Carbon storage potential in size-density fractions from semi-natural grassland ecosystems with different productivities over varying soil depths.

Researchers have increasingly recognised a profound need for more information on SOC stocks in the soil and the factors governing their stability and dynamics. Many questions still remain unanswered about the interplay between changes in plant communities and the extent to which changes in aboveground productivity affect the carbon dynamics in soils through changes in its quantity and quality. Therefore, the main aim of this research was to examine the SOC accumulation potential of semi-natural grasslands of different productivities and determine the distribution of SOM fractions over varying soil depth intervals (0-10, 10-20, 20-30 30-50 50-80 and 80+cm). SOM fractionation was considered as a relative measure of stability to separate SOM associated with clay minerals from SOM of specific light densities less than 2 g cm(-3) (size-density fractionation). Two clay-associated fractions (CF1, <1 µm; and CF2, 1-2 µm) and two light fractions (LF1, <1.8 g cm(-3); and LF2, 1.8-2.0 g cm(-3)) were separated. The stability of these fractions was characterised by their carbon hot water extractability (CHWE) and stable carbon isotope composition. In the semi-natural grasslands studied, most OC was stored in the top 30 cm, where turnover is rapid. Effects of low productivity grasslands became only significantly apparent when fractional OC contributions of total SOM was considered (CF1 and LF1). In deeper soil depths OC was largely attributed to the CF1 fraction of low productivity grasslands. We suggest that the majority of OM in deeper soil depth intervals is microbially-derived, as evidenced by decreasing C/N ratios and decreasing δ(13)C values. The hot water extraction and natural δ(13)C abundance, employed here allowed the characterisation of SOM stabilisation properties, however how climatic changes affect the fate of OM within different soil depth intervals is still unknown.

Spatial variability and temporal trends in water-use efficiency of European forests.

The increasing carbon dioxide (CO2 ) concentration in the atmosphere in combination with climatic changes throughout the last century are likely to have had a profound effect on the physiology of trees: altering the carbon and water fluxes passing through the stomatal pores. However, the magnitude and spatial patterns of such changes in natural forests remain highly uncertain. Here, stable carbon isotope ratios from a network of 35 tree-ring sites located across Europe are investigated to determine the intrinsic water-use efficiency (iWUE), the ratio of photosynthesis to stomatal conductance from 1901 to 2000. The results were compared with simulations of a dynamic vegetation model (LPX-Bern 1.0) that integrates numerous ecosystem and land-atmosphere exchange processes in a theoretical framework. The spatial pattern of tree-ring derived iWUE of the investigated coniferous and deciduous species and the model results agreed significantly with a clear south-to-north gradient, as well as a general increase in iWUE over the 20th century. The magnitude of the iWUE increase was not spatially uniform, with the strongest increase observed and modelled for temperate forests in Central Europe, a region where summer soil-water availability decreased over the last century. We were able to demonstrate that the combined effects of increasing CO2 and climate change leading to soil drying have resulted in an accelerated increase in iWUE. These findings will help to reduce uncertainties in the land surface schemes of global climate models, where vegetation-climate feedbacks are currently still poorly constrained by observational data.

Reduced climate sensitivity of carbon, oxygen and hydrogen stable isotope ratios in tree-ring cellulose of silver fir (Abies alba Mill.) influenced by background SO2 in Franconia (Germany, Central Europe).

The climate sensitivity of carbon (δ(13)C), oxygen (δ(18)O) and hydrogen (δ(2)H) isotope signatures in tree-ring cellulose of Abies alba Mill. from a marginally industrialized area of Franconia (Germany) was analysed for the last 130 years. All isotopes preserve climatic signals up to c. 1950 AD. After 1950 we observe a clear reduction in climate sensitivity of δ(13)C and δ(2)H while δ(18)O - climate relations remain well pronounced. Nevertheless statistical tests implied that SO2 background emissions of West Germany had influenced isotope signatures long before 1950. The relationships between isotope values and concentrations of SO2, dust, O3 and NO2 at the regional level during the period 1979-2006 indicate that δ(13)C and δ(18)O were influenced primarily by SO2. The impact of SO2 on δ(2)H was negligible, but the observed reduction of climate sensitivity may be caused by synergic influences. The results have significant implications if isotope signatures from tree-rings from anthropogenic influenced regions are used to reconstruct past climate.

