Isotope Discrimination Research Articles

Chiral isotopic fractionation in lemon essential oil: A tool for authenticity assessment?

The present research aimed to retrieve key information about the genuineness of Sicilian lemon essential oils by evaluating simultaneously the chiral and isotopic data of target terpene components. With respect to previous literature references, where chiral recognition and isotope discrimination were performed by distinct gas chromatographic methods, this study aimed to develop a single analytical approach. To overcome limitations associated to monodimensional gas chromatographic approaches, an enantio‑selective multidimensional gas chromatographic approach coupled to isotopic ratio mass spectrometry and to parallel single quadrupole detection (Es-MDGC-C-IRMS/qMS) was developed. Thanks to the features of this system, enantiomeric excesses and target δ13C of the chiral and achiral components were evaluated in a single gas chromatographic run, allowing to reduce total time analysis, as well the consumption of electricity, solvents and samples. Moreover, due to the capability to baseline separate the enantiomeric couples, further considerations were done about the specific δ13C value of the target separated enantiomers. Dealing with the genuine lemon oils analysed, a different δ13C value was found between the enantiomers of the same chiral component, namely (-) and (+) of α and β-pinene, suggesting a different isotopic fractionation related to a specific biosynthetic pathway. This research aimed to evaluate the reasons behind this behaviour, paving the way to newer considerations in the field of authenticity assessment.

The extent of nitrogen isotopic fractionation in rumen bacteria is associated with changes in rumen nitrogen metabolism.

Nitrogen use efficiency is an important index in ruminants and can be indirectly evaluated through the N isotopic discrimination between the animal and its diet (Δ15Nanimal-diet). The concentration and source of N may determine both the extent of the N isotopic discrimination in bacteria and N use efficiency. We hypothesised that the uptake and release of ammonia by rumen bacteria will affect the natural 15N enrichment of the bacterial biomass over their substrates (Δ15Nbacteria-substrate) and thereby further impacting Δ15Nanimal-diet. To test this hypothesis, two independent in vitro experiments were conducted using two contrasting N sources (organic vs inorganic) at different levels either in pure rumen bacteria culture incubations (Experiment #1) or in mixed rumen cultures (Experiment #2). In Experiment #1, tryptone casein or ammonium chloride were tested at low (1 mM N) and high (11.5 mM N) concentrations on three rumen bacterial strains (Fibrobacter succinogenes, Eubacterium limosum and Xylanibacter ruminicola) incubated in triplicate in anaerobic batch monocultures during 48h. In Experiment #2 mixed rumen cultures were incubated during 120 h with peptone or ammonium chloride at five different levels of N (1.5, 3, 4.5, 6 and 12-mM). In experiment #1, Δ15Nbacteria-substrate was lowest when the ammonia-consumer bacterium Fibrobacter succinogenes was grown on ammonium chloride, and highest when the proteolytic bacterial strain Xylanibacter ruminicola was grown on tryptone. In experiment #2, Δ15Nbacteria-substrate was lower with inorganic (ammonium chloride) vs organic (peptone) N source. A strong negative correlation between Δ15Nbacteria-substrate and Rikenellaceae_RC9_gut_group, a potential fibrolytic rumen bacterium, was detected. Together, our results showed that Δ15Nbacteria-substrate may change according to the balance between synthesis of microbial protein from ammonia versus non-ammonia N sources and confirm the key role of rumen bacteria as modulators of Δ15Nanimal-diet.

Carbon isotope discrimination (so-called δ<sup>13</sup>C) measured on grape juice is an accessible tool to monitor vine water status in production conditions

Assessment of vine water status is needed to understand the effect of environmental factors and management practices on dry-farmed and irrigated vineyards. Among plant-based indicators, carbon isotope discrimination (δ13C) is easily accessible, reliable, and inexpensive. As it provides a post-hoc assessment of vine water status during the berry ripening period, it can be useful for assessing the results of vineyard management practices during the season, and to map water status in the vineyard to aid in future precision management. Possible applications and limitations of this technique for practical vineyard management are discussed in this article.

