15N Isotope Research Articles

Hydrogen isotope labeling unravels origin of soil-bound organic contaminant residues in biodegradability testing

Biodegradability testing in soil helps to identify safe synthetic organic chemicals but is still obscured by the formation of soil-bound ‘non-extractable’ residues (NERs). Present-day methodologies using radiocarbon or stable (13C, 15N) isotope labeling cannot easily differentiate soil-bound parent chemicals or transformation products (xenoNERs) from harmless soil-bound biomolecules of microbial degraders (bioNERs). Hypothesizing a minimal retention of hydrogen in biomolecules, we here apply stable hydrogen isotope – deuterium (D) – labeling to unravel the origin of NERs. Soil biodegradation tests with D- and 13C-labeled 2,4-D, glyphosate and sulfamethoxazole reveal consistently lower proportions of applied D than 13C in total NERs and in amino acids, a quantitative biomarker for bioNERs. Soil-bound D thus mostly represents xenoNERs and not bioNERs, enabling an efficient quantification of xenoNERs by just measuring the total bound D. D or tritium (T) labeling could thus improve the value of biodegradability testing results for diverse organic chemicals forming soil-bound residues.

Maternal NO2 exposure and fetal growth restriction: Hypoxia transmission and lncRNAs-proinflammation-mediated abnormal hematopoiesis

Epidemiological studies show a strong correlation between air pollution and fetal growth restriction (FGR), but existing results are controversial due to inherent limitations, such as causality of specific pollutants, developmental origin, and maternal-fetal transmission. To address this controversy, we first conducted a retrospective analysis of 28,796 newborns and revealed that maternal nitrogen dioxide (NO2) exposure during the second trimester was positively associated with FGR, with an adjusted odds ratio of 1.075 (95% confidence interval: 1.020-1.133) per 10 μg/m3 NO2 increase for small for gestational age. Then, by establishing an animal model of prenatal NO2 exposure, we confirmed its adverse effects on embryonic growth and hematopoiesis in the yolk sac and fetal liver, primarily affecting the differentiation of hematopoietic stem and progenitor cells and erythroid maturation. By applying internal exposure analyses coupled with 15N isotope tracing, we found that maternal NO2 inhalation induced acquired methemoglobinemia through its byproducts and placental hypoxia in pregnant mice. Importantly, by combining transcriptional profiling, bioinformatics analysis, and RNA binding protein immunoprecipitation (RIP)/chromatin immunoprecipitation (CHIP), we clarified that placental-fetal hypoxia transmission activated hypoxia-inducible factors, disturbed hematopoiesis through the hypoxia-inducible factor 1β-long noncoding RNAs-CCAAT/enhancer binding protein alpha-proinflammatory signaling pathway, ultimately contributing to FGR progression. These findings provide insights for risk prevention and clinical intervention to promote child well-being in NO2-polluted areas.

Warm growing season activates microbial nutrient cycling to promote fertilizer nitrogen uptake by maize

The influence of nitrogen (N) inputs on soil microbial communities and N uptake by plants is well-documented. Seasonal variations further impact these microbial communities and their nutrient-cycling functions, particularly within multiple cropping systems. Nevertheless, the combined effects of N fertilization and growing seasons on soil microbial communities and plant N uptake remain ambiguous, thereby constraining our comprehension of the optimal growing season for maximizing crop production. In this study, we employed 15N isotope labeling, high-throughput sequencing, and quantitative polymerase chain reaction (qPCR) techniques to investigate the effects of two distinct growing seasons on microbial communities and maize 15N uptake ratios (15NUR). Our results showed that the warm growing season (26.6 °C) increased microbial diversity, reduced network complexity but enhanced stability, decreased microbial associations, and increased modularization compared to the cool growing season (23.1 °C). Additionally, the warm growing season favored oligotrophic species and increased the abundance of microbial guilds and functional genes related to N, phosphorus, and sulfur cycling. Furthermore, alterations in the characteristics of soil microbial keystone taxa were closely linked to variations in maize 15NUR. Overall, our findings demonstrate significant seasonal variations in soil microbial diversity and functioning, with maize exhibiting higher 15NUR during the warm growing season of the double cropping system.