Intra-seasonal dynamics in metabolic processes of 13C/12C and 18O/16O in components of Scots pine twigs from southern Siberia interpreted with a conceptual framework based on the Carbon Metabolism Oscillatory Model.

BACKGROUND: Carbon isotope data from conifer trees play an important role in research on the boreal forest carbon reservoir in the global carbon cycle. Carbon isotopes are routinely used to study interactions between the environment and tree growth. Moreover, carbon isotopes became an essential tool for the evaluation of carbon assimilation and transport from needles into reserve pools, as well as the allocation of stored assimilates within a tree. The successful application and interpretation of carbon isotopes rely on the coherence of isotopic fractionation modeling. This study employs a new Carbon Metabolism Oscillatory Model (CMOM) to interpret the experimental data sets on metabolic seasonal dynamics of 13C/12 C and 18O/16O ratios measured in twig components of Scots pine growing in southern Siberia (Russia). RESULTS: The dynamics of carbon isotopic variables were studied in components of Pinus sylvestris L. in light and in dark chambers during the vegetation period from 14 June to 28 July 2006. At the beginning of this period water-soluble organic matter, mostly labile sugars (including sucrose as the main component) and newly formed bulk needle material, displayed relatively "light" δ13C values (depletion in 13 C). Then, 13 C content increased again with noticeable "depletion" events in the middle of the growth period. A gradual 13 C accumulation took place in the second half of the vegetation period. Similar effects were observed both in the light and in the dark with some temporal shifts. Environmental factors did not influence the δ13C values. A gradual 12C-depletion effect was noticed in needles of the previous year. The δ13C values of sucrose and proteins from needle biomass altered independently from each other in the light chamber. A distinct negative correlation between δ13C and δ18O values was revealed for all studied variables. CONCLUSIONS: The abrupt 13C depletion recorded by all tested trees for the period from June to July provides clear evidence of the transition from the dominant role of reserve carbohydrate pool (RCP) during the first half of the growth season to the preferable current year carbohydrate pool (CCP) consumption by new needles during its second half. The investigation of the isotopic signatures of Pinus sylvestris L. emphasizes the pivotal role of the intra-seasonal dynamics in carbon metabolism through the transport of assimilates from autotrophic (needles) to heterotrophic (twigs) organs of the studied trees. This provides an explanation for changes of carbon isotopic values observed within the growth season. The CMOM-based results support the hypothesis of the integration of three carbohydrate pools by photosynthesizing cells. The fluctuations of the carbon isotope ratios in different carbohydrate pools underlie various physiological processes in the tree metabolism. The possible mechanisms and pathways of formation of these carbohydrate pools are further discussed. Hence, CMOM provides a reasonable explanation for the absence of the impact of environmental conditions on the needle isotopic variables, the 12C-depletion effects and the use of RCP in needles. The model explains the negative connections between δ13C and δ18O values in all studied variables.

A new serial pooling method of shifted tree ring blocks to construct millennia long tree ring isotope chronologies with annual resolution.

The study presents a new serial pooling method of shifted tree ring blocks for the building of isotope chronologies. This method combines the advantages of traditional 'serial' and 'intertree' pooling, and can be recommended for the construction of sub-regional long isotope chronologies with sufficient replication, and on annual resolution, especially for the case of extremely narrow tree rings. For Scots pines (Pinus sylvestris L., Khibiny Low Mountains, NW Russia) and Silver firs (Abies alba Mill., Franconia, Southern Germany), serial pooling of five consecutive tree rings seems appropriate because the species- and site-specific particularities lead to blurs of climate linkages in their tree rings for the period up to ca. five years back. An equivalent to a five-year running means that curve gained on the base annual data sets of single trees can be derived from the analysis of yearly shifted five-year blocks of consecutive tree rings, and therefore, with approximately 20% of the expense. Good coherence of delta(13)C- and delta(18)O-values between calculated means of annual total rings or late wood data and means of five-year blocks of consecutive total tree rings analysed experimentally on most similar material confirms this assumption.

Routine hydrogen isotope measurement of cellulose nitrate by high-temperature pyrolysis--reference materials and precision.

