Autotrophic carbon sources for heterotrophic bacterioplankton in a floodplain lake of central Amazon

The relative contribution of autotrophic carbon sources (aquatic macrophytes, flooded forest, phytoplankton) for heterotrophic bacterioplankton was evaluated in a floodplain lake of the Central Amazon. Stable carbon isotopes (¹³C) were used as tracers. Values of ¹³C of different autotrophic sources were compared to those of dissolved organic carbon (DOC) and those of bacterially produced CO₂.The percentage of carbon derived from C₄ macrophytes for bacterially produced CO₂ was the highest, on average 89%. The average ¹³C value of CO₂ from bacterial respiration was –18.5 ± 3.3. Considering a fractionation of CO₂ of 3 by bacterial respiration, ¹³C value was –15.5, near C₄ macrophyte ¹³C value (–13.1).The average value of total DOC ¹³C was –26.8 ± 2.4. The percentage of C₄ macrophytes carbon for total DOC was on average 17%. Considering that bacteria consume mainly carbon from macrophytes, the dominance of C₃ plants for total DOC probably reflects a faster consumption of the former source, rather than a major contribution of the latter source.Heterotrophic bacterioplankton in the floodplain may be an important link in the aquatic food web, transferring the carbon from C₄ macrophytes to the consumers.     

Stable isotope and radiocarbon compositions of methane emitted from tropical rice paddies and swamps in Southern Thailand

Stable isotopes (¹³C, D) and radiocarbon weremeasured in methane bubbles emitted from rice paddies and swamps in southernThailand. Methane emitted from the Thai rice paddies was enriched in¹³C (mean ¹³C; –51.5 ±7.1 and–56.5 ± 4.6 for mineral soil and peat soil paddies,respectively)relative to the reported mean value of methane from temperate rice paddies(– 63 ± 5). Large seasonal variation was observed in¹³C(32) in the rice paddies, whereas variationinD was much more smaller (20), indicating that variation in¹³C is due mainly to changes in methane production pathways.Values of ¹³C were lower in swamps (–66.1 ±5.1)than in rice paddies. The calculated contribution of acetate fermentation from¹³C value was greater in rice paddies (mineral soils:62–81%, peat soils: 57–73%) than in swamps (27–42%). Din methane from Thai rice paddies (–324± 7 (n=46)) isrelativelyhigher than those from 14 stations in Japanese rice paddies ranging from–362 ± 5 (Mito: n=2) to –322 ± 8(Okinawa: n=3), due tohigher D in floodwaters. ¹⁴C content in methane produced fromThai rice paddies (127±1 pMC) show higher ¹⁴Cactivity compared with previous work in paddy fields and those from Thai swamps(110±2 pMC).     

Contribution of methanol to the production of methane and its <Superscript>13</Superscript>C-isotopic signature in anoxic rice field soil

Conversion of methanol to CH₄ has a large isotope effect so that a small contribution of methanol-dependent CH₄ production may decrease the ¹³CH₄ of total CH₄ production. Therefore, we investigated the role of methanol for CH₄ production. Methanol was not detectable above 10 M in anoxic methanogenic rice field soil. Nevertheless, addition of ¹³C-labeled methanol (99% enriched) resulted in immediate accumulation of ¹³CH₄. Addition of 0.1 M ¹³C-methanol resulted in increase of the ¹³CH₄ from –47 to –6 within 2 h, followed by a slow decrease. Addition of 1 M ¹³C-methanol increased ¹³CH₄ to +500 within 4 h, whereas 10 M increased ¹³CH₄ to +2500 and continued to increase. These results indicate that the methanol concentrations in situ, which diluted the ¹³C-methanol added, were 0.1 M and that the turnover of methanol contributed only about 2% to total CH₄ production at 0.1 M. However, contribution increased up to 5 and 17% when 1 and 10 M methanol were added, respectively. Anoxic rice soil that was incubated at different temperatures between 10 and 37 °C exhibited maximally 2–6% methanol-dependent methanogenesis about 1–2 h after addition of 1 M ¹³C-methanol. Only at 50 °C, contribution of methanol to CH₄ production reached a maximum of 10%. After longer (7–10 h) incubation, however, contribution generally was only 2–4%. Methanol accumulated in the soil when CH₄ production was inhibited by chloroform. However, the accumulated methanol accounted for only up to 0.7 and 1.2% of total CH₄ production at 37 and 50 °C, respectively. Collectively, our results show that methanol-dependent methanogenesis was operating in anoxic rice field soil but contributed only marginally to total CH₄ production and the isotope effect observed at both low and high temperature.     