Deep Rooting as an Indicator of Deep Soil Water and N Uptake in Potato (Solanum tuberosum L.)

AbstractBreeding for potato deep roots can increase water and nitrogen uptake by potatoes and it can be an option to maintain stable yields with decreased inputs. This study investigates the relationship between potato root characteristics, water stress resistance and deep soil nitrogen uptake, accessing variations among cultivars and nitrogen fertilization levels. Thirteen potato cultivars were grown during 2018 and 2020 at a semi-field root phenotyping platform in Denmark. Root growth was monitored via minirhizotron tubes down to 1.8 m soil depth. Drought treatment started in the mid-June and deep soil nitrogen uptake was tracked via 15N isotope application at 1.3–1.4 m soil depth during tuber formation. Water stress resilience was identified using 13C natural discrimination process in plants. Tuber samples were analyzed for 15N and 13C content. While drought affected potato yield (not always significantly), it did not affect nitrogen uptake. Root length and distribution varied among varieties, with deeper roots (down to 1.30 m) observed in August. Statistical differences (p < 0.05) in root length, yield and nitrogen uptake were found among varieties. Cultivars with longer growing season exhibited larger, deeper roots and increased nitrogen uptake from deep soil. High correlation (R = 0.8) between deep roots and 15N uptake was observed for all varieties. Deeper roots are contributing to deep soil nitrogen uptake, but 13C content in tubers is not a reliable indicator of water stress resilience. Despite this, the study suggests the potential for breeding potatoes with deep roots to achieve stable yields, considering differences in water and nitrogen uptake among varieties.

Plastic ingestion in aquatic insects: Implications of waterbirds and landfills and association with stable isotopes

Wetlands provide numerous ecosystem services including freshwater purification. Nonetheless, their functionality is continuously impacted by many pollutants. Plastics are considered as an emerging threat for these ecosystems, but only recently have studies began to focus on plastic and microplastic (MP) contamination in wetlands, especially in biota. This study aims to investigate the abundance of MPs in two ubiquitous aquatic insect taxa (i.e. Corixidae (Hemiptera) and Chironomidae (Diptera)) collected in twelve zones within Mediterranean wetlands belonging to three basins located in Andalusia (south-west Spain). We compared MP contamination across basins and tested the proximity to landfills and presence of colonial waterbirds [i.e. white storks (Ciconia ciconia) and gulls (Larus michahellis and L. fuscus)] on MP abundance in aquatic insects. We also performed stable isotope analyses of nitrogen and carbon (δ15N and δ13C) to evaluate the potential association between MP abundance and isotopic values. We detected 571 suspected MPs (mostly blue fibers) in insects of different developmental stages (i.e., larvae, pupae, nymphs and adults). Polyesters and polypropylene were the most frequent polymers detected. The generalized linear mixed models indicated that MP abundance decreased with increasing distance from landfills; but it also increased in sites with birds that fed on landfills and roost in wetlands. When controlling for landfill effects, sites in the smallest basin (Guadalete) had lower MP contamination than those in Odiel-Tinto and the much larger (>15×) Guadalquivir. Moreover, we found a negative association between MPs items/g (or mean MPs) and 15N isotopes in adult corixids. Our findings showed that MP pollution is present in all the study areas, including strictly protected wetlands. The use of aquatic insects for biomonitoring of MP pollution can help identify priority areas for management actions to mitigate plastic pollution.

