Water limitation intensity shifts carbon allocation dynamics in Scots pine mesocosms.

Background and aims: Tree species worldwide suffer from extended periods of water limitation. These conditions not only affect the growth and vitality of trees but also feed back on the cycling of carbon (C) at the plant-soil interface. However, the impact of progressing water loss from soils on the transfer of assimilated C belowground remains unresolved. Methods: Using mesocosms, we assessed how increasing levels of water deficit affect the growth of Pinus sylvestris saplings and performed a 13C-CO2 pulse labelling experiment to trace the pathway of assimilated C into needles, fine roots, soil pore CO2, and phospholipid fatty acids of soil microbial groups. Results: With increasing water limitation, trees partitioned more biomass belowground at the expense of aboveground growth. Moderate levels of water limitation barely affected the uptake of 13C label and the transit time of C from needles to the soil pore CO2. Comparatively, more severe water limitation increased the fraction of 13C label that trees allocated to fine roots and soil fungi while a lower fraction of 13CO2 was readily respired from the soil. Conclusions: When soil water becomes largely unavailable, C cycling within trees becomes slower, and a fraction of C allocated belowground may accumulate in fine roots or be transferred to the soil and associated microorganisms without being metabolically used. Supplementary Information: The online version contains supplementary material available at 10.1007/s11104-023-06093-5.

Impact of salinity gradient, water pollution and land use types on greenhouse gas emissions from an urbanized estuary.

Estuaries have been recognized as one of the major sources of greenhouse gases (GHGs) in aquatic systems; yet we still lack insights into the impact of both anthropogenic and natural factors on the dynamics of GHG emissions. Here, we assessed the spatiotemporal dynamics and underlying drivers of the GHG emissions from the Scheldt Estuary with a focus on the effects of salinity gradient, water pollution, and land use types, together with their interaction. Overall, we found a negative impact of salinity on carbon dioxide (CO2) and nitrous oxide (N2O) emissions which can be due to the decrease of both salinity and water quality when moving upstream. Stronger impact of water pollution on the GHG emissions was found at the freshwater sites upstream compared to saline sites downstream. In particular, when water quality of the sites reduced from good, mainly located in the mouth and surrounded by arable sites, to polluted, mainly located in the upstream and surrounded by urban sites, CO2 emissions from the sites doubled while N2O emissions tripled. Similarly, the effects of water pollution on methane (CH4) emissions became much stronger in the freshwater sites compared to the saline sites. These decreasing effects from upstream to the mouth were associated with the increase in urbanization as sites surrounded by urban areas released on average almost two times more CO2 and N2O than sites surrounded by nature and industry areas. Applied machine learning methods also revealed that, in addition to salinity effects, nutrient and organic enrichment stimulated the GHG emissions from the Scheldt Estuary. These findings highlight the importance of the interaction between salinity, water pollution, and land use in order to understand their influences on GHG emissions from dynamic estuarine systems.

Cassava-maize intercropping systems in southern Nigeria: Radiation use efficiency, soil moisture dynamics, and yields of component crops.

Efficient utilization of incident solar radiation and rainwater conservation in rain-fed smallholder cropping systems require the development and adoption of cropping systems with high resource use efficiency. Due to the popularity of cassava-maize intercropping and the food security and economic importance of both crops in Nigeria, we investigated options to improve interception of photosynthetically active radiation (IPAR), radiation use efficiency (RUE), soil moisture retention, and yields of cassava and maize in cassava-maize intercropping systems in 8 on-farm researcher-managed multi-location trials between 2017 and 2019 in different agro-ecologies of southern Nigeria. Treatments were a combination of (1) maize planting density (low density at 20,000 maize plants ha-1 versus high density at 40,000 maize plants ha-1, intercropped with 12,500 cassava plants ha-1); (2) fertilizer application and management targeting either the maize crop (90 kg N, 20 kg P and 37 kg K ha-1) or the cassava crop (75 kg N, 20 kg P and 90 kg K ha-1), compared with control without fertilizer application. Cassava and maize development parameters were highest in the maize fertilizer regime, resulting in the highest IPAR at high maize density. The combined intercrop biomass yield was highest at high maize density in the maize fertilizer regime. Without fertilizer application, RUE was highest at low maize density. However, the application of the maize fertilizer regime at high maize density resulted in the highest RUE, soil moisture content, and maize grain yield. Cassava storage root yield was higher in the cassava fertilizer regime than in the maize fertilizer regime. We conclude that improved IPAR, RUE, soil moisture retention, and grain yield on nutrient-limited soils of southern Nigeria, or in similar environments, can be achieved by intercropping 40,000 maize plants ha-1 with 12,500 cassava plants ha-1 and managing the system with the maize fertilizer regime. However, for higher cassava storage root yield, the system should be managed with the cassava fertilizer regime.

