Carbon nutrition of mature green orchid <Emphasis Type="Italic">Serapias strictiflora</Emphasis> and its mycorrhizal fungus <Emphasis Type="Italic">Epulorhiza</Emphasis> sp.

We studied the nutritional modes of the orchid Serapias strictiflora and its mycorrhizal fungus Epulorhiza sp. using the differences in carbon isotopic composition (δ¹³C) of C₃ orchid and C₄ maize tissues. We found that if cultivated in substrate lacking any organic compounds, the mycorrhizal extraradical mycelia (δ¹³C = −26.3 ± 0.2 ‰) developed well, despite being fully dependent on nutrition from orchid roots (δ¹³C = −28.6 ± 0.1 ‰). If the mycorrhizal fungus had additional access to and colonized decaying maize roots (δ¹³C = −14.6 ± 0.1 ‰), its isotopic composition (δ¹³C = −21.6 ± 0.4 ‰) reflected a mixture of biotrophy and saprotrophy. No statistically significant differences in δ¹³C of new storage tubers were found between Epulorhiza-associated orchids with (δ¹³C = -28.2 ± 0.1 ‰) and without access to maize roots (δ¹³C = −28.6 ± 0.2 ‰). We conclude that autotrophy is the predominant nutritional mode of mature S. strictiflora plants and that they supply their mycorrhizal fungus with substantial amount of carbon (69 ± 3 % of the fungus demand), even if the fungus feeds saprotrophically.     

Autochthonous origin of semi-labile dissolved organic carbon in a large monomictic lake (Lake Biwa): carbon stable isotopic evidence

Semi-labile dissolved organic carbon (DOC) plays an important role in the transport and hypolimnetic remineralization of carbon in large freshwater lakes. However, sources of semi-labile DOC in lakes remain unclear. This study used a carbon stable isotope approach to examine relative contributions of autochthonous and allochthonous sources to semi-labile DOC. Vertical and seasonal variations in the concentration and carbon stable isotope ratio (δ¹³C) of DOC were determined in large (surface area 674 km²; maximum depth 104 m), monomictic Lake Biwa. A sharp vertical gradient of δ¹³C of DOC (δ¹³C-DOC) during the stratification period [mean ± standard error (SE) −25.5 ± 0.1 and −26.0 ± 0.0‰ in the epi- and hypolimnion, respectively] indicated the accumulation of ¹³C-rich DOC in the epilimnion. Vertical mixing explained the intermediate values of δ¹³C-DOC (−25.7 ± 0.0‰) measured throughout the water column during the overturn period. Both DOC concentration and δ¹³C-DOC decreased in the hypolimnion during stratification, indicating selective remineralization of ¹³C-rich DOC. Using a two-component mixing model, we estimated the δ¹³C value of semi-labile DOC to be −22.2 ± 0.3‰, which was close to the δ¹³C of particulate organic carbon collected in the epilimnion during productive seasons (−22.7 ± 0.7‰) but much higher than the δ¹³C-DOC in river waters (−26.5 ± 0.1‰). Semi-labile DOC appeared to be mainly autochthonous in origin, produced by planktonic communities during productive seasons. The spatiotemporal uncoupling between production and remineralization of semi-labile DOC implies that hypolimnetic oxygen consumption may be affected by pelagic primary production during productive seasons of the preceding year.     

The Use of the abundance ratio of 13C and 12C isotopes for characteristic of the origin of ethyl alcohol

During alcohol fermentation, the carbon isotope composition of ethyl alcohol produced depended on the substrate used and was characterized by the value of delta 13C equal to -24.7 +/- 0.8/1000 (wheat grain), -22 +/- 0.1/1000 (rye grain), -22 +/- 0.5/1000 (products of wood hydrolysis), -15.3 +/- 0.3/1000 (maize grain) and -10 +/- 0.1/1000 (sugar cane). The isotope composition of carbon of ethyl alcohol obtained during catalytic hydroxylation of ethylene has a delta 13C of -30.6 +/- 0.3/1000. The possibility of quantitative determination of specific components in mixtures of ethanol samples with various isotope compositions (chemical synthesis and alcohol fermentation of raw material from C3- or C4-plants) was shown.     