γδ T-cell receptor repertoire in human peripheral blood and thymus

T-cell clones expressing the T-cell receptor (Tcr) were generated from peripheral blood lymphocytes (PBLs) and from a thymus sample. In the panel of ten thymus-derived clones, four Tcr phenotypes [as defined by the reaction of monoclonal antibodies (mAbs) directed against known V and V regions] were identified. All the clones lacked expression of the V3 V region, while seven clones were V1+ . V1 was found in combination with V9 or with undefined VVregions. In addition, two other Tcr phenotypes were identified on these clones: V9⁺ V1^– V3^– and V9^– V1^– V3^– One of the clones expressed CD4 and another was CD8positive. The remaining clones were CD4^– CD8^–. In the panel of 76 PBL-derived, Tcr-bearing clones, five Tcr phenotypes could be identified. In contrast to the thymus-derived clones, 30% of the clones were V3⁺ whereas V1 was expressed by a minority of the clones only. One clone was CD4-positive and approximately 30% of the clones were CD8-positive. Four of the five mAb-defined Tcr phenotypes could be identified on both thymus and PBL-derived T-cell clones. However, biochemical analysis of the Tcrs demonstrates differences in the usage of Ct- and C2-encoded y chains by T cells derived from the thymus and PBLs. The results therefore indicate that, at the clonal level, similarities and differences exist between the Tcr repertoires expressed in the thymus and by PBLs. Furthermore, they indicate that combinatorial Tcr heterogeneity is larger than has so far been described. The receptor diversity, combined with the potential of Tcr+ cells to express CD4 or CD8, indicates that these cells are a heterogeneous population that might mediate a number of immune functions.     

Intracellular sites of the synthesis of sweet potato mitochondrial F1ATPase subunits

Summary Five subunits (-, -, -, - and -subunits) of the six -and -subunits) in the F₁ portion (F₁ATPase) of sweet potato (Ipomoea batatas) mitochondrial adenosine triphosphatase were isolated by an electrophoretic method. The - and -subunits were not distinguishable immunologically but showed completely different tryptic peptide maps, indicating that they were different molecular species. In vitro protein synthesis with isolated sweet potato root mitochondria produced only the -subunit when analyzed with anti-sweet potato F₁ATPase antibody reacting with all the subunits except the -subunit. Sweet potato root poly(A)⁺RNA directed the synthesis of six polypeptides which were immunoprecipitated by the antibody: two of them immunologically related to the -subunit and the others to the - and -subunits. We conclude that the -subunit of the F₁ATPase is synthesized only in the mitochondria and the -, - and -subunits are in the cytoplasm.     

Analysis of primary food sources and trophic relationships of aquatic animals in a mangrove-fringed estuary,Khung Krabaen Bay (Thailand) using dual stable isotope techniques

Stable carbon (¹³C) and nitrogen (¹⁵N) isotopes were used to elucidate primary food sources and trophic relationships of organisms in Khung Krabaen Bay and adjacent offshore waters. The three separate sampling sites were mangroves, inner bay and offshore. The ¹³C values of mangrove leaves were –28.2 to –29.4, seagrass –10.5, macroalgae –14.9 to –18.2, plankton –20.0 to –21.8, benthic detritus –15.1 to –26.3, invertebrates –16.5 to –26.0, and fishes –13.4 to –26.3. The ¹⁵N values of mangrove leaves were 4.3 to 5.7, seagrass 4.3, macroalgae 2.2 to 4.4, plankton 5.7 to 6.4 , benthic detritus 5.1 to 5.3, invertebrates 7.2 to 12.2 , and fishes 6.3 to 15.9. The primary producers had distinct ¹³C values. The ¹³C values of animals collected from mangroves were more negative than those of animals collected far from shore. The primary carbon sources that support food webs clearly depended on location. The contribution of mangroves to food webs was confined only to mangroves, but a mixture of macroalgae and plankton was a major carbon source for organisms in the inner bay area. Offshore organisms clearly derived their carbon through the planktonic food web. The ¹⁵N values of consumers were enriched by 3–4 relative to their diets. The ¹⁵N data suggests that some of aquatic animals had capacity to change their feeding habits according to places and availability of foods and as a result, individuals of the same species could be assigned to different trophic levels at different places.     

