Natural Abundance Stable Isotope Research Articles

Temporal Variation in Community Composition of Root Associated Endophytic Fungi and Carbon and Nitrogen Stable Isotope Abundance in Two Bletilla Species (Orchidaceae).

Mycorrhizae are an important energy source for orchids that may replace or supplement photosynthesis. Most mature orchids rely on mycorrhizae throughout their life cycles. However, little is known about temporal variation in root endophytic fungal diversity and their trophic functions throughout whole growth periods of the orchids. In this study, the community composition of root endophytic fungi and trophic relationships between root endophytic fungi and orchids were investigated in Bletilla striata and B. ochracea at different phenological stages using stable isotope natural abundance analysis combined with molecular identification analysis. We identified 467 OTUs assigned to root-associated fungal endophytes, which belonged to 25 orders in 10 phyla. Most of these OTUs were assigned to saprotroph (143 OTUs), pathotroph-saprotroph (63 OTUs) and pathotroph-saprotroph-symbiotroph (18 OTUs) using FunGuild database. Among these OTUs, about 54 OTUs could be considered as putative species of orchid mycorrhizal fungi (OMF). For both Bletilla species, significant temporal variation was observed in the diversity of root endophytic fungi. The florescence and emergence periods had higher fungal community richness of total species and endemic species than did other periods. Both Bletilla species were dominated by Agaricomycetes and Basidiomycota fungi throughout the whole year; however, their abundances varied between two Bletilla species and among phenological stages. Meanwhile, the ranges of 13C and 15N natural abundance were also highly dynamic across all growth stages of Bletilla species. Compared with the surrounding autotrophic plants, significant 13C enrichments (ε13C) were found across all phenological stages, while significant 15N enrichment in the florescence period and strong 15N depletion during the fruiting period were found for both Bletilla species. We can deduce that both Bletilla species obtained carbon from root endophytic fungi during the whole year. Additionally, the temporal varying tendency of root endophytic fungal diversity was consistent with 13C enrichments, which was also accord with the nutritional requirement of plant.

An isotopic study of abiotic nitrite oxidation by ligand-bound manganese (III)

Redox transformations of nitrogen (N) play a critical role in determining its speciation and biological availability, thus defining the magnitude and extent of productivity in many ecosystems. A range of important nitrogen transformations often co-occur in regions hosting other redox-active elements, including sulfur, iron, and manganese (Mn), especially along sharp redox gradients within aquatic sediments. This proximity in “redox real estate” produces conditions under which multi-element interactions and coupled cycling are thermodynamically favored. While previous work has reported anoxic nitrification linked to the presence of manganese (Mn) oxides in sediments, a clear connection between the cycling of Mn and N has remained elusive. Soluble Mn(III), which is stabilized via ligand-complexation, has recently been shown to represent the dominant dissolved Mn species in many environments. Here, we examined the reactivity of ligand-stabilized Mn(III) with nitrite, using natural abundance stable nitrogen and oxygen isotopes to explore reaction dynamics under a range of conditions. Oxidation of nitrite to nitrate by Mn(III)-pyrophosphate proceeded abiotically under both oxygen replete and nitrogen-purged conditions. Kinetics and isotope systematics of this reaction were measured over a range of pH (5–8), with reaction rates decreasing with increasing pH. Under all treatments, an inverse kinetic isotope effect of −19.9 ± 0.7‰ was observed for N, remarkably similar to previously documented fractionation by nitrite-oxidizing organisms. Experiments using 18O-labeled water confirmed that the source of the additional oxygen atom was from water. These findings suggest that nitrite oxidation in environments hosting abundant ligand-bound Mn(III), including porewaters, estuaries, and coastal waters, may be facilitated in part by abiotic reactions with Mn, even under functionally anoxic conditions.