Increasing calcium scarcity along Afrotropical forest succession.

Secondary forests constitute an increasingly important component of tropical forests worldwide. Although cycling of essential nutrients affects recovery trajectories of secondary forests, the effect of nutrient limitation on forest regrowth is poorly constrained. Here we use three lines of evidence from secondary forest succession sequences in central Africa to identify potential nutrient limitation in regrowing forests. First, we show that atmospheric phosphorus supply exceeds demand along forest succession, whereas forests rely on soil stocks to meet their base cation demands. Second, soil nutrient metrics indicate that available phosphorus increases along the succession, whereas available cations decrease. Finally, fine root, foliar and litter stoichiometry show that tissue calcium concentrations decline relative to those of nitrogen and phosphorus during succession. Taken together, these observations suggest that calcium becomes an increasingly scarce resource in central African forests during secondary succession. Furthermore, ecosystem calcium storage shifts from soil to woody biomass over succession, making it a vulnerable nutrient in the wake of land-use change scenarios that involve woody biomass export. Our results thus call for a broadened focus on elements other than nitrogen and phosphorus regarding tropical forest biogeochemical cycles and identify calcium as a scarce and potentially limiting nutrient in an increasingly disturbed and dynamic tropical forest landscape.

Greenhouse gas dynamics in an urbanized river system: influence of water quality and land use.

Rivers act as a natural source of greenhouse gases (GHGs). However, anthropogenic activities can largely alter the chemical composition and microbial communities of rivers, consequently affecting their GHG production. To investigate these impacts, we assessed the accumulation of CO2, CH4, and N2O in an urban river system (Cuenca, Ecuador). High variation of dissolved GHG concentrations was found among river tributaries that mainly depended on water quality and land use. By using Prati and Oregon water quality indices, we observed a clear pattern between water quality and the dissolved GHG concentration: the more polluted the sites were, the higher were their dissolved GHG concentrations. When river water quality deteriorated from acceptable to very heavily polluted, the mean value of pCO2 and dissolved CH4 increased by up to ten times while N2O concentrations boosted by 15 times. Furthermore, surrounding land-use types, i.e., urban, roads, and agriculture, could considerably affect the GHG production in the rivers. Particularly, the average pCO2 and dissolved N2O of the sites close to urban areas were almost four times higher than those of the natural sites while this ratio was 25 times in case of CH4, reflecting the finding that urban areas had the worst water quality with almost 70% of their sites being polluted while this proportion of nature areas was only 12.5%. Lastly, we identified dissolved oxygen, ammonium, and flow characteristics as the main important factors to the GHG production by applying statistical analysis and random forests. These results highlighted the impacts of land-use types on the production of GHGs in rivers contaminated by sewage discharges and surface runoff.

Low N2O and variable CH4 fluxes from tropical forest soils of the Congo Basin.

Globally, tropical forests are assumed to be an important source of atmospheric nitrous oxide (N2O) and sink for methane (CH4). Yet, although the Congo Basin comprises the second largest tropical forest and is considered the most pristine large basin left on Earth, in situ N2O and CH4 flux measurements are scarce. Here, we provide multi-year data derived from on-ground soil flux (n = 1558) and riverine dissolved gas concentration (n = 332) measurements spanning montane, swamp, and lowland forests. Each forest type core monitoring site was sampled at least for one hydrological year between 2016 - 2020 at a frequency of 7-14 days. We estimate a terrestrial CH4 uptake (in kg CH4-C ha-1 yr-1) for montane (-4.28) and lowland forests (-3.52) and a massive CH4 release from swamp forests (non-inundated 2.68; inundated 341). All investigated forest types were a N2O source (except for inundated swamp forest) with 0.93, 1.56, 3.5, and -0.19 kg N2O-N ha-1 yr-1 for montane, lowland, non-inundated swamp, and inundated swamp forests, respectively.