Plasticity of Plantago lanceolata L. in Adaptation to Extreme Environmental Conditions.

This study aimed at characterizing some adaptive changes in Plantago lanceolata L. exposed to harsh conditions of a desert-like environment generating physiological stress of limited water availability and exposure to strong light. It was clearly shown that the plants were capable of adapting their root system and vascular tissues to enable efficient vegetative performance. Soil analyses, as well as nitrogen isotope discrimination data show that P. lanceolata leaves in a desert-like environment had better access to nitrogen (nitrite/nitrate) and were able to fix it efficiently, as compared to the plants growing in the surrounding forest. The arbuscular mycorrhiza was also shown to be well-developed, and this was accompanied by higher bacterial frequency in the root zone, which might further stimulate plant growth. A closer look at the nitrogen content and leaf veins with a higher number of vessels and a greater vessel diameter made it possible to define the changes developed by the plants populating sandy habitats as compared with the vegetation sites located in the nearby forest. A determination of the photosynthesis parameters indicates that the photochemical apparatus in P. lanceolata inhabiting the desert areas adapted slightly to the desert-like environment and the time of day, with some changes of the reaction center (RC) size (photosystem II, PSII), while the plants' photochemical activity was at a similar level. No differences between the two groups of plants were observed in the dissipation of light energy. The exposure of plants to harsh conditions of a desert-like environment increased the water use efficiency (WUE) value in parallel with possible stimulation of the β-carboxylation pathway.

Herbarium specimens reveal century-long trait shifts in poison ivy due to anthropogenic CO2 emissions.

Previous experimental studies have shown that poison ivy (Toxicodendron radicans; Anacardicaceae) responds to elevated CO2 with increased leaf production, water-use efficiency, and toxicity (allergenic urushiol). However, long-term field data suggest no increase in poison ivy abundance over time. Using herbarium specimens, we examined whether poison ivy and other species shifted leaf traits under natural conditions with increasing atmospheric CO2 (pCO2 ) over the past century. We measured stomatal density, leaf area, leaf N, leaf C:N, leaf carbon isotope discrimination (Δleaf ), and intrinsic water-use efficiency (iWUE) from 327 specimens collected from 1838 to 2020 across Pennsylvania. We compared poison ivy's responses to two evolutionarily related tree species, Toxicodendron vernix and Rhus typhina (Anacardiacae) and one ecological analog, Parthenocissus quinquefolia (Vitaceae), a common co-occurring liana. Stomatal density significantly decreased (P < 0.05) in poison ivy and the ecologically similar liana P. quinquefolia over the past century, but did not change in the related trees T. vernix and R. typhina. None of these species showed significant trends in changes in leaf N or C:N. Surprisingly, in poison ivy, but not the other species, Δleaf increased with increased pCO2 , corresponding to significant declines in iWUE over time. In contrast to the results of short-term experimental studies, iWUE decreased in poison ivy over the last century. Trait responses to pCO2 varied by species. Herbarium specimens suggest that realized long-term plant physiological responses to increased CO2 may not be reflected in short-term experimental growth studies, highlighting the value of collections.

Extrusion of lupines with or without addition of reducing sugars: Effects on the formation of Maillard reaction compounds, partition of nitrogen and Nε-carboxymethyl-lysine, and performance of dairy cows

