Stratification of δ13C values of leaves in Amazonian rain forests

Summary The contribution of soil respiration to the photosynthesis of the shade flora in the Amazon forest was evaluated by measuring the ¹³C values of leaves collected at different levels in two forest communities. Canopy leaves have an average ¹³C of-30.5 in the podsol forest and-28.7 in the laterite forest. Leaves from plants in the lower forest strata have a significantly lower value of-35.2 in the podsol forest and-34.3 in the laterite forest.Mailing address of the first author: Before May 31, 1980: Department of Biological Sciences, Stanford University, Stanford, California 94305 USA. After May 31: Centro de Ecologia, IVIC Aptdo. 1827. Caracas, Venezuela     

Differences in the C age, δC and δO of Holocene tufa and speleothem in the Dinaric Karst

We studied Holocene speleothems and tufa samples collected in numerous caves and rivers in the Dinaric Karst of Croatia, Slovenia, Bosnia and Herzegovina, as well as Serbia and Montenegro. Differences in the formation process of tufa and speleothems are discussed in the context of their isotopic composition (¹⁴C, ¹³C and ¹⁸O), as well as the chemistry of surface water (rivers, lakes) and drip water (in caves). The physical and chemical parameters monitored in the surface water (tufa precipitation) and drip water (speleothem precipitation) show that more stable conditions accompany speleothem rather than tufa formation. This is particularly obvious in the water temperature variations (2-22°C in surface water and 7-12°C in drip water) and in saturation index variation (3-11 in surface water and 1-6 in drip water). The range of ¹⁴C ages recorded by Holocene speleothems (∼12?000 yr) is wider by several thousands years than that of Holocene tufa samples (∼6000 yr). δ¹³C values for tufa samples range from −12‰ to −6‰ and for speleothem samples from −12‰ to +3‰ reflecting higher soil carbon and/or vegetation impact on the process of tufa than on speleothem formation. The differences in δ¹⁸O values of tufa and speleothem samples from different areas reflect different temperature conditions and differing isotopic composition in the water. The study shows that speleothems from the Dinaric Karst can be used as global palaeoclimatic records, whereas tufa records changes in the local palaeoenvironment.     

Reliability of δ13C and δ15N in faeces for reconstructing savanna herbivore diet

We tested the reliability of herbivore faecal δ¹³C and δ¹⁵N values for reconstructing diet through review of an extensive database derived from a 3-year study of ungulates in South Africa's Kruger National Park. Faeces are a useful material for stable isotope studies of diet because they record dietary turnover at very short time scales, and because sampling is non-invasive. However, the validity of faecal isotope proxies may be questioned because they represent only undigested food remains. Results from Kruger Park confirm that free-ranging browsers have faecal δ¹³C consistent with C₃ feeding, grazer faeces are C₄, and mixed-feeder faeces intermediate. Although the respective ranges do not overlap, there is significant variation in faecal δ¹³C of browsers and grazers (～2.0–4.0‰) across space and through time. We demonstrate that most (～70%) of this variation can be ascribed to corresponding patterns of variation in the δ¹³C of C₃ and C₄ plants, respectively, re-enforcing the fidelity of faecal isotope proxies for diet but highlighting a need for mixing models that control for variations in plant δ¹³C in order to achieve accurate diet reconstructions. Predictions for the effects of climate (rainfall) and ecophysiology on ¹⁵N-abundance variations in mammals do not persist in faeces. Rather, faecal δ¹⁵N tracks changes in plant δ¹⁵N, with further fractionation occurring primarily due to variations in dietary protein (reflected by %N). Controlling for these effects, we show that a dual-isotope multiple source mixing model (Isosource) can extend diet reconstructions for African savanna herbivores beyond simplified C₃/C₄ distinctions, although further understanding of variations in mammal δ¹⁵N are needed for greater confidence in this approach.     

