Effects of N-application on utilization of 15N and 13C and quality in two wheat cultivars

We studied N uptake and distribution in wheat, and the incorporation of nitrogen and carbon into gluten and non-gluten proteins using a double-labelling approach with ¹⁵N and ¹³C. Doses of N-fertilizer were split and applied at emergence, onset of stem elongation, and heading at rates of 280/140/140 mg N pot^?1, respectively simulating 90/45/45 kg N ha^?1. Five different combinations of N-fertilizations containing no or 10 % ¹⁵N were performed. The recovery of ¹⁵N added at the stages emergence, stem elongation or heading were 42, 60, and 64 %. Application of ¹⁵N at all three stages yielded in 51 % recovery. Remobilisation of straw N was greater for Golia. The ¹⁵N concentration in gluten proteins of Golia show higher values than Gönen. The ratio of ¹⁵N gluten/¹⁵N non-gluten proteins of Golia were higher, which implies a lower non-gluten protein activity during grain filling. The ¹³C concentration in gluten and non-gluten proteins did not differ between both cultivars.     

Effects of N-application on utilization of 15N and 13C and quality in two wheat cultivars

We studied N uptake and distribution in wheat, and the incorporation of nitrogen and carbon into gluten and non-gluten proteins using a double-labelling approach with 15N and 13C. Doses of N-fertilizer were split and applied at emergence, onset of stem elongation, and heading at rates of 280/140/140 mg N pot(-1), respectively simulating 90/45/45 kg N ha(-1). Five different combinations of N-fertilizations containing no or 10 % 15N were performed. The recovery of 15N added at the stages emergence, stem elongation or heading were 42, 60, and 64 %. Application of 15N at all three stages yielded in 51 % recovery. Remobilisation of straw N was greater for Golia. The 15N concentration in gluten proteins of Golia show higher values than Gonen. The ratio of 15N gluten/15N non-gluten proteins of Golia were higher, which implies a lower non-gluten protein activity during grain filling. The 13C concentration in gluten and non-gluten proteins did not differ between both cultivars.     

The effect of different fish feed compositions on δ13C and δ15N signatures of sea bass and its potential value for tracking mariculture-derived nutrients

Increasing demand for fish and seafood calls for an expansion of aquaculture production. At the same time, the status of the marine environment must not be jeopardised. Stable isotopes are potential markers for tracking feed-based nutrient flows from aquaculture into marine biota. Here, we demonstrate how four experimental diets (main protein components: fishmeal, soya protein concentrate, wheat gluten, and Jatropha kernel meal) and a commercial diet induce characteristic δ¹³C and δ¹⁵N signals in sea bass (Dicentrarchus labrax) during nine weeks of laboratory feeding under replicate conditions. The plant-protein-based diets containing wheat gluten and soya, and the commercial feed consistently induced the largest isotopic differentiation of the fish, both from the feed source and from the pre-experimental condition. The large difference of the fish on plant-protein-based diets compared to the range of natural isotopic variation in the macrozoobenthos of the North Sea lends support to the idea that plant-based feeds are suitable for tracing mariculture-derived organic matter under practical conditions. The commercial feed had a similar effect as the experimental feeds and would be a cost-effective option for an offshore aquaculture experiment.     

Maternal obesity does not influence human milk protein 15N natural isotope abundance

ABSTRACT

Obesity increases protein metabolism with a potential effect on nitrogen isotope fractionation. The aim of this study was to test the influence of obesity on human milk extracted protein ¹⁵N natural isotope abundance (NIA) at one month post-partum and to compare human milk extracted protein ¹⁵N NIA and bulk infant hair ¹⁵N NIA. This cross-sectional observational study involved 16 obese mothers (body mass index (BMI)?≥?30?kg?m^?2 before pregnancy) matched with 16 normal-weight mothers (18.5?kg?m^?2?≤?BMI?<?25?kg?m^?2) for age and pregnancy characteristics. Human milk extracted protein and bulk infant hair ¹⁵N NIA were determined by isotope ratio monitoring by mass spectrometry interfaced to an elemental analyser (IRM-EA/MS). No significant difference was found in human milk protein ¹⁵N NIA values between obese and normal-weight mothers (8.93?±?0.48?‰ vs. 8.95?±?0.27?‰). However, human milk protein ¹⁵N NIA was significantly lower than bulk infant hair ¹⁵N NIA: 8.94?±?0.38?‰ vs. 9.66?±?0.69?‰, respectively. On the basis of these results, it is concluded that human milk protein ¹⁵N NIA measured at one month post-partum is not influenced by maternal obesity. These findings suggest that ¹⁵N NIA may be exploited to study metabolism without considering maternal obesity as a confounder.     