Stable Isotope Analysis of Greenhouse Gases Requires Analyte Preconcentration.

Nitrous oxide (N2O) is an important trace gas contributing to global warming and depletion of ozone in the stratosphere. Its increasing abundance is caused mainly by anthropogenic sources, such as application of fertilizers in agriculture or emissions from industry. To understand the N2O global budget, its sources and sinks need to be well-described and quantified. In this project, a new method for N2O source appointment was developed that can help with this task. The method is based on analysis of the eight most abundant isotopic molecules of N2O, using quantum cascade laser absorption spectroscopy (QCLAS). The applicability of the method towards the N2O biogeochemical cycle was demonstrated on a prominent N2O source (bacterial denitrification) and the most important N2O sink (UV photolysis) on samples prepared in laboratory experiments. An extension of the QCLAS method to natural samples can be achieved by hyphenation with a preconcentration technique that increases concentration of the analyte and standardizes the sample matrix. This article provides an overview of currently applied preconcentration techniques in the field of greenhouse-gas analysis and a description of the preconcentration device TREX that will be employed in future projects with the developed QCLAS method.

Global maps of soil temperature.

Research in global change ecology relies heavily on global climatic grids derived from estimates of air temperature in open areas at around 2 m above the ground. These climatic grids do not reflect conditions below vegetation canopies and near the ground surface, where critical ecosystem functions occur and most terrestrial species reside. Here, we provide global maps of soil temperature and bioclimatic variables at a 1-km2 resolution for 0-5 and 5-15 cm soil depth. These maps were created by calculating the difference (i.e. offset) between in situ soil temperature measurements, based on time series from over 1200 1-km2 pixels (summarized from 8519 unique temperature sensors) across all the world's major terrestrial biomes, and coarse-grained air temperature estimates from ERA5-Land (an atmospheric reanalysis by the European Centre for Medium-Range Weather Forecasts). We show that mean annual soil temperature differs markedly from the corresponding gridded air temperature, by up to 10°C (mean = 3.0 ± 2.1°C), with substantial variation across biomes and seasons. Over the year, soils in cold and/or dry biomes are substantially warmer (+3.6 ± 2.3°C) than gridded air temperature, whereas soils in warm and humid environments are on average slightly cooler (-0.7 ± 2.3°C). The observed substantial and biome-specific offsets emphasize that the projected impacts of climate and climate change on near-surface biodiversity and ecosystem functioning are inaccurately assessed when air rather than soil temperature is used, especially in cold environments. The global soil-related bioclimatic variables provided here are an important step forward for any application in ecology and related disciplines. Nevertheless, we highlight the need to fill remaining geographic gaps by collecting more in situ measurements of microclimate conditions to further enhance the spatiotemporal resolution of global soil temperature products for ecological applications.

In-depth analysis of N2O fluxes in tropical forest soils of the Congo Basin combining isotope and functional gene analysis.

Primary tropical forests generally exhibit large gaseous nitrogen (N) losses, occurring as nitric oxide (NO), nitrous oxide (N2O) or elemental nitrogen (N2). The release of N2O is of particular concern due to its high global warming potential and destruction of stratospheric ozone. Tropical forest soils are predicted to be among the largest natural sources of N2O; however, despite being the world's second-largest rainforest, measurements of gaseous N-losses from forest soils of the Congo Basin are scarce. In addition, long-term studies investigating N2O fluxes from different forest ecosystem types (lowland and montane forests) are scarce. In this study we show that fluxes measured in the Congo Basin were lower than fluxes measured in the Neotropics, and in the tropical forests of Australia and South East Asia. In addition, we show that despite different climatic conditions, average annual N2O fluxes in the Congo Basin's lowland forests (0.97 ± 0.53 kg N ha-1 year-1) were comparable to those in its montane forest (0.88 ± 0.97 kg N ha-1 year-1). Measurements of soil pore air N2O isotope data at multiple depths suggests that a microbial reduction of N2O to N2 within the soil may account for the observed low surface N2O fluxes and low soil pore N2O concentrations. The potential for microbial reduction is corroborated by a significant abundance and expression of the gene nosZ in soil samples from both study sites. Although isotopic and functional gene analyses indicate an enzymatic potential for complete denitrification, combined gaseous N-losses (N2O, N2) are unlikely to account for the missing N-sink in these forests. Other N-losses such as NO, N2 via Feammox or hydrological particulate organic nitrogen export could play an important role in soils of the Congo Basin and should be the focus of future research.