Carbon and nitrogen isotope composition of vegetation on King George Island,maritime Antarctic

We report abundance of ¹³C and ¹⁵N contents in terrestrial plants (mosses, lichens, liverworts, algae and grasses) from the area of Barton Peninsula (King George Island, maritime Antarctic). The investigated plants show a wide range of δ¹³C and δ¹⁵N values between −29.0 and −20.0‰ and between −15.3 and 22.8‰, respectively. The King George Island terrestrial plants show species specificity of both carbon and nitrogen isotope compositions, probably due to differences in plant physiology and biochemistry, related to their sources and in part to water availability. Carbon isotope compositions of Antarctic terrestrial plants are typical of the C₃ photosynthetic pathway. Lichens are characterized by the widest carbon isotope range, from −29.0 to −20.0‰. However, the average δ¹³C value of lichens is the highest (−23.6 ± 2.8‰) among King George Island plants, followed by grasses (−25.6 ± 1.7‰), mosses (−25.9 ± 1.6‰), liverworts (−26.3 ± 0.5‰) and algae (−26.3 ± 1.2‰), partly related to habitats controlled by water availability. The δ¹⁵N values of moss samples range widest (−9.0 to 22.8‰, with an average of 4.6 ± 6.6‰). Lichens are on the average most depleted in ¹⁵N (mean = −7.4 ± 6.4‰), whereas algae are most enriched in ¹⁵N (10.0 ± 3.3‰). The broad range of nitrogen isotope compositions suggest that the N source for these Antarctic terrestrial plants is spatially much variable, with the local presence of seabird colonies being particularly significant.     

An isotopic method for testing the influence of leaf litter quality on carbon fluxes during decomposition

During microbial breakdown of leaf litter a fraction of the C lost by the litter is not released to the atmosphere as CO₂ but remains in the soil as microbial byproducts. The amount of this fraction and the factors influencing its size are not yet clearly known. We performed a laboratory experiment to quantify the flow of C from decaying litter into the soil, by means of stable C isotopes, and tested its dependence on litter chemical properties. Three sets of ¹³C-depleted leaf litter (Liquidambar styraciflua L., Cercis canadensis L. and Pinus taeda L.) were incubated in the laboratory in jars containing ¹³C-enriched soil (i.e. formed C4 vegetation). Four jars containing soil only were used as a control. Litter chemical properties were measured using thermogravimetry (Tg) and pyrolysis–gas chromatography/mass spectrometry–combustion interface–isotope ratio mass spectrometry (Py–GC/MS–C–IRMS). The respiration rates and the δ¹³C of the respired CO₂ were measured at regular intervals. After 8 months of incubation, soils incubated with both L. styraciflua and C. canadensis showed a significant change in δ¹³C (δ¹³C_final = −20.2 ± 0.4‰ and −19.5 ± 0.5‰, respectively) with respect to the initial value (δ¹³C_initial = −17.7 ± 0.3‰); the same did not hold for soil incubated with P. taeda (δ¹³C_final:−18.1 ± 0.5‰). The percentages of litter-derived C in soil over the total C loss were not statistically different from one litter species to another. This suggests that there is no dependence of the percentage of C input into the soil (over the total C loss) on litter quality and that the fractional loss of leaf litter C is dependent only on the microbial assimilation efficiency. The percentage of litter-derived C in soil was estimated to be 13 ± 3% of total C loss.     

Bat breath reveals metabolic substrate use in free-ranging vampires

We analysed the stable carbon isotope ratio in exhaled CO₂ (δ¹³C_breath) of free-ranging vampires to assess the type of metabolized substrate (endogenous or exogenous substrate) and its origin, i.e. whether the carbon atoms came from a C₄ food web (grass and cattle) or the C₃ food web in which they were captured (a rainforest remnant and its mammals). For an improved understanding of factors influencing the δ¹³C_breath of vampires, we conducted feeding experiments with captive animals. The mean δ¹³C_breath of starved bats was depleted in ¹³C in relation to the diet by 4.6‰ (n = 10). Once fed with blood, δ¹³C_breath levelled off within a short time approximately 2.2‰ above the stable carbon isotope signature of the diet. The median time required to exchange 50% of the carbon atoms in exhaled CO₂ with carbon atoms from the ingested blood was 18.6 min (mean 29.5 ± 19.0 min, n = 5). The average δ¹³C of wing membrane and fur in free-ranging vampire bats suggested that bats almost exclusively foraged for cattle blood during the past weeks. The δ¹³C_breath of the same bats averaged −19.1‰. Given that all free-ranging vampires were starving and that the δ¹³C of cattle was more in enriched in ¹³C by 5–6‰ than the δ¹³C_breath of vampires, we conclude that the vampire bats of our study metabolised fat that was predominantly built from carbon atoms originating from cattle blood. Since δ¹³C of wing membrane and fur integrates over weeks and months respectively and δ¹³C_breath over hours and days, we also conclude that vampire bats of the studied population consistently ignored rainforest mammals and chose cattle as their prey during and prior to our study.     