The extrusion of leguminous seeds induces the formation of Maillard reaction compounds (MRC) as a product of protein advanced glycation and oxidation, which lowers protein degradability in the rumen. However, the quantitative relationship between the parameters of pretreatment (i.e., addition of reducing sugars) and extrusion, and the formation of MRC has not been established yet. Moreover, the fate of the main stable MRC, Nε-carboxymethyl-lysine (CML), in the excretory routes has never been investigated in ruminants. We aimed to test the effects of the temperature of extrusion of white lupines with or without addition of reducing sugars on the formation of MRC, crude protein (CP) degradability in the rumen, N use efficiency for milk production (milk N/N intake), and performance of dairy cows. Two experiments with a replicated 4 × 4 Latin square design were conducted simultaneously with 16 (3 rumen-cannulated) multiparous Holstein cows to measure indicators of ruminal CP degradability (ruminal NH3 concentration, branched-chain volatile fatty acids), metabolizable protein supply (plasma essential AA concentration), N use efficiency (N isotopic discrimination), and dairy performance. In parallel, apparent total-tract digestibility of dry matter, organic matter, neutral detergent fibers, N, total Lys and CML, and partition of N and CML were measured with 4 cows in both experiments. The diets consisted on a DM basis of 20% raw or extruded lupines and 80% basal mixed ration of corn silage, silage and hay from permanent grasslands, pelleted concentrate, and a vitaminized mineral mix. Expected output temperatures of lupine extrusion were 115°C, 135°C, and 150°C, without and with the addition of reducing sugars before extrusion. The extrusion numerically reduced the in vitro ruminal CP degradability of the lupines, and consequently increased the predicted supply of CP to the small intestine. Nitrogen balance and urinary N excretion did not differ among dietary treatments in either experiment. Milk yield and N use efficiency for milk production increased with extrusion of lupines at 150°C without addition of reducing sugars compared with raw lupines. Nitrogen isotopic discrimination between dietary and animal proteins (the difference between δ15N in plasma and δ15N in the diet) were lower with lupines extruded at 150°C without and with addition of reducing sugars. Regardless of sugar addition, milk true protein yield was not affected, but milk urea concentration and fat:protein ratio were lower with lupines extruded at 150°C than with raw lupines. In the CML partition study, we observed that on average 26% of the apparently digested CML was excreted in urine, and a much lower proportion (0.63% on average) of the apparently digested CML was secreted in milk, with no differences among dietary treatments. In conclusion, we showed that the extrusion of white lupines without or with addition of reducing sugars numerically reduced enzymatic CP degradability, with limited effects on N partition, but increased milk yield and N use efficiency at the highest temperature of extrusion without addition of reducing sugars.

Fatty acid isotopic composition in Atlantic pollock is not influenced by environmentally relevant dietary fat concentrations.

The application of fatty acid (FA) isotopic analysis has great potential in elucidating food web structure, but it has not experienced the same wide-spread use as amino acid isotopic analyses. The failure to adopt FA isotopic methods is almost certainly linked to a lack of reliable information on trophic fractionation of FA, particularly in higher predators. In this work, we attempt to address this shortfall, through comparison of FA δ13C values in captive Atlantic pollock (Pollachius virens) liver and their known diets. Since catabolism is likely the main cause of fractionation and it may vary with dietary fat content, we investigated the impact of dietary fat concentration on isotopic discrimination in FA. We fed Atlantic pollock three formulated diets with similar FA isotopic compositions but different fat concentrations (5-9% of diet), representative of the range found in natural prey, for 20weeks. At the conclusion of the study, δ13C values of liver FA were very similar to the FA within the corresponding diets, with most discrimination factors < 1. For all FA except 22:6n-3, dietary fat had no effect on discrimination factors. Only for 22:6n-3 did fish fed the highest fat diet have lower δ13C values than the diet consumed. Thus, these FA-specific discrimination factors can be applied to evaluate diets in marine fish consuming natural diets and will serve as additional and valuable biomarkers in fish feeding ecology.

Variations of deep water uptake and water use efficiency indicated divergence in tree growth stability

Insights into forest growth stability in response to intensified climate change and disturbances is critical to maintain the continued flow and stability of forest services. However, the underlying drivers for tree growth stability remain poorly understood. In this study, we quantified the spatiotemporal pattern in tree growth stability of Chinese pine (Pinus tabulaeformis) plantations and its relationship to diverse driving factors (climate, stand structure, ecophysiological traits, soil properties, and topography) across a great climatic gradient in northern China in the period of 1979–2018. We calculated the coefficient of variation of tree ring chronology (TRIcv) to characterize tree growth stability and revealed that the TRIcv increased significantly with increasing multiyear mean aridity index (MAI) and vapor pressure deficit (VPD) spatially. Temporally, TRIcv displayed increasing trend in 25 of 40 study sites. Constrained redundancy analysis revealed that TRIcv was strongly mediated by water availability, water uptake from different soil layers, wood density, tree height, leaf stable carbon isotope discrimination (δ13C), and leaf nutrients. The ratio of water uptake from 60 to 80 cm soil layer and leaf δ13C were the two most important ecohydrological-related factors determining TRIcv, highlighting that the water uptake strategy and leaf water use efficiency play critical role in reshaping the tree growth stability of Chinese pine to cope with the intensifying climate change.