Spatial and temporal variations of greenhouse gas emissions from a waste stabilization pond: Effects of sludge distribution and accumulation.

Due to regular influx of organic matter and nutrients, waste stabilization ponds (WSPs) can release considerable quantities of greenhouse gases (GHGs). To investigate the spatiotemporal variations of GHG emissions from WSPs with a focus on the effects of sludge accumulation and distribution, we conducted a bathymetry survey and two sampling campaigns in Ucubamba WSP (Cuenca, Ecuador). The results indicated that spatial variation of GHG emissions was strongly dependent on sludge distribution. Thick sludge layers in aerated ponds and facultative ponds caused substantial CO2 and CH4 emissions which accounted for 21.3% and 78.7% of the total emissions from the plant. Conversely, the prevalence of anoxic conditions stimulated the N2O consumption via complete denitrification leading to a net uptake from the atmosphere, i.e. up to 1.4±0.2 mg-N m-2 d-1. Double emission rates of CO2 were found in the facultative and maturation ponds during the day compared to night-time emissions, indicating the important role of algal respiration, while no diel variation of the CH4 and N2O emissions was found. Despite the uptake of N2O, the total GHG emissions of the WSP was higher than constructed wetlands and conventional centralized wastewater treatment facilities. Hence, it is recommended that sludge management with proper desludging regulation should be included as an important mitigation measure to reduce the carbon footprint of pond treatment facilities.

Mixed Effects of Soil Compaction on the Nitrogen Cycle Under Pea and Wheat.

Soil compaction caused by highly mechanized agriculture can constrain soil microbial diversity and functioning. Physical pressure on the soil decreases macropores and thereby limits oxygen diffusion. The associated shift from aerobic to anaerobic conditions can reduce nitrification and promote denitrification processes, leading to nitrogen (N) losses and N depletion that affect plant productivity. High soil moisture content during trafficking can exacerbate the negative effects of soil compaction. However, the extent to which soil moisture amplifies the effects of compaction on the soil microbiome and its control over N cycling is not well understood. Using a controlled greenhouse experiment with two different crops (pea and wheat), we compared the effects of compaction at three different soil moisture levels on soil physicochemical properties, microbial diversity, and the abundance of specific N species and quantification of associated microbial functional groups in the N cycle. Soil compaction increased bulk density from 15% (light compaction) to 25% (severe compaction). Compaction delayed germination in both crops and reduced yield by up to 60% for pea and 40% for wheat. Compaction further induced crop-specific shifts in microbial community structures. After compaction, the relative abundance of denitrifiers increased along with increased nitrate (NO3 -) consumption and elevated nitrous oxide (N2O) concentrations in the soil pores. Conversely, the relative abundance of nitrifiers remained stable under compaction, but potentially decelerated nitrification rates, resulting in ammonium (NH4 +) accumulation in the soil. This study showed that soil compaction effects are proportional to the initial soil moisture content, which could serve as a good indicator of compaction severity on agricultural fields. However, the impact of soil compaction on crop performance and on microbial communities and functions associated with the N cycle were not necessarily aligned. These findings demonstrate that not only the soil physical properties but also various biological indicators need to be considered in order to provide more precise recommendations for developing sustainable farming systems.

Livestock enclosures in drylands of Sub-Saharan Africa are overlooked hotspots of N2O emissions.

Sub-Saharan Africa (SSA) is home to approximately » of the global livestock population, which in the last 60 years has increased by factors of 2.5-4 times for cattle, goats and sheep. An important resource for pastoralists, most livestock live in semi-arid and arid environments, where they roam during the day and are kept in enclosures (or bomas) during the night. Manure, although rich in nitrogen, is rarely used, and therefore accumulates in bomas over time. Here we present in-situ measurements of N2O fluxes from 46 bomas in Kenya and show that even after 40 years following abandonment, fluxes are still ~one magnitude higher than those from adjacent savanna sites. Using maps of livestock distribution, we scaled our finding to SSA and found that abandoned bomas are significant hotspots for atmospheric N2O at the continental scale, contributing ~5% of the current estimate of total anthropogenic N2O emissions for all of Africa.