Isotopic signature of nitrate in two contrasting watersheds of Brush Brook,Vermont, USA

We used the dual isotope method to study differences in nitrate export in two subwatersheds in Vermont, USA. Precipitation, soil water and streamwater samples were collected from two watersheds in Camels Hump State Forest, located within the Green Mountains of Vermont. These samples were analyzed for the δ¹⁵N and δ¹⁸O of NO₃⁻. The range of δ¹⁵N–NO₃⁻ values overlapped, with precipitation −4.5‰ to +2.0‰ (n = 14), soil solution −10.3‰ to +6.2‰ (n = 12) and streamwater +0.3‰ to +3.1‰ (n = 69). The δ¹⁸O of precipitation NO₃⁻ (mean 46.8 ± 11.5‰) was significantly different (P < 0.001) from that of the stream (mean 13.2 ± 4.3‰) and soil waters (mean 14.5 ± 4.2‰) even during snowmelt periods. Extracted soil solution and streamwater δ¹⁸O of NO₃⁻ were similar and within the established range of microbially produced NO₃⁻, demonstrating that NO₃⁻ was formed by microbial processes. The δ¹⁵N and δ¹⁸O of NO₃⁻ suggests that although the two tributaries have different seasonal NO₃⁻ concentrations, they have a similar NO₃⁻ source.     

Contributions of C3 and C4 plants to higher trophic levels in an Amazonian savanna

We studied the energy flow from C₃ and C₄ plants to higher trophic levels in a central Amazonian savanna by comparing the carbon stable-isotope ratios of potential food plants to the isotope ratios of species of different consumer groups. All C₄ plants encountered in our study area were grasses and all C₃ plants were bushes, shrubs or vines. Differences in δ¹³C ratios among bushes (xˉ = −30.8, SD = 1.2), vines (xˉ = −30.7, SD = 0.46) and trees (xˉ = −29.7, SD = 1.5) were small. However the mean δ¹³C ratio of dicotyledonous plants (xˉ = −30.4, SD = 1.3) was much more negative than that of the most common grasses (xˉ = −13.4, SD = 0.27). The insect primary consumers had δ¹³C ratios which ranged from a mean of −29.5 (SD = 0.47) for the grasshopper Tropidacris collaris to a mean of −14.7 (SD = 0.56) for a termite (Nasutitermes sp.), a range similar to that of the vegetation. However, the common insectivorous and omnivorous vertebrates had intermediate values for δ¹³C, indicating that carbon from different autotrophic sources mixes rapidly as it moves up the food chain. Despite this mixing, the frogs and lizards generally had higher values of δ¹³C (xˉ = −21.7, SD = 1.6; xˉ = −21.9, SD = 1.8, respectively) than the birds (xˉ = −24.8, SD = 1.8) and the only species of mammal resident in the savanna (xˉ = −25.4), indicating that they are generally more dependent on, or more able to utilise, food chains based on C₄ grasses. Received: 7 May 1998 / Accepted: 30 November 1998     

Nocturnal and seasonal patterns of carbon isotope composition of leaf dark-respired carbon dioxide differ among dominant species in a semiarid savanna