Assessing the subsequent effect of nitrogen released from tobacco-waste on maize crop using a 15N isotope technique

The investigation of the residual effect of nitrogen (N) released from tobacco-waste (TW) using isotope techniques will provide valuable data for sustainable organic farming. For this aim, a pot experiment was conducted using the ¹⁵N isotope technique. The experiment was based on a completely randomised design with four replications and was conducted on a calcareous ustochrepts soil. TW at levels of 0, 10, 20, 30 and 40 t ha^?1 and N fertiliser as (NH₄)₂SO₄ at levels of 0, 20, 40, 60 and 80 kg N ha^?1 were used for the Bezostaja-1 wheat variety. Concerning mineral N fertilisation with 20 and 80 kg N ha^?1, additional treatments with ¹⁵N-labelled (NH₄)₂SO₂ (10 at.% exc.) have been applied. Following harvesting wheat plants, the Pioneer 3377 maize variety was used to see the residual effect of TW. After harvesting, dry matter yields were recorded and total N concentrations were determined. ¹⁵N determinations and calculations were also made for ¹⁵N treatments separately. TW had a significant residual effect on the growth of corn plant under the pot condition. Increasing rates of TW significantly increased the dry matter yield of corn plant following wheat from 3.31 t ha^?1 (at control) to 7.89 t ha^?1 (at the TW treatment of 40 t ha^?1). The ¹⁵N values derived from the ¹⁵N fertiliser decreased with increasing TW application. The average values of N derived from N fertiliser (Ndff) varied from 2.14 to 3.09% at the rates of 20 and 80 kg N ha^?1, respectively. However, N derived from TW (Ndftw) significantly increased from 16.93 to 24.59% (at 20 kg N ha^?1), and it also increased from 23.06 to 28.15% (at 80 kg N ha^?1) with increasing TW applications from 20 to 40 t ha^?1, respectively.     

Relationships between Nitrogen Translocation and Accumulation in Growing Plant Parts of Grain Legumes

Abstract

Grain legumes absorbed mineral ¹⁵N at every stage of development and transported it directedly (just as symbiotically fixed ¹⁵N₂) into the plant parts growing at the respective stage. The ¹⁵N accumulation in the grains after long-lasting ¹⁵N supply can be ascribed, for the major part, to a secondary ¹⁵N translocation after a temporary incorporation into older plant parts (leaves, stem). Inhibition experiments with antibiotics revealed no direct relation between the accumulation of amino acid ¹⁵N in growing pods and seeds and the protein synthesis in this target organs. It may include, however, processes of (active ?) uptake and transport with a possible contribution of carrier systems specific for distinct amino acids.     

Release of nitrous oxide and dinitrogen from a transition bog under drained and rewetted conditions due to denitrification: results from a [15N]nitrate–bromide double-tracer study

