_nitratevalues between +5 and +8 and¹⁸O_nitrate values generallybelow +15. Correlations between nitrateconcentrations, ¹⁵N_nitratevalues, and N fluxes suggest that nitrate inwaste water constituted a major, and nitrate inmanure a minor additional source of riverinenitrate. Atmospheric nitrate deposition ornitrate-containing fertilizers were not asignificant source of riverine nitrate inwatersheds with significant agricultural andurban land use. Although complementary studiesindicate that in-stream denitrification wassignificant in all rivers, the isotopiccomposition of riverine nitrate sampled at theoutlet of the 16 watersheds did not provideevidence for denitrification in the form ofelevated ¹⁵N_nitrate and¹⁸O_nitrate values. Relativelylow isotopic enrichment factors for nitrogenand oxygen during in-stream denitrification andcontinuous admixture of nitrate from theabove-described sources are thought to beresponsible for this finding.     

Estimation of symbiotic dinitrogen fixation in alder forest by the method based on natural15N abundance

Annual N₂-fixation in virgin forest ecosystems has been measured using a¹⁵N natural abundance (¹⁵N) procedure. This method was compared to a¹⁵N labelled fertilizer isotopic dilution method. For young alders (5–6 years old), ¹⁵N of leaves gave results in good agreement with the isotopic dilution of fertilizer method. Since ¹⁵N variability was expected according to plant physiology, for alder trees, leaves were collected at various heights after the end of the growing season, and, to take account of isotopic variations coming from derived inputs, ¹⁵N of leaves of a large number of other plants in the same are were measured to give control values. Following this procedure, the ¹⁵N method gave reliable evaluation of the nitrogen supply, by through N₂-fixation, to alders, which were found to maintain high nitrogen fixing capacity in a sequence ranging from first stage of establishment of climactic formation. Moreover, the same method is reported to discriminate various origins ofAlnus glutinosa grown in natural conditions, possibly in relation to the genetic diversity of this species.     

Short-term patterns in water and nitrogen acquisition by two desert shrubs following a simulated summer rain

A field experiment was conducted at the Jornada Long-Term Ecological Research (LTER) site in the Chihuahuan Desert of New Mexico to compare the rapidity with which the shrubs Larrea tridentata and Prosopis glandulosa utilized water, CO² and nitrogen (N) following a simulated summer rainfall event. Selected plants growing in a roughly 50-m² area were assigned to treatment and control groups. Treatment plants received the equivalent of 3 cm of rain, while no supplemental water was added to the control plants. Xylem water potential (_x) and net assimilation rate (A_net) were evaluated one day before and one and three days after watering. To monitor short-term N uptake, soils around each plant were labeled with eight equally distant patches of enriched ¹⁵N before watering. Each tracer patch contained 20 ml of 20 mM ¹⁵ NH₄ ¹⁵NO³ (99 atom%) solution applied to the soil at 20 cm from the center of the plant at soil depths of 10 and 20 cm. Nitrogen uptake, measured as leaf ¹⁵N, was evaluated at smaller time intervals and for a longer period than those used for _x and A^net. Both A^net and ^x exhibited a significant recovery in watered vs. control Larrea plants within 3 days after the imposition of treatment, but no such recovery was observed in Prosopis in that period. Larrea also exhibited a greater capacity for N uptake following the rain. Leaf ¹⁵N was five-fold greater in watered compared to unwatered Larrea plants within 2 days after watering, while foliar ¹⁵N was not significantly different between the watered and unwatered Prosopis plants during the same period. Lack of a significant change in root ¹⁵ NO^{– 3} uptake kinetics of Larrea, even three days after watering, indicated that the response of Larrea to a wetting pulse may have been due to a greater capacity to produce new roots. The differential ability of these potential competitors in rapidly acquiring pulses of improved soil resources following individual summer rainfall events may have significant implications for the dynamic nature of resource use in desert ecosystems.     

Preliminary studies of the impact of excreted N on cycling and uptake of N in pasture systems using natural abundance stable isotopic discrimination

The possibility of using natural abundance techniques to determine N transformations and flows after deposition of cattle dung has been examined. These preliminary results showed that ¹⁵N in dung was greater than in plants growing in association with particular pats. This, and other observational information, indicated that dung pats of different ages were being examined. There were significant variations in plant ¹⁵N signatures within and between species grown in association and away from the dung. It was probable that variation in plant ¹⁵N was brought about by changes in soil mineral N pools after transfer of N derived from the dung. This resulted in different ¹⁵N signatures in Trifolium repens (because of changes in N utilization from soil or atmospheric pools), in Lolium perenne (because of changes in ¹⁵N in soil mineral N), but not in Ranunculus repens (because the majority of active roots were outside the range of immediate influence of the deposited dung). The differences in ¹⁵N allowed the development of hypotheses for changes in soil N pools and the acquisition of N by plants from soil, dung or atmospheric sources.     

Pulse schemes for the measurement of3 JC′Cγ and3 JNCγ scalar couplings in 15N,13C uniformly labeled proteins

Pulse sequences are presented for the measurement of³J_CC and³J_NC scalar couplings for allC containing residues in¹⁵N,¹³C uniformly labeled proteins. The methodsdescribed are based on quantitative J correlation spectroscopy pioneered byBax and co-workers [Bax et al. (1994) Methods Enzymol., 239, 79–105].The combination of ³J_CC and³J_NC scalar coupling constants allows theassignment of discrete rotameric states about the ₁ torsion angle in cases where such states exist or, alternatively,facilitates the establishment of noncanonical ₁conformations or the presence of rotameric averaging. The methods areapplied to a 1.5 mM sample of staphylococcal nuclease.