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Diethylamine (DEA) is a known precursor to some of the most toxic chemical warfare agents (CWA) as both V-agents and Fourth Generation Agents (FGAs) contain the dialkylamine functionality. DEA is also a readily available commercial substance with extensive use in the chemical industry for legitimate purposes. Because of this potential dual use, it is desirable to develop methods to trace the origin of dialkylamines if they are used for illicit purposes. We herein demonstrate that it is possible to differentiate six commercial batches of DEA using position-specific isotope analysis (PSIA) by both ²H and ¹³C NMR. Using a high-field NMR spectrometer together with a cryogenic ²H probe, we have produced ²H-{¹H} NMR data with high accuracy and precision. The PSIA by NMR results show that the intramolecular ²H ratios of all six DEAs are significantly different while two of the six DEAs have unique ¹³C ratios. Further, the intramolecular isotopic variations can be used to link the DEAs to different suppliers. The two nuclei separately contribute to isotopic profiles of the DEAs. However, combining the two techniques provides a higher-resolved isotopic profile that can differentiate all DEAs and thus be useful in forensic investigations of illegal use of chemical weapons.

Tropical forests substantially influence the terrestrial carbon sink. Their contributions to the forest carbon sink may increase due to the stimulation of photosynthesis by rising atmospheric CO₂ (C_a); however, the magnitude of this effect is poorly quantified for tropical canopy trees.

We measured the ratio of two deuterium isotopomers of glucose derived from tree rings to estimate how photosynthetic efficiency (photorespiration-to-photosynthesis ratio) has responded to C_a rise at a centennial scale. Wood samples were obtained from Toona ciliata trees from three climatically distinct forests in Asia and Australia. We applied Bayesian mixed effect models to test how the isotopomer ratio changes with C_a, tree diameter (as a proxy for crown exposure), temperature, and precipitation.

Across all sites, long-term C_a rise increased photosynthetic efficiency, likely due to increased photosynthesis and the concurrent suppression of photorespiration. Increasing tree size reduced photosynthetic efficiency, likely due to reduced leaf internal CO₂ at higher irradiance and stronger hydraulic limitation. Associations of photosynthetic efficiency with temperature and precipitation were inconclusive.

Our study reveals a centennial-scale association between photosynthetic efficiency and increasing C_a in canopy trees and provides a new and independent line of evidence for C_a-induced stimulation of photosynthetic efficiency in tropical forests.

Aims: Plant inputs are the primary organic carbon source that transforms into soil organic matter (SOM) through microbial processing. One prevailing view is that lignin plays a major role in the accumulation of SOM. This study investigated lignin decomposition using wood from different genotypes of Populus tremula as the model substrate. The genotypes naturally varied in lignin content and composition, resulting in high and low lignin substrates.

Methods: The wood was inoculated with fresh soil and decomposition was interpreted through mass loss and CO2 produced during a 12-month lab incubation. Detailed information on the decomposition patterns of lignin was obtained by Two-dimensional Nuclear magnetic resonance (2D NMR) spectroscopy on four occasions during the incubations.

Results: The lignin content per se did not affect the overall decomposition and ~ 60% of the mass was lost in both substrates. In addition, no differences in oxidative enzyme activity could be observed, and the rate of lignin decomposition was similar to that of the carbohydrates. The 2D NMR analysis showed the oxidized syringyl present in the initial samples was the most resistant to degradation among lignin subunits as it followed the order p-hydroxybenzoates > syringyl > guaiacyl > oxidized syringyl. Furthermore, the degradability of β–O–4 linkages in the lignin varied depending on the subunit (syringyl or guaiacyl) it is attached to.

Conclusions: Our study demonstrates that lignin contains fractions that are easily degradable and can break down alongside carbohydrates. Thus, the initial differences in lignin content per se do not necessarily affect magnitude of SOM accumulation.

