The sensitivity of root water uptake to cold root temperature follows species-specific upper elevational distribution limits of temperate tree species.

Physiological water stress induced by low root temperatures might contribute to species-specific climatic limits of tree distribution. We investigated the low temperature sensitivity of root water uptake and transport in seedlings of 16 European tree species which reach their natural upper elevation distribution limits at different distances to the alpine treeline. We used 2H-H2O pulse-labelling to quantify the water uptake and transport velocity from roots to leaves in seedlings exposed to constant 15°C, 7°C or 2°C root temperature, but identical aboveground temperatures between 20°C and 25°C. In all species, low root temperatures reduced the water transport rate, accompanied by reduced stem water potentials and stomatal conductance. At 7°C root temperature, the relative water uptake rates among species correlated positively with the species-specific upper elevation limits, indicating an increasingly higher sensitivity to lower root zone temperatures, the lower a species' natural elevational distribution limit. Conversely, 2°C root temperature severely inhibited water uptake in all species, irrespective of the species' thermal elevational limits. We conclude that low temperature-induced hydraulic constraints contribute to the cold distribution limits of temperate tree species and are a potential physiological cause behind the low temperature limits of plant growth in general.

A novel 13 C pulse-labelling method to quantify the contribution of rhizodeposits to soil respiration in a grassland exposed to drought and nitrogen addition.

Rhizodeposition plays an important role in below-ground carbon (C) cycling. However, quantification of rhizodeposition in intact plant-soil systems has remained elusive due to methodological issues. We used a 13 C-CO2 pulse-labelling method to quantify the contribution of rhizodeposition to below-ground respiration. Intact plant-soil cores were taken from a grassland field, and in half, shoots and roots were removed (unplanted cores). Both unplanted and planted cores were assigned to drought and nitrogen (N) treatments. Afterwards, shoots in planted cores were pulse labelled with 13 C-CO2 and then clipped to determine total below-ground respiration and its δ13 C. Simultaneously, δ13 C was measured for the respiration of live roots, soils with rhizodeposits, and unplanted treatments, and used as endmembers with which to determine root respiration and rhizodeposit C decomposition using two-source mixing models. Rhizodeposit decomposition accounted for 7-31% of total below-ground respiration. Drought reduced decomposition of both rhizodeposits and soil organic carbon (SOC), while N addition increased root respiration but not the contribution of rhizodeposit C decomposition to below-ground respiration. This study provides a new approach for the partitioning of below-ground respiration into different sources, and indicates that decomposition of rhizodeposit C is an important component of below-ground respiration that is sensitive to drought and N addition in grassland ecosystems.

Drought decreases incorporation of recent plant photosynthate into soil food webs regardless of their trophic complexity.

Theory suggests that more complex food webs promote stability and can buffer the effects of perturbations, such as drought, on soil organisms and ecosystem functions. Here, we tested experimentally how soil food web trophic complexity modulates the response to drought of soil functions related to carbon cycling and the capture and transfer below-ground of recent photosynthate by plants. We constructed experimental systems comprising soil communities with one, two or three trophic levels (microorganisms, detritivores and predators) and subjected them to drought. We investigated how food web trophic complexity in interaction with drought influenced litter decomposition, soil CO2 efflux, mycorrhizal colonization, fungal production, microbial communities and soil fauna biomass. Plants were pulse-labelled after the drought with 13 C-CO2 to quantify the capture of recent photosynthate and its transfer below-ground. Overall, our results show that drought and soil food web trophic complexity do not interact to affect soil functions and microbial community composition, but act independently, with an overall stronger effect of drought. After drought, the net uptake of 13 C by plants was reduced and its retention in plant biomass was greater, leading to a strong decrease in carbon transfer below-ground. Although food web trophic complexity influenced the biomass of Collembola and fungal hyphal length, 13 C enrichment and the net transfer of carbon from plant shoots to microbes and soil CO2 efflux were not affected significantly by varying the number of trophic groups. Our results indicate that drought has a strong effect on above-ground-below-ground linkages by reducing the flow of recent photosynthate. Our results emphasize the sensitivity of the critical pathway of recent photosynthate transfer from plants to soil organisms to a drought perturbation, and show that these effects may not be mitigated by the trophic complexity of soil communities, at least at the level manipulated in this experiment.

Protura are unique: first evidence of specialized feeding on ectomycorrhizal fungi in soil invertebrates.

