Photosynthetic carbon uptake by marine phytoplankton: comparison of the stable (13C) and radioactive (14C) isotope methods

The ¹⁴C and the ¹³C methods are routinely used for measuringphytoplankton production. However, few systematic comparisonsof estimates using the two approaches have been conducted. Thepresent comparison is based on 257 pairs of samples, representing1 year of monthly sampling at 10 depths in the euphotic zone.Data, obtained by the same operators following a standard protocol,were collected at three different stations on the Scotian Shelf(Northwestern Atlantic Ocean). Overall agreement between thetwo methods was good (r=0.827). However, relative differencesbetween the two sets of estimates were not randomly distributedin time and space. Three factors (photic depth, station andsampling date), identified to explain the observed differences,were included in a three-way analysis of variance (ANOVA) usingrelative differences as the covariate. Following this ANOVA,the whole set was split into three subsets. For the two subsetswhere the above identified factors did not have any significanteffect, the distribution of relative differences was narrowerthan these for the whole data set. Significant effects of thethree factors persisted for the third subset and relative differencesexhibited wide variations. Possible explanations for the observeddifferences include (i) the volume of incubation bottles, (ii)the incubation temperature and (iii) the absence of measurementsof dark uptake.     

Distribution characteristics of soil humus fractions stable carbon isotope natural abundance (delta 13C) in paddy field under long-term ridge culture

A 16-year field experiment was conducted in a ridge culture paddy field in the hilly region of Sichuan Basin, aimed to investigate the distribution characteristics of stable carbon isotope natural abundance (delta 13C) in soil humus fractions. The soil organic carbon (SOC) content in the paddy field under different cultivation modes ranked in the order of wide ridge culture > ridge culture > paddy and upland rotation. In soil humus substances (HS), humin (HU) was the main composition, occupying 21% - 30% of the total SOC. In the extracted soil carbon, humic acid (HA) dominated, occupying 17% - 21% of SOC and 38% - 65% of HS. The delta 13C value of SOC ranged from -27.9 per thousand to -25.6 per thousand, and the difference of the delta 13C value between 0-5 cm and 20-40 cm soil layers was about 1.9 per thousand. The delta 13C value of HA under different cultivation modes was 1 per thousand - 2 per thousand lower than that of SOC, and more approached to the delta 13C value of rapeseed and rice residues. As for fulvic acid (FA), its delta 13C value was about 2 per thousand and 4 per thousand higher than that of SOC and HA, respectively. The delta 13C value of HU in plough layer (0-20 cm) and plow layer (20-40 cm) ranged from -23.7 per thousand - -24.9 per thousand and -22.6 per thousand - -24.2 per thousand, respectively, reflecting the admixture of young and old HS. The delta 13C value in various organic carbon fractions was HU>FA>SOC>rapeseed and rice residues>HA. Long-term rice planting benefited the increase of SOC content, and cultivation mode played an important role in affecting the distribution patterns of soil humus delta 13C in plough layer and plow layer.     

Using stable isotope natural abundances (delta 15N and delta 13C) to integrate the stress responses of wild barley (Hordeum spontaneum C. Koch.) genotypes

To integrate the complex physiological responses of plants tostress, natural abundances (     

Intrapopulation variation in gray wolf isotope (delta(15)N and delta(13)C) profiles: implications for the ecology of individuals

Trophic relationships among organisms in terrestrial boreal ecosystems define ecological communities and are important in determining dynamics of energy flow and ecosystem function. We examined trophic relationships between the gray wolf (Canis lupus) and 18 mammalian species from the boreal forest of central Saskatchewan, Canada, using delta(13)C and delta(15)N stable isotope values measured in guard hair samples. Variance in isotope values for wolves and other carnivores was investigated as a proxy for variation in diet among individuals. Isosource, an isotopic source partitioning model, quantified the relative range in proportions of five most-likely prey items in the diets of wolves. The distribution of feasible contributions from each source was dominated by elk (Cervus elaphus; mean: 48%, range:11-75%), followed by white-tailed deer (Odocoileus virginianus; mean: 21%, range: 0-54%), moose (Alces alces; mean:14%, range: 0-41%), beaver (Castor canadensis; mean: 8%, range:0-25%) and snowshoe hare (Lepus americanus; mean: 8%, range: 0-24%). Despite social foraging, our results indicate highly variable diets among individuals and we discuss this in terms of individual versus group ecology of boreal wolves.     