Understanding the composition of organic phosphorus (P) in soils is relevant to various disciplines, from agricultural sciences to ecology. Despite past efforts, the precise nature of soil organic P remains an enigma, especially that of the orthophosphate monoesters, which dominate 31P NMR spectra of NaOH-EDTA extracts of soils worldwide. The monoester region often exhibits an unidentified, broad background believed to represent high molecular weight (MW) P. We investigated this monoester background using 1D 31P NMR and 2D 1H[sbnd]31P NMR, as well as 31P transverse relaxation (T2) measurements to calculate its intrinsic linewidth and relate it to MW. Analyzing seven soils from different ecosystems, we observed linewidths of 0.5 to 3 Hz for resolved monoester signals and the background, indicating that it consists of many, possibly >100, sharp signals associated with small (<1.5 kDa) organic P molecules. This result was further supported by 2D 1H[sbnd]31P NMR spectra revealing signals not resolved in the 1D spectra. Our findings align with 31P NMR studies detecting background signals in soil-free samples and modern evidence that alkali-soluble soil organic matter consists of self-assemblies of small organic compounds mimicking large molecules.

• Recently, we reported estimates of anaplerotic carbon flux through the oxidative pentose phosphate pathway (OPPP) in chloroplasts into the Calvin–Benson cycle. These estimates were based on intramolecular hydrogen isotope analysis of sunflower leaf starch. However, the isotope method is believed to underestimate the actual flux at low atmospheric CO₂ concentration (C_a).
• Since the OPPP releases CO₂ and reduces NADP⁺, it can be expected to affect leaf gas exchange under both rubisco- and RuBP-regeneration-limited conditions. Therefore, we expanded Farquhar-von Caemmerer–Berry models to account for OPPP metabolism. Based on model parameterisation with values from the literature, we estimated OPPP-related effects on leaf carbon and energy metabolism in the sunflowers analysed previously.
• We found that flux through the plastidial OPPP increases both above and below C_a ≈ 450 ppm (the condition the plants were acclimated to). This is qualitatively consistent with our previous isotope-based estimates, yet gas-exchange-based estimates are larger at low C_a.
• We discuss our results in relation to regulatory properties of the plastidial and cytosolic OPPP, the proposed variability of CO₂ mesophyll conductance, and the contribution of day respiration to the A/C_i curve drop at high C_a. Furthermore, we critically examine the models and parameterisation and derive recommendations for follow-up studies.

As the central carbon uptake pathway in photosynthetic cells, the Calvin–Benson cycle is among the most important biochemical cycles for life on Earth. A carbon flux of anaplerotic origin (i.e. through the chloroplast-localized oxidative branch of the pentose phosphate pathway) into the Calvin–Benson cycle was proposed recently.

Here, we measured intramolecular deuterium abundances in leaf starch of Helianthus annuus grown at varying ambient CO2 concentrations, Ca. Additionally, we modelled deuterium fractionations expected for the anaplerotic pathway and compared modelled with measured fractionations.

We report deuterium fractionation signals at H1 and H2 of starch glucose. Below a Ca change point, these signals increase with decreasing Ca consistent with modelled fractionations by anaplerotic flux. Under standard conditions (Ca = 450 ppm corresponding to intercellular CO2 concentrations, Ci, of 328 ppm), we estimate negligible anaplerotic flux. At Ca = 180 ppm (Ci = 140 ppm), more than 10% of the glucose-6-phosphate entering the starch biosynthesis pathway is diverted into the anaplerotic pathway.

In conclusion, we report evidence consistent with anaplerotic carbon flux into the Calvin–Benson cycle in vivo. We propose the flux may help to: maintain high levels of ribulose 1,5-bisphosphate under source-limited growth conditions to facilitate photorespiratory nitrogen assimilation required to build-up source strength; and counteract oxidative stress.

Stable isotopes at natural abundance are key tools to study physiological processes occurring outside the temporal scope of manipulation and monitoring experiments. Whole-molecule carbon isotope ratios (13C/12C) enable assessments of plant carbon uptake yet conceal information about carbon allocation. Here, we identify an intramolecular 13C/12C signal at tree-ring glucose C-5 and C-6 and develop experimentally testable theories on its origin. More specifically, we assess the potential of processes within C3 metabolism for signal introduction based (inter alia) on constraints on signal propagation posed by metabolic networks. We propose that the intramolecular signal reports carbon allocation into major metabolic pathways in actively photosynthesizing leaf cells including the anaplerotic, shikimate, and non-mevalonate pathway. We support our theoretical framework by linking it to previously reported whole-molecule 13C/12C increases in cellulose of ozone-treated Betula pendula and a highly significant relationship between the intramolecular signal and tropospheric ozone concentration. Our theory postulates a pronounced preference for leaf cytosolic triose-phosphate isomerase to catalyse the forward reaction in vivo (dihydroxyacetone phosphate to glyceraldehyde 3-phosphate). In conclusion, intramolecular 13C/12C analysis resolves information about carbon uptake and allocation enabling more comprehensive assessments of carbon metabolism than whole-molecule 13C/12C analysis.