Phenotypic variation in leaf photosynthetic traits, leaf area index, and carbon discrimination of field-grown wheat genotypes and their relationship with yield performance in Mediterranean environments.

Leaf water potential, gas exchange, and chlorophyll fluorescence exhibited significant differences among genotypes, high environmental effects, but low heritability. The highest-yielding and drought-tolerant genotypes presented superior harvest index and grain weight, compared to drought-susceptible ones. Physiological phenotyping can help identify useful traits related to crop performance under water-limited conditions. A set of fourteen bread wheat genotypes with contrasting grain yield (GY) was studied in eight Mediterranean environments in Chile, resulting from the combination of two sites (Cauquenes and Santa Rosa), two water conditions (rainfed-WL and irrigated-WW), and four growing seasons (2015-2018). The objectives were to (i) evaluate the phenotypic variation of leaf photosynthetic traits after heading (anthesis and grain filling) in different environments; (ii) analyze the relationship between GY and leaf photosynthetic traits and carbon isotope discrimination (Δ13C); and (iii) identify those traits that could have a greater impact in the determination of tolerant genotypes under field conditions. Agronomic traits exhibited significant genotypic differences and genotype × environment (GxE) interaction. The average GY under the WW condition at Santa Rosa was 9.2Mgha-1 (range 8.2-9.9Mgha-1) and under the WL condition at Cauquenes was 6.2Mgha-1 (range 3.7-8.3Mgha-1). The GY was closely related to the harvest index (HI) in 14 out of 16 environments, a trait exhibiting a relatively high heritability. In general terms, the leaf photosynthetic traits presented low GxE interaction, but high environmental effects and low heritability, except for the chlorophyll content. The relationships between GY and leaf photosynthetic traits were weaker when performed across genotypes in each environment, indicating low genotypic effects, and stronger when performed across environments for each genotype. The leaf area index and Δ13C also presented high environmental effects and low heritability, and their correlations with GY were influenced by environmental effects. The highest-yielding and drought-tolerant genotypes presented superior HI and grain weight, but no clear differences in leaf photosynthetic traits or Δ13C, compared to drought-susceptible ones. It seems that the phenotypic plasticity of agronomic and leaf photosynthetic traits is very important for crop adaptation to Mediterranean environments.

The Effect of Increasing Irrigation Rates on the Carbon Isotope Discrimination of Apple Leaves

13C discrimination (Δ13C) has been used in research as an indicator of water availability in crops; however, few data are available concerning fruit trees. The aim of this study was to examine the effect of irrigation on the Δ13C values of apple leaves. We assumed that Δ13C would increase with irrigation intensity. The Δ13C of apple trees (Malus domestica) cv. ‘Red Jonaprince’ was determined in the years 2019–2022. Leaf samples were collected in spring, in June, at the beginning of the irrigation campaign, and in autumn, in September, following the harvest. The irrigation doses were applied to replenish the water consumption, 0% (ET0), 50% (ET50), 75% (ET75), and 100% (ET100), of the calculated evapotranspiration (ET) levels. In November, the leaves collected from different positions on the shoots were sampled, assuming the Δ13C signature would reflect the changes occurring in the water supply during their growth. The irrigation rates had a significant effect on the Δ13C of the leaves when the data for the spring and summer months were pooled. On average, Δ13C increased from 20.77‰ and 20.73‰ for ET0 and ET50, respectively, to 20.80‰ and 20.95‰ for ET75 and ET100, respectively. When the data obtained for the spring and summer months were analysed separately, the effect of irrigation was weak (p &lt; 0.043). The Δ13C value was always higher for treatment ET100 than treatment ET0, for individual experimental years and terms; however, the differences were minor and mostly insignificant. The leaf position had a strong significant effect on Δ13C; the values gradually decreased from the leaves growing from two-years-old branches (22.50‰) to the youngest leaves growing at the top of the current year’s shoots (21.07‰). This order was similar for all the experimental years. The results of the experiment suggest that 13C discrimination in apples is affected not only by water availability during growth, but also by the use of C absorbed in previous years.