Stereochemistry of protected ornithine side chains in gramicidin S derivatives and their resistance to N-methylation

Derivatives of gramicidin S (GS) and its mono- and di-d-cyclohexylalanine (d-Cha) analogs possessing various protecting groups on Orn side chains were prepared. ¹H NMR spectra of the unsymmetrically protected analogs [Orn(X)²,Orn(X)²,d-Cha⁴]GS were similar to the composites of the spectra of the symmetrical derivatives [Orn(X)²,2,d-Cha⁴,4]GS and [Orn(X)²,2]GS, revealing the proximity of the protecting groups of NH of Orn residues at the 2 and 2 positions to the side chains of d-Phe (or d-Cha) residues at the 4 and 4 positions, respectively. The results indicated the presence of H-bonds between the NH of Orn and the carbonyl of d-Phe residues in the i i + 2 sense and not in i i – 3, which was also supported by the ROESY analysis. The substantially strong H-bonds can explain the observed resistance of the urethane NH of the Orn side chains in the GS derivatives to the N-methylation with CH³I–Ag²O in DMF.     

Evidence suggesting energy-dependent formaldehyde transport in an RuMP-type methylotroph (T15)

Formaldehyde accumulation ratios ([¹⁴CH₂O]_i/[¹⁴CH₂O]_o) as high as 12-fold were measured in anaerobic, CH₃OH-energized, whole cell suspensions of the ribulose monophosphate (RuMP)-type methylotrophic strain T15. Uptake kinetics were extremely rapid, enabling the attainment of equilibrium in only 10–30 s. Transport appears to be energy-dependent and associated with the protonmotive force (pmf). Anaerobic incubation with 5 M carbonyl p-(trifluoromethoxy)-phenylhydrazone (FCCP) led to 70%–90% reduction of the accumulation ratio. Though not as pronounced, diminished uptake was also observed in the presence of 140 M nigericin, 161 M valinomycin and 90 mM KSCN, commensurate with their effects on pmf. Accumulation of CH₂O as a function of external pH followed a trend more similar to that of pmf than either pH or . Preventing energization by incubation with 100 M N,N-dicyclohexylcarbodiimide (DCCD) led to nearly 80% inhibition of CH₂O transport. Over short time periods it was possible to chase accumulated ¹⁴CH₂O from previously loaded cells by collapsing pmf; however, this technique also indicated that significant ¹⁴CH₂O incorporation began to occur within 3 min.Abbreviations FCCP Carbonyl cyanide p-(trifluoromethyoxy)-phenylhydrazone - DCCD N,N-dicyclohexylcarbodiimide - RuMP ribulose monophosphate - TPP⁺ tetra[U-¹⁴C]phenylphosphonium - pmf protonmotive force     

Leaf 15N abundance of subarctic plants provides field evidence that ericoid,ectomycorrhizal and non-and arbuscular mycorrhizal species access different sources of soil nitrogen