Radiocarbon Chronology and Isotope Data of Ust-Tartasskiye Kurgany Mound 51, the Baraba Forest-Steppe

Results of radiocarbon dating of items from mound No. 51 of Ust-Tartasskiye Kurgany burial ground are presented. This is a key site of the Sargat culture in Baraba. Characteristics of samples and pretreatment procedure are provided. Twenty-four radiocarbon dates were generated. Radiocarbon ages correlate with biological ages of the deceased persons. Bayesian KDE chronology modeling suggests a short-term intense use of the site for burying those who died between the 3rd and 1st centuries BC. Based on MCMC-modeling, a conclusion is reached about two periods in the use of the space allotted for graves between 200–40 BC. Burials of the “first period” (~25 %) could have been repeatedly made before 150 BC. Most burials (~75 %) were likely arranged between 150–120 BC. The last burial in the mound (no earlier than 110 BC) is No. 13. Minor differences in 15N isotope apparently evidence various diets of males and females at the second period.

Copper–nickel–MOF/nickel foam catalysts grown in situ for efficient electrochemical nitrate reduction to ammonia

Reducing nitrate (NO3−) in an aqueous solution to ammonia under ambient conditions can provide a green and sustainable NH3‐synthesis technology and mitigate global energy and pollution issues. In this work, a CuNi0.75–1,3,5‐benzenetricarboxylic acid/nickel foam (CuNi0.75–MOF/NF) catalyst grown in situ was prepared via a one‐pot method as an efficient cathode material for electrocatalytic nitrate reduction reaction (NO3RR). The CuNi0.75–MOF/NF catalyst exhibited excellent electrocatalytic NO3RR performance at −1.0 V versus a reversible hydrogen electrode, achieving an outstanding faradaic efficiency of 95.88 % and an NH3 yield of 51.78 mg h−1 cm−2. The 15N isotope labeling experiments confirmed that the sole source of N in the electrocatalytic NO3RR was the NO3− in the electrolyte. The reaction pathway for the electrocatalytic NO3RR was derived by in situ Fourier transform infrared spectroscopy and in situ differential electrochemical mass spectrometry. Density functional theory calculations revealed that the Ni element in the CuNi0.75–MOF/NF catalyst had excellent O–H activation ability and strong *H adsorption capacity. These *H species were transferred from the Ni sites to the *NO adsorption intermediates located on the Cu sites, providing a continuous supply of *H to Cu, thereby promoting the formation of *NOH intermediates and enhancing the hydrogenation process of the electrocatalytic NO3RR.

Dietary Niche Variation in an Invasive Omnivore: The Effects of Habitat on Feral Pig Resource Use in Hawai'i.

Invasive omnivores may have profound impacts on ecological communities through diet selection, particularly when their functional roles differ from those in their native range. While the threat of feral pigs (Sus scrofa) to native plant communities in Hawai'i is well known, their trophic dynamics and the drivers of variation in their diet remain understudied. We investigated the feral pig trophic niche on Hawai'i Island using stable isotopes (13C and 15N) and Bayesian mixing models to identify drivers of variation in resource use. We also reconstructed intra-individual variability for six subsampled individuals to understand temporal variation in resource use and individual diet specialization. Our results revealed that feral pigs on Hawai'i Island exhibit a broad trophic niche characterized by diverse diets, with substantial overlap in resource use across districts and habitats. Differences in dietary composition in the transition from forest to open habitat were driven primarily by a decline in invertebrates and an increasing reliance on resources enriched in 15N, which may reflect a shift in protein sources with habitat. Pigs in forested areas exhibited a smaller trophic niche than those in open habitats, largely driven by differences in feeding strategies and resource availability. Diets for subsampled individuals varied little, suggesting feral pig resource-use strategies in Hawai'i tend to be relatively stable through time. Individual niche width was relatively narrow compared to that of feral pigs in Hawai'i at large, indicating the relatively wide feral pig dietary niche is characterized by substantial intraspecific diet specialization, likely as a result of strong intraspecific competition. Understanding the drivers of feral pig resource use offers key information for management strategies aimed at mitigating their ecological impacts in imperiled systems like Hawai'i.