The determination of isotope ratios of non-exchangeable hydrogen in tree-ring cellulose is commonly based on the nitration of wood cellulose followed by online high-temperature pyrolysis and isotope ratio mass spectrometry measurement of cellulose nitrate samples. The application of this method requires a proper calibration using appropriate reference materials whose delta(2)H values have been reliably normalized to the V-SMOW/SLAP scale. In our study, we achieve this normalization by a direct alternating measurement of reference waters (V-SMOW and SLAP) and three cellulose nitrates chosen as reference materials. For that purpose, both water and solid organic samples are introduced into the pyrolysis reactor by silver capsule injection. The analytical precision of the water measurement using the capsule method is +/-1.5 per thousand. The hydrogen isotopic composition of three cellulose nitrate standards measured ranges from -106.7 to -53.9 per thousand. The standard deviation of the calculated means from five measurement periods of those standards is better than 1 per thousand. Twenty-four different measurements of the hydrogen isotope composition of cellulose nitrate were evaluated in order to assess the precision of the described method. We obtained an analytical precision of +/-3.0 per thousand as representative for the 95% confidence interval applicable for routine measurements of cellulose nitrate samples. Evidence was found for significant differences in the behavior of cellulose nitrate and PE foil during the pyrolitic conversion that emphasizes the need for a proper calibration of the routine measurements. This calibration can only be successful if the reference materials used have a very similar chemical composition and undergo the same preparation procedure as the samples.

13C discriminations of Pinus sylvestris vs. Pinus ponderosa at a dry site in Brandenburg (eastern Germany): 100-year growth comparison.

The carbon isotope composition (delta(13)C, per thousand) and discrimination (Delta, per thousand) of old grown North American Pinus ponderosa Dougl. Ex P. et C. Laws. and European Pinus sylvestris L. were determined using trees grown under almost identical growing conditions in a mixed stand in Bralitz, Northeast Germany. Single-tree delta(13)C analyses of tree-ring cellulose of both species were carried out at a yearly resolution for the period 1901-2001 and the results compared with growth (basal area increment). Annual mean delta(13)C values for P. ponderosa ranged from-21.6 per thousand to-25.2 per thousand and for P. sylvestris from-21.4 per thousand to-24.4 per thousand. Accordingly, (13)C discrimination (Delta) showed higher values for P. ponderosa throughout the investigation period. Five characteristic periods of Delta were identified for both the tree species, reflecting positive and negative influences of environmental factors. Good growing conditions such as after-thinning events had a positive effect on Delta, reflecting higher values, while poor conditions like aridity and air pollution had a negative influence, reflecting lower values. The dynamics of Delta were likewise reflected in the growth (basal area increment, BAI). Higher (13)C discrimination values of P. ponderosa led to higher BAIs of P. ponderosa in comparison with P. sylvestris. Correlation function analyses confirmed that P. sylvestris was more dependent on precipitation than P. ponderosa, which showed a closer relationship with temperature. The results confirm that under predominantly dry growing conditions, P. ponderosa showed better growth performance than P. sylvestris, indicating better common intrinsic water-use efficiency and, therefore, higher rates of net photosynthesis at a given transpiration. In view of the prospect of climate change, the results are very significant for assessing both trees' physiological properties and, hence, their potential for coping with future growing conditions.

Wood cellulose preparation methods and mass spectrometric analyses of delta13C, delta18O, and nonexchangeable delta2H values in cellulose, sugar, and starch: an interlaboratory comparison.

Interlaboratory comparisons involving nine European stable isotope laboratories have shown that the routine methods of cellulose preparation resulted in data that generally agreed within the precision of the isotope ratio mass spectrometry (IRMS) method used: +/-0.2 per thousand for carbon and +/-0.3 per thousand for oxygen. For carbon, the results suggest that holocellulose is enriched up to 0.39 per thousand in 13C relative to the purified alpha-cellulose. The comparisons of IRMS measurements of carbon on cellulose, sugars, and starches showed low deviations from -0.23 to +0.23 per thousand between laboratories. For oxygen, IRMS measurements varied between means from -0.39 to 0.58 per thousand, -0.89 to 0.42 per thousand, and -1.30 to 1.16 per thousand for celluloses, sugars, and starches, respectively. This can be explained by different effects arising from the use of low- or high-temperature pyrolysis and by the variation between laboratories in the procedures used for drying and storage of samples. The results of analyses of nonexchangeable hydrogen are very similar in means with standard deviations between individual methods from +/-2.7 to +/-4.9 per thousand. The use of a one-point calibration (IAEA-CH7) gave significant positive offsets in delta2H values up to 6 per thousand. Detailed analysis of the results allows us to make the following recommendations in order to increase quality and compatibility of the common data bank: (1) removal of a pretreatment with organic solvents, (2) a purification step with 17% sodium hydroxide solution during cellulose preparation procedure, (3) measurements of oxygen isotopes under an argon hood, (4) use of calibration standard materials, which are of similar nature to that of the measured samples, and (5) using a two-point calibration method for reliable result calculation.