The C isotope composition of leaf dark-respired CO₂ (δ¹³C_l) integrates short-term metabolic responses to environmental change and is potentially recorded in the isotopic signature of ecosystem-level respiration. Species differences in photosynthetic pathway, resource acquisition and allocation patterns, and associated isotopic fractionations at metabolic branch points can influence δ¹³C_l, and differences are likely to be modified by seasonal variation in drought intensity. We measured δ¹³C_l in two deep-rooted C₃ trees (Prosopis velutina and Celtis reticulata), and two relatively shallow-rooted perennial herbs (a C₃ dicot Viguiera dentata and a C₄ grass Sporobolus wrightii) in a floodplain savanna ecosystem in southeastern Arizona, USA during the dry pre-monsoon and wet monsoon seasons. δ¹³C_l decreased during the nighttime and reached minimum values at pre-dawn in all species. The magnitude of nocturnal shift in δ¹³C_l differed among species and between pre-monsoon and monsoon seasons. During the pre-monsoon season, the magnitude of the nocturnal shift in δ¹³C_l in the deep-rooted C₃ trees P. velutina (2.8 ± 0.4‰) and C. reticulata (2.9 ± 0.2‰) was greater than in the C₃ herb V. dentata (1.8 ± 0.4‰) and C₄ grass S. wrightii (2.2 ± 0.4‰). The nocturnal shift in δ¹³C_l in V. dentata and S. wrightii increased to 3.2 ± 0.1‰ and 4.6 ± 0.6‰, respectively, during the monsoon season, but in C₃ trees did not change significantly from pre-monsoon values. Cumulative daytime net CO₂ uptake was positively correlated with the magnitude of the nocturnal decline in δ¹³C_l across all species, suggesting that nocturnal δ¹³C_l may be controlled by ¹³C/¹²C fractionations associated with C substrate availability and C metabolite partitioning. Nocturnal patterns of δ¹³C_l in dominant plant species in the semiarid savanna apparently have predictable responses to seasonal changes in water availability, which is important for interpreting and modeling the C isotope signature of ecosystem-respired CO₂.     

Variations in leaf β13C along a vertical profile of irradiance in a temperate Japanese forest

The vertical profile of stable carbon isotope ratios (δ¹³C) of leaves was analyzed for 13 tree species in a cool-temperate deciduous forest in Japan. The vertical distribution of long-term averaged δ¹³C in atmospheric CO₂ (δ_a) was estimated from δ¹³C of dry matter from NADP-malic enzyme type C₄ plant (Zea mays L. var. saccharata Sturt.) grown at a tower in the forest for 32␣days, assuming constant Δ value (3.3‰) in Z. mays against height. The δ_a value obtained from δ¹³C in Z.␣mays was lowest at the forest floor (−9.30 ± 0.03‰), increased with height, and was almost constant above 10␣m (−7.14 ± 0.14‰). Then leaf Δ values for the tree species were calculated from tree leaf δ¹³ C andδ_a. Mean leaf Δ values for the three tall deciduous species (Fraxinus mandshurica, Ulmus davidiana, and Alnus hirsuta) were significantly different among three height levels in the forest: 23.1 ± 0.7‰ at the forest floor (understory), 21.4 ± 0.5‰ in lower canopy, and 20.5 ± 0.3‰ in upper canopy. The true difference in tree leaf Δ among the forest height levels might be even greater, because Δ in Z. mays probably increased with shading by up to ∼‰. The difference in tree leaf Δ among the forest height levels would be mainly due to decreasing intercellular CO₂ (C _i) with the increase in irradiance. Potential assimilation rate for the three tree species probably increased with height, since leaf nitrogen content on an area basis for these species also increased with height. However, the increase in stomatal conductance for these tree species would fail to meet the increase in potential assimilation rate, which might lead to increasing the degree of stomatal limitation in photosynthesis with height. Received: 30 September 1995 / Accepted: 25 October 1996     

Carbon isotope composition as a tool to control the quality of herbs and medicinal plants

Isotope screening is a simple test for determining the photosynthetic pathway used by plants. The scope of this work was to classify the photosynthetic type of some herbs and medicinal plants through studies of the carbon isotope composition (δ¹³C). Also, we propose the use of carbon isotope composition as a tool to control the quality of herbs and medicinal plants. For studies of δ¹³C, δ¹³C‰ = [R (sample)/R (standard) − 1] × 10⁻³, dry leaves powdered in cryogenic mill were analyzed in a mass spectrometer coupled with an elemental analyzer for determining the ratio R = ¹³CO₂/¹²CO₂. In investigation of δ¹³C of 55 species, 23 botanical families, and 44 species possessed a C₃ photosynthetic type. Six species found among the botanical families Euphorbiaceae and Poaceae were C₄ plants, and 5 species found among the botanical families Agavaceae, Euphorbiaceae, and Liliaceae possessed CAM-type photosynthesis. Carbon isotope composition of plants can be used as quality control of herbs and medicinal plants, allowing the identification of frauds or contaminations. Also, the information about the photosynthetic type found for these plants can help in introducing and cultivating exotic and wild herbs and medicinal plants.     