Investigating drivers of free-living diazotroph activity in paddy soils across China

Microbially mediated N fixation is widespread in rice paddy ecosystems and crucial in maintaining soil fertility. However, our understanding of the factors determining the distribution of free-living diazotrophic microorganisms that perform this process in paddy fields is limited. This study investigated the spatial distribution and factors influencing presence and potential activity of free-living microorganisms capable of N2 fixation in addition to dissimilatory nitrate reduction to ammonium (DNRA), anaerobic ammonium oxidation (anammox), and denitrification in 50 paddy soils across China. Using 15N isotope tracing in laboratory incubations and microbial community analysis via metagenomics, we demonstrate that paddy soils may represent a previously underappreciated hotspot for N2 fixation with mean potential rates of 24.4 ± 17.8 nmol N g−1 h−1, 10-fold higher than DNRA (2.55 ± 0.4 nmol N g−1 h−1), and could counterbalance a portion of N2 losses through anammox and denitrification (9.24 ± 1.1 nmol N g−1 h−1). Site longitude and organic carbon (C) concentrations, as well as the diazotrophic community composition, were the dominant abiotic and biotic factors accounting for regional variations in potential N2 fixation rates. The N2 metabolic pathways predicted from the metagenome-assembled genomes (MAGs) revealed significant co-occurrence of the diazotroph marker gene nifH with denitrification-associated genes (nirS/K and nosZ) and organic C oxidation-related genes (yiaY and galM). Furthermore, enzymes involved in organic C oxidation, particularly glycoside hydrolases and glycosyltransferases, were not only phenotypically correlated with free-living N2 fixation rates but were also identified in nifH-containing MAGs, indicating the heterotrophic capabilities of diazotrophs in paddy soils. Collectively, our results underscore the substantial contribution of free-living N2 fixation to soil N fertility in paddy fields, and highlight the importance of coupling organic C oxidation with nitrate reduction to enhance N2 fixation.

Nitrate Enhanced Sulfamethoxazole Degradation by 222 nm Far-UVC Irradiation: Role of Reactive Nitrogen Species.

The application of 222 nm far-UVC irradiation for degrading organic micropollutants in water shows promise. Nitrate (NO3-), found in nearly all water bodies, can significantly impact the performance of 222 nm far-UVC-driven systems. This work was the first to investigate the effect of NO3- on sulfamethoxazole (SMX) photodegradation at 222 nm, finding that NO3- significantly enhances SMX degradation in different dissociated forms. Besides the hydroxyl radical (•OH), reactive nitrogen species (RNS) also played important roles in SMX degradation. With increasing NO3- concentration, the RNS contribution to SMX degradation decreased from 25.7 to 8.6% at pH 3 but increased from 1.5 to 24.7% at pH 7, since the deprotonated SMX with electron-rich groups reacted more easily with RNS. The transformation mechanisms of SMX involving isomerization, bond cleavage, hydroxylation, nitrosation, and nitration processes were proposed. 15N isotope labeling experiments showed that the RNS-induced nitrated products even became the major products of SMX in the 222 nm far-UVC/NO3- system at pH 7 and exhibited a higher toxicity than SMX itself. Further research is necessary to avoid or eliminate these toxic byproducts. This study provides valuable insights for guiding the utilization of 222 nm far-UVC for treating antibiotics in NO3--containing water.

Microbial nitrogen cycling in Microcystis colonies and its contribution to nitrogen removal in eutrophic Lake Taihu, China

Cyanobacterial blooms induced by excessive loadings of nitrogen (N) and other nutrients are a severe ecological problem in aquatic ecosystems. Previous studies of N removal have primarily focused on sediment-water interface, yet the role of cyanobacterial colonies has recently been attracting more research attention. In this study, N cycling processes were quantified for cyanobacterial colonies (primarily Microcystis colonies) and their contribution to N removal was estimated for a large, shallow eutrophic lake in China, Lake Taihu. Various N cycling processes were determined via stable 15N isotope, together with 16S rRNA gene sequencing and quantitative microbial element cycling (QMEC) chip. Denitrification was found to be the most prominent process, estimated to be 36.63, 9.85, 3.35, and 3.15 times higher than dissimilatory nitrate reduction to ammonium (DNRA), nitrification, ammonium (NH4+) uptake and nitrate (NO3−) uptake rates, respectively. Denitrifiers accounted for a large part of the bacterial taxa (35.50 ± 24.65%), and the nirS gene was the most abundant among N cycling-related genes, with (2.54 ± 0.51) × 109 copies g−1Microcystis colonies. A field investigation revealed a positive correlation between the potential denitrification rate and the Chl-a concentration (mostly derived from Microcystis colonies). Based on a multiple stepwise regression model and historical data from 2007 to 2015 for Lake Taihu, the total amount of N removed via denitrification by Microcystis colonies was estimated at 171.72 ± 49.74 t yr−1; this suggests that Microcystis colonies have played an important role in N removal in Lake Taihu since the drinking water crisis in 2007. Overall, this study revealed the importance of denitrification within Microcystis colonies for N removal in eutrophic lakes, like Lake Taihu.