13C isotope fractionation during rhizosphere respiration of C3 and C4 plants

Stable carbon isotopes are used extensively to partition total soil CO₂ efflux into root-derived rhizosphere respiration or autotrophic respiration and soil-derived heterotrophic respiration. However, it remains unclear whether CO₂ from rhizosphere respiration has the same δ¹³C value as root biomass. Here we investigated the magnitude of ¹³C isotope fractionation during rhizosphere respiration relative to root biomass in six plant species. Plants were grown in a carbon-free sand-perlite medium inoculated with microorganisms from a farm soil for 62 days inside a greenhouse. We measured the δ¹³C value of rhizosphere respiration using a closed-circulation 48-hour CO₂ trapping method during 40~42 and 60~62 days after sowing. We found a consistent depletion in ¹³C (0.9~1.7‰) of CO₂ from rhizosphere respiration relative to root biomass in three C₃ species (Glycine max L. Merr., Helianthus annuus L. and Triticum aestivum L.), but a relatively large depletion in ¹³C (3.7~7.0‰) in three C₄ species (Amaranthus tricolor L., Sorghum bicolor (L.) Moench and Zea mays L. ssp. mays). Overall, our results indicate that CO₂ from rhizosphere respiration is more ¹³C-depleted than root biomass. Therefore, accounting for this ¹³C fractionation is required for accurately partitioning total soil CO₂ efflux into root-derived and soil-derived components using natural abundance stable carbon isotope methods.     

Determination of isotope fractionation factors and quantification of carbon flow by stable carbon isotope signatures in a methanogenic rice root model system

Methanogenic processes can be quantified by stable carbon isotopes, if necessary modeling parameters, especially fractionation factors, are known. Anoxically incubated rice roots are a model system with a dynamic microbial community and thus suitable to investigate principal geochemical processes in anoxic natural systems. Here we applied an inhibitor of acetoclastic methanogenesis (methyl fluoride), calculated the thermodynamics of the involved processes, and analyzed the carbon stable isotope signatures of CO₂, CH₄, propionate, acetate and the methyl carbon of acetate to characterize the carbon flow during anaerobic degradation of rice roots to the final products CO₂ and CH₄. Methyl fluoride inhibited acetoclastic methanogenesis and thus allowed to quantify the fractionation factor of CH₄ production from H₂/CO₂. Since our model system was not affected by H₂ gradients, the fractionation factor could alternatively be determined from the Gibbs free energies of hydrogenotrophic methanogenesis. The fractionation factor of acetoclastic methanogenesis was also experimentally determined. The data were used for successfully modeling the carbon flow. The model results were in agreement with the measured process data, but were sensitive to even small changes in the fractionation factor of hydrogenotrophic methanogenesis. Our study demonstrates that stable carbon isotope signatures are a proper tool to quantify carbon flow, if fractionation factors are determined precisely.     

Impact of deficit irrigation on water use efficiency and carbon isotope composition (delta13C) of field-grown grapevines under Mediterranean climate

The objective of this study was to evaluate the effect of deficit irrigation on intrinsic water use efficiency (A/g(s)) and carbon isotope composition (delta13C) of two grapevine cultivars (Moscatel and Castel?o), growing in a commercial vineyard in SW Portugal. The study was done in two consecutive years (2001 and 2002). The treatments were full irrigation (FI), corresponding to 100% of crop evapotranspiration (ETc), rain-fed (no irrigation, NI), and two types of deficit irrigation (50% ETc): (i) by supplying the water either to one side of the root system or to the other, which is partial rootzone drying (PRD), or (ii) dividing the same amount of water by the two sides of the root system, the normal deficit irrigation (DI). The water supplied to the PRD treatment alternated sides approximately every 15 d. The values of predawn leaf water potential (Psi(pd)) and the cumulative integral of Psi(pd) (S(Psi)) during the season were lower in 2001 than in the 2002 growing season. Whereas differences in Psi(pd) and S(Psi) between PRD and DI were not significantly different in 2001, in 2002 (a dryer year) both cultivars showed lower values of S(Psi) in the PRD treatment as compared with the DI treatment. This suggests that partial rootzone drying may have a positive effect on water use under dryer conditions, either as a result of better stomatal control and/or reduced vigour. The effects of the water treatments on delta13C were more pronounced in whole grape berries and pulp than in leaves. The delta13C of pulp showed the best correlation with intrinsic water use efficiency (A/g(s)) as well as with S(Psi). In spite of the better water status observed in PRD compared with DI in the two cultivars in 2002, no statistical differences between the two treatments were observed in A/g(s) and delta13C. On the other hand, they showed a higher delta13C compared with FI. In conclusion, it is apparent that the response to deficit irrigation varies with the environmental conditions of the particular year, the driest conditions exacerbating the differences among treatments. The highest values of delta13C found in the pulp of NI vines in Castel?o compared with Moscatel suggest different sensitivities to water deficits in the two cultivars, as was empirically observed.     