Influence of stand structure on carbon-13 of vegetation, soils, and canopy air within deciduous and evergreen forests in Utah, United States

Carbon isotope ratios (δ¹³C) were studied in evergreen and deciduous forest ecosystems in semi-arid Utah (Pinus contorta, Populus tremuloides, Acer negundo and Acer grandidentatum). Measurements were taken in four to five stands of each forest ecosystem differing in overstory leaf area index (LAI) during two consecutive growing seasons. The δ¹³C_leaf (and carbon isotope discrimination) of understory vegetation in the evergreen stands (LAI 1.5–2.2) did not differ among canopies with increasing LAI, whereas understory in the deciduous stands (LAI 1.5–4.5) exhibited strongly decreasing δ¹³C_leaf values (increasing carbon isotope discrimination) with increasing LAI. The δ¹³C values of needles and leaves at the top of the canopy were relatively constant over the entire LAI range, indicating no change in intrinsic water-use efficiency with overstory LAI. In all canopies, δ¹³C_leaf decreased with decreasing height above the forest floor, primarily due to physiological changes affecting c _i/c _a (> 60%) and to a minor extent due to δ¹³C of canopy air (< 40%). This intra-canopy depletion of δ¹³C_leaf was lowest in the open stand (1‰) and greatest in the denser stands (4.5‰). Although overstory δ¹³C_leaf did not change with canopy LAI, δ¹³C of soil organic carbon increased with increasing LAI in Pinus contorta and Populus tremuloides ecosystems. In addition, δ¹³C of decomposing organic carbon became increasingly enriched over time (by 1.7–2.9‰) for all deciduous and evergreen dry temperate forests. The δ¹³C_canopy of CO₂ in canopy air varied temporally and spatially in all forest stands. Vertical canopy gradients of δ¹³C_canopy, and [CO₂]_canopy were larger in the deciduous Populus tremuloides than in the evergreen Pinu contorta stands of similar LAI. In a very wet and cool year, ecosystem discrimination (Δ_e) was similar for both deciduous Populus tremulodies (18.0 ± 0.7‰) and evergreen Pinus contorta (18.3 ± 0.9‰) stands. Gradients of δ¹³C_canopy and [CO₂]_canopy were larger in denser Acer spp. stands than those in the open stand. However, ¹³C enrichment above and photosynthetic draw-down of [CO₂]_canopy below tropospheric baseline values were larger in the open than in the dense stands, due to the presence of a vigorous understory vegetation. Seasonal patterns of the relationship δ¹³C_canopy versus 1/[CO₂]_canopy were strongly influenced by precipitation and air temperature during the growing season. Estimates of Δ_e for Acer spp. did not show a significant effect of stand structure, and averaged 16.8 ± 0.5‰ in 1933 and 17.4 ± 0.7‰ in 1994. However, Δ_e varied seasonally with small fluctuations for the open stand (2‰), but more pronounced changes for the dense stand (5‰). Received: 15 April 1996 / Accepted: 19 October 1996     

New organic reference materials for carbon- and nitrogen-stable isotope ratio measurements provided by Center for Ecological Research,Kyoto University,and Institute of Biogeosciences,Japan Agency for Marine-Earth Science and Technology

We determined both carbon and nitrogen isotope ratios of ten organic reference materials (CERKU-01 to CERKU-10) in the Center for Ecological Research (CER), Kyoto University, and three organic reference materials (BG-A, BG-P, and BG-T) in the Institute of Biogeosciences (BioGeos), Japan Agency for Marine-Earth Science and Technology (JAMSTEC), using an internationally recommended calibration method of two-point anchoring. The reference materials cover δ¹³C_VPDB range of −34.92 to −9.45‰ and δ¹⁵N_Air range of −5.22 to 22.71‰ and can be used to measure isotope ratios of naturally occurring substances.     