The natural abundance of the nitrogen isotope 15, ¹⁵N, was analysed in leaves of 23 subarctic vascular plant species and two lichens from a tree-line heath at 450 m altitude and a fellfield at 1150 m altitude close to Abisko in N. Sweden, as well as in soil, rain and snow. The aim was to reveal if plant species with different types of mycorrhizal fungi also differ in their use of the various soil N sources. The dwarf shrubs and the shrubs, which in combination formed more than 65% of the total above-ground biomass at both sites, were colonized by ericoid or ectomycorrhizal fungi. Their leaf ¹⁵N was between–8.8 and–5.5 at the heath and between–6.1 and –3.3 at the fellfield. The leaf ¹⁵N of non- or arbuscular mycorrhizal species was markedly different, ranging from –4.1 to –0.4 at the heath, and from –3.4 to+2.2 at the fellfield. We conclude that ericoid and ectomycorrhizal dwarf shrubs and shrubs utilize a distinct N source, most likely a fraction of the organic N in fresh litter, and not complexed N in recalcitrant organic matter. The latter is the largest component of soil total N, which had a ¹⁵N of –0.7 at the heath and +0.5 at the fellfield. Our field-based data thus support earlier controlled-environment studies and studies on the N uptake of excised roots, which have demonstrated protease activity and amino acid uptake by ericoid and ectomycorrhizal tundra species. The leaves of ectomycorrhizal plants had slightly higher ¹⁵N (fellfield) and N concentration than leaves of the ericoids, and Betula nana, Dryas octopetala and Salix spp. also showed NO _inf3 ^sup- reductase activity. These species may depend more on soil inorganic N than the ericoids. The ¹⁵N of non- or arbuscular mycorrhizal species indicates that the ¹⁵N of inorganic N available to these plants was higher than that of average fresh litter, probably due to high microbial immobilization of inorganic N. The ¹⁵N of NH _inf4 ^sup+ -N was +12.3 in winter snow and +1.9 in summer rain. Precipitation N might be a major contributer in species with poorly developed root systems, e.g. Lycopodium selago. Our results show that coexisting plant species under severe nutrient limitation may tap several different N sources: NH _inf4 ^sup+ , NO _inf3 ^sup- and organic N from the soil, atmospheric N₂, and N in precipitation. Ericoid and ectomycorrhizal fungi are of major importance for plant N uptake in tundra ecosystems, and mycorrhizal fungi probably exert a major control on plant ¹⁵N in organic soils.     

Distribution of greenhouse gases, nitrite, and δ13C of dissolved inorganic carbon in Lake Biwa: Implications for hypolimnetic metabolism

The vertical distribution of dissolved greenhouse gases (CH₄,CO₂ and N₂O), NO ,and ¹³C of CO₂ in Lake Biwa during a stagnantperiod was precisely determined. CO₂ as well as NO was accumulated in the hypolimnion, whereas NO and CH₄concen\-trations were generally higher in theepilimnion than in the hypolimnion. In August, NO andCH₄ were ephemerally accumulated at the thermocline. Theconcentration of CH₄ always exceeded equilibrium with respectto air/water exchange. N₂O was rather uniformly distributed inboth time and space, and remained near equilibrium with respect toair/water exchange. All of these observations are similar to otherstratified, oligotrophic lakes, in which the hypolimnia were welloxygenated. The ¹³C of CO₂ became morenegative with increasing depth, and showed a strong negativecorrelation with apparent oxygen utilization. From the data, the ¹³Cvalue of organic matter decomposed into CO₂ inthe hypolimnion was calculated by isotope mass-balance, and found tobe in a similar range to ¹³C of phytoplankton and benthic algaeand distinctively higher than ¹³C of both terrestrial andsedimentary organic matters. This suggests that autochthonous organicmatter was the major source of CO₂.     

Comparisons of δ13C values in leaves of aquatic macrophytes from different habitats in Britain and Finland; some implications for photosynthetic processes in aquatic plants

Summary The ¹³C values of submerged aquatic plants from contrasting but relatively defined habitats, and the ¹³C values of emergent, floating and submerged leaves of dimorphic aquatic plants, were measured. In many instances the ¹³C values of dissolved inorganic carbon in the water were also measured. Plant ¹³C values in the vicinity of-40 to-50 were found in rapidly flowing spring waters with carbonate ¹³C values of-16 to-21, consistent with the notion that species such as Fontinalis antipyretica almost exclusively assimilate free CO₂ via RuP₂ carboxylase. Plant ¹³C values in the vicinity of-10 to-15 in sluggish water with carbonate ¹³C values of about-5 were observed, consistent with the notion that boundary layer diffusion and/or HCO₃ ^- uptake may determine the ¹³C value of submerged aquatic plants in these circumstances. Comparisons of ¹³C values of the same or related species growing in waters of similar carbonate ¹³C value but different flow rates confirmed this view; more negative ¹³C values were frequently associated with plants in fast moving water. In Britain, but not in Finland, the ¹³C values of submerged leaves of dimorphic plants were almost invariably more negative than in aerial leaves. The ¹³C value of carbonate from chalk streams and in acid springs indicate substantial inputs of respiratory CO₂, as opposed to atmospheric carbon. The contributions of these variations in ¹³C of the carbon source, and of isotope fractionation in diffusion, to the ¹³C value of submerged parts of dimorphic plants is discussed.     