Distribution of the heavy isotope of carbon (13C) in biological systems

Causes conditioning fractionation of carbon isotopes in biological systems are considered. Concepts of E. M. Galimov are discussed. According to these concepts distribution of carbon isotopes between biomolecules and their fragments is quaziequilibrium, i. e. it differs from the equilibrium one by a constant multiplier, which is the same for all the biomolecules of an organism but different for various organisms. These concepts have no theoretical grounds and do not agree with the experimental evidence available. An analysis of experimental data, as well as theoretical considerations, indicate that the observed differences in isotope composition of metabolytes and their fragments in the living organisms are conditioned by the kinetic isotope effects of carbon at the stages of their enzymic transformations and by the portion of substances participating in the reaction. It means that these differences do not depend directly on the constants characterizing the equilibrium distribution of carbon isotopes between corresponding compounds and between different groups inside their molecules.     

集约化生产对农田土壤碳氮含量及δ13C和δ15N同位素丰度的影响

集约化生产下农田土壤碳、氮含量变化是衡量土壤肥力持久性的重要指标.对常规水稻-蚕豆轮作地、露地蔬菜地、3年塑料大棚地和10年以上塑料大棚地的土壤pH、电导率(EC)、土壤有机碳(SOC)和总氮(TN)含量及δ13C和δ15N同位素丰度进行测定,研究了集约化生产程度对土壤特性的影响.结果表明:与水稻-蚕豆轮作地相比,露地蔬菜地、3年塑料大棚地和10年以上塑料大棚地0 ～20 cm耕层土壤pH分别降低1.1、0.8和0.7,而土壤EC分别是水稻-蚕豆轮作地的4.2、4.9和5.2倍;土壤碳、氮含量随塑料大棚地生产年限的增加总体上呈先增大后减小的趋势.与水稻-蚕豆轮作地相比,10年以上塑料大棚地0～20、20～40、40 ～60、60 ～ 80、80 ～ 100 cm土层的土壤SOC含量分别下降了54％、46％、60％、63％和59％,土壤TN含量分别下降了53％、53％、71％、82％和85％.农田集约化生产程度显著影响土壤SOC、TN含量和δ13C、δ15N丰度,土壤δ13C丰度与SOC含量呈显著负相关.土壤δ13C丰度可作为评价农田土壤碳循环受人为干扰强度的指标.     

Differential C isotope discrimination by fungi during decomposition of C(3)- and C(4)-derived sucrose

Stable isotope analysis is a major tool used in ecosystem studies to establish pathways and rates of C exchange between various ecosystem components. Little is known about isotopic effects of many such components, especially microbes. Here we report on the discovery of an unexpected pattern of C isotopic discrimination by basidiomycete fungi with far-reaching consequences for our understanding of isotopic processing in ecosystems where these microbes mediate material transfers across trophic levels. We measured fractionation effects on three ecologically relevant basidiomycete species under controlled laboratory conditions. Sucrose derived from C(3) and C(4) plants is fractionated differentially by these microbes in a taxon-specific manner. The differentiation between mycorrhizal and saprotrophic fungi observed in the field by others is not explained by intrinsic discrimination patterns. Fractionation occurs during sugar uptake and is sensitive to the nonrandom distribution of stable isotopes in the sucrose molecule. The balance between respiratory physiology and fermentative physiology modulates the degree of fractionation. These discoveries disprove the assumption that fungal C processing does not significantly alter the distribution of stable C isotopes and provide the basis for a reevaluation of ecosystem models based on isotopic evidence that involve C transfer across microbial interfaces. We provide a mechanism to account for the observed differential discrimination effects.     