• Stable isotope abundances convey valuable information about plant physiological processes and underlying environmental controls. Central gaps in our mechanistic understanding of hydrogen isotope abundances impede their widespread application within the plant and biogeosciences.
• To address these gaps, we analysed intramolecular deuterium abundances in glucose of Pinus nigra extracted from an annually resolved tree-ring series (1961–1995).
• We found fractionation signals (i.e. temporal variability in deuterium abundance) at glucose H¹ and H² introduced by closely related metabolic processes. Regression analysis indicates that these signals (and thus metabolism) respond to drought and atmospheric CO₂ concentration beyond a response change point. They explain ≈ 60% of the whole-molecule deuterium variability. Altered metabolism is associated with below-average yet not exceptionally low growth.
• We propose the signals are introduced at the leaf level by changes in sucrose-to-starch carbon partitioning and anaplerotic carbon flux into the Calvin–Benson cycle. In conclusion, metabolism can be the main driver of hydrogen isotope variation in plant glucose.

Peatlands store carbon in the form of dead organic residues. Climate change and human impact impose risks on the sustainability of the peatlands carbon balance due to increased peat decomposition. Here, we investigated molecular changes in the upper peat layers (0-40 cm), inferred from high-resolution vertical depth profiles, from a boreal peatland using two-dimensional 1H-13C nuclear magnetic resonance (NMR) spectroscopy, and comparison to δ13C, δ15N, and carbon and nitrogen content. Effects of hydrological conditions were investigated at respective sites: natural moist, drainage ditch, and natural dry. The molecular characterization revealed preferential degradation of specific side-chain linkages of xylan-type hemicelluloses within 0-14 cm at all sites, indicating organic matter losses up to 25%. In contrast, the xylan backbone, galactomannan-type hemicelluloses, and cellulose were more resistant to degradation and accumulated at the natural moist and drainage site. δ13C, δ15N, and carbon and nitrogen content did not correlate with specific hemicellulose structures but reflected changes in total carbohydrates. Our analysis provides novel insights into peat carbohydrate decomposition and indicates substantial organic matter losses in the acrotelm due to the degradation of specific hemicellulose structures. This suggests that variations in hemicellulose content and structure influence peat stability, which may have important implications with respect to climate change.

Wildfire is the main disturbance in most boreal forests. In the prolonged absence of wildfire, ecosystem retrogression occurs, which is characterized by reduced productivity, plant biomass and belowground process rates. Previous evidence suggests that phosphorus (P) decreases during retrogression, but the mechanisms involved remain poorly understood. Here we use 1-D 31P and 2-D, 1H-31P NMR to characterize changes in humus P composition across a 5000 year post-fire chronosequence in northern Sweden, to understand why P availability declines during long term fire absence. Against expectations, humus P composition varied only modestly with increasing time since fire. Using a method to back-calculate the in situ soil organic P speciation, we found that it was dominated by biologically active compounds such as RNA (41%), phospholipids (28%) and DNA (22%). The concentration of DNA and pyrophosphate was 19% and 29% lower, respectively, on infrequently burnt than recently burnt islands, and the concentration of DNA, phospholipids and nucleotides was positively correlated with net primary productivity (NPP). Given the lack of evidence for the accumulation of “recalcitrant” P or a geochemical P sink, reductions in P availability during retrogression may be associated with impaired P cycling through slower decomposition rates, and increasing humus depth separating surface humus from P-rich mineral soil. Our findings align with observed negative relationships between NPP and organic P concentration across other chronosequences. They also suggest that changing fire regimes in the boreal zone could indirectly affect the P cycle through changes in NPP and soil microflora rather than through changes in humus P composition.