The unknown third - Hydrogen isotopes in tree-ring cellulose across Europe.

This is the first Europe-wide comprehensive assessment of the climatological and physiological information recorded by hydrogen isotope ratios in tree-ring cellulose (δ2Hc) based on a unique collection of annually resolved 100-year tree-ring records of two genera (Pinus and Quercus) from 17 sites (36°N to 68°N). We observed that the high-frequency climate signals in the δ2Hc chronologies were weaker than those recorded in carbon (δ13Cc) and oxygen isotope signals (δ18Oc) but similar to the tree-ring width ones (TRW). The δ2Hc climate signal strength varied across the continent and was stronger and more consistent for Pinus than for Quercus. For both genera, years with extremely dry summer conditions caused a significant 2H-enrichment in tree-ring cellulose. The δ2Hc inter-annual variability was strongly site-specific, as a result of the imprinting of climate and hydrology, but also physiological mechanisms and tree growth. To differentiate between environmental and physiological signals in δ2Hc, we investigated its relationships with δ18Oc and TRW. We found significant negative relationships between δ2Hc and TRW (7 sites), and positive ones between δ2Hc and δ18Oc (10 sites). The strength of these relationships was nonlinearly related to temperature and precipitation. Mechanistic δ2Hc models performed well for both genera at continental scale simulating average values, but they failed on capturing year-to-year δ2Hc variations. Our results suggest that the information recorded by δ2Hc is significantly different from that of δ18Oc, and has a stronger physiological component independent from climate, possibly related to the use of carbohydrate reserves for growth. Advancements in the understanding of 2H-fractionations and their relationships with climate, physiology, and species-specific traits are needed to improve the modelling and interpretation accuracy of δ2Hc. Such advancements could lead to new insights into trees' carbon allocation mechanisms, and responses to abiotic and biotic stress conditions.

A novel approach for the homogenization of cellulose to use micro-amounts for stable isotope analyses.

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.

δ13C-variations of leaves in forests as an indication of reassimilated CO2 from the soil.

An attempt has been made to evaluate the contribution of soil respired CO2 to the total assimilation of a forest tree, by heeding the 13C-concentrations of CO2 from the free atmosphere and from mineralization processes within the soil respectively. An expression has been derived, according to which the assimilated fraction of CO2 from the soil at a particular height of a tree is given by the δ13C-value of the corresponding leaves, δ13C of atmospheric CO2, δ13C of soil respired CO2 and the physiological state of the leaves expressed as the ratio of total respiration over gross photosynthesis and internal over external CO2-concentration. In the particular case investigated, a δ13C-difference of 5‰ has been determined from bottom to top of a beech tree which results in a CO2 contribution from the soil of about 22% for the lower forest strata, while the total contribution of soil respired CO2 accounts for about 5% of the overall assimilation.

Oxygen isotope fractionation during respiration for different temperatures of T. utilis and E. coli K12.

The temperature dependence of the oxygen isotope fractionation factor during respiration has been examined for two different microorganisms, namely Torulopsis utilis and Escherichia coli K12 representing a yeast and a bacterium, respectively. The investigation covered a temperature range of 18 degrees C, that is from 16 degrees C to 34 degrees C for T. utilis and from 19 degrees C to 37 degrees C for E. coli K12. Within this temperature range the fractionation factor of T. utilis increases by 0.18% an insignificant change (delta delta 10 degrees C = 0.063%; r = 0.067), whereas with E. coli K12 an increase of 1.12% has been observed (delta delta 10 degrees C = 0.6%; r = 0.55).