N2 fixation dominates nitrogen cycling in a mangrove fiddler crab holobiont

Mangrove forests are among the most productive and diverse ecosystems on the planet, despite limited nitrogen (N) availability. Under such conditions, animal-microbe associations (holobionts) are often key to ecosystem functioning. Here, we investigated the role of fiddler crabs and their carapace-associated microbial biofilm as hotspots of microbial N transformations and sources of N within the mangrove ecosystem. 16S rRNA gene and metagenomic sequencing provided evidence of a microbial biofilm dominated by Cyanobacteria, Alphaproteobacteria, Actinobacteria, and Bacteroidota with a community encoding both aerobic and anaerobic pathways of the N cycle. Dinitrogen (N2) fixation was among the most commonly predicted process. Net N fluxes between the biofilm-covered crabs and the water and microbial N transformation rates in suspended biofilm slurries portray these holobionts as a net N2 sink, with N2 fixation exceeding N losses, and as a significant source of ammonium and dissolved organic N to the surrounding environment. N stable isotope natural abundances of fiddler crab carapace-associated biofilms were within the range expected for fixed N, further suggesting active microbial N2 fixation. These results extend our knowledge on the diversity of invertebrate-microbe associations, and provide a clear example of how animal microbiota can mediate a plethora of essential biogeochemical processes in mangrove ecosystems.

Mycoheterotrophic plants living on arbuscular mycorrhizal fungi are generally enriched in 13C, 15N and 2H isotopes

Abstract Fully mycoheterotrophic plants are thought to obtain carbon exclusively from their root‐associated fungal partners. The general enrichment of these plants in the heavy isotopes 13C and 15N suggests that fungi are the main nutrient source for these plants. Yet, the majority of studies have targeted mycoheterotrophic plants associated with ectomycorrhizal, orchid mycorrhizal and saprotrophic fungi, while mycoheterotrophic plants living on arbuscular mycorrhizal fungi remain understudied. Here, we sampled 13 species of arbuscular mycorrhizal fully mycoheterotrophic plants from five families and co‐occurring autotrophic reference plants growing in forests of tropical South America, tropical South East Asia and temperate Australasia. We measured stable isotope natural abundances (δ13C, δ15N, δ2H and δ18O), determined total nitrogen concentrations and used high‐throughput DNA sequencing to characterize the arbuscular mycorrhizal fungal communities associated with the sampled mycoheterotrophic plants. We observed a general enrichment in 13C and 15N isotopes across mycoheterotrophic plant families and geographic regions. We confirm cases where no 15N enrichment is present, but we show that in general arbuscular mycoheterotrophic plants are enriched in 15N. Moreover, we demonstrate for the first time that these plants are significantly enriched in 2H but not in 18O in relation to their autotrophic references. The fungal communities targeted by the mycoheterotrophs mainly consist of Glomeraceae and show strong association with the isotopic signatures and geographic origin of the plants. Synthesis. Our findings enlarge the limited knowledge on the multi‐element stable isotopic signatures of mycoheterotrophic plants living on arbuscular mycorrhizal fungi. We show that these plants are enriched in 13C and 2H as expected due to their mycoheterotrophic nutrition, and that in general they are also enriched in 15N, despite some exceptions. Variation in stable isotope signatures is likely influenced by plant taxonomy, geography and fungal community composition.

Isotope applications to soil science at the University of Alberta — an historical perspective