Fractionation of stable carbon isotopes by photoautotrophically growing anoxygenic purple and green sulfur bacteria

Fractionation of stable carbon isotopes ¹²C and ¹³C by three pure cultures of photoautotrophic purple sulfur bacteria (Ectothiorhodospira shaposhnikovii, Lamprocystis purpureus, and Thiocapsa sp.) (PSB) and the green sulfur bacterium Prosthecochloris sp. (GSB) was investigated in 13–15-day experiments. The cultivation was carried out in a luminostat (2000 lx) on mineral media with 1–1.5 g/l NaHCO₃ (inoculum) with the subsequent transfer to the medium with up to 10 g/l NaHCO₃. For PSB, the difference in the quantitative characteristics of the isotopic composition of suspended carbon (including bacterial cells) and mineral carbon of the medium (Δ¹³C = δ¹³C_substrate − δ¹³C_biomass) changed from 15.0 to 34.3‰. For GSB, the range of Δ¹³C changes was significantly less (18.3–22.7‰). These data suggested the possibility of a pool of soluble mineral carbon in PSB cells. The pool of intracellular mineral carbon was calculated; depending on the PSB species and growth stage, it varied from 0 to 68% of the total cell carbon. The α coefficients reflecting the carbon isotope fractionation by PSB and GBS and calculated from the changes of the bicarbonate carbon isotopic composition in the medium depending on its consumption were 1.029 ± 0.003 and 1.019 ± 0.001, respectively. These α values did not depend on the growth rate. CO₂ fixation on ribulose-bisphosphate was shown to be the major factor determining the carbon isotope fractionation by PSB; at the stage of CO₂ penetration into the cell, fractionation was insignificant. In GSB, fractionation occurred mostly at CO₂ penetration into the cell, while it was insignificant at the stage of carbon dioxide fixation in the reverse TCA cycle. Analysis of the isotopic data of the photosynthesis by PSB and GSB in meromictic lakes also revealed that in PSB-dominated natural communities suspended organic matter was more enriched with light ¹³C (Δ¹³C = 23.4−24.6‰) than in the communities with more active GSB (Δ¹³C = 10.2−14.0‰)     

Challenges using stable isotopes for estimating trophic levels in marine amphipods

In marine food web studies, stable isotopes of nitrogen (δ¹⁵N) and carbon (δ¹³C) are widely used to estimate organisms’ trophic levels (TL) and carbon sources, respectively. For smaller organisms, whole specimens are commonly analyzed. However, this “bulk method” involves several pitfalls since different tissues may fractionate stable isotopes differently. We compared the δ¹⁵N and δ¹³C values of exoskeleton versus soft tissue, in relation to whole specimens, of three common Arctic amphipods in Svalbard waters: the benthic Anonyx nugax, the sympagic (ice-associated) Gammarus wilkitzkii and the pelagic Themisto libellula. The δ¹⁵N values of the exoskeletons were significantly lower than those of the soft tissues for A. nugax (10.5 ± 0.7‰ vs. 15.7 ± 0.7‰), G. wilkitzkii (3.3 ± 0.3‰ vs. 8.3 ± 0.4‰) and T. libellula (8.6 ± 0.3‰ vs.10.8 ± 0.3‰). The differences in δ¹³C values between exoskeletons and soft tissues were insignificant, except for A. nugax (−21.2 ± 0.2‰ vs. −20.3 ± 0.2‰, respectively). The δ¹⁵N values of whole organisms were between those of the exoskeletons and the soft tissues, being similarly enriched in ¹⁵N as the exoskeletons (except G. wilkitzkii) and depleted in ¹⁵N by 1.2–3.7‰ compared to the soft tissues. The δ¹⁵N-derived TLs of the soft tissues agreed best with the known feeding preferences of the three amphipods, which suggest a potential underestimation of 0.5–1.0 TL when stable isotope analyses are performed on whole crustaceans with thick exoskeletons. The insignificant or small differences in δ¹³C values among exoskeletons, soft tissues and whole specimens, however, suggest low probability for misinterpretations of crustaceans’ primary carbon source in marine ecosystems with distinctly different δ¹³C-carbon sources.     

Diet: tissue stable isotope fractionation of carbon and nitrogen in blood plasma and whole blood of male reindeer Rangifer tarandus