BACKGROUND: Ectomycorrhizal fungi (ECM) play a central role in nutrient cycling in boreal and temperate forests, but their role in the soil food web remains little understood. One of the groups assumed to live as specialised mycorrhizal feeders are Protura, but experimental and field evidence is lacking. We used a combination of three methods to test if Protura are specialized mycorrhizal feeders and compared their trophic niche with other soil invertebrates. Using pulse labelling of young beech and ash seedlings we analysed the incorporation of 13C and 15N into Acerentomon gallicum. In addition, individuals of Protura from temperate forests were collected for the analysis of neutral lipid fatty acids and natural variations in stable isotope ratios. RESULTS: Pulse labelling showed rapid incorporation of root-derived 13C, but no incorporation of root-derived 15N into A. gallicum. The transfer of 13C from lateral roots to ectomycorrhizal root tips was high, while it was low for 15N. Neutral lipid fatty acid (NLFA) analysis showed high amounts of bacterial marker (16:1ω7) and plant marker (16:0 and 18:1ω9) fatty acids but not of the fungal membrane lipid 18:2ω6,9 in A. gallicum. Natural variations in stable isotope ratios in Protura from a number of temperate forests were distinct from those of the great majority of other soil invertebrates, but remarkably similar to those of sporocarps of ECM fungi. CONCLUSIONS: Using three in situ methods, stable isotope labelling, neutral lipid fatty acid analysis and natural variations of stable isotope ratios, we showed that Protura predominantly feed on mycorrhizal hyphae via sucking up hyphal cytoplasm. Predominant feeding on ectomycorrhizal mycelia by Protura is an exception; the limited consumption of ECM by other soil invertebrates may contribute to carbon sequestration in temperate and boreal forests.

Seasonal variations drive short-term dynamics and partitioning of recently assimilated carbon in the foliage of adult beech and pine.

13 CO2 pulse-labelling experiments were performed in situ on adult beeches (Fagus sylvatica) and pines (Pinus pinaster) at different phenological stages to study seasonal and interspecific short-term dynamics and partitioning of recently assimilated carbon (C) in leaves. Polar fraction (PF, including soluble sugars, amino acids and organic acids) and starch were purified from foliage sampled during a 10-d chase period. C contents, isotopic compositions and 13 C dynamics parameters were determined in bulk foliage, PF and starch. Decrease in 13 C amount in bulk foliage followed a two-pool exponential model highlighting 13 C partitioning between 'mobile' and 'stable' pools, the relative proportion of the latter being maximal in beech leaves in May. Early in the growing season, new foliage acted as a strong C sink in both species, but although young leaves and needles were already photosynthesizing, the latter were still supplied with previous-year needle photosynthates 2 months after budburst. Mean 13 C residence times (MRT) were minimal in summer, indicating fast photosynthate export to supply perennial organ growth in both species. In late summer, MRT differed between senescing beech leaves and overwintering pine needles. Seasonal variations of 13 C partitioning and dynamics in field-grown tree foliage are closely linked to phenological differences between deciduous and evergreen trees.

The fate of recently fixed carbon after drought release: towards unravelling C storage regulation in Tilia platyphyllos and Pinus sylvestris.

Carbon reserves are important for maintaining tree function during and after stress. Increasing tree mortality driven by drought globally has renewed the interest in how plants regulate allocation of recently fixed C to reserve formation. Three-year-old seedlings of two species (Tilia platyphyllos and Pinus sylvestris) were exposed to two intensities of experimental drought during ~10 weeks, and 13 C pulse labelling was subsequently applied with rewetting. Tracking the 13 C label across different organs and C compounds (soluble sugars, starch, myo-inositol, lipids and cellulose), together with the monitoring of gas exchange and C mass balances over time, allowed for the identification of variations in C allocation priorities and tree C balances that are associated with drought effects and subsequent drought release. The results demonstrate that soluble sugars accumulated in P. sylvestris under drought conditions independently of growth trends; thus, non-structural carbohydrates (NSC) formation cannot be simply considered a passive overflow process in this species. Once drought ceased, C allocation to storage was still prioritized at the expense of growth, which suggested the presence of 'drought memory effects', possibly to ensure future growth and survival. On the contrary, NSC and growth dynamics in T. platyphyllos were consistent with a passive (overflow) view of NSC formation.

Summer drought alters carbon allocation to roots and root respiration in mountain grassland.

Drought affects the carbon (C) source and sink activities of plant organs, with potential consequences for belowground C allocation, a key process of the terrestrial C cycle. The responses of belowground C allocation dynamics to drought are so far poorly understood. We combined experimental rain exclusion with (13)C pulse labelling in a mountain meadow to analyse the effects of summer drought on the dynamics of belowground allocation of recently assimilated C and how it is partitioned among different carbohydrate pools and root respiration. Severe soil moisture deficit decreased the ecosystem C uptake and the amounts and velocity of C allocated from shoots to roots. However, the proportion of recently assimilated C translocated belowground remained unaffected by drought. Reduced root respiration, reflecting reduced C demand under drought, was increasingly sustained by C reserves, whilst recent assimilates were preferentially allocated to root storage and an enlarged pool of osmotically active compounds. Our results indicate that under drought conditions the usage of recent photosynthates is shifted from metabolic activity to osmotic adjustment and storage compounds.