15N natural abundance studies in Australian commercial sugarcane

The measurement of natural ¹⁵N abundance is a well-established technique for the identification and quantification of biological N₂ fixation in plants. Associative N₂ fixing bacteria have been isolated from sugarcane and reported to contribute potentially significant amounts of N to plant growth and development. It has not been established whether Australian commercial sugarcane receives significant input from biological N₂ fixation, even though high populations of N₂ fixing bacteria have been isolated from Australian commercial sugarcane fields and plants. In this study, ¹⁵N measurements were used as a primary measure to identify whether Australian commercial sugarcane was obtaining significant inputs of N via biological N₂ fixation. Quantification of N input, via biological N₂ fixation, was not possible since suitable non-N₂ fixing reference plants were not present in commercial cane fields. The survey of Australian commercially grown sugarcane crops showed the majority had positive leaf ¹⁵N values (73% >3.00, 63% of which were >5.00), which was not indicative of biological N₂ fixation being the major source of N for these crops. However, a small number of sites had low or negative leaf ¹⁵N values. These crops had received high N fertiliser applications in the weeks prior to sampling. Two possible pathways that could result in low ¹⁵N values for sugarcane leaves (other than N₂ fixation) are proposed; high external N concentrations and foliar uptake of volatilised NH₃. The leaf ¹⁵N value of sugarcane grown in aerated solution culture was shown to decrease by approximately 5 with increasing external N concentration (0.5–8.0 mM), with both NO₃ ^– and NH₄ ⁺ nitrogen forms. Foliar uptake of atmospheric NH₃ has been shown to result in depleted leaf ¹⁵N values in many plant species. Acid traps collected atmospheric N with negative ¹⁵N value (–24.45±0.90) from above a field recently surface fertilised with urea. The ¹⁵N of leaves of sugarcane plants either growing directly in the soil or isolated from soil in pots dropped by 3.00 in the same field after the fertiliser application. Both the high concentration of external N in the root zone (following the application of N-fertilisers) and/or subsequent foliar uptake of volatilised NH₃ could have caused the depleted leaf ¹⁵N values measured in the sugarcane crops at these sites.     

Temporal variability in <Superscript>13</Superscript>C of respired CO<Subscript>2</Subscript> in a pine and a hardwood forest subject to similar climatic conditions

Temporal variability in the ¹³C of foliage (¹³C_F), soil (¹³C_S) and ecosystem (¹³C_R) respired CO₂ was contrasted between a 17.2-m tall evenly aged loblolly pine forest and a 35-m tall unevenly aged mature second growth mixed broadleaf deciduous forest in North Carolina, USA, over a 2-year period. The two forests are located at the Duke Forest within a kilometer of each other and are subject to identical climate and have similar soil types. The ¹³C_F, collected just prior to dawn, was primarily controlled by the time-lagged vapor pressure deficit (VPD) in both stands; it was used for calculating the ratio of intercellular to ambient CO₂ (Ci/Ca). A remarkable similarity was observed in the relationship between Ci/Ca and time-lagged VPD in these two forests despite large differences in hydraulic characteristics. This similarity emerged as a result of physiological adjustments that compensated for differences in plant hydraulic characteristics, as predicted by a recently proposed equilibrium hypothesis, and has implications to ecophysiological models. We found that in the broadleaf forest, the ¹³C of forest floor CO₂ efflux dominated the ¹³C_R, while in the younger pine forest, the ¹³C of foliage respired CO₂ dominated ¹³C_R. This dependence resulted in a more variable ¹³C_R in the pine forest when compared to the broadleaf forest due to the larger photosynthetic contribution. Given the sensitivity of the atmospheric inversion models to ¹³C_R, the results demonstrate that these models could be improved by accounting for stand characteristics, in addition to previously recognized effects of moisture availability, when estimating ¹³C_R.     