Rapid online equilibration method to determine the D/H ratios of non-exchangeable hydrogen in cellulose.

An improved method for the determination of deuterium-to-hydrogen (D/H) ratios of non-exchangeable hydrogen in cellulose is presented. The method is based on the equilibration reaction of the hydroxyl hydrogen of cellulose and water vapour of known isotopic composition. The equilibrated cellulose is pyrolysed and the total D/H ratio determined by subsequent online isotope ratio mass spectrometry (IRMS). With a mass balance system the D/H ratio of non-exchangeable hydrogen is recalculated after an empirical calibration has been performed, yielding a mean exchangeability of 0.239 and an equilibrium fractionation factor of 1.082 between the hydroxyl hydrogen of cellulose and water hydrogen at 110 degrees C. Equilibration takes 10 min per sample. Results obtained by this online equilibration method agree very well with values obtained by the nitration technique (R2 = 0.941). The uncertainty of the equilibration method is +/-4 per thousand resulting from a single standard deviation of +/-2.8 per thousand for the equilibration determined by standard cellulose and 2.8 per thousand from the variable exchangeability of the hydroxyl hydrogen in cellulose due to crystalline areas. The latter uncertainty may be lowered by minimising the crystallinity of the cellulose. Advantages of this new technique are (i) the considerably reduced sample amount required (as low as 0.2 mg, ideally 0.5 mg compared with 20 mg for the conventional nitration technique); (ii) an approximately 100-fold reduced process time; and (iii) no need for the hazardous chemicals used in the nitration technique.

Microwave-supported preparation of alpha-cellulose for analysis of delta13C in tree rings.

The microwave technique is found to be very applicable for the preparation of alpha-cellulose from wood samples and can be recommended for analyzing the stable carbon isotopes in tree rings. At a reaction temperature of 80 degrees C, the extraction time can be decreased from 36 h to 15 min. Microwave-supported prepared cellulose contains more amorphous constituents, resulting in a relatively higher reactivity and amenability for a following nitration with regard to determination of nonexchangeable hydrogen. The delta18O values of microwave-enhanced extracted cellulose remain significantly lighter than reference values, possibly as a result of an increased oxygen isotope exchange between bleaching solution and cellulose under conditions of high energy input. Therefore, this technique cannot be recommended for oxygen isotope analyses in wood cellulose.

Carbon isotope analyses of cellulose using two different on-line techniques (elemental analysis and high-temperature pyrolysis)--a comparison.

Even though the recent development in on-line methods for the stable isotope determination in cellulose has led to a significant increase in sample throughput and decrease in sample preparation expenditure, there still is a large potential for optimizing the analytical procedures by simultaneously measuring the isotope ratios of two or even more elements. Therefore, the main objective of this study was to answer the question whether high-temperature pyrolysis (HTP) is a suitable and reliable technique for the determination of the carbon isotopic composition of cellulose simultaneously during the well-known conventional oxygen isotope analysis. This study shows that HTP of cellulose is a technique that can produce reasonable delta(13)C values, matching the requirements of most research problems related to paleoclimatology. The reproducibility of the delta values for (13)C/(12)C is better than 0.2 per thousand. Some deficiencies of the method are related to the incomplete conversion of the organic carbon in the sample to carbon monoxide. A clear isotope effect seems to be related to the non-statistical conversion of the carbon in the cellulose to CO. The extent of this effect appears to be controlled by the relative proportion of crystallized and amorphous matter in the cellulose structure. Those deficiencies can be eliminated by using an appropriate normalization and by applying the principles of identical treatment for reference materials and samples. In general, a very good agreement is achieved for carbon isotope values determined by HTP and elemental analysis (EA).