For the past 70 yr, researchers in the Soil Science/Renewable Resources Department at the University of Alberta have used isotopes to study topics of ecological importance. This review highlights the soil isotope research conducted within our department over this time, including an historical overview of studies of interest. Analytical techniques and advances in instrumentation are discussed, focusing on the measurement of light stable isotope ratios (i.e., for C, H, N, S, and O) using isotope ratio mass spectrometry (IRMS). Early soil isotope work (1950–2000s) focused on agricultural soils and soil fertility issues. These studies included the use of radioactive isotopes such as 14C and 35S, and (or) artificially enriched stable isotopes including 15N-labelled fertilizers. More recently (2000–present), the scope of research widened to include natural-abundance stable isotope ratio studies as higher-sensitivity IRMS systems became more prevalent. Current isotope research topics include N biogeochemistry in natural and managed ecosystems, land management effects on greenhouse gas emissions, carbon cycling in northern landscapes, paleo-reconstruction in peatlands, carbon sequestration in boreal forests, and biodegradation of petroleum hydrocarbons. Further technological progress also enabled new techniques such as compound-specific IRMS analysis, including δ13C and δ2H measurements of soil n-alkanes and phospholipid fatty acids. In conclusion, current IRMS instrumentation presents unparalleled opportunities for multidisciplinary research to track carbon, plant nutrients, and pollutants as they move through soils.

Weaning and stunting affect nitrogen and carbon stable isotope natural abundances in the hair of young children

Natural abundances of stable nitrogen and carbon isotopes (δ15N and δ13C) can vary with both dietary intake and metabolic (specifically catabolic) state. In low-income countries, weaning is a period of dietary transition from milk to plant-based foods and a high-risk period for malnutrition. We explored how diet and malnutrition impact hair δ15N and δ13C in young children by an observational, cross-sectional study in Cox’s Bazar District, Bangladesh [255 children, 6–59 months with 19.6% wasted (7.1% severely) and 36% stunted (9.8% severely)]. Hair δ15N and δ13C exhibited exponential decreases with age, with the loss of one trophic level (3.3‰ and 0.8‰, respectively) from 6 to 48 months, which we associate with the shift from exclusive breastfeeding to complete weaning. After adjustment for age and breastfeeding status, hair isotopic values were unaffected by wasting but lower in severe stunting (−0.45‰ to −0.6‰, P < 0.01). In this population of young children, whose isotopic values in hair primarily depended on age, we failed to observe any effect of wasting, likely due to opposite, compensating effects between dietary and metabolic changes involved. In contrast, we evidenced low δ15N and δ13C values in severely stunted children that likely indicate chronic exposure to diets low in animal products.

Stable carbon isotope analyses offer insights into net carbon degradation of maize silages in anaerobic batch fermentations

Carbon degradation indicates the efficiency of anaerobic digestion processes. Common carbon degradation determination methods define gross carbon degradation (C deggross) of substrate and inoculum inseparably. The aim of this study was to test an isotope-based method defining solely substrate-based net carbon degradation (C degnet) on maize silage. As the natural abundance of stable isotopes in agricultural substrates vary, the method’s applicability was tested on (i) different maize silages sampled from agricultural farms, (ii) maize silage in fresh (MSfresh) and impaired storage (MSimpaired) conditions.Experiments included six maize silages digested in a total of 19 lab-scale batch reactors, analyzed for digestion parameters, stable isotopes, gross and net carbon degradation. MSimpaired showed significantly different stable carbon isotope composition at the start of the experiments, compared to MSfresh. Both methods indicated quality losses in MSimpaired. Results showed significantly higher C degnet values, ranging from 58.4% to 86.5%, compared to deggross values, ranging from 23.1% to 48.7%. This indicated the applicability of an isotope-based method C degnet to assess net carbon degradation of maize silages more detailed by excluding the masking effect of the inoculum. The isotope-based net carbon degradation method was found applicable on maize silages from (i) different farms and (ii) in different storage qualities.

Fate of bioavailable nutrients released to a stream during episodic effluent releases from a municipal wastewater treatment lagoon.