Estimates of diet derived from stable isotope analyses are sensitive to the accuracy of corrections made for diet-tissue fractionation. In particular, diet-tissue fractionation in reindeer Rangifer tarandus may be expected to differ significantly from the generic values often used in stable isotope dietary calculations, given the known values obtained from other ungulates and the complex digestive system and nutrient recycling characteristic of the species. We fed domestic reindeer a homogenous artificial diet of known isotopic value in order to directly determine diet-tissue isotopic fractionation of carbon and nitrogen, the main elements used in stable isotope dietary analyses. Diet-tissue fractionation values for blood plasma were +3.5 ± 0.1‰ (δ¹³C) and +4.2 ± 0.3‰ (δ¹⁵N). Diet-tissue fractionation values for whole blood were +3.7 ± 0.2‰ (δ¹³C) and +2.5 ± 0.3‰ (δ¹⁵N). Metabolic turnover rates were clearly sufficient for complete tissue replacement over the period of artificial feeding for blood plasma, but may not have been so for whole blood, especially for δ¹⁵N. Our values, except for whole blood δ¹⁵N, differ considerably from the generic values often used in dietary studies and interspecific comparisons of trophic niche. The results underline the importance of obtaining as specific as possible fractionation values for the species, tissue, and in some cases sex and physiological status of animals under examination, and the potential problems associated with assuming ‘generic’ fractionation values when comparing species, especially where digestive processes are dissimilar.     

Contributions of Different Organic Carbon Sources to <Emphasis Type="Italic">Daphnia</Emphasis> in the Pelagic Foodweb of a Small Polyhumic Lake: Results from Mesocosm DI<Superscript>13</Superscript>C-Additions

Abstract Freshwater ecosystems derive organic carbon from both allochthonous and autochthonous sources. We studied the relative contributions of different carbon sources to zooplankton in a small, polyhumic, steeply stratified lake, using six replicate surface-to-sediment enclosures established during summer and autumn 2004. We added ¹³C-enriched bicarbonate to the epilimnion of half the enclosures for three weeks during each season and monitored carbon stable isotope ratios of DIC, DOC, POC and Daphnia, along with physical, chemical and biological variables. During summer, ¹³C-enriched DIC (δ¹³C up to 44 ± 7.2‰) was soon taken up by phytoplankton (δ¹³C up to −5.1 ± 13.6‰) and was transmitted to Daphnia (δ¹³C up to −1.7 ± 7.2‰), demonstrating consumption of phytoplankton. In contrast, during autumn, ¹³C-enriched DIC (δ¹³C up to 56.3 ± 9.8‰) was not transmitted to Daphnia, whose δ¹³C became progressively lower (δ¹³C down to −45.6 ± 3.3‰) concomitant with decreasing methane concentration. Outputs from a model suggested phytoplankton contributed 64–84% of Daphnia diet during summer, whereas a calculated pelagic carbon mass balance indicated only 30–40% could have come from phytoplankton. Although autumn primary production was negligible, zooplankton biomass persisted at the summer level. The model suggested methanotrophic bacteria contributed 64–87% of Daphnia diet during autumn, although the calculated carbon mass balance indicated a contribution of 37–112%. Thus methanotrophic bacteria could supply virtually all the carbon requirement of Daphnia during autumn in this lake. The strongly ¹³C-depleted Daphnia values, together with the outputs from the models and the calculated carbon mass balance showed that methanotrophic bacteria can be a greater carbon source for Daphnia in lakes than previously suspected.     

Does Bienertia cycloptera with the single-cell system of C(4) photosynthesis exhibit a seasonal pattern of delta (13)C values in nature similar to co-existing C (4) Chenopodiaceae having the dual-cell (Kranz) system?

Family Chenopodiaceae is an intriguing lineage, having the largest number of C₄ species among dicots, including a number of anatomical variants of Kranz anatomy and three single-cell C₄ functioning species. In some previous studies, during the culture of Bienertia cycloptera Bunge ex Boiss., carbon isotope values (δ¹³C values) of leaves deviated from C₄ to C₃−C₄ intermediate type, raising questions as to its mode of photosynthesis during growth in natural environments. This species usually co-occurs with several Kranz type C₄ annuals. The development of B. cycloptera morphologically and δ¹³C values derived from plant samples (cotyledons, leaves, bracts, shoots) were analyzed over a complete growing season in a salt flat in north central Iran, along with eight Kranz type C₄ species and one C₃ species. For a number of species, plants were greenhouse-grown from seeds collected from the site, in order to examine leaf anatomy and C₄ biochemical subtype. Among the nine C₄ species, the cotyledons of B. cycloptera, and of the Suaeda spp. have the same respective forms of C₄ anatomy occurring in leaves, while cotyledons of members of tribe Caroxyloneae lack Kranz anatomy, which is reflected in the δ¹³C values found in plants grown in the natural habitat. The nine C₄ species had average seasonal δ¹³C values of −13.9‰ (with a range between species from −11.3 to −15.9‰). The measurements of δ¹³C values over a complete growing season show that B. cycloptera performs C₄ photosynthesis during its life cycle in nature, similar to Kranz type species, with a seasonal average δ¹³C value of −15.2‰. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Use of saguaro fruit by white-winged doves: isotopic evidence of a tight ecological association