Muscle delta13C change in Nile tilapia (Oreochromis niloticus): effects of growth and carbon turnover

The contribution of growth and turnover to the muscle delta(13)C change process was investigated using mathematical models which associate delta(13)C change to time of intake of a new diet or increase in body mass. Two groups of Nile tilapia (Oreochromis niloticus) were fed on diets based on C3 (delta(13)C=-25.64+/-0.06 per thousand) or C4 (delta(13)C=-16.01+/-0.06 per thousand) photosynthetic cycle plants to standardize the muscle delta(13)C. After establishing the carbon isotopic equilibrium, fish (mean mass 24.12+/-6.79 g) then received the other treatment diet until a new carbon isotopic equilibrium could be established, characterizing T1 (C3-C4) and T2 (C4-C3) treatments. No significant differences were observed in fish productive performance. Good fits were obtained for the models that associated the delta(13)C change to time, resulting in carbon half-life values of 23.33 days for T1 and 25.96 days for T2. Based on values found for the muscle delta(13)C change rate from growth (0.0263 day(-1) and 0.0254 day(-1)) and turnover (0.0034 day(-1) and 0.0013 day(-1)), our results indicate that most of the delta(13)C change could be attributed to growth. The application of model that associated the delta(13)C change to body mass increase seems to produce results with no apparent biological explanation. The delta(13)C change rate could directly reflect the daily ration and growth rate, and consequently the isotopic change rates of carbon and other tissue elements can be properly used to assess different factors that may interfere in nutrient utilization and growth.     

湖泊水生植物稳定碳同位素分馏机制与应用研究进展

水生植物是维持湖泊生态系统健康发展的重要基础与支撑。与陆地植物相比,水生植物稳定碳同位素(δ¹³C)的分馏过程较复杂,为系统的了解水生植物δ¹³C的分馏过程及其在湖泊现代生态系统和古环境研究中的应用,调研了国内外相关研究进展。总结已有研究表明,水生植物的δ¹³C主要与其吸收利用的碳源和水体环境要素等密切相关,影响因素较复杂,而且目前在利用水生植物的δ¹³C研究不同时间尺度上湖泊环境变化时所选用的载体存在差异。植物不同组分样品的选择对解释水生植物δ¹³C与环境之间的关系至关重要。因此,以太湖典型水生植物为研究实例,探讨了水生植物不同组分稳定碳同位素的分布特征及其对环境因子的响应差异,结果显示水生植物不同组分的δ¹³C存在一定的差异,而且对环境的响应也不同,未来在利用水生植物δ¹³C研究湖泊环境变化时对于水生植物样品组分的选择应谨慎考虑。     

Comparison of radioactive and stable isotope tracer techniques for measuring photosynthesis: 13C and 14C uptake by marine phytoplankton

The stable carbon isotope ¹³C has been used in the open oceanto estimate the inorganic carbon uptake by phytoplankton andthis technique has been compared with the ¹³C tracer method.An overall correlation coefficient of 0.806 and a regressionslope of 1.29 were calculated from 50 sample pairs gatheredduring three cruises in widely different oceanic areas rangingin production rates from 0.01 to 6 mgC m^–3 h^–1.However, significant differences between the two methods wereapparent for cruises located in nutrient-depleted areas. Possibleexplanations lie either in a volume effect, the high silicatecontent of the ¹⁴C solution which could stimulate the ¹⁴C uptakeor in errors associated with the particulate carbon measurementswhich are necessary to convert specific uptake rates to absoluteuptake rates and to yield compatible units for the comparison,in laboratory cultures the ¹⁴C technique overestimated the netparticulate carbon increase by — 16%. ⁺Present address: Laboratoire marin CNRS-IFREMER, L'Houmeau,17137 Nieul-sur-Mer, France. ^*This paper is the result of a study made at the Group for AquaticPrimary Productivity (GAP) First International Workshop heldat the Limnological Institute, University of Konstanz, in April1982.     

Minor stable carbon isotope fractionation between respired carbon dioxide and bulk soil organic matter during laboratory incubation of topsoil