Large-acceptance isotope identification array FAZIA: Status and R&D activities for upgrade

The FAZIA apparatus is a multi-detector array designed to identify a wide range of charge and mass of reaction products in heavy-ion collisions in the Fermi energy domain. The basic module of FAZIA is the block, consisting of 16 three-layer telescopes. The first two layers are highly homogeneously doped Si detectors with thicknesses of 300μm and 500μm, respectively, and the third layer is a 10 cm thick CsI(Tl) scintillator read out by a photodiode. The detector signals are extracted in real time based on the digital signal processing implemented on the FPGAs. The recent experiments demonstrated that the charge could be discriminated up to more than Z = 54 using the ΔE-E technique and the pulse shape analysis. In addition, the isotopic discrimination has been achieved up to Z ∼ 25 with the ΔE-E technique and up to Z ∼ 20 with the pulse shape analysis in the silicon layer. Recently, there have been activities for the FAZIA detector upgrade to use thicker and thinner silicon layers for enlarging the kinematic coverage. In addition, R&D is in progress to make the front-end electronics board more compact and versatile. This paper is dedicated to the description of the current status of the FAZIA detector and some highlights of the R&D activities for the upgrade.

Tissue‐ and diet‐dependent stable carbon and nitrogen isotope discrimination: a calibration study in a captive shorebird species

In ecology, stable‐isotope ratios are widely used to determine diets of organisms and reconstruct food webs. This is usually done by analyzing the stable‐isotope ratios of nitrogen (δ15N), which increase with increasing trophic level, and those of carbon (δ13C), which correlate with the δ13C value of food source(s) and generally differ between terrestrial and marine food sources. Assimilation of food changes stable‐isotope ratios, resulting in different values between the food source and its consumer. These differences are known as isotope trophic discrimination factors and, if known, can be used to determine from the stable‐isotope ratios in the consumer's tissue what the consumer has been eating. What is often ignored is that discrimination factors can differ between consumer's food sources and also between tissue types. Therefore, we performed a controlled feeding study in red knots Calidris canutus to determine discrimination factors between different food sources and red knot tissues. We kept two groups of red knots in captivity on a stable diet, one group feeding on mudsnails and the other on Trouvit pellets, for several months, during which the birds molted their feathers. We analyzed δ13C and δ15N in both food sources and in five red knot tissues (blood cells, blood plasma and three feather types) and subsequently calculated the isotope discrimination factors. We confirmed that the discrimination factors differed between tissues, and also between diets. Our values deviated from general averages reported in reviews on a wide range of animals/birds, but were very similar to values from previous red knot and dunlin studies. We therefore think that our discrimination factors can be used in future stable isotope studies, not only on red knots, but also on other marine shorebird species and plea for careful consideration of using the right discrimination factors.Keywords: δ13C, δ15N, discrimination factor, red knot, shorebird, stable isotope

Round goby (Neogobius melanostomus) δ13C/δ15N discrimination values and comparisons of diets from gut content and stable isotopes in Oneida Lake.