Denitrification is well known being the most important nitrate-consuming process in water-logged peat soils, whereby the intermediate compound nitrous oxide (N₂O) and the end product dinitrogen (N₂) are ultimately released. The present study was aimed at evaluating the release of these gases (due to denitrification) from a nutrient-poor transition bog ecosystem under drained and three differently rewetted conditions at the field scale using a ¹⁵N-tracer approach ([¹⁵N]nitrate application, 30?kg N ha^?1) and a common closed-chamber technique. The drained site is characterized by a constant water table (WT) of –30?cm (here referred to as D30), while rewetted sites represent a constant WT of –15?cm, a constant WT of 0?cm (i.e. waterlogged), and an initial WT of 0?cm (which decreased slightly during the experiment), respectively, (here referred to as R15, R0, and R0_d, respectively). The highest N₂O emissions were observed at D30 (291?µg N₂O–N m^?2 h^?1) as well as at R0_d (665?µg N₂O–N m^?2 h^?1). At the rewetted peat sites with a constant WT (i.e. R15 and R0), considerably lower N₂O emissions were observed (maximal 37?µg N₂O–N m^?2 h^?1). Concerning N₂ only at the initially water-logged peat site R0_d considerable release rates (up to 3110?µg N₂–N m^?2 h^?1) were observed, while under drained conditions (D30) no N₂ emission and under rewetted conditions with a constant WT (R15 and R0) significantly lower N₂ release rates (maximal 668?µg N₂–N m^–2 h^?1) could be detected. In addition, it has been found that natural WT fluctuations at rewetted peat sites, in particular a rapid drop down of the WT, can induce high emission rates for both N₂O and N₂.     

Feeding level and diet quality influence trophic shift of c and n isotopes in nile tilapia (oreochromis niloticus (L.))

Many scientists use naturally occurring stable isotopes to reconstruct the diets of animals. However, isotopic ratios may be affected not only by the composition of the diet but also by the amount of food consumed. Thus, an experiment using tilapia (Oreochromis niloticus) was carried out to test the influence of feeding level on δ ¹³C and δ ¹⁵N of fish given a semi-synthetic wheat gluten/wheat starch based diet. In addition, the effect of diet quality was tested by comparing tilapia given this feed with tilapia fed a fish meal/wheat meal based diet. Forty-four tilapia were reared individually. After a prefeeding phase, fish were randomly assigned to five groups, four on the semi-synthetic diet at different feeding levels and one group on the fish meal/wheat meal based diet fed at the equivalent of the highest level of the semi-synthetic diet. The experiment lasted eight weeks. Proximate composition, gross energy content and δ ¹³C and δ ¹⁵N values were determined in feed and fish, for δ ¹³C separately in the lipids and the lipid-free matter. δ ¹³C in the lipids and the lipid-free matter and δ ¹⁵N of tilapia fed the semi-synthetic diet decreased significantly with increasing feeding rate. The absolute values of the trophic shift in fish fed the semi-synthetic wheat based diet were significantly higher than in fish fed the fish meal/wheat meal based diet. The different δ ¹³C and δ ¹⁵N values in tilapia fed the same diet at different feeding levels and the influence of feed quality on the trophic shift add to the uncertainty involved in the use of stable isotopes in ecological research.     

Measuring nitrification in sediments – comparison of two techniques and three 15NO measurement methods

Nitrification is a crucial process in sediment nitrogen cycling. We compared two ¹⁵N tracer-based nitrification measurement techniques (isotope pairing technique (IPT) combined with ¹⁵N nitrate pool dilution and ¹⁵N ammonium oxidation) and three different ¹⁵N analyses from bottom water nitrate (ammonia diffusion, denitrifier and SPINMAS) in a sediment mesocosm. The ¹⁵N nitrate pool dilution technique combined with IPT can be used to quantify the in situ nitrification, but the minimum detection limit for the total nitrification is higher than that in the ¹⁵N ammonium oxidation technique. The ¹⁵N ammonium oxidation technique, however, is not applicable for sediments that have high ammonium content. If nitrate concentration and the amount of ¹⁵N label in the sample are low, the ¹⁵N nitrate analysis should be done with the denitrifier method. In higher ¹⁵N concentrations, the less sensitive SPINMAS method can also be applied. The ammonia diffusion method is not suitable for bottom water ¹⁵N nitrate analyses.     