Municipal wastewater lagoons are common across North America and, unlike larger mechanical wastewater treatment plants, typically release nutrient-rich effluent directly to rivers in intermittent pulses. However, little is known about the fate of nutrients from these episodic events, which may happen under varying hydrologic or thermal conditions. We assessed fate of nitrogen (N) and phosphorus (P) from lagoon effluent during three releases to Deadhorse Creek, Manitoba, Canada. Using net nutrient uptake lengths and natural abundance stable isotope ratios of dissolved inorganic nitrogen (DIN) and primary producers, we found that DIN was processed during the summer releases though the dominant mechanism was unclear. However, nitrate was largely exported in autumn. Primary producers assimilated lagoon N but did not appear to reduce DIN concentrations. The longitudinal pattern of soluble reactive phosphorus (SRP) varied between releases and in summer 2019 the stream became a net source of SRP despite concomitant processing of DIN. We hypothesize that low demand for P in Deadhorse Creek, as suggested by upstream SRP > 0.05 mg P L-1, and nutrient ratios indicative of N limitation, reduced instream processing of P. Furthermore, our results indicated that cool or high flow conditions may result in the export of much of the lagoon nutrient load downstream. Our findings suggest the processes that transform wastewater nutrients are overwhelmed during effluent releases. Managers should consider increasing effluent dilution via continuous release of effluent rather than pulsed delivery. However, management of upstream nutrient supply may also be needed when relying upon the self-purifying capacity of rivers.

Controls on the Isotopic Composition of Nitrite (\u03b415N and \u03b418O) during Denitrification in Freshwater Sediments

The microbial reduction of nitrate, via nitrite into gaseous di-nitrogen (denitrification) plays a major role in nitrogen removal from aquatic ecosystems. Natural abundance stable isotope measurements can reveal insights into the dynamics of production and consumption of nitrite during denitrification. In this study, batch experiments with environmental bacterial communities were used to investigate variations of concentrations and isotope compositions of both nitrite and nitrate under anoxic conditions. To this end, denitrification experiments were carried out with nitrite or nitrate as sole electron acceptors at two substrate levels respectively. For experiments with nitrate as substrate, where the intermediate compound nitrite is both substrate and product of denitrification, calculations of the extent of isotope fractionation were conducted using a non-steady state model capable of tracing chemical and isotope kinetics during denitrification. This study showed that nitrogen isotope fractionation was lower during the use of nitrite as substrate (ε = −4.2 and −4.5‰ for both treatments) as compared to experiments where nitrite was produced as an intermediate during nitrate reduction (ε = −10 and −15‰ for both treatments). This discrepancy might be due to isotopic fractionation within the membrane of denitrifiers. Moreover, our results confirmed previously observed rapid biotic oxygen isotope exchange between nitrite and water.

Strain improvement of Escherichia coli K-12 for recombinant production of deuterated proteins

Deuterium isotope labelling is important for structural biology methods such as neutron protein crystallography, nuclear magnetic resonance and small angle neutron scattering studies of proteins. Deuterium is a natural low abundance stable hydrogen isotope that in high concentrations negatively affect growth of cells. The generation time for Escherichia coli K-12 in deuterated medium is substantially increased compared to cells grown in hydrogenated (protiated) medium. By using a mutagenesis plasmid based approach we have isolated an E. coli strain derived from E. coli K-12 substrain MG1655 that show increased fitness in deuterium based growth media, without general adaptation to media components. By whole-genome sequencing we identified the genomic changes in the obtained strain and show that it can be used for recombinant production of perdeuterated proteins in amounts typically needed for structural biology studies.

Distribution in natural abundances of stable isotopes of nitrate and retained sludge in a nitrifying bioreactor: Drastic changes in isotopic signatures

This study determined the spatial and temporal changes in natural abundance of stable isotopes (δ13C, δ15N, and δ18O) with regard to nitrate (NO3−) and retained sludge in a nitrifying bioreactor. The bioreactor was continuously fed with synthetic wastewater including ammonium for 61 days at 20 °C. After the start-up period of the bioreactor, the NO3− concentration in the effluent gradually increased. The stable isotopes (δ15N and δ18O) of NO3− in the effluent also increased in a phase of incomplete nitrification. The profile experiments showed that the concentration and stable isotopes of NO3− changed simultaneously along the wastewater flow in the bioreactor. The stable isotope analysis revealed that nitrification efficiency seems to be strongly related to the δ15N of NO3−. Moreover, the δ13C and δ15N of the retained sludge drastically changed along the reactor length, from −26‰ to −18‰ and from 5‰ to 30‰, respectively, after 61 days of operation. The isotopic composition of the retained sludge might be affected by the isotope ratios (δ15N and δ18O) of NO3− in the bioreactor. Therefore, the isotope signatures of the retained sludge seem to closely reflect process performance such as nitrification efficiency throughout the operational period. Our findings suggest that the spatial distribution of the isotopic composition of the retained sludge can be used to detect process occurrence within the bioreactor over time.

Plant water uptake along a diversity gradient provides evidence for complementarity in hydrological niches

Biodiversity enhances a variety of ecosystem processes, and yet the underlying mechanisms through which these relationships occur remain a critical knowledge gap. Here, we used the natural abundance of stable isotopes to measure depth of water uptake in five common grassland species (Asclepias tuberosa, Lespedeza capitata, Liatris aspera, Schizachyrium scoparium and Sorghastrum nutans) growing across an experimental grassland diversity gradient. Using this approach, we addressed the following questions: 1) does the depth‐specific provenance of water uptake differ among species and/or do interspecific differences in water source manifest with increasing community diversity? 2) Does the isotopic niche space occupied by plants change with increasing diversity? 3) Is plasticity in water uptake depth across a diversity gradient associated with functional plant responses? We found that the depth of soil water used by plants was inherently different among species when grown in monocultures. All species used less shallow soil water and more intermediate‐depth soil water in mixed assemblages than in monocultures, resulting in similar interspecific differences in water source across the diversity gradient. However, plasticity in the locations of water used were positively associated with increases in plant growth in higher diversity treatments. These results indicate that plasticity in water‐use may contribute to positive biodiversity–productivity relationships commonly observed in temperate grasslands.

Picky carnivorous plants? Investigating preferences for preys' trophic levels - a stable isotope natural abundance approach with two terrestrial and two aquatic Lentibulariaceae tested in Central Europe.

Stable isotope two-source linear mixing models are frequently used to calculate the nutrient-uptake efficiency of carnivorous plants from pooled prey. This study aimed to separate prey into three trophic levels as pooled prey limits statements about the contribution of a specific trophic level to the nutrition of carnivorous plants. Phytoplankton were used as an autotrophic reference for aquatic plants as the lack of suitable reference plants impedes calculation of their efficiency. Terrestrial (Pinguicula) and aquatic (Utricularia) carnivorous plants alongside autotrophic reference plants and potential prey from six sites in Germany and Austria were analysed for their stable isotope natural abundances (δ15N, δ13C). A two-source linear mixing model was applied to calculate the nutrient-uptake efficiency of carnivorous plants from pooled prey. Prey preferences were determined using a Bayesian inference isotope mixing model. Phytophagous prey represented the main contribution to the nutrition of Pinguicula (approx. 55 %), while higher trophic levels contributed a smaller amount (diverse approx. 27 %, zoophagous approx. 17 %). As well as around 48 % nitrogen, a small proportion of carbon (approx. 9 %) from prey was recovered in the tissue of plants. Aquatic Utricularia australis received 29 % and U. minor 21 % nitrogen from zooplankton when applying phytoplankton as the autotrophic reference. The separation of prey animals into trophic levels revealed a major nutritional contribution of lower trophic level prey (phytophagous) for temperate Pinguicula species. Naturally, prey of higher trophic levels (diverse, zoophagous) are rarer, resulting in a smaller chance of being captured. Phytoplankton represents an adequate autotrophic reference for aquatic systems to estimate the contribution of zooplankton-derived nitrogen to the tissue of carnivorous plants. The autonomous firing of Utricularia bladders results in the additional capture of phytoplankton, calling for new aquatic references to determine the nutritional importance of phytoplankton for aquatic carnivorous plants.