Experimental drought reduces the transfer of recently fixed plant carbon to soil microbes and alters the bacterial community composition in a mountain meadow.

Drought affects plants and soil microorganisms, but it is still not clear how it alters the carbon (C) transfer at the plant-microbial interface. Here, we tested direct and indirect effects of drought on soil microbes and microbial turnover of recent plant-derived C in a mountain meadow. Microbial community composition was assessed using phospholipid fatty acids (PLFAs); the allocation of recent plant-derived C to microbial groups was analysed by pulse-labelling of canopy sections with (13) CO2 and the subsequent tracing of the label into microbial PLFAs. Microbial biomass was significantly higher in plots exposed to a severe experimental drought. In addition, drought induced a shift of the microbial community composition, mainly driven by an increase of Gram-positive bacteria. Drought reduced belowground C allocation, but not the transfer of recently plant-assimilated C to fungi, and in particular reduced tracer uptake by bacteria. This was accompanied by an increase of (13) C in the extractable organic C pool during drought, which was even more pronounced after plots were mown. We conclude that drought weakened the link between plant and bacterial, but not fungal, C turnover, and facilitated the growth of potentially slow-growing, drought-adapted soil microbes, such as Gram-positive bacteria.

Biocrusts enhance soil organic carbon stability and regulate the fate of new-input carbon in semiarid desert ecosystems.

Given their global prevalence, dryland (including hyperarid, arid, semiarid, and dry subhumid regions) ecosystems are critical for supporting soil organic carbon (SOC) stocks, with even small changes in such SOC pools affecting the global carbon (C) cycling. Biocrusts play an essential role in supporting C cycling in semiarid ecosystems. However, the influence of biocrusts and their successional stages on SOC and its fraction contents, as well as their role in regulating new input C into SOC fractions remain largely unknown. In this study, we collected continuous samples of bare soil (BS) and three successional stages of biocrust soils (cyanobacterial (CC), low-cover moss (LM), and high-cover moss (HM)) at 0-5 cm depth every month for one year in a semiarid desert ecosystem. We analyzed SOC changes among the samples and their fraction contents including: labile organic C (LOC) (composed of microbial biomass C (MBC), dissolved organic C (DOC), and easily oxidized organic C (EOC)) and recalcitrant organic C (ROC) fractions, soil nutrient content including: ammonium (NH4+-N), nitrate (NO3--N), and available phosphorus (AP), and soil temperature and moisture. We also conducted a 13C pulse-labelling experiment in the field to accurately quantify the effects of biocrust successional stage on exogenous C allocation to SOC fractions. Our results showed that the three successional stages of biocrust (CC-LM-HM) increased SOC and ROC contents by an average of 5.3 ± 3.6 g kg-1 and 4.0 ± 3.0 g kg-1, respectively; and the MBC, DOC, and EOC contents increased by an average of 41.7 ± 24.8 mg kg-1, 28.7 ± 12.6 mg kg-1, and 1.2 ± 0.6 g kg-1, respectively, compared to that of BS. These increases were attributed to an increase in photosynthetic pigment content, higher nutrient levels, and more suitable microclimates (e.g., higher moisture and more moderate temperature) during biocrust succession. More importantly, SOC stability was greatly improved with biocrust succession from cyanobacteria to moss, as evidenced by the reduction in soil EOC:SOC and EOC:ROC ratios by an average of 50 ± 34 % and 99 ± 67 %, respectively, while the ROC:SOC ratio increased by 33 ± 16 % with biocrust succession compared to those of BS. The biocrust SOC, DOC, and MBC 13C contents at different stages were on average 0.096 ± 0.034 mg kg-1, 0.010 ± 0.005 mg kg-1, and 0.014 ± 0.005 mg kg-1 higher than those of BS. Similarly, the allocation of new-input C among the DOC and MBC at different biocrust stages (19 ± 10 %) was significantly higher than that of BS (9 ± 6 %). New-input C into the biocrusts was fixed by microbes (43 ± 18 %) within ∼10 days and converted into other forms of C (85 ± 5 %) after 80 days. Our study provides a new perspective on how biocrusts support C cycling in semiarid desert ecosystems by mediating new C inputs into diverse fractional contents, and highlights the significance of biocrust successional stages in maintaining soil C stocks and stability in the dryland soil system.

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.

Species selection determines carbon allocation and turnover in Miscanthus crops: Implications for biomass production and C sequestration.