Structural basis for calcium and phosphatidylserine regulation of phospholipase C δ1

Many membrane-associated enzymes, including those of the phospholipase C (PLC) superfamily, are regulated by specific interactions with lipids. Previously, we have shown that the C2 domain of PLC δ1 is required for phosphatidylserine (PS)-dependent enzyme activation and that activation requires the presence of Ca(2+). To identify the site of interaction and the role of Ca(2+) in the activation mechanism, we mutagenized three highly conserved Ca(2+) binding residues (Asp-653, Asp-706, and Asp-708) to Gly in the C2 domain of PLC δ1. The PS-dependent Ca(2+) binding affinities of the mutant enzymes D653G, D706G, and D708G were reduced by 1 order of magnitude, and the maximal level of Ca(2+) binding was reduced to half of that of the native enzyme. The level of Ca(2+)-dependent PS binding was also reduced in the mutant enzymes. Under basal conditions, the Ca(2+) dependence and the maximal level of hydrolysis of phosphatidylinositol 4,5-bisphosphate were not altered in the mutants. However, the Ca(2+)-dependent PS stimulation was severely defective. PS reduces the K(m) of the native enzyme almost 20-fold, but far less for the mutants. Replacing Asp-653, Asp-706, and Asp-708 simultaneously with glycine in the C2 domain of PLC δ1 leads to a complete and selective loss of the stimulation and binding by PS. These results show that D653, D706, and D708 are required for Ca(2+) binding in the C2 domain and demonstrate a mechanism by which C2 domains can mediate regulation of enzyme activity by specific lipid ligands.     

The environmental water of the middle Eocene Arctic: Evidence from δD, δ18O and δ13C within specific compounds

The extensive vertical exposure (> 150 m) of terrestrial sediments on Axel Heiberg Island, which contain thick fossiliferous lignites, presents an exceptional opportunity to follow the establishment and re-establishment of Arctic Metasequoia forests during the middle Eocene. We compared δD values in n-alkanes of chain length 23, 25, 27 and 29 with δ¹⁸O values in phenylglucosazone (P-G) derived from α-cellulose; we also analyzed %-abundance of ferns, gymnosperms and angiosperms using pollen and spores isolated from each lignite. Our results showed that forest composition was altered upon uplift, as gymnosperms became more abundant within the relatively well-drained upland sediments. This was also reflected in the small (1‰), but significant, increase in the δ¹³C value of TOM from lowland to upland environments. However, neither the δD values of n-alkanes nor the δ¹⁸O in P-G were statistically different in the upland sediments, as compared to the lowland sediments; from this we inferred that the oxygen isotope signature of environmental water available to the forests for plant growth was relatively uniform throughout the time of the fossil forests. The δD value of environmental water implied by both n-alkanes and P-G ranged from ? 168 to ? 131% and was considerably enriched compared to all environmental water samples available from the modern Arctic region (< ? 180%). In addition to indicating a warmer Eocene Arctic, subject to meteoric transport patterns different from today's, these results argue against the presence of an Eocene polar ice cap.     

Combined use of δ¹³C, δ18O and δ15N tracks nitrogen metabolism and genotypic adaptation of durum wheat to salinity and water deficit

? Accurate phenotyping remains a bottleneck in breeding for salinity and drought resistance. Here the combined use of stable isotope compositions of carbon (δ13C), oxygen (δ1?O) and nitrogen (δ1?N) in dry matter is aimed at assessing genotypic responses of durum wheat under different combinations of these stresses. ? Two tolerant and two susceptible genotypes to salinity were grown under five combinations of salinity and irrigation regimes. Plant biomass, δ13C, δ1?O and δ1?N, gas-exchange parameters, ion and N concentrations, and nitrate reductase (NR) and glutamine synthetase (GS) activities were measured. ? Stresses significantly affected all traits studied. However, only δ13C, δ1?O, δ1?N, GS and NR activities, and N concentration allowed for clear differentiation between tolerant and susceptible genotypes. Further, a conceptual model explaining differences in biomass based on such traits was developed for each growing condition. ? Differences in acclimation responses among durum wheat genotypes under different stress treatments were associated with δ13C. However, except for the most severe stress, δ13C did not have a direct (negative) relationship to biomass, being mediated through factors affecting δ1?O or N metabolism. Based upon these results, the key role of N metabolism in durum wheat adaptation to salinity and water stress is highlighted.     