Distribution and Utilization of 15N in Cowpeas Injected into the Stem Under Influence of Water Deficit

Abstract

Investigations were carried out on Vigna unguiculata L. Walp. to estimate the distribution and utilization of ¹⁵N in different organs after stem injection during vegetative, flowering and pod filling stage. During flowering effects of water deficit were also examined. In well watered plants, within 4 days after injection, 65% of ¹⁵N accumulated in leaves. This was drastically reduced to 42% by water deficit. ¹⁵N accumulation in stems increased under water deficit. The translocation of ¹⁵N from the stem base to roots were not altered by water deficit. During pod filling 62% of recovered ¹⁵N in the plants had accumulated in seeds, 24% in leaves and 11% in stems within 4 days, whereas the uptake of nitrogen in pod walls and roots remained low (2%). These results demonstrate that the method of injecting very small quantities (1 mg/plant) of ¹⁵N into the stem base allows an exact and detailed quantitative assessment of N translocation/distribution with regard to different organs, different growth stages and different treatments.     

Influence of dietary fibre on gluten proteins structure – a study on model flour with application of FT‐Raman spectroscopy

Dietary fibres are regarded as the source of polysaccharides and antioxidants such as polyphenols. However, addition of dietary fibre to bread causes significant reduction in its quality. The bread quality is connected with the structure of gluten proteins. For this reason, Fourier transform Raman spectroscopy was applied to determine changes in structure of gluten proteins modified by seven dietary fibres. The fibres were added to model flour reconstituted with wheat gluten and wheat starch. The model flour was used to provide gluten proteins of definite structure. The obtained results showed that six out of seven fibres caused similar changes in β‐turn structures. The appearance of the band at 1642 cm⁻¹ and the shift toward lower wavenumbers of the band at 1670 cm⁻¹ in the difference spectra indicated hydrogen bonding of carbonyl groups in β‐turns leading to protein folding/aggregation. Addition of fibre preparations caused also changes in conformation of disulfide bridges (S–S), corresponding to transformation to trans‐gauche‐gauche and trans‐gauche‐trans conformations at the expense of the stable gauche‐gauche‐gauche conformation. The S–S bonds in less stable conformations were formed inside the protein complex as well as between protein complexes in the form of β‐structures. Generally, the observed changes in gluten proteins after addition of dietary fibres were results of interactions between fibre polysaccharides and gluten proteins rather than between polyphenols and gluten proteins. Copyright © 2015 John Wiley & Sons, Ltd.     

The Effect of Diet Quality on δ13C and δ15N in the Tissues of Locusts,Locusta migratoria L

Abstract

Locust nymphs were raised from hatching to adult locusts on either seedling wheat (C₃) or maize (C₄), to determine whether relative enrichments/depletions of ¹⁵N and ¹³C within body tissues are influenced by diet. The maize contained less hexose sugars and protein per gram than wheat.

The isotopic spacing between the food and the whole insect was found to differ between the two diets. The lower quality maize diet showed an overall +5.1‰ enrichment in δ¹⁵N compared to + 2.8‰ for wheat, possibly due to increased fractionation due to protein recycling.

The maize diet resulted in increased depletion in lipid and trehalose and depletion in chitin relative to diet. The results for both δ¹⁵N and δ¹³C suggest that substrate recycling was occurring on the low quality maize diet. Therefore diet quality determines the enrichment/depletion in δ¹⁵N and δ¹³C within organisms.     

Development and application of 15N-tracer substances for measuring the whole-body protein turnover rates in the human,especially in neonates: a review

Our research group of the Children's Hospital of the University of Rostock (Rostock group) has long-time experience in ¹⁵N-labelling and in using yeast protein and its hydrolysates for tracer kinetic studies to evaluate parameters of the whole-body protein metabolism in premature infants. The particular advantage of applying an economically convenient, highly ¹⁵N-enriched, and completely labelled yeast protein for evaluating protein turnover rates is the fact that the ¹⁵N dose is spread among all proteinogenic amino acids. The absorption has been improved by hydrolysing [¹⁵N]yeast protein with thermitase into a mixture of amino acids, dipeptides and tripeptides so that faecal analysis becomes unnecessary when determining turnover rates. The review shows that, in contrast to the application of single ¹⁵N-labelled amino acids with resulting overestimation of protein turnover rates, the ¹⁵N-labelled yeast protein thermitase hydrolysate represents the amino acid metabolism more closely without causing amino acid imbalances. The ¹⁵N-labelled yeast protein thermitase hydrolysate leads to the estimation of reliable protein turnover rates, particularly in premature infants.     