Growing Miscanthus species and hybrids has received strong scientific and commercial support, with the majority of the carbon (C) modelling predictions having focused on the high-yield, sterile and noninvasive hybrid Miscanthus × giganteus. However, the potential of other species with contrasting phenotypic and physiological traits has been seldom explored. To better understand the mechanisms underlying C allocation dynamics in these bioenergy crops, we pulse-labelled (13CO2) intact plant-soil systems of Miscanthus × giganteus (GIG), Miscanthus sinensis (SIN) and Miscanthus lutarioriparius (LUT) and regularly analysed soil respiration, leaves, stems, rhizomes, roots and soils for up to 190 days until leaf senescence. A rapid isotopic enrichment of all three species was observed after 4 h, with the amount of 13C fixed into plant biomass being inversely related to their respective standing biomass prior to pulse-labelling (i.e., GIG < SIN < LUT). However, both GIG and LUT allocated more photoassimilates in the aboveground biomass (leaves+stems = 78 % and 74 %, respectively) than SIN, which transferred 30% of fixed 13C in its belowground biomass (rhizomes+roots). Although less fixed 13C was recovered from the soils (<1 %), both rhizospheric and bulk soils were signficantly more enriched under SIN and LUT than under GIG. Importantly, the soils under SIN emitted less CO2, which suggests it could be the best choice for reaching C neutrality. These results from this unique large-scale study indicate that careful species selection may hold the success for reaching net GHG mitigation.

Sugarcane/soybean intercropping with reduced nitrogen addition promotes photosynthesized carbon sequestration in the soil.

Introduction: Sugarcane/soybean intercropping with reduced nitrogen (N) addition has improved soil fertility and sustainable agricultural development in China. However, the effects of intercropping pattern and N fertilizer addition on the allocation of photosynthesized carbon (C) in plant-soil system were far less understood. Methods: In this study, we performed an 13CO2 pulse labeling experiment to trace C footprints in plant-soil system under different cropping patterns [sugarcane monoculture (MS), sugarcane/soybean intercropping (SB)] and N addition levels [reduced N addition (N1) and conventional N addition (N2)]. Results and discussion: Our results showed that compared to sugarcane monoculture, sugarcane/soybean intercropping with N reduced addition increased sugarcane biomass and root/shoot ratio, which in turn led to 23.48% increase in total root biomass. The higher root biomass facilitated the flow of shoot fixed 13C to the soil in the form of rhizodeposits. More than 40% of the retained 13C in the soil was incorporated into the labile C pool [microbial biomass C (MBC) and dissolved organic C (DOC)] on day 1 after labeling. On day 27 after labeling, sugarcane/soybean intercropping with N reduced addition showed the highest 13C content in the MBC as well as in the soil, 1.89 and 1.14 times higher than the sugarcane monoculture, respectively. Moreover, intercropping pattern increased the content of labile C and labile N (alkaline N, ammonium N and nitrate N) in the soil. The structural equation model indicated that the cropping pattern regulated 13C sequestration in the soil mainly by driving changes in labile C, labile N content and root biomass in the soil. Our findings demonstrate that sugarcane/soybean intercropping with reduced N addition increases photosynthesized C sequestration in the soil, enhances the C sink capacity of agroecosystems.

Foliar water uptake as a source of hydrogen and oxygen in plant biomass.

Introductory biology lessons around the world typically teach that plants absorb water through their roots, but, unfortunately, absorption of water through leaves and subsequent transport and use of this water for biomass formation remains a field limited mostly to specialists. Recent studies have identified foliar water uptake as a significant net water source for terrestrial plants. The growing interest in the development of a new model that includes both foliar water uptake (in liquid form) and root water uptake to explain hydrogen and oxygen isotope ratios in leaf water and tree rings demands a method for distinguishing between these two water sources. Therefore, in this study, I have devised a new labelling method that utilizes two different water sources, one enriched in deuterium (HDO + D2O; δD = 7.0 × 10 4‰, δ18O = 4.1‰) and one enriched in oxygen-18 (H218O; δD = -85‰, δ18O = 1.1 × 104‰), to simultaneously label both foliar-absorbed and root-absorbed water and quantify their relative contributions to plant biomass. Using this new method, I here present evidence that, in the case of well-watered Cryptomeria japonica D. Don, hydrogen and oxygen incorporated into new leaf cellulose in the rainy season derives mostly from foliar-absorbed water (69% from foliar-absorbed water and 31% from root-absorbed water), while that of new root cellulose derives mostly from root-absorbed water (20% from foliar-absorbed water and 80% from root-absorbed water), and new branch xylem is somewhere in between (55% from foliar-absorbed water and 45% from root-absorbed water). The dual-labelling method first implemented in this study enables separate and simultaneous labelling of foliar-absorbed and root-absorbed water and offers a new tool to study the uptake, transport and assimilation processes of these waters in terrestrial plants.

Nitrogen addition and defoliation alter belