Isotopically characterised N2 O reference materials for use as community standards.

RATIONALE: Information on the isotopic composition of nitrous oxide (N2 O) at natural abundance supports the identification of its source and sink processes. In recent years, a number of mass spectrometric and laser spectroscopic techniques have been developed and are increasingly used by the research community. Advances in this active research area, however, critically depend on the availability of suitable N2 O isotope Reference Materials (RMs). METHODS: Within the project Metrology for Stable Isotope Reference Standards (SIRS), seven pure N2 O isotope RMs have been developed and their 15 N/14 N, 18 O/16 O, 17 O/16 O ratios and 15 N site preference (SP) have been analysed by specialised laboratories against isotope reference materials. A particular focus was on the 15 N site-specific isotopic composition, as this measurand is both highly diagnostic for source appointment and challenging to analyse and link to existing scales. RESULTS: The established N2 O isotope RMs offer a wide spread in delta (δ) values: δ15 N: 0 to +104‰, δ18 O: +39 to +155‰, and δ15 NSP : -4 to +20‰. Conversion and uncertainty propagation of δ15 N and δ18 O to the Air-N2 and VSMOW scales, respectively, provides robust estimates for δ15 N(N2 O) and δ18 O(N2 O), with overall uncertainties of about 0.05‰ and 0.15‰, respectively. For δ15 NSP , an offset of >1.5‰ compared with earlier calibration approaches was detected, which should be revisited in the future. CONCLUSIONS: A set of seven N2 O isotope RMs anchored to the international isotope-ratio scales was developed that will promote the implementation of the recommended two-point calibration approach. Particularly, the availability of δ17 O data for N2 O RMs is expected to improve data quality/correction algorithms with respect to δ15 NSP and δ15 N analysis by mass spectrometry. We anticipate that the N2 O isotope RMs will enhance compatibility between laboratories and accelerate research progress in this emerging field.

Effects of short-term freezing on nitrous oxide emissions and enzyme activities in a grazed pasture soil after bovine-urine application.

Nitrous oxide (N2O) emissions from soils applied with livestock excreta have been widely reported previously. The highest N2O emissions from soils are also often reported during thawing periods in cold regions where soil freezing is common. However, the combined effects of cow urine application and freeze-thaw events on N2O emissions and the related enzyme activities are still not clear. Thus, we simulated a freeze-thaw event at -3 °C for 7 days, and then increased to 3 °C for 46 days using intact soil cores with cow urine (392 kg N ha-1). We compared the factors influencing the magnitudes of N2O emissions through soil microbial processes with and without the freeze-thaw event. Dicyandiamide (DCD), an inhibitor of nitrification, was added to investigate the significance of nitrification on N2O emissions. The N2O emission rates from the urine-applied soils peaked to approximately 1000 µg N2O-N m-2 h-1 immediately after the soils thawed. Soil freezing with urine application was significantly higher cumulative N2O emissions (537 mg N2O-N m-2), compared to non-frozen soils with urine (247 mg N2O-N m-2) during the incubation period (54 days). The effect of DCD application on N2O emissions was not clear during the freeze-thaw event, although nitrate production rates were reduced. After the freezing event, soil moisture (water-filled pore space) was significantly higher in the non-frozen soils compared to the frozen soils, due to a 9% decline in bulk density of frozen soils. Additionally, the impact of thawing on urease and denitrification enzyme activities was influenced by the urine application. Urine application increased the urease activity, while the freezing event decreased the magnitudes. The physical changes in the soils were also important controlling factors of the N2O emissions from cow urine-applied soils in cold regions.

Forest understory plant and soil microbial response to an experimentally induced drought and heat-pulse event: the importance of maintaining the continuum.

Drought duration and intensity are expected to increase with global climate change. How changes in water availability and temperature affect the combined plant-soil-microorganism response remains uncertain. We excavated soil monoliths from a beech (Fagus sylvatica L.) forest, thus keeping the understory plant-microbe communities intact, imposed an extreme climate event, consisting of drought and/or a single heat-pulse event, and followed microbial community dynamics over a time period of 28 days. During the treatment, we labeled the canopy with (13) CO2 with the goal of (i) determining the strength of plant-microbe carbon linkages under control, drought, heat and heat-drought treatments and (ii) characterizing microbial groups that are tightly linked to the plant-soil carbon continuum based on (13) C-labeled PLFAs. Additionally, we used 16S rRNA sequencing of bacteria from the Ah horizon to determine the short-term changes in the active microbial community. The treatments did not sever within-plant transport over the experiment, and carbon sinks belowground were still active. Based on the relative distribution of labeled carbon to roots and microbial PLFAs, we determined that soil microbes appear to have a stronger carbon sink strength during environmental stress. High-throughput sequencing of the 16S rRNA revealed multiple trajectories in microbial community shifts within the different treatments. Heat in combination with drought had a clear negative effect on microbial diversity and resulted in a distinct shift in the microbial community structure that also corresponded to the lowest level of label found in the PLFAs. Hence, the strongest changes in microbial abundances occurred in the heat-drought treatment where plants were most severely affected. Our study suggests that many of the shifts in the microbial communities that we might expect from extreme environmental stress will result from the plant-soil-microbial dynamics rather than from direct effects of drought and heat on soil microbes alone.