Climatic implications of δ13C and δ18O ratios from C3 and C4 plants growing in a tropical montane habitat in southern India

The stable carbon and oxygen isotope ratios in cellulose of C3 and C4 plants growing on the surface of a montane peat bog in the Nilgiri hills, southern India, were measured. The mean monthly δ¹³C values in cellulose of both C3 and C4 plants are found to be significantly related to rainfall, while the δ¹⁸O values are sensitive to changes in maximum temperature and relative humidity of the region. Further, higher δ¹⁸O values were observed in C4 plants compared to C3 plants, suggesting that C4 plants are probably less sensitive to relative humidity as compared to C3 plants and are able to photosynthesize even during drier conditions. The plant isotope-climate correlations thus established can be used for reconstructing the past temperature and rainfall conditions of the tropics from the isotopic ratios of peat deposits, derived from a mixture of C3 and C4 plants in the region.     

Estimation of canopy average mesophyll conductance using δ(13) C of phloem contents

Conductance to CO(2) inside leaves, known as mesophyll conductance (g(m)), imposes large limitations on photosynthesis. Because g(m) is difficult to quantify, it is often neglected in calculations of (13)C photosynthetic discrimination. The 'soluble sugar method' estimates g(m) via differences between observed photosynthetic discrimination, calculated from the δ(13)C of soluble sugars, and discrimination when g(m) is infinite. We expand upon this approach and calculate a photosynthesis-weighted average for canopy mesophyll conductance ((c) g(m)) using δ(13)C of stem phloem contents. We measured gas exchange at three canopy positions and collected stem phloem contents in mature trees of three conifer species (Pseudotsuga menziesii, Thuja plicata and Larix occidentalis). We generated species-specific and seasonally variable estimates of (c)g(m) . We found that (c)g(m) was significantly different among species (0.41, 0.22 and 0.09 mol m(-2) s(-1) for Larix, Pseudotsuga and Thuja, respectively), but was similar throughout the season. Ignoring respiratory and photorespiratory fractionations ((c)Δ(ef)) resulted in ≈30% underestimation of (c)g(m) in Larix and Pseudotsuga, but was innocuous in Thuja. Substantial errors (~1-4‰) in photosynthetic discrimination calculations were introduced by neglecting (c)g(m) and (c)Δ(ef) . Our method is easy to apply and cost-effective, captures species variation and would have captured seasonal variation had it existed. The method provides an average canopy value, which makes it suitable for parameterization of canopy-scale models of photosynthesis, even in tall trees.     

Seasonal Variation of δ13C of Four Tree Species: A Biological Integrator of Environmental Variables

Foliar δ13C values, an indicator of long-term intercellular carbon dioxide concentration and, thus,of long-term water use efficiency (WUE) in plants, were measured for Pinus massoniana Lamb., P. elliottii Engelm., Cunninghamia laceolata (Lamb.) Hook., and Schima superba Gardn. et Champ. in a restored forest ecosystem in the Jiazhu River Basin. Seasonal variation and the relationship between the foliar δ13Cvalues of the four species and environmental factors (monthly total precipitation, monthly average air temperature, relative humidity, atmospheric pressure, and monthly total solar radiation and evaporation)were investigated. The monthly δ13C values and WUE of the four species increased with increasing precipitation, air temperature, solar radiation, and evaporation, whereas δ13C values of the four species decreased with increasing relative humidity and atmospheric pressure. Despite significant differences in δ13C seasonal means for the four species, our results demonstrate a significant convergence in the responses of δ13C values and WUE to seasonal variations in environmental factors among the species investigated and that the δ13C signature for each species gives a strong indication of environmental variables.     