A study on anomalous yield drop behaviour in modified beta titanium alloys

ABSTRACT

The yield drop phenomenon observed in the Ti–15V-3Al–3Sn-3Cr (Ti–15–3) beta-titanium alloy and its anomalous behaviour in the boron and carbon added Ti–15–3 alloys have been studied. While the base and the carbon containing alloys exhibit yield drop, the boron containing alloy with smaller grain size than base alloy does not appear to show this phenomenon. Tensile tests were interrupted at different stress levels followed by analyses of slip lines and sub-structural characteristics using scanning and transmission electron microscopes to understand this anomalous yield point phenomenon. Infrared thermal imaging technique was used to map the strain localisation and the spatiotemporal evolution of deformation along the gauge length of the specimens during the tensile tests. Deformation in these alloys initiates only in a few grains. Pile-up of dislocations in these grains subsequently triggers the formation of dislocations in other grains and their rapid multiplications. The spreading of deformation by the generation of dislocations from pile up dislocations in one grain to neighbouring un-deformed grains and their rapid multiplication to new regions influence the yield drop phenomenon and its characteristics. It is shown in this study that microscopic instability in the grain level is a necessary, but not the sufficient condition for the manifestation of macroscopic instability during tensile deformation in polycrystalline materials. The presence of boride particles at grain boundaries restricts the slip transfer across the grains as well as the spreading of deformation to new regions, which causes the suppression of yield drop in the boron containing alloy.     

Latitudinal distribution of nitrogen isotopic composition in suspended particulate organic matter in tropical/subtropical seas

Natural nitrogen isotopic composition (δ¹⁵N) of suspended particulate organic matter (POM) and nitrogen fixation rates via ¹⁵N₂ assay were measured in surface waters along 120° E from 30° N to 30° S in the Asian marginal seas (the East/South China Seas and the Sulu/Celebes/Java Seas) and the northeastern Indian Ocean in November–December 2005 and March 2006. The POM δ¹⁵N values ranged from?1.8 to 12.2‰ with an average of 3.6‰ and showed a decreasing trend towards the equator in both hemispheres. In parallel, the measured N₂ fixation rates showed an increase from the subtropical to the tropical seas. This implies that a higher contribution of ¹⁵N-depleted POM was derived from enhanced N₂ fixation. Water temperature and the stability of water column were partly responsible for the observed variations in nitrogen fixation. The large-scale spatial variations in suspended POM δ¹⁵N and N₂ fixation rates suggest that the suspended POM δ¹⁵N may be a potential indicator of nitrogen fixation in surface waters in tropical/subtropical seas.     

15N natural abundance of non-fixing woody species in the Brazilian dry forest (caatinga)

Foliar δ¹⁵N values are useful to calculate N₂ fixation and N losses from ecosystems. However, a definite pattern among vegetation types is not recognised and few data are available for semi-arid areas. We sampled four sites in the Brazilian caatinga, along a water availability gradient. Sites with lower annual rainfall (700 mm) but more uniform distribution (six months) had δ¹⁵N values of 9.4 and 10.1 ‰, among the highest already reported, and significantly greater than those (6.5 and 6.3 ‰) of sites with higher rainfall (800 mm) but less uniform distribution (three months). There were no significant differences at each site among species or between non-fixing legume and non-legume species, in spite of the higher N content of the first group. Therefore, they constitute ideal reference plants in estimations of legume N₂ fixation. The higher values could result from higher losses of ¹⁵N depleted gases or lower losses of enriched ¹⁵N material.     