Interpretations of gradients in δ13C value in thick photosynthetic tissues of plants with Crassulacean acid metabolism

In Ceropegia dichotoma, Crassula argentea, Esheveria colorata, Kalanchoë beharensis, Opuntia ficus-indica, Sansveria stuckyi and Opuntia inermis the carbon-isotope ratio ( ¹³C) of tissues close to the epidermis is 2–4.3 more negative than those in the centre of the leaf or cladode. The greatest change in ¹³C value occurs between the epidermal layer and the layer of mesophyll tissue immediately underneath. Analysis of major metabolic and structural components in successive layers of Crassula argentea grown under controlled environmental conditions conducive to Crassulacean acid metabolism confirmed that much of the variation in ¹³C values of bulk carbon is caused by differences in chemical composition. Thus the steep gradient in ¹³C value at the epidermis reflects, in part, the contribution of more-negative ¹³C values of lipids in these tissues. Moreover, during nocturnal CO₂ fixation the amount of malic acid synthesised decreases with depth and the ¹³C value of the methanol-soluble fraction is less negative with distance away from the upper epidermis. These results are consistent with diffusion limitation to CO₂ uptake in these thick leaf tissues, which also contributes to the observed gradients in ¹³C value.Abbreviation CAM Crassulacean acid metabolism - FW fresh weight - PEPCase phospoenolpyruvate carboxylase - Rubisco ribulose-1,5-bisphosphate carboxylase-oxygenase - ¹³C carbon-isotope ratio This work was supported by grants from the North Carolina Biotechnology Centre to Duke University, National Science Foundation (NSF) grant DCB90-06830, Department of Energy, Division of Energy Biosciences grant DE-FG05-89 ER 14005, and NSF grant BSR 87-06429 to Duke University Phytotron.     

Polar bears make little use of terrestrial food webs: evidence from stable-carbon isotope analysis

Summary The mean stable-carbon isotope ratios (¹³C) for polar bear (Ursus maritimus) tissues (bone collagen –15.7, muscle –17.7, fat –24.7) were close to those of the same tissues from ringed seals (Phoca hispida) (–16.2, –18.1, and –26.1, respectively), which feed exclusively from the marine food chain. The ¹³C values for 4 species of fruits to which polar bears have access when on land in summer ranged from –27.8 to –26.2, typical of terrestrial plants in the Arctic. An animal's ¹³C signature reflects closely the ¹³C signature of it's food. Accordingly, the amount of food that polar bears consume from terrestrial food webs appears negligible, even though some bears spend 1/3 or more of each year on land during the seasons of greatest primary productivity.     

Carbon isotope fractionation of methyl bromideduring agricultural soil fumigations

The isotopic composition ofmethyl bromide (CH₃Br) has been suggestedto be a potentially useful tracer forconstraining the global CH₃Br budget. Inorder to determine the carbon isotopiccomposition of CH₃Br emitted from the mostsignificant anthropogenic application(pre-plant fumigation) we directly measured the¹³C of CH₃Br released duringcommercial fumigation. We also measured theisotopic fractionation associated withdegradation in agricultural soil under typicalfield fumigation conditions. The isotopiccomposition of CH₃Br collected in soilseveral hours after injection of the fumigantwas –44.5 and this value increased to –20.7over the following three days. The mean kineticisotope effect (KIE) associated withdegradation of CH₃Br in agricultural soil(12) was smaller than the reported value formethylotrophic bacterial strain IMB-1, isolatedfrom previously fumigated agricultural soil,but was similar to methylotrophic bacterialstrain CC495, isolated from a pristine forestlitter zone. Using this fractionationassociated with the degradation of CH₃Brin agricultural soil and the mean¹³C of the industriallymanufactured CH₃Br (–54.4), we calculatethat the agricultural soil fumigation sourcehas a carbon isotope signature that ranges from–52.8 to –42.0. Roughly 65% ofindustrially manufactured CH₃Br is usedfor field fumigations. The remaining 35% isused for structural and post-harvestfumigations with a minor amount used duringindustrial chemical manufacturing. Assumingthat the structural and post-harvest fumigationsources of CH₃Br are emitted withoutsubstantial fractionation, we calculate thatthe ¹³C of anthropogenicallyemitted CH₃Br ranges from –53.2 to –47.5.     