Light limitation and partial mycoheterotrophy in rhizoctonia-associated orchids.

Partially mycoheterotrophic (PMH) plants obtain organic molecules from their mycorrhizal fungi in addition to carbon (C) fixed by photosynthesis. Some PMH orchids associated with ectomycorrhizal fungi have been shown to flexibly adjust the proportion of organic molecules obtained from fungi according to the habitat's light level. We hypothesise that Neottia ovata and Ophrys insectifera, two orchids associated with saprotrophic rhizoctonia fungi, are also able to increase uptake of organic molecules from fungi as irradiance levels decrease. We continuously measured light availability for individuals of N. ovata and O. insectifera at a grassland and a forest during orchid flowering and fruiting. We repeatedly sampled leaves of N. ovata, O. insectifera and autotrophic reference species for stable isotope natural abundances (δ13C, δ15N, δ2H, δ18O) and C and N concentrations. We found significant 13C enrichment in both orchids relative to autotrophic references at the forest but not the grassland, and significant 2H enrichment at both sites. The 13C enrichment in O. insectifera was linearly correlated with the habitat's irradiance levels. We conclude that both species can be considered as PMH and at least in O. insectifera, the degree of partial mycoheterotrophy can be fine-tuned according to light availability. However, exploitation of mycorrhizal fungi appears less flexible in saprotroph-associated orchids than in orchids associated with ectomycorrhizal fungi.

Urine effects on grass and legume nitrogen isotopic composition: Pronounced short-term dynamics of δ15N.

Nitrogen stable isotope (15N) natural abundance is widely used to study nitrogen cycling. In grazed ecosystems, urine patches are hot-spots of nitrogen inputs, losses, and changes in δ15N. Understanding δ15N dynamics in urine-affected vegetation is therefore crucial for accurate inferences from 15N natural abundance in grasslands. We hypothesized that leaf δ15N following urine deposition varies with time and plant functional group. Specifically, we expected (i) short-term decreases in δ15N due to foliar absorption of 15N-depleted volatilized ammonia, (ii) followed by increases in δ15N due to uptake of 15N-enriched soil inorganic nitrogen, and (iii) that the magnitude of these changes is less in legumes than in grasses. The latter should be expected because ammonia absorption depends on leaf nitrogen concentration, which is higher in legumes than grasses, and because biological nitrogen fixation will modify the influence of urine-derived nitrogen on δ15N in legumes. We applied cattle urine to a mixture of Lolium perenne and Trifolium repens in a pot experiment. Nitrogen concentration and δ15N were determined for successive leaf cohorts and bulk biomass either 17 (early) or 32 (late) days after urine application. Early after urine application, leaves of L. perenne were 15N-depleted compared to control plants (δ15N 0.1 vs. 5.8‰, respectively), but leaves of T. repens were not (-1.1 vs. -1.1‰, respectively). Later, both species increased their δ15N, but T. repens (4.5‰) less so than L. perenne (5.9‰). Vegetation sampled within and outside urine patches in the field further supported these results. Our findings confirm that foliar ammonia uptake can substantially decrease grass foliar δ15N, and that in both grass and legume the direction of the δ15N response to urine changes over time. Temporal dynamics of plant δ15N at urine patches therefore need to be explicitly addressed when 15N natural abundance is used to study nitrogen cycling in grazed grasslands.

Estimating contributions of pelagic and benthic pathways to consumer production in coupled marine food webs.