Gut content analyses have found that round gobies (Neogobius melanostomus) are highly dependent on dreissenid mussels but stable isotope analysis has often suggested that the dreissenid contribution is lower. However, estimation of dietary contributions with stable isotopes relies on accurate discrimination factors (fractionation factors). To test if discrimination values commonly used in aquatic food web studies are suitable for round gobies, we collected round gobies from Oneida Lake, raised them for 63 days under four different diets (Chironomus plumosus, Mytilus chilensis, Dreissenia spp., Euphausia superba) and measured the change in white muscle δ13C and δ15N. Gobies were also collected throughout Oneida Lake for gut content and stable isotope analysis. Diets changed as round gobies grew, with small round gobies (17-42mm) feeding mostly on cladocera and chironomids, intermediate sized gobies (43-94mm) transitioning from chironomid to dreissenid consumption, and larger gobies (95-120mm) predominantly consuming dreissenids, similar to findings in other studies. Discrimination factors were obtained by fitting a commonly used asymptotic regression equation describing changes in fish δ13C and δ15N as a function of time and diet stable isotope ratios. The discrimination factor determined for δ13C (-0.4‰ ± 0.32, SE) was lower than the "standard" value of 0.4‰, while that of δ15N (4.0‰ ± 0.32, SE) was higher than the standard value of 3.4‰. Turnover rates for both δ13C and δ15N were estimated as 0.02 ‰*day-1. The use of experimentally determined discrimination factors rather than "standard" values resulted in model estimates that agree more closely with the observed increasing importance of dreissenids in gut content of larger gobies. Our results suggest that the importance of dreissenid mussels inferred from stable isotope studies may be underestimated when using standard isotopic discrimination values.

Addressing the Relationship between Leaf Nitrogen and Carbon Isotope Discrimination from the Three Levels of Community, Population and Individual.

The carbon, nitrogen and water cycles of terrestrial ecosystems are important biogeochemical cycles. Addressing the relationship of leaf nitrogen (N) and carbon isotope discrimination (Δ) will enhance the understanding of the links between these three cycles in plant leaves because Δ can reflect time-integrated leaf-level water-use efficiency (WUE) over the period when the leaf material is produced. Previous studies have paid considerable attention to the relationship. However, these studies have not effectively eliminated the interference of environmental factors, inter-species, and inter-individual differences in this relationship, so new research is necessary. To minimize these interferences, the present work explored the relationship at the three levels of community, population, and plant individual. Three patterns of positive, negative and no relationship were observed across communities, populations, and individuals, which is dependent on environmental conditions, species, and plant individuals. The results strongly suggested that there is no general pattern for the relationship between leaf N and Δ. Furthermore, the results indicated that there is often no coupling between leaf-level long-term WUE and leaf N in the metabolic process of carbon, N and water in leaves. The main reason for the lack of this relationship is that most plants do not invest large amounts of nitrogen into photosynthesis. In addition, the present study also observed that, for most plant species, leaf N was not related to photosynthetic rate, and that variations in photosynthetic rates are mainly driven by stomatal conductance.

Nonlinear Growth and Physiological Responses of White Spruce at North American Arctic Treeline

AbstractMuch is still unknown about the growth and physiological responses of trees to global change at the northern treeline. We combined tree‐ring width data with century‐long stable carbon and oxygen isotope records to investigate growth and physiological responses of white spruce at two treeline sites in the Canadian Arctic to concurrent increases in temperature, atmospheric CO2concentration (ca), and decline in sea ice extent over the past century. The tree‐ring records were assessed during three periods with contrasting climatic conditions: (a) the early 20th century warming, (b) the 1940–1970 cooling period, and (c) the anthropogenic late 20th century warming period. We found opposing growth trends between the two sites, but similar carbon isotope discrimination (Δ13C) and intrinsic water‐use efficiency (iWUE) trajectories. While tree growth (defined as basal area increment) increased at the site nearer to the Arctic Ocean during the 20th century following the rise in temperature and sea ice loss, growth declined after 1950 at the more interior site. At both sites, Δ13C slightly increased over these periods. However, trees showed a nonlinear response to increasedca, shifting after 1970 from a passive stomatal response (i.e., no changes iniWUE) to an active response (i.e., a moderate ∼12% increase iniWUE). Further, our isotope‐based findings do not support the idea that temperature‐induced drought stress caused the divergent growth trends at our treeline sites. This study thus highlights nonlinear and complex physiological and growth adjustments to concomitant changes in temperature, sea ice extent, andcaover the last century at the northern treeline.