Effects of biochar on the transformation and utilization of nitrogen fertilizer in the black soil region of Northeast China

Nitrogen (N) fertilizer is often used in production practice to effectively maintain crop productivity; however, low nitrogen use efficiency (Nue) has always been a problem. Specifically, the transformation and utilization of nitrogen fertilizer by biochar and the driving mechanisms remain unclear. We used four biochar application rates (0, 3750, 7500, and 11,250 kg·ha−1) and analyzed the effects of biochar on nitrogen fertilizer utilization, residue, and loss over three years using 15N isotope tracer technology. The results showed that (1) biochar improved the nitrogen use efficiency of maize plants, reduced total nitrogen loss, and increased the maize yield. Compared to the control treatment in the same year, the application of 7500 kg·ha−1 biochar increased the nitrogen use efficiency by 24.27 %, 27.77 %, and 35.82 %, and the yield increased by 21.1 %, 26.7 %, and 24.5 %, respectively. (2) Biochar increased the proportion of mineral nitrogen supplied by fertilizer in the mineral nitrogen pool. The application of 7500 kg·ha−1 biochar increased mineral nitrogen by 3.05 %, 3.22 %, and 3.8 %, respectively, compared to the control treatments in the same year. Biochar promoted the transformation of nitrogen in the 0–40 cm soil layer to three different soil nitrogen pools, especially the organic nitrogen pool. (3) Biochar significantly improved the soil bacterial community and increased the abundances of N transformation functional genes. The redundancy analysis (RDA) showed that the gdhA mineralization gene was the driving factor of nitrogen fertilizer transformation, contributing 43.6 % of the variance. In summary, the application of 7500 kg·ha−1 of biochar for two consecutive years was conducive to maintaining farmland soil fertility, while its use would not be recommended for more than three consecutive years.

The Combined Application of Urea and Fulvic Acid Regulates Apple Tree Carbon and Nitrogen Metabolism and Improves Anthocyanin Biosynthesis

Improving apple peel color has been an important objective in apple production. To better understand the effect and mechanism of the combined application of urea and FA (fulvic acid) regulation of anthocyanin biosynthesis, a field experiment was performed in 2022 and 2023, respectively, under five treatments of urea + FA (CK, urea only; FA50, urea + 50 kg ha−1 FA; FA100, urea + 100 kg ha−1 FA; FA150, urea + 150 kg ha−1 FA; FA200, urea + 200 kg ha−1 FA), using isotope (13C and 15N) marking to analyze the changes in carbon (C) and nitrogen (N) nutrient distribution as well as anthocyanin biosynthesis in fruits. We observed that, under FA application conditions, anthocyanin content in the peel was elevated in both years, with increases of 15.98~52.88% in 2022 and 15.93~52.94% in 2023. The best promotion effects were observed under FA150 treatment. Apart from the expression levels of anthocyanin biosynthesis-related genes and transcription factors in the apple peel, this positive effect on anthocyanin content induced by FA addition was also found to be associated with the optimization of C and N distribution in leaves and fruits. On the one hand, the application of FA not only enhanced leaf photosynthetic-related indexes (such as Pn, Gs, and Rubisco activity) and influenced (increased) S6PDH, SPS, and SS activities in leaves, but also elevated fruit sugar metabolism-related enzyme (SDH, SS-c, AI, and NI) activity and upregulated fruit stalk sugar transporter (MdSOT1, MdSOT3, MdSUT1 and MdSUT4) gene expression, which ultimately promoted the synthesis and the leaf to fruit transport of photosynthates, thus promoting 13C-photosynthate accumulation in fruits. On the other hand, FA application elevated leaves’ N metabolism-related enzyme (GS and GOGAT) activity and optimized 15N distribution in leaves and fruits. Moreover, we also observed that FA application altered the fate of N fertilizer in apple orchards, showed an elevation in apple tree 15NUE and soil 15N residuals and showed a decrease in soil 15N loss. In summary, the appropriate application of FA150 (urea + 150 kg ha−1) synergistically optimized C and N nutrient distribution, and promoted anthocyanin biosynthesis in apple trees.