Pelagic and benthic systems usually interact, but their dynamics and production rates differ. Such differences influence the distribution, reproductive cycles, growth rates, stability and productivity of the consumers they support. Consumer preferences for, and dependence on, pelagic or benthic production are governed by the availability of these sources of production and consumer life history, distribution, habitat, behavioural ecology, ontogenetic stage and morphology. Diet studies may demonstrate the extent to which consumers feed on prey in pelagic or benthic environments. But they do not discriminate benthic production directly supported by phytoplankton from benthic production recycled through detrital pathways. The former will track the dynamics of phytoplankton production more closely than the latter. We develop and apply a new analytical method that uses carbon (C) and sulphur (S) natural abundance stable isotope data to assess the relative contribution of pelagic and benthic pathways to fish consumer production. For 13 species of fish that dominate community biomass in the northern North Sea (estimated >90% of total biomass), relative modal use of pelagic pathways ranged from <25% to >85%. Use of both C and S isotopes as opposed to just C reduced uncertainty in relative modal use estimates. Temporal comparisons of relative modal use of pelagic and benthic pathways revealed similar ranking of species dependency over 4years, but annual variation in relative modal use within species was typically 10%-40%. For the total fish consumer biomass in the study region, the C and S method linked approximately 70% and 30% of biomass to pelagic and benthic pathways, respectively. As well as providing a new method to define consumers' links to pelagic and benthic pathways, our results demonstrate that a substantial proportion of fish biomass, and by inference production, in the northern North Sea is supported by production that has passed through transformations on the seabed.

Serum Nitrogen and Carbon Stable Isotope Ratios Meet Biomarker Criteria for Fish and Animal Protein Intake in a Controlled Feeding Study of a Women's Health Initiative Cohort

Natural abundance stable isotope ratios are candidate biomarkers of dietary intake that have not been evaluated in a controlled feeding study in a US population. Our goals were to evaluate dietary associations with serum carbon (CIR), nitrogen (NIR), and sulfur (SIR) isotope ratios in postmenopausal women, and to evaluate whether statistical models of dietary intake that include multiple isotopes and participant characteristics meet criteria for biomarker evaluation. Postmenopausal women from the Women's Health Initiative (n=153) were provided a 2-wk controlled diet that approximated each individual's habitual food intake. Dietary intakes of animal protein, fish/seafood, red meat, poultry, egg, dairy, total sugars, added sugars, sugar-sweetened beverages (SSBs), and corn products were characterized during the feeding period with the use of the Nutrition Data System for Research (NDS-R). The CIR, NIR, and SIR were measured in sera collected from fasting women at the beginning and the end of the feeding period. Linear models based on stable isotope ratios and participant characteristics predicted dietary intake. The criterion used for biomarker evaluation was R2≥0.36, based on the study's power to detect true associations with R2≥0.50. The NIR was associated with fish/seafood intake and met the criterion for biomarker evaluation (R2=0.40). The CIR was moderately associated with intakes of red meat and eggs, but not to the criterion for biomarker evaluation, and was not associated with intake of sugars (total, added, or SSB). A model of animal protein intake based on the NIR, CIR, and participant characteristics met the criterion for biomarker evaluation (R2=0.40). Otherwise, multiple isotopes did not improve models of intake, and improvements from including participant characteristics were modest. Serum stable isotope ratios can, with participant characteristics, meet biomarker criteria as measures of fish/seafood and animal protein intake in a sample of postmenopausal women. This trial was registered at clinicaltrials.gov as NCT00000611.

Stable isotope natural abundances (δ13C and δ15N) and carbon-water relations as drought stress mechanism response of taro (Colocasia esculenta L. Schott)