We report the use of stable isotope and crop content analyses to quantify the use of saguaro (Carnegiea gigantea) nectar and fruit by migratory desert white-winged doves (Zenaida asiatica mearsnii). Saguaro resources had characteristically ¹³C-enriched CAM values (δ¹³C=–12.8±0.7‰ SD VPDB and –13.1±0.5‰ SD VPDB for nectar and fruit, respectively) relative to other food plants used by doves (δ¹³C_C3=–24.9±3.3‰ SD VPDB). The water contained in saguaro nectar and fruit was deuterium enriched (δD=19.6±2.0‰ SD VSMOW and 48.4±1.6‰ SD VSMOW for nectar and fruit, respectively) relative to other water sources (ranging from –41 to –19‰ VSMOW). During the fruiting season, there was a positive correlation between δ¹³C in dove liver tissues and percent of saguaro in crop contents. A two-point mixing model indicated that during the peak of saguaro fruit use, most of the carbon incorporated in dove tissues was from saguaro. Desert white-winged doves appear to be saguaro specialists. Averaged over the period when doves were resident, saguaro comprised about 60% of the total carbon incorporated into dove tissues. Tissue δ¹³C and δD of body water showed a significant positive correlation, indicating that doves were using saguaro as a source of both nutrients and water. However, at the peak of saguaro utilization, the doves’ body-water δD was more positive (by about 20‰) than saguaro fruit water. We hypothesize that this enrichment is due to fractionated evaporative water losses by doves. Using dove carbon isotope data and a two end-point mixing model we estimate that, on average, doves consume the equivalent of 128 saguaro fruits per season; each fruit contains on average 26.0±14.8 g SD of pulp (wet mass) of which 19.4 g is water. Stable isotopes have been used to produce qualitative re-constructions of animal diets. Our study shows that they can be used to provide quantitative estimates of the flow of nutrients from resources into consumers as well. Received: 30 September 1999 / Accepted: 23 March 2000     

Wetting and drying cycles drive variations in the stable carbon isotope ratio of respired carbon dioxide in semi-arid grassland

In semi-arid regions, where plants using both C₃ and C₄ photosynthetic pathways are common, the stable C isotope ratio (δ¹³C) of ecosystem respiration (δ¹³C_R) is strongly variable seasonally and inter-annually. Improved understanding of physiological and environmental controls over these variations will improve C cycle models that rely on the isotopic composition of atmospheric CO₂. We hypothesized that timing of precipitation events and antecedent moisture interact with activity of C₃ and C₄ grasses to determine net ecosystem CO₂ exchange (NEE) and δ¹³C_R. Field measurements included CO₂ and δ¹³C fluxes from the whole ecosystem and from patches of different plant communities, biomass and δ¹³C of plants and soils over the 2000 and 2001 growing seasons. NEE shifted from C source to sink in response to rainfall events, but this shift occurred after a time lag of up to 2 weeks if a dry period preceded the rainfall. The seasonal average of δ¹³C_R was higher in 2000 (−16‰) than 2001 (20‰), probably due to drier conditions during the 2000 growing season (79.7 mm of precipitation from April up to and including July) than in 2001 (189 mm). During moist conditions, δ¹³C averaged −22‰ from C₃ patches, −16‰ from C₄ patches, and −19‰ from mixed C₃ and C₄ patches. However, during dry conditions the apparent spatial differences were not obvious, suggesting reduced autotrophic activity in C₄ grasses with shallow rooting depth, soon after the onset of dry conditions. Air and soil temperatures were negatively correlated with δ¹³C_R; vapor pressure deficit was a poor predictor of δ¹³C_R, in contrast to more mesic ecosystems. Responses of respiration components to precipitation pulses were explained by differences in soil moisture thresholds between C₃ and C₄ species. Stable isotopic composition of respiration in semi-arid ecosystems is more temporally and spatially variable than in mesic ecosystems owing to dynamic aspects of pulse precipitation episodes and biological drivers.