Description: 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% for carbon and +/- 0.3% for oxygen. For carbon, the results suggest that holocellulose is enriched up to 0.39% in C-13 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% between laboratories. For oxygen, IRMS measurements varied between means from -0.39 to 0.58%, -0.89 to 0.42%, and -1.30 to 1.16% 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%. The use of a one-point calibration (IAEA-CH7) gave significant positive offsets in delta H-2 values up to 6%. 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.

Description: Climate reconstructions using stable isotopes from tree-rings are steadily increasing. The investigations concentrate mostly on cellulose due to its high stability. In recent years the available amount of cellulose has steadily decreased, mainly because micro-structures of plant material have had to be analyzed. Today, the amounts of cellulose being studied are frequently in the milligram and often in the microgram range. Consequently, homogeneity problems with regard to the stable isotopes of carbon and oxygen from cellulose have occurred and these have called for new methods in the preparation of cellulose for reliable isotope analyses. Three different methods were tested for preparing isotopically homogenous cellulose, namely mechanical grinding, freezing by liquid nitrogen with subsequent milling and ultrasonic breaking of cellulose fibres. The best precision of isotope data was achieved by freeze-milling and ultrasonic breaking. However, equipment for freeze-milling is expensive and the procedure is labour-intensive. Mechanical grinding resulted in a rather high loss of material and it is also labour-intensive. The use of ultrasound for breaking cellulose fibres proved to be the best method in terms of rapidity of sample throughput, avoidance of sample loss, precision of isotope results, ease of handling, and cost.

Description: Twentieth-century warming could lead to increases in the moisture-holding capacity of the atmosphere, altering the hydrological cycle and the characteristics of precipitation. Such changes in the global rate and distribution of precipitation may have a greater direct effect on human well-being and ecosystem dynamics than changes in temperature itself. Despite the co-variability of both of these climate variables, attention in long-term climate reconstruction has mainly concentrated on temperature changes. Here we present an annually resolved oxygen isotope record from tree-rings, providing a millennial-scale reconstruction of precipitation variability in the high mountains of northern Pakistan. The climatic signal originates mainly from winter precipitation, and is robust over ecologically different sites. Centennial-scale variations reveal dry conditions at the beginning of the past millennium and through the eighteenth and early nineteenth centuries, with precipitation increasing during the late nineteenth and the twentieth centuries to yield the wettest conditions of the past 1,000 years. Comparison with other long-term precipitation reconstructions indicates a large-scale intensification of the hydrological cycle coincident with the onset of industrialization and global warming, and the unprecedented amplitude argues for a human role.

Description: We present the first European network of tree ring delta(13)C and delta(18)O, containing 23 sites from Finland to Morocco. Common climate signals are found over broad climatic-ecological ranges. In temperate regions we find positive correlations with summer maximum temperatures and negative correlations with summer precipitation and Palmer Drought Severity Indices (PDSI) with no obvious speciesspecific differences. Regional delta(13)C and delta(18)O chronologies share high common variance in year-to-year variations. Long-term variations, however, exhibit differences that may reflect spatial variability in environmental forcings, age trends and/or plant physiological responses to increasing atmospheric CO(2) concentration. Rotated principal component analysis (RPCA) and climate field correlations enable the identification of four sub-regions in the delta(18)O network - northern and eastern Central Europe, Scandinavia and the western Mediterranean. Regional patterns in the delta(13)C network are less clear and are timescale dependent. Our results indicate that future reconstruction efforts should concentrate on delta(18)O data in the identified European regions.

Description: The analysis of delta C-13 and delta O-18 in tree-ring archives offers retrospective insights into environmental conditions and ecophysiological processes. While photosynthetic carbon isotope discrimination and evaporative oxygen isotope enrichment are well understood, we lack information on how the isotope signal is altered by downstream metabolic processes. In Pinus sylvestris, we traced the isotopic signals from their origin in the leaf water (delta O-18) or the newly assimilated carbon (delta C-13), via phloem sugars to the tree-ring, over a time-scale that ranges from hours to a growing season. Seasonally, variable C-13 enrichment of sugars related to phloem loading and transport did lead to uncoupling between delta C-13 in the tree-ring, and the c(i)/c(a) ratio at the leaf level. In contrast, the oxygen isotope signal was transferred from the leaf water to the tree-ring with an expected enrichment of 27 parts per thousand, with time-lags of approximately 2 weeks and with a 40% exchange between organic oxygen and xylem water oxygen during cellulose synthesis. This integrated overview of the fate of carbon and oxygen isotope signals within the model tree species P. sylvestris provides a novel physiological basis for the interpretation of delta C-13 and delta O-18 in tree-ring ecology.

Description: This dendroclimatological research is based on two close pine forests (Pinus sylvestris and Pinus uncinata) located at the Northern Iberian System (Spain), and three tree-ring variables (ring widths, δ13C and δ18O). The climate-tree growth system was assessed at local and regional scales using three climate datasets. Calibration of tree-ring records with climate showed a diversity of information recorded in the different variables, such as a general response to temperature and precipitation of current growing period, and an important contribution of previous year conditions understood as the use of food reserves. The analysis of the stability of climate-tree growth relationships throughout the twentieth century showed a shift of those climatic variables to which trees responded and results suggested an enhancement of reserve use on current tree growth. The results obtained in this research made clear a physiological adaptation of trees to changing climate. The results provided hints that the recent warming coupled to slight precipitation decay are forcing growth of studied trees to a higher stress status and to a higher climate-growth synchronisation. These instabilities also have implications on future dendroclimatic reconstructions performed with trees growing under changing environments.

Description: In this study we have analyzed the variability of tree-ring widths and stable isotopes (δ13C and δ18O) of a single sessile oak tree (Quercus petraea (Matt.) Liebl.) since these parameters are critical in reconstructing the environment, particularly climatic conditions. Tree rings were separated into early- and latewood (EWt; LWt), tree ring (TRt), and transfer tree ring (TTRt, the latter being the latewood plus the earlywood of the subsequent year. Mean sensitivity, simple correlation, partial correlation and autocorrelation analyses were applied to describe data and relationships. Although this research focused on a single tree, the results compared well with average site data. Widths and δ18O values showed generally low autocorrelation for all tree-ring components, whereas δ13C revealed highly significant autocorrelations for most tree-ring components. Mean sensitivity of the standardized values turned out to be high for δ18O, marginally lower for width and the lowest for δ13C. Correlation analyses have proven that the relationships within the tree-ring widths or within the isotope parameters are much stronger than across widths and isotope parameters. The study demonstrates the unique potential of all measured tree-ring data to be used as climate proxies.