Taro (Colocasia esculenta L. Schott) is an important staple food crop in tropical and developing countries, having high water requirements. The purpose of this study was to evaluate the feasibility of using carbon and nitrogen isotopic composition (δ13C and δ15N) as a physiological indicator of taro response to drought, and elucidation of the relationship between the water use efficiency (WUE) under drought conditions and carbon isotope discrimination (Δ13C). As an alternative to WUE determination, obtained by measuring plant growth and water loss during an entire vegetative cycle, we have used Δ13C to determine the tolerance of C3 taro plants to drought. Seven taro accessions from Madeira, Canary Islands and the Secretariat of the Pacific Community (Fiji) collections were grown under greenhouse conditions and subjected to different watering regimes during a one-year cycle. Total plant biomass (TPB), WUE and δ15N were determined at the whole-plant level (WP). Corms and shoots were evaluated separately for nitrogen content (N), δ13C, Δ13C and δ15N. WUE showed positive correlation with TPB (r = 0.4) and negative with Δ13C (r = −0.3); Corm δ15N showed positive correlations with WP δ15N (r = 0.6) and corm N (r = 0.3). Accordingly, the taro plants with enhanced WUE exhibited low Δ13C and δ15N values as a physiological response to drought stress. The approach used in the present study has developed new tools that could be used in further research on taro response to environmental stresses.

Isotopic evidence of biotrophy and unusual nitrogen nutrition in soil-dwelling Hygrophoraceae.

SummarySeveral lines of evidence suggest that the agaricoid, non‐ectomycorrhizal members of the family Hygrophoraceae (waxcaps) are biotrophic with unusual nitrogen nutrition. However, methods for the axenic culture and lab‐based study of these organisms remain to be developed, so our current knowledge is limited to field‐based investigations. Addition of nitrogen, lime or organophosphate pesticide at an experimental field site (Sourhope) suppressed fruiting of waxcap basidiocarps. Furthermore, stable isotope natural abundance in basidiocarps were unusually high in 15N and low in 13C, the latter consistent with mycorrhizal nutritional status. Similar patterns were found in waxcap basidiocarps from diverse habitats across four continents. Additional data from 14C analysis of basidiocarps and 13C pulse label experiments suggest that these fungi are not saprotrophs but rather biotrophic endophytes and possibly mycorrhizal. The consistently high but variable δ15N values (10–20‰) of basidiocarps further indicate that N acquisition or processing differ from other fungi; we suggest that N may be derived from acquisition of N via soil fauna high in the food chain.

Nitrogen source track and associated isotopic dynamic characteristic in a complex ecosystem: A case study of a subtropical watershed, China.

By identifying the main sources of nitrate (NO3-) can obtain useful information to support the management of NO3- pollution, particularly in subtropical catchments with shallow drinking water wells. This study used water chemistry and dual stable isotopes δ15N and δ18O methods to assess seasonal and spatial variations of NO3- in precipitation, surface water, and groundwater in an agricultural and forest subtropical catchment in Jiangxi Province, China. The maximum concentrations of nitrate-nitrogen (NO3--N) and ammonium-nitrogen (NH4+-N) were 10.4 and 10.8 mg L-1in samples collected from 221 rainfall events from 2011 to 2013. About 4.4% and 12.3% NH4+-N concentrations of surface water and groundwater exceeded the thresholds of 1.0 and 0.2 mg L-1. The NO3--N concentrations in surface water were closely correlated with NH4+-N concentrations in surface water and groundwater (r = -0.71 and r = -0.71, P < 0.05). The concentrations of NH4+-N and NO3--N were significantly higher in a fishery pond and nearby drinking wells than in other monitoring points. Annual exports of NO3--N and NH4+-N were 4.06 × 104 and 8.14 × 103 kg yr-1, respectively and NO3--N is the main form of N loss. The δ15N values ranged from 0‰ to 20‰ in surface water and groundwater, and the δ18O values ranged from 0‰ to 15‰ and 1‰-13‰, respectively. Dual stable isotope natural abundance distribution and water chemistry [NO3-]/[Cl-] molar ratio information suggested that manure and sewage and soil N were the main sources of NO3- in surface water and manure and sewage in groundwater in summer and winter. In spring, water occurred denitrification and ammonium fertilizer, manure and sewage were the main sources of NO3- in surface water and groundwater which sampling points were closer residential area and fish ponds than paddy field and local farmers used more Manure. Manure applications should be reasonable around drinking water wells to protect the drinking water quality.