A Novel Cancer Therapy Using Proton Beam: An Integrated Proton-Dynamic Therapy, iPDT

[Background and Aims]A super-aged society, the prime example of which is Japan, is being flooded with increasing in numerous numbers of aged cancer patients, who often take anti-coagulant medicines to prevent the attacks of ischemic heart diseases and/ or strokes. Since they show bleeding tendency induced by the medicines, it’s hard for them to receive surgical operations. Thus, low invasive cancer treatments without bleedings are abundantly desired. In this study, the applicant will establish a new proton therapy, in which a resonance nuclear reaction (RNR) induced by collision of a proton with a stable nitrogen isotope (15N), can be a clue to injure cancer-cells specifically. The LET of the product ions (12C6+, 4He2+) of the 15N RNR is above 50 times in comparison with the LET of proton. To establish the cancer-specific 15N delivery, 15N-aminolevulinic acid (15N-ALA) will be used. This reagent has been already used and obtained regulatory approvals as a diagnostic medicine for the location of the glioblastoma in Japan.[Methods]In this study, a pair of rat gastric epithelial cells, one is normal cell RGM1, and another is cancerous characteristics cells RGK1 were used. The authors treated these cells with both 1mM 15N-ALA and 14N-ALA. Thereafter, we measured and compared porphyrin fluorescent intensity of these cells. Moreover, these cells will be irradiated with 4 nc proton beam generated by an electrostatic accelerator, then 4.43 MeV gamma-rays will be measured to demonstrate the amount of 15N collected specifically in the tumour cells. After these proton irradiations, cells were stained with hoechest33258 and dapi to observe cellular apoptosis.[Results] Cancer cells accumulated 9 folds of 15N in comparison with normal epithelial cells. After 3hours of proton irradiation, both cancer cells and normal cells were not injured. However, almost all cancer cells were died with apoptosis while normal cells showed few cellular injuries after 24 hours.[Conclusions] Cancer cells were completely killed by RNR involved with imM 15N-ALA and 4nc proton irradiation whereas this dose of proton showed few effects of ionizing radiation on normal cells. Thus, iPDT should be an ideal treatment for cancer. Moreover, the quantitative analysis of 15N in the tumour cells can be a new pre-diagnosis for iPDT.

High biological N fixation potential dominated by heterotrophic diazotrophs in alpine permafrost rivers on the Qinghai‒Tibet Plateau