Ecophysiological responses of Nothofagus obliqua forests to recent climate drying across the Mediterranean-Temperate biome transition in south-central Chile.

The forests of south-central Chile are facing a drying climate and a megadrought that started in 2010. This study addressed the physiological responses of five Nothofagus obliqua stands across the Mediterranean-Temperate gradient (35.9 ° -40.3° S) using carbon isotope discrimination (Δ13 C) and intrinsic water use efficiency (iWUE) in tree rings during 1967-2017. Moreover, δ18O was evaluated in the northernmost site to better understand the effects of the megadrought in this drier location. These forests have become more efficient in their use of water. However, trees from the densest stand are discriminating more against 13C, probably due to reduced photosynthetic rates associated with increasing competition. The strongest associations between climate and Δ13C were found in the northernmost stand, suggesting that warmer and drier conditions could have reduced 13C discrimination. Tree growth in this site has not decreased, and δ18O was negatively related to annual rainfall. However, a shift in this relationship was found since 2007, when both precipitation and δ18O decreased, while correlations between δ18O and growth increased. This implies that tree growth and δ18O are coupled in recent years, but precipitation is not the cause, suggesting that trees probably changed their water source to deeper and more depleted pools. Our research demonstrates that forests are not reducing their growth in central Chile, mainly due to a shift towards the use of deeper water sources. Despite a common climate trend across the gradient, there is a non-uniform response of N. obliqua forests to climate drying, being their response site specific. Keywords: Tree rings, stable isotopes, tree physiology, climate gradient, megadrought, climate change.

Sulfur isotopic discrimination factors differ among avian tissues and diets: Insights from a case study in Gentoo Penguins

Abstract The use of stable isotopes of sulfur (δ34S) to infer avian diets, foraging habitats, and movements is relatively uncommon, resulting in a lack of information on patterns of δ34S incorporation in avian tissue. In a controlled study of Gentoo Penguins (Pygoscelis papua), we found that diet-tissue isotopic discrimination factors (Δ34Sdiet-tissue) differed among egg components and feathers synthesized from a common diet, ranging from –0.4 to –1.7‰. We also found that methodical choices such as lipid extraction and prey tissue selection influenced calculated Δ34Sdiet-tissue values. Specifically, Δ34Sdiet-tissue values were lower (i.e., more negative) when calculated using whole fish relative to fish muscle and lipid-extraction biased egg yolk, but not fish tissue, δ34S values. The Δ34Sdiet-tissue values obtained for Gentoo Penguins fed a marine fish diet were generally lower than those reported for freshwater fish consumption by Double-crested Cormorants (Phalacrocorax auritus), the only other bird species in which Δ34Sdiet-tissue has been quantified. We found support for the hypothesis that tissue Δ34Sdiet-tissue values are inversely related to dietary δ34S values in birds, similar to what has been observed in mammals. Given this relationship, the discrimination factors reported here for Gentoo Penguins may be broadly applicable to other avian species with a similar marine diet. Finally, we provide recommendations for future studies seeking to quantify Δ34Sdiet-tissue in avian tissues and guidance to allow for greater application of sulfur stable isotope analysis in ornithological research.

A 195-Year Growing Season Relative Humidity Reconstruction Using Tree-Ring Cellulose δ13C in the Upper Tarim River Basin, NW China