What can we learn from N2 O isotope data? - Analytics, processes and modelling.

The isotopic composition of nitrous oxide (N2 O) provides useful information for evaluating N2 O sources and budgets. Due to the co-occurrence of multiple N2 O transformation pathways, it is, however, challenging to use isotopic information to quantify the contribution of distinct processes across variable spatiotemporal scales. Here, we present an overview of recent progress in N2 O isotopic studies and provide suggestions for future research, mainly focusing on: analytical techniques; production and consumption processes; and interpretation and modelling approaches. Comparing isotope-ratio mass spectrometry (IRMS) with laser absorption spectroscopy (LAS), we conclude that IRMS is a precise technique for laboratory analysis of N2 O isotopes, while LAS is more suitable for in situ/inline studies and offers advantages for site-specific analyses. When reviewing the link between the N2 O isotopic composition and underlying mechanisms/processes, we find that, at the molecular scale, the specific enzymes and mechanisms involved determine isotopic fractionation effects. In contrast, at plot-to-global scales, mixing of N2 O derived from different processes and their isotopic variability must be considered. We also find that dual isotope plots are effective for semi-quantitative attribution of co-occurring N2 O production and reduction processes. More recently, process-based N2 O isotopic models have been developed for natural abundance and 15 N-tracing studies, and have been shown to be effective, particularly for data with adequate temporal resolution. Despite the significant progress made over the last decade, there is still great need and potential for future work, including development of analytical techniques, reference materials and inter-laboratory comparisons, further exploration of N2 O formation and destruction mechanisms, more observations across scales, and design and validation of interpretation and modelling approaches. Synthesizing all these efforts, we are confident that the N2 O isotope community will continue to advance our understanding of N2 O transformation processes in all spheres of the Earth, and in turn to gain improved constraints on regional and global budgets.

Century-long apparent decrease in intrinsic water-use efficiency with no evidence of progressive nutrient limitation in African tropical forests.

Forests exhibit leaf- and ecosystem-level responses to environmental changes. Specifically, rising carbon dioxide (CO2 ) levels over the past century are expected to have increased the intrinsic water-use efficiency (iWUE) of tropical trees while the ecosystem is gradually pushed into progressive nutrient limitation. Due to the long-term character of these changes, however, observational datasets to validate both paradigms are limited in space and time. In this study, we used a unique herbarium record to go back nearly a century and show that despite the rise in CO2 concentrations, iWUE has decreased in central African tropical trees in the Congo Basin. Although we find evidence that points to leaf-level adaptation to increasing CO2 -that is, increasing photosynthesis-related nutrients and decreasing maximum stomatal conductance, a decrease in leaf δ13 C clearly indicates a decreasing iWUE over time. Additionally, the stoichiometric carbon to nitrogen and nitrogen to phosphorus ratios in the leaves show no sign of progressive nutrient limitation as they have remained constant since 1938, which suggests that nutrients have not increasingly limited productivity in this biome. Altogether, the data suggest that other environmental factors, such as increasing temperature, might have negatively affected net photosynthesis and consequently downregulated the iWUE. Results from this study reveal that the second largest tropical forest on Earth has responded differently to recent environmental changes than expected, highlighting the need for further on-ground monitoring in the Congo Basin.

Mobilization of aged and biolabile soil carbon by tropical deforestation.

In the mostly pristine Congo Basin, agricultural land-use change has intensified in recent years. One potential and understudied consequence of this deforestation and conversion to agriculture is the mobilization and loss of organic matter from soils to rivers as dissolved organic matter. Here, we quantify and characterize dissolved organic matter sampled from 19 catchments of varying deforestation extent near Lake Kivu over a two-week period during the wet season. Dissolved organic carbon from deforested, agriculturally-dominated catchments was older (14C age: ~1.5kyr) and more biolabile than from pristine forest catchments. Ultrahigh-resolution mass spectrometry revealed that this aged organic matter from deforested catchments was energy-rich and chemodiverse, with higher proportions of nitrogen- and sulfur-containing formulae. Given the molecular composition and biolability, we suggest that organic matter from deforested landscapes is preferentially respired upon disturbance, resulting in elevated in-stream concentrations of carbon dioxide. We estimate that while deforestation reduces the overall flux of dissolved organic carbon by ~56%, it does not significantly change the yield of biolabile dissolved organic carbon. Ultimately, the exposure of deeper soil horizons through deforestation and agricultural expansion releases old, previously stable, and biolabile soil organic carbon into the modern carbon cycle via the aquatic pathway.