Measurement of two- and three-bond 13C−1H J couplings to the Cδ carbons of leucine residues in staphylococcal nuclease

Summary A new ¹H-detected 3D NMR experiment is described that permits quantitative measurement of two- and three-bond ¹³C–¹H couplings in proteins with selectively ¹³C-enriched methyl sites. The method is demonstrated for staphylococcal nuclease selectively [5,5 ¹³C]-labeled in all 11 leucine positions and ligated with thymidine 3,5-biphosphate and Ca²⁺. Two- and three-bond ¹³C methyl-proton couplings are reported and, together with the measured three-bond J_CC in uniformly ¹³C-enriched staphylococcal nuclease, the ²- and the stereospecific assignments of the C methyl group with respect to the prochiral -protons were determined. The same residues that were previously found to have high degrees of internal mobility on the basis of ¹³C relaxation times have measured coupling constants that are indicative of motional averaging.     

Using stable hydrogen and oxygen isotope measurements of feathers to infer geographical origins of migrating European birds

Successful application of stable-hydrogen isotope measurements (D_f) of feathers to track origins of migratory birds and other wildlife requires a fundamental understanding of the correlation between D_f and deuterium patterns in rainfall (D_p) over continental scales. A strong correlation between D_p and D_f has been confirmed for birds and insects in North America, but not yet for other continents. Here, we compare D_f data from resident European birds to new D_p basemaps for Europe. Three maps, representing growing-season and mean annual D_p estimates from an elevation-explicit, detrended interpolation model and growing-season D_p estimates from simple Kriging, all indicate that strong isotope gradients occur across Europe with a general depletion occurring in a northeast direction. The feather data, representing 141 individuals of 25 avian species from 38 sites, ranged from –131 to –38. Regression analysis showed that strong correlations existed between both mean annual and growing-season D_p estimated by detrended interpolation and D_f of non-aquatic and non-corvid birds (r²=0.66 and 0.65, respectively). We also examined mean annual and growing-season ¹⁸O_p vs. ¹⁸O_f for our samples. Both oxygen regressions were similar (r²=0.56 and 0.57, respectively) but poorer than for deuterium. Our study reveals that D measurements of feathers from migratory birds in Europe may be used to track their origin and movements, and so provide a powerful investigative tool for avian migration research in Europe.     

Black alder (Alnus Glutinosa (L.) Gaertn.) trees mediate methane and nitrous oxide emission from the soil to the atmosphere

Three-year-old seedlings of black alder (Alnus glutinosa (L.) Gaertn.), a common European wetland tree species, were grown in native soil taken from an alder swamp. Fluxes of methane (CH₄) and nitrous oxide (N₂O) between the tree stem and the atmosphere were determined under controlled conditions. Both CH₄ and N₂O were emitted through the bark of the stem into the atmosphere when the root zone exhibited higher-than-ambient CH₄ and N₂O gas mixing ratios. Flooding of the soil caused a decreased N₂O emission but an increased CH₄ efflux from the stem. Immediately after flooding of the soil, N₂O was emitted from the seedlings' bark at a rate of 350 mol N₂O m^-2 h^-1 whereas CH₄ flux could not be detected. After more than 40 days of flooding CH₄ fluxes up to 3750 mol CH₄ m^-2 h^-1 from the stem were measured, while N₂O emission had decreased below the limit of detection. Gas efflux decreased with increasing stem height and correlated with gas mixing ratios in the soil, indicating diffusion through the aerenchyma as the major path of gas transport. From these results it is assumed that woody species with aerenchyma can serve as conduits for soil-derived trace gases into the atmosphere, to date only shown for herbaceous plants. This, yet unidentified, woody plant pathway contributes to the total greenhouse gas source strength of wetlands.     

Trophic interactions of Antarctic seals as determined by stable isotope signatures

The diets and trophic interactions among Weddell, crabeater, Ross, and leopard seals in the eastern Ross Sea, Antarctica, were investigated by the use of stable isotope techniques during the 1999–2000 summer seasons. The ¹³C and ¹⁵N values in seal serum clearly distinguished the three Antarctic pack-ice seal species at different trophic positions (Weddell>Ross>crabeater). These patterns appeared to reflect a close linkage to their known foraging ecology and diving behaviors, and agreed well with their presumed dietary diversity. The more enriched ¹³C and ¹⁵N values in male Weddell seals than those in females suggested differences in foraging preferences between them. Significant differences in ¹⁵N were also found among different age groups of Weddell seals. A strong correlation between the C:N ratios and serum cholesterol was probably due to extremely high cholesterol levels in phocids. Comparisons of isotope data with harbor seals revealed distinct differences between Antarctic phocids and the northern seal species.