Reconstruction of relative humidity changes in the upper Tarim River using carbon isotopic tree-ring chronology bridges the gap in historical observations on the Tarim River Basin in Arid Central Asia. Populus euphratica Olivier (P. euphratica), growing in the Tarim River Basin of Xinjiang, is an excellent record of past climate change. Based on precise dating, we analysed alpha-cellulose stable carbon isotopes in four cores of P. euphratica taken from the Alaer region of the upper Tarim River Basin. The four stable carbon isotope series records were corrected by the “pin method” and then combined into a carbon isotopic discrimination (Δ13C) series by the “numerical mix method”. The discrimination (Δ13C) series were clearly correlated with the mean relative humidity (RHAS) in April–September of the growing season (n = 60, r = −0.78, p &lt; 0.001), and according to the climate response analysis, we designed a simple regression equation to reconstruct the mean relative humidity (RHAS) in April–September from 1824 to 2018 on the Alaer region. The reconstructed sequence showed mainly dry periods in the last 195 years, 1857–1866 and 1899–1907, while primarily wet periods from 1985 to 2016. Due to increased global warming and human activities, the climate shifted from “warm–dry” to “warm–wet” in the mid-to-late 1980s, when there were signs of a shift from “warm–wet” to “warm–dry” in the 2010s, with an increasing trend towards aridity. The RHAS series of Alaer compares well to other hydroclimate series’ surrounding the research area, and the spatial correlation analysis indicates that the reconstructed series has good regional representativeness. On an interdecadal scale, the revamped RHAS series is positively correlated with the Atlantic Multidecadal Oscillation (AMO) and negatively correlated with the North Atlantic Oscillation (NAO), reflecting the influence of westerly circulation on regional wet and dry variability. At the same time, the RHAS may also be influenced by The Pacific Decadal Oscillation (PDO).

Determination of respiration and photosynthesis fractionation factors for atmospheric dioxygen inferred from a vegetation–soil–atmosphere analogue of the terrestrial biosphere in closed chambers

Abstract. The isotopic composition of dioxygen in the atmosphere is a global tracer which depends on the biosphere flux of dioxygen toward and from the atmosphere (photosynthesis and respiration) as well as exchanges with the stratosphere. When measured in fossil air trapped in ice cores, the relative concentration of 16O, 17O, and 18O of O2 can be used for several applications such as ice core dating and past global productivity reconstruction. However, there are still uncertainties about the accuracy of these tracers as they depend on the integrated isotopic discrimination of different biological processes of dioxygen production and uptake, for which we currently have very few independent estimates. Here we determined the respiration and photosynthesis fractionation factors for atmospheric dioxygen from experiments carried out in a replicated vegetation–soil–atmosphere analogue of the terrestrial biosphere in closed chambers with growing Festuca arundinacea. The values for 18O discrimination during soil respiration and dark respiration in leaves are equal to -12.3±1.7 ‰ and -19.1±2.4 ‰, respectively. In these closed biological chambers, we also found a value attributed to terrestrial photosynthetic isotopic discrimination equal to +3.7±1.3 ‰. This last estimate suggests that the contribution of terrestrial productivity in the Dole effect may have been underestimated in previous studies.

Aggregational differentiation of soil-respired CO2 and its δ13C variation across land-use types

Land-use type can affect CO2 emission fluxes, but whether it has effects on the δ13C of respired CO2 (δ13CO2), and whether these effects vary with the heterogeneity of the soil microenvironment remain unknown. In this study, long-term soil incubation experiments and soil fractionation were combined to determine the effects of land-use type (upland, paddy and woodland) on soil-respired CO2 and δ13CO2 with different aggregate fractions under three temperature conditions. This study extended the thermal adaptation of soil respiration to the soil microenvironmental scale, i.e., the scale of smaller aggregates than bulk soil. An exponential decay function (R = a × e−bt) was fitted to the temporal soil respiration data, separately for each combination of soil aggregate, land-use type and incubation temperature. At each incubation stage, the woodland exhibited the highest average soil respiration rates. The exponential decay trend of δ13CO2 with time was influenced by the land-use type and incubation temperature; the shift from enriched to depleted δ13CO2 values first appeared in woodland. Interestingly, carbon isotopic discrimination between the substrate and respired CO2 (Δ13C, δ13CO2 – δ13CSOC) increased with aggregate size only in woodland, especially at 30 °C. Overall, aggregational differentiation of soil-respired CO2 and its δ13C was found only in the woodland, and warming intensified these differences, suggesting that it would be difficult to generalize the integrative effects of warming on soil respiration and its isotopic fractionation at the microscale from a single ecosystem study. Our findings highlight that land-use types affect the feedback of soil carbon pool to future global warming at the soil microenvironmental scale (aggregate), and are of great significance for improving carbon emission prediction in terrestrial ecosystems.