Greenhouse gas fluxes over managed grasslands in Central Europe.

Central European grasslands are characterized by a wide range of different management practices in close geographical proximity. Site-specific management strategies strongly affect the biosphere-atmosphere exchange of the three greenhouse gases (GHG) carbon dioxide (CO2 ), nitrous oxide (N2 O), and methane (CH4 ). The evaluation of environmental impacts at site level is challenging, because most in situ measurements focus on the quantification of CO2 exchange, while long-term N2 O and CH4 flux measurements at ecosystem scale remain scarce. Here, we synthesized ecosystem CO2 , N2 O, and CH4 fluxes from 14 managed grassland sites, quantified by eddy covariance or chamber techniques. We found that grasslands were on average a CO2 sink (-1,783 to -91 g CO2  m-2  year-1 ), but a N2 O source (18-638 g CO2 -eq. m-2  year-1 ), and either a CH4 sink or source (-9 to 488 g CO2 -eq. m-2  year-1 ). The net GHG balance (NGB) of nine sites where measurements of all three GHGs were available was found between -2,761 and -58 g CO2 -eq. m-2  year-1 , with N2 O and CH4 emissions offsetting concurrent CO2 uptake by on average 21 ± 6% across sites. The only positive NGB was found for one site during a restoration year with ploughing. The predictive power of soil parameters for N2 O and CH4 fluxes was generally low and varied considerably within years. However, after site-specific data normalization, we identified environmental conditions that indicated enhanced GHG source/sink activity ("sweet spots") and gave a good prediction of normalized overall fluxes across sites. The application of animal slurry to grasslands increased N2 O and CH4 emissions. The N2 O-N emission factor across sites was 1.8 ± 0.5%, but varied considerably at site level among the years (0.1%-8.6%). Although grassland management led to increased N2 O and CH4 emissions, the CO2 sink strength was generally the most dominant component of the annual GHG budget.

Strong Coupling of Shoot Assimilation and Soil Respiration during Drought and Recovery Periods in Beech As Indicated by Natural Abundance δ13C Measurements.

Drought down-regulates above- and belowground carbon fluxes, however, the resilience of trees to drought will also depend on the speed and magnitude of recovery of these above- and belowground fluxes after re-wetting. Carbon isotope composition of above- and belowground carbon fluxes at natural abundance provides a methodological approach to study the coupling between photosynthesis and soil respiration (SR) under conditions (such as drought) that influence photosynthetic carbon isotope discrimination. In turn, the direct supply of root respiration with recent photoassimilates will impact on the carbon isotope composition of soil-respired CO2. We independently measured shoot and soil CO2 fluxes of beech saplings (Fagus sylvatica L.) and their respective δ13C continuously with laser spectroscopy at natural abundance. We quantified the speed of recovery of drought stressed trees after re-watering and traced photosynthetic carbon isotope signal in the carbon isotope composition of soil-respired CO2. Stomatal conductance responded strongly to the moderate drought (-65%), induced by reduced soil moisture content as well as increased vapor pressure deficit. Simultaneously, carbon isotope discrimination decreased by 8‰, which in turn caused a significant increase in δ13C of recent metabolites (1.5-2.5‰) and in δ13C of SR (1-1.5‰). Generally, shoot and soil CO2 fluxes and their δ13C were in alignment during drought and subsequent stress release, clearly demonstrating a permanent dependence of root respiration on recently fixed photoassimilates, rather than on older reserves. After re-watering, the drought signal persisted longer in δ13C of the water soluble fraction that integrates multiple metabolites (soluble sugars, amino acids, organic acids) than in the neutral fraction which represents most recently assimilated sugars or in the δ13C of SR. Nevertheless, full recovery of all aboveground physiological variables was reached within 4 days - and within 7 days for SR - indicating high resilience of (young) beech against moderate drought.