Biological nitrogen (N) fixation is a pivotal N source in N-deficient ecosystems. The Qinghai‒Tibet Plateau (QTP) region, which is assumed to be N limited and suboxic, is an ideal habitat for diazotrophs. However, the diazotrophic communities and associated N fixation rates in these high-altitude alpine permafrost QTP rivers remain largely unknown. Herein, we examined diazotrophic communities in the sediment and biofilm of QTP rivers via the nitrogenase (nifH) gene sequencing and assessed their N fixing activities via a 15N isotope incubation assay. Strikingly, anaerobic heterotrophic diazotrophs, such as sulfate- and iron-reducing bacteria, had emerged as dominant N fixers. Remarkably, the nifH gene abundance and N fixation rates increased with altitude, and the average nifH gene abundance (2.57 ± 2.60 × 108 copies g−1) and N fixation rate (2.29 ± 3.36 nmol N g−1d−1) surpassed that documented in most aquatic environments (nifH gene abundance: 1.31 × 105 ∼ 2.57 × 108 copies g−1, nitrogen fixation rates: 2.34 × 10−4 ∼ 4.11 nmol N g−1d−1). Such distinctive heterotrophic diazotrophic communities and high N fixation potential in QTP rivers were associated with low-nitrogen, abundant organic carbon and unique C:N:P stoichiometries. Additionally, the significant presence of psychrophilic bacteria within the diazotrophic communities, along with the enhanced stability and complexity of the diazotrophic networks at higher altitudes, clearly demonstrate the adaptability of diazotrophic communities to extreme cold and high-altitude conditions in QTP rivers. We further determined that altitude, coupled with organic carbon and phosphorus, was the predominant driver shaping diazotrophic communities and their N-fixing activities. Overall, our study reveals high N fixation potential in N-deficient QTP rivers, which provides novel insights into nitrogen dynamics in alpine permafrost rivers.

Plant traits mediate foliar uptake of deposited nitrogen by mature woody plants.

Increased atmospheric nitrogen (N) deposition significantly disturbs ecosystem N cycle. Although foliar interception and uptake of N deposition can provide an important alternative N supply to forest ecosystems, the mechanisms regulating foliar N uptake from wet deposition are not fully understood. Here, we selected 19 woody species with a wide range of plant traits from different functional groups and conducted a 15N isotope labelling experiment through brushing 15NH4 + and 15NO3 - solution on canopy leaves. Our findings demonstrate that leaves can directly absorb N from wet deposition within a few hours. The average leaf 15N recoveries were 10% and 28% under 15NH4 + and 15NO3 - treatments across species, respectively, while twig N recoveries were only 1%-7% of leaf N recoveries. Differences in foliar N uptake efficiency among species were closely associated with leaf traits but were little influenced by meteorological conditions or soil nutrient status. Specifically, plants with higher leaf N concentration, larger specific leaf area and lower wax concentration exhibited higher leaf N recovery. Our results indicated that tree canopies could directly absorb N from atmospheric deposition. We highlight the critical role of leaf traits in determining canopy foliar N uptake, which may consequently influence plant competition under elevated N deposition.

Enhancement of nitrogen removal in constructed wetlands through boron mud and elemental sulfur addition: Regulation of sulfur and oxygen cycling

Constructed wetlands (CWs) utilizing elemental sulfur (S0) as an electron donor are effective for nitrate (NO3–-N) removal from low C/N wastewater. However, the limited bioavailability of chemical S0 (chem-S0) due to low water solubility and inadequate mass transfer impedes complete denitrification. In this study, boron mud waste and S0 (S0&BM) were processed and combined to convert chem-S0 to biological S0 (bio-S0) in CWs. The employment of S0&BM improved NO3–-N and total nitrogen (TN) removal rates by 15.36 % and 10.55 % compared to S0 alone. The 15N isotope tracing experiment unveiled the pathways of N2O production and demonstrated that S0&BM facilitated full denitrification, resulting in a higher percentage of nitrogen being degraded in a non-polluting form (54.70 %). Releasing trace amounts of boron and magnesium in S0&BM mitigated the excessive acidification caused by the sulfur autotrophic process and promoted biofilm formation, thereby pronouncing vertical oxygen fluctuations. Meanwhile, oxygen vacancies generated in the S0&BM promoted the closed-loop sulfur cycle, starting from the oxidation of chem-S0 to sulfate (SO42-), the important process of SO42--reducing bacteria using partial organic compounds as electron donors to reduce SO42- to sulfide (S2-) was realized, and finally the S2- was oxidized to bio-S0 with better properties. S0&BM enriched functional microbial communities in the biofilm related to nitrogen and sulfur transformation. Functional gene analysis showed an increase in the narG, nirS, norB, and nosZ genes supporting denitrification. The dsrA and sqr genes responsible for the sulfur cycle exhibited the highest abundance. This realization allowed for the utilization of waste materials for treatment, enhancing the energy efficiency of carbon reduction and pollution reduction in CWs.