A common assumption in paleoenvironmental reconstructions using soils is that the carbon isotope composition of soil-respired CO₂ is equivalent to the carbon isotope composition of bulk soil organic matter (SOM). However, the occurrence of a non-zero per mil carbon isotope enrichment factor between CO₂ and SOM (\(\varepsilon_{{{\text{CO}}_{ 2} - {\text{SOM}}}}\)) during soil respiration is the most widely accepted explanation for the down-profile increase in SOM δ¹³C values commonly observed in well-drained soils. In order to shed light on this apparent discrepancy, we incubated soil samples collected from the top 2 cm of soils with pure C₃ vegetation and compared the δ¹³C values of soil-respired CO₂ to the δ¹³C values of bulk SOM. Our results show near-zero \(\varepsilon_{{{\text{CO}}_{ 2} - {\text{SOM}}}}\) values (?0.3 to 0.4 ‰), supporting the use of paleosol organic matter as a proxy for paleo soil-respired CO₂. Significantly more negative \(\varepsilon_{{{\text{CO}}_{ 2} - {\text{SOM}}}}\) values are required to explain the typical δ¹³C profiles of SOM in well-drained soils. Therefore our results also suggest that typical SOM δ¹³C profiles result from either (1) a process other than carbon isotope fractionation between CO₂ and SOM during soil respiration or (2) \(\varepsilon_{{{\text{CO}}_{ 2} - {\text{SOM}}}}\) values that become increasingly negative as SOM matures.     

Respiratory carbon metabolism following illumination in intact French bean leaves using (13)C/(12)C isotope labeling

The origin of the carbon atoms in the CO(2) respired by French bean (Phaseolus vulgaris) leaves in the dark has been studied using (13)C/(12)C isotopes as tracers. The stable isotope labeling was achieved through a technical device that uses an open gas-exchange system coupled online to an elemental analyzer and linked to an isotope ratio mass spectrometer. The isotopic analysis of the CO(2) respired in the dark after a light period revealed that the CO(2) was labeled, but the labeling level decreased progressively as the dark period increased. The pattern of disappearance depended on the amount of carbon fixed during the labeling and indicated that there were several pools of respiratory metabolites with distinct turnover rates. We demonstrate that the carbon recently assimilated during photosynthesis accounts for less than 50% of the carbon in the CO(2) lost by dark respiration and that the proportion is not influenced by leaf starvation in darkness before the labeling. Therefore, most of the carbon released by dark respiration after illumination does not come from new photosynthates.     

Insights into the fate of a 13C labelled phenol pulse for stable isotope probing (SIP) experiments

Stable isotope probing (SIP) using DNA or RNA as a biomarker has proven to be a useful method for attributing substrate utilisation to specific microbial taxa. In this study we followed the transfer of a (13)C(6)-phenol pulse in an activated sludge micro-reactor to examine the resulting distribution of labelled carbon in the context of SIP. Most of the added phenol was metabolically converted within the first 100 min after (13)C(6)-phenol addition, with 49% incorporated into microbial biomass and 6% respired as CO(2). Less than 1% of the total (13)C labelled carbon supplied was incorporated into microbial RNA and DNA, with RNA labelling 6.5 times faster than DNA. The remainder of the added (13)C was adsorbed and/or complexed to suspended solids within the sludge. The (13)C content of nucleic acids increased beyond the initial consumption of the (13)C-phenol pulse. This study confirms that RNA labels more efficiently than DNA and reveals that only a small proportion of a pulse is incorporated into nucleic acids. Evidence of continued (13)C incorporation into nucleic acids suggests that cross-feeding of the SIP substrate was rapid. This highlights both the benefits of using a biomarker that is rapidly labelled and the importance of sampling within appropriate timescales to avoid or capture the effects of cross-feeding, depending on the goal of the study.     

Seasonal carbon allocation to arbuscular mycorrhizal fungi assessed by microscopic examination, stable isotope probing and fatty acid analysis

Background and Aim

Climate change models are limited by lack of baseline data, in particular carbon (C) allocation to – and dynamics within – soil microbial communities. We quantified seasonal C-assimilation and allocation by plants, and assessed how well this corresponds with intraradical arbuscular mycorrhizal fungal (AMF) storage and structural lipids (16:1ω5 NLFA and PLFA, respectively), as well as microscopic assessments of AMF root colonization.

Methods

Coastal Hypochoeris radicata plants were labeled with ¹³CO₂ in February, July and October, and ¹³C-allocation to fine roots and NLFA 16:1ω5, as well as overall lipid contents and AM colonization were quantified.

Results

C-allocation to fine roots and AMF storage lipids differed seasonally and mirrored plant C-assimilation, whereas AMF structural lipids and AM colonization showed no seasonal variation, and root colonization exceeded 80 % throughout the year. Molecular analyzes of the large subunit rDNA gene indicated no seasonal AMF community shifts.

Conclusions

Plants allocated C to AMF even at temperatures close to freezing, and fungal structures persisted in roots during times of low C-allocation. The lack of seasonal differences in PLFA and AM colonization indicates that NLFA analyses should be used to estimate fungal C-status. The implication of our findings for AM function is discussed.