Metabolic Fate of the Carboxyl Groups of Malate and Pyruvate and their Influence on δ(13)C of Leaf-Respired CO2 during Light Enhanced Dark Respiration.

The enhanced CO2 release of illuminated leaves transferred into darkness, termed "light enhanced dark respiration (LEDR)", is often associated with an increase in the carbon isotope ratio of the respired CO2 (δ(13)CLEDR). The latter has been hypothesized to result from different respiratory substrates and decarboxylation reactions in various metabolic pathways, which are poorly understood so far. To provide a better insight into the underlying metabolic processes of δ(13)CLEDR, we fed position-specific (13)C-labeled malate and pyruvate via the xylem stream to leaves of species with high and low δ(13)CLEDR values (Halimium halimifolium and Oxalis triangularis, respectively). During respective label application, we determined label-derived leaf (13)CO2 respiration using laser spectroscopy and the (13)C allocation to metabolic fractions during light-dark transitions. Our results clearly show that both carboxyl groups (C-1 and C-4 position) of malate similarly influence respiration and metabolic fractions in both species, indicating possible isotope randomization of the carboxyl groups of malate by the fumarase reaction. While C-2 position of pyruvate was only weakly respired, the species-specific difference in natural δ(13)CLEDR patterns were best reflected by the (13)CO2 respiration patterns of the C-1 position of pyruvate. Furthermore, (13)C label from malate and pyruvate were mainly allocated to amino and organic acid fractions in both species and only little to sugar and lipid fractions. In summary, our results suggest that respiration of both carboxyl groups of malate (via fumarase) by tricarboxylic acid cycle reactions or by NAD-malic enzyme influences δ(13)CLEDR. The latter supplies the pyruvate dehydrogenase reaction, which in turn determines natural δ(13)CLEDR pattern by releasing the C-1 position of pyruvate.

Bias in estimated online leaf carbon isotope discrimination due to woody tissues.

Branch or shoot chamber measurements integrate over both foliar and woody tissue carbon dioxide (CO2) fluxes and their associated influences on the carbon isotopic composition of atmospheric/headspace CO2. Here, we quantified the bias introduced by woody tissue carbon isotope fluxes on apparent leaf (13)C discrimination (Δ(13)Capparent) estimates, using laser spectroscopy under controlled conditions. CO2 efflux from woody tissues of defoliated beech saplings in the dark was strongly related to temperature (R(2) = 0.78), which served as the basis to model light-dependent woody tissue photosynthesis (R(2) = 0.72). We then quantified the contributions of leaf and woody tissues to leaf Δ(13)Capparent of foliated beech saplings in the light. Unbiased foliar Δ(13)C was 1.1 to 4.9‰ lower than leaf Δ(13)Capparent, depending on photosynthetic rates of woody tissues. Therefore, we strongly recommend accounting for isotope-related bias due to woody tissues when estimating leaf Δ(13)Capparent based on branch or shoot chamber measurements.

Soil H2¹8O labelling reveals the effect of drought on C¹8OO fluxes to the atmosphere.

Above- and belowground processes in plants are tightly coupled via carbon and water fluxes through the soil-plant-atmosphere system. The oxygen isotopic composition of atmospheric CO2 and water vapour (H2Ov) provides a valuable tool for investigating the transport and cycling of carbon and water within this system. However, detailed studies on the coupling between ecosystem components and environmental drivers are sparse. Therefore, we conducted a H2 (18)O-labelling experiment to investigate the effect of drought on the speed of the link between below- and aboveground processes and its subsequent effect on C(18)OO released by leaves and soils. A custom-made chamber system, separating shoot from soil compartments, allowed separate measurements of shoot- and soil-related processes under controlled conditions. Gas exchange of oxygen stable isotopes in CO2 and H2Ov served as the main tool of investigation and was monitored in real time on Fagus sylvatica saplings using laser spectroscopy. H2(18)O-labelling showed that drought caused a slower transport of water molecules from soil to shoot, which was indicated by its direct derivation from independently measured concentrations and (18)O/(16)O ratios of CO2 and H2Ov, respectively. Furthermore, drought reduced the (18)O equilibrium between H2O and CO2 at the shoot level, resulting in less-enriched C(18)OO fluxes from leaf to atmosphere compared with control plants. Compared with the shoot, (18)O equilibrium was not instantaneous in the soil and no drought effect was apparent.