Metagenomic insights into nitrogen cycling functional gene responses to nitrogen fixation and transfer in maize-peanut intercropping.

The fixation and transfer of biological nitrogen from peanuts to maize in maize-peanut intercropping systems play a pivotal role in maintaining the soil nutrient balance. However, the mechanisms through which root interactions regulate biological nitrogen fixation and transfer remain unclear. This study employed a 15N isotope labelling method to quantify nitrogen fixation and transfer from peanuts to maize, concurrently elucidating key microorganisms and genera in the nitrogen cycle through metagenomic sequencing. The results revealed that biological nitrogen fixation in peanut was 50 mg and transfer to maize was 230 mg when the roots interacted. Moreover, root interactions significantly increased nitrogen content and the activities of protease, dehydrogenase (DHO) and nitrate reductase in the rhizosphere soil. Metagenomic analyses and structural equation modelling indicated that nrfC and nirA genes played important roles in regulating nitrogen fixation and transfer. Bradyrhizobium was affected by soil nitrogen content and DHO, indirectly influencing the efficiency of nitrogen fixation and transfer. Overall, our study identified key bacterial genera and genes associated with nitrogen fixation and transfer, thus advancing our understanding of interspecific interactions and highlighting the pivotal role of soil microorganisms and functional genes in maintaining soil ecosystem stability from a molecular ecological perspective.

Stable isotopes reveal trophic ontogeny in Cisco (Coregonus artedi).

Cisco (Coregonus artedi) are a widespread, cold-water zooplanktivore native to North America. Although Cisco are generally referred to as an "obligate zooplanktivore," there is some evidence that the species exhibits considerable variability in trophic niche. Here, we assessed how Cisco body size relates to trophic position, that is, trophic ontogeny. We analysed 13C and 15N isotopes from Cisco ranging from 127 to 271 mm in body length (n = 66) from Trout Lake, Vilas County, Wisconsin, USA. 15N isotopes showed smaller Cisco had a trophic position of ~3, which steadily increased to ~3.5 for larger Cisco. Further, 13C isotope signatures showed Cisco transitioned to be more pelagically reliant (lower 13C signatures). Using gillnet catch data, we found that larger Cisco were using deeper habitats than smaller Cisco. Our results support that Cisco have significant variability in trophic niche even though they are traditionally thought of as an obligate planktivore. Overall, we emphasize that researchers should be cautious when generalizing Cisco trophic function, particularly when considering the broader food web.

Unveiling Charge-Pair Salt-Bridge Interaction Between GAGs and Collagen Protein in Cartilage: Atomic Evidence from DNP-Enhanced ssNMR at Natural Isotopic Abundance.

The interactions between glycosaminoglycans (GAGs) and proteins are essential in numerous biochemical processes that involve ion-pair interactions. However, there is no evidence of direct and specific interactions between GAGs and collagen proteins in native cartilage. The resolution of solid-state NMR (ssNMR) can offer such information but the detection of GAG interactions in cartilage is limited by the sensitivity of the experiments when 13C and 15N isotopes are at natural abundance. In this communication, this limitation is overcome by taking advantage of dynamic nuclear polarization (DNP)-enhanced magic-angle spinning (MAS) experiments to obtain two-dimensional (2D) 15N-13C and 13C-13C correlations on native samples at natural abundance. These experiments unveiled inter-residue correlations in the aliphatic regions of the collagen protein previously unobserved. Additionally, our findings provide direct evidence of charge-pair salt-bridge interactions between negatively charged GAGs and positively charged arginine (Arg) residues of collagen protein. We also identified potential hydrogen bonding interactions between hydroxyproline (Hyp) and GAGs, offering atomic insights into the biochemical interactions within the extracellular matrix of native cartilage. Our approach may provide a new avenue for the structural characterization of other native systems.