The carbon origin of arbuscular mycorrhizal fungi estimated from δ13C values of individual spores

 The origin of carbon in the spores of arbuscular mycorrhizal (AM) fungi was quantified based on their obligate symbiosis with C₃ and C₄ plants showing clearly different δ¹³C values. The δ¹³C values of individual spores of the AM fungus Gigaspora margarita were analyzed. In monoculture pots of a C₃ or a C₄ plant species, spore δ¹³C values were ca. 3.5‰ lower than those of host roots. In coculture pots of a C₃ and a C₄ plant species, spore δ¹³C values varied between those of the roots of C₃ and C₄ plants, and increased linearly from the C₃ to the proximity of the C₄ plant (P<0.01). This reflects the higher δ¹³C values in C₄ plants than in C₃ plants. Thus the carbon origin of G. margarita spores changed with growth state and combination of host plants. In the presence of fresh plant residue instead of living host plants, spore δ¹³C values did not vary with distance from the residue. This finding supports the current view that AM fungi are obligate symbionts. Accepted: 12 February 1999     

Analysis of δ13C of CO2 distinguishes between microbial respiration of added C4-sucrose and other soil respiration in a C3-ecosystem

The main aim of this study was to test various hypotheses regarding the changes in ¹³C of emitted CO₂ that follow the addition of C₄-sucrose to the soil of a C₃-ecosystem. It forms part of an experimental series designed to assess whether or not the contributions from C₃-respiration (root and microbial) and C₄-respiration (microbial) to total soil respiration can be calculated from such changes. A series of five experiments, three on sieved (root-free) mor-layer material, and two in the field with intact mor-layer (and consequently with active roots), were performed. Both in the experiments on sieved mor-layer and the field experiments, we found a C₄-sucrose-induced increase in C₃-respiration that accounted for between 30% and 40% of the respiration increase 1 h after sucrose addition. When the course of C₃-, C₄- and total respiration was followed in sieved material over four days following addition of C₄-sucrose, the initially increased respiration of C₃-C was transient, passing within less than 24 h. In a separate pot experiment, neither ectomycorrhizal Pinus sylvestrisL. roots nor non-mycorrhizal roots of this species showed respiratory changes in response to exogenous sucrose. No shift in the ¹³C of the evolved CO₂ after adding C₃-sucrose to sieved mor-layer material was found, confirming that the sucrose-induced increase in respiration of endogenous C was not an artefact of discrimination against ¹³C during respiration. Furthermore, we conclude that the C₄-sucrose induced transient increase in C₃-respiration is most likely the result of accelerated turnover of C in the microbial biomass. Thus, neither respiration of mycorrhizal roots, nor processes discriminating against ¹³C were likely sources of error in the field. The estimated ¹³C of evolved soil CO₂ in three field experiments lay between –25.2 and –23.6. The study shows that we can distinguish between CO₂ evolved from microbial mineralisation of added C₄-sucrose, and CO₂ evolved from endogenous carbon sources (roots and microbial respiration).     

Seasonal Variation of δ^13C of Four Tree Species： A Biological Integrator of Environmental Variables

Foliar δ^13C values, an indicator of long-term intercellular carbon dioxide concentration and, thus, of long-term water use efficiency （WUE） in plants, were measured for Pinus massoniana Lamb., P. elliottii Engelm., Cunninghamia laceolata （Lamb.） Hook., and Schima superba Gardn. et Champ. in a restored forest ecosystem in the Jiazhu River Basin. Seasonal variation and the relationship between the foliar δ^13C values of the four species and environmental factors （monthly total precipitation, monthly average air temperature, relative humidity, atmospheric pressure, and monthly total solar radiation and evaporation） were investigated. The monthly δ^13C values and WUE of the four species increased with increasing precipitation, air temperature, solar radiation, and evaporation, whereas δ^13C values of the four species decreased with increasing relative humidity and atmospheric pressure. Despite significant differences in δ^13C seasonal means for the four species, our results demonstrate a significant convergence in the responses of δ^13C values and WUE to seasonal variations in environmental factors among the species investigated and that the δ^13C signature for each species gives a strong indication of environmental variables.     