Source attribution of agriculture-related deposition by using total nitrogen and δ15N in epiphytic lichen tissue,bark and deposition water samples in Germany

Compared with physico-chemical deposition measurement methods, lichens are able to identify the long-term overall effects of high N pollution concentrations in the air. In addition, the natural abundances of the stable isotope of N, ¹⁵N, are being widely used in research on N cycling in ecosystems. They can also be used as instruments for source attribution. In this study, epiphytic lichens were tested to determine whether their respective N content and δ¹⁵N ratios can be used to estimate N deposition rates and to locate various sources of N compounds. Epiphytic lichen and bark samples were collected from around various deposition measurement field stations at different sites in the western part of Germany. The N content of epiphytic lichens reflects the species-specific, agriculture-related circumstances of N deposition at various sites in Germany. At the same time, δ¹⁵N signatures of the different investigated epiphytic lichen species and bark samples are highly depleted in ¹⁵N under high ammonium deposition. The different surface types of lichens and barks exhibit different concentrations of N and δ¹⁵N ratios, despite being exposed to similar N deposition rates. The verification of highly negative δ¹⁵N ratios at sites with local and regional emitters shows that source attribution is possible by comparing different δ¹⁵N signatures in areas with a wide range of different N deposition types and the corresponding differences in δ¹⁵N among various source N pools. Especially nitrophytic lichens can support the on-site instrumentation measuring N deposition by qualification and quantification.     

Nd-Fe-B/FeCo多层纳米复合膜的结构和磁性

制备了Nd₂₈Fe₆₆B₆/Fe₅₀Co₅₀多层纳米复合磁性薄膜，对溅射态和650℃退火处理15 min试样的相成分分析和微结构的观察显示，溅射态薄膜呈非晶态，经650℃退火处理15 min后，薄膜主要相成分为硬磁性Nd₂Fe₁₄B相和软磁性相FeCo(110)相.Nd₂Fe₁₄B相呈柱状，其易磁化c轴垂直于膜面，尺寸约10 nm.在硬磁性相和软磁性相之间存在少量富Nd相和非晶态，富Nd相大小约7 nm.磁性测量和分析表明，1)该系列薄膜退火态具有垂直于膜面的磁晶各向异性.2)对于固定厚度(10 nm)层Nd-Fe-B和不同厚度(t_FeCo=1—100 nm)层FeCo多层纳米复合膜,剩磁随软磁相FeCo 厚度的增加快速增加，而矫顽力则减小.当t_FeCo=5 nm时，最大磁能积达到200 kJ/m³. 3)硬磁相Nd-Fe-B层和软磁相FeCo层之间交换耦合导致剩磁和磁能积增强. 关键词： Nd-Fe-B/FeCo多层纳米复合膜 交换耦合 磁各向异性     

Spatial variations in larch needle and soil δ15N at a forest–grassland boundary in northern Mongolia

The spatial patterns of plant and soil δ¹⁵N and associated processes in the N cycle were investigated at a forest–grassland boundary in northern Mongolia. Needles of Larix sibirica Ledeb. and soils collected from two study areas were analysed to calculate the differences in δ¹⁵N between needle and soil (Δδ¹⁵N). Δδ¹⁵N showed a clear variation, ranging from ?8?‰ in the forest to ?2?‰ in the grassland boundary, and corresponded to the accumulation of organic layer. In the forest, the separation of available N produced in the soil with ¹⁵N-depleted N uptake by larch and ¹⁵N-enriched N immobilization by microorganisms was proposed to cause large Δδ¹⁵N, whereas in the grassland boundary, small Δδ¹⁵N was explained by the transport of the most available N into larch. The divergence of available N between larch and microorganisms in the soil, and the accumulation of diverged N in the organic layer control the variation in Δδ¹⁵N.     

30 years of production of highly enriched nitrogen-15 in the GDR

Almost 30 years ago, the stable isotope ¹⁵N had covered a wide field of application in science and technology. With the preparation of the first smaller amounts of ¹⁵N in 1958, an intensive work of research and development started for the production, analysis, and application of ¹⁵N in the GDR. In this publication, activities of research and development for ¹⁵N production using the principle of chemical exchange in the system NO _x /HNO₃ from a laboratory scale to the introduction into chemical industry are described and new projects reported.