Uptake of organic matter by the roots of sweet potato: Analysis of the δ14C value of plants

Summary To determine whether sweet potato (Ipomoea batatas (L.) Poir.) takes up organic matter through the roots from the medium, the concentrations of natural¹⁴C (¹⁴C) in plant organic matter, atmospheric CO₂ and compost applied to media were examined under soil and sand culture conditions. In these experiments, three kinds of composts of different ¹⁴C were used. CO₂ derived from the mineralization of compost was continuously pumped out from the pots and its direct uptake by the leaves was prevented.¹⁴C of plant parts harvested after the 43 days experimental period were affected by the ¹⁴C of the compost in the treatments where the compost of rice straw was applied, and which suggested that a significant amount of plant carbon was derived from the compost.     

Temporal records of δ13C and δ15N in North Pacific pinnipeds: inferences regarding environmental change and diet

Sea lion and seal populations in Alaskan waters underwent various degrees of decline during the latter half of the twentieth century and the cause(s) for the declines remain uncertain. The stable carbon (¹³C/¹²C) and nitrogen (¹⁵N/¹⁴N) isotope ratios in bone collagen from wild Steller sea lions (Eumetopias jubatus), northern fur seals (Callorhinus ursinus) and harbor seals (Phoca vitulina) from the Bering Sea and Gulf of Alaska were measured for the period 1951-1997 to test the hypothesis that a change in trophic level may have occurred during this interval and contributed to the population declines. A significant change in '¹⁵N in pinniped tissues over time would imply a marked change in trophic level. No significant change in bone collagen '¹⁵N was found for any of the three species during the past 47 years in either the Bering Sea or the Gulf of Alaska. However, the ¹⁵N in the Steller sea lion collagen was significantly higher than both northern fur seals and harbor seals. A significant decline in '¹³C (almost 2 ‰ over the 47 years) was evident in Steller sea lions, while a declining trend, though not significant, was evident in harbor seals and northern fur seals. Changes in foraging location, in combination with a trophic shift, may offer one possible explanation. Nevertheless, a decrease in '¹³C over time with no accompanying change in '¹⁵N suggests an environmental change affecting the base of the foodweb rather than a trophic level change due to prey switching. A decline in the seasonal primary production in the region, possibly resulting from decreased phytoplankton growth rates, would exhibit itself as a decline in '¹³C. Declining production could be an indication of a reduced carrying capacity in the North Pacific Ocean. Sufficient quantities of optimal prey species may have fallen below threshold sustaining densities for these pinnipeds, particularly for yearlings and subadults who have not yet developed adequate foraging skills.     

Research article: small-scale spatial variation of δ13C and δ15N isotopes in Antarctic carbon sources and consumers

Regional food web studies that fail to account for small-scale isotopic variability can lead to a mismatch between an organism’s inferred and true trophic position. Misinterpretation of trophic status may result, substantially limiting spatial and temporal comparability of food web studies. We sampled several carbon sources and consumers in a nested design to assess the variability of food web members across small spatial scales (100 s of m to several km) in regions around the Windmill Islands and Vestfold Hills in East Antarctica. For carbon sources, δ¹³C in sea ice POM was particularly variable between locations (km apart) and between sites (100 s of m apart) with replicate samples varying by up to 16‰. Macroalgae δ¹³C was less variable (replicate samples ranging up to 6.9‰ for the red alga Iridaea cordata), yet still differed between locations. Sediment POM and pelagic POM were the least variable, displaying minimal differences between locations or sites for δ¹³C and δ¹⁵N. Three out of eight consumers were significantly different between locations for δ¹³C, and five out of eight for δ¹⁵N, with the fish Trematomus bernacchii the most variable for both δ¹³C and δ¹⁵N. At smaller scales, the amphipod Paramorea walkeri showed significant variation between sites in δ¹³C but not in δ¹⁵N. We attribute small-scale variability to the dynamic physical environment for carbon sources in coastal systems and a close coupling of diet to habitat for consumers. We highlight the need to account for small-scale spatial variation in sampling designs for regional food web studies.     

Ontogenetic shift in crayfish δ13C as a measure of land-water ecotonal coupling

Although ontogenetic changes in the carbon isotope ratios of marine fauna have been well studied those of freshwater organisms have not. As a result, we may have a less than adequate assessment of the incorporation of allochthonous detritus into freshwater foodwebs. This study found a ¹³C range of 9 for crayfish (Orconectes virilis) from oligotrophic Canadian Shield lakes. Much (60–83%) of this variability was explained by body size. A simple isotopic mixing model suggests that by their third year of life, crayfish in these lakes rely more substantially upon terrestrial detritus than epilithic algae for energy.     

Use of δ13C values to determine vegetation selectivity in East African herbivores

Summary The quantitative plant species composition of the rumen contents of a large number of individuals from eight East African herbivores was determined by direct visual analysis. All plant species were classified as either C₃ or C₄, and an estimated ₁₃C for the rumen sample was calculated. This estimated value was compared to a measured value determined directly from rumen subsample. The two methods of determining quantitative C₃ and C₄ composition differed by less than 1%, and the isotopic analysis has the advantage of being rapid and totally objective.The isotopic analysis allowed us to differentiate between grazers and browsers and to determine the quantitative dependence of each animal on C₃ and C₄ photosynthetic types. Kongoni, wildebeest, cattle, and sheep were nearly pure grazers on the Athi Kapiti Plains; and the Grant's gazelle were predominantly browsers. Thompson's gazelle, goast and impala were intermediate. The species most dependent upon browse showed a marked and rapid shift to grass within a few days following rain. This isotopic method may have general utility in the study of East African ecology.     

Patterns of δ^13C and δ^15N in wolverine Gulo gulo tissues from the Brooks Range,Alaska

Knowledge of carnivore diets is essential to understand how carnivore populations respond demographically to variations in prey abundance. Analysis of stable isotopes is a useful complement to traditional methods of analyzing carnivore diets. We used data on δ^13C and δ^15N in wolverine tissues to investigate patterns of seasonal and annual diet variation in a wolverine Gulo gulo population in the western Brooks Range, Alaska, USA. The stable isotope ratios in wolverine tissues generally reflected that of terrestrial carnivores, corroborating previous diet studies on wolverines. We also found variation in δ^13C and δ^15N both between muscle samples collected over several years and between tissues with different assimilation rates, even after correcting for isotopic fractionation. This suggests both annual and seasonal diet variation. Our results indicate that data on δ^13C and δ^15N holds promise for qualitative assessments of wolverine diet changes over time. Such temporal variation may be important indicators of ecological responses to environmental perturbations, and we suggest that more refined studies of stable isotopes may be an important tool when studying temporal change in diets of wolverines and similar carnivores [ Current Zoology 55 （3）： 188- 192, 2009].     

Differences in δ15N and δ13C between embryonic and maternal tissues of the ovoviviparous bluntnose sixgill shark Hexanchus griseus

Stable nitrogen (δ¹⁵N) and carbon (δ¹³C) isotope ratios from muscle, liver and yolk were analysed from the mother and embryos of an ovoviviparous shark, Hexanchus griseus. Embryonic liver and muscle had similar δ¹⁵N and δ¹³C ratios or were depleted in heavy isotopes, compared to the same maternal somatic and reproductive yolk tissues, but no relationship existed between δ¹⁵N or δ¹³C and embryo length, as expected, because a switch to placental nourishment is lacking in this species. This study expands the understanding of maternal nourishment and embryonic stable isotope differences in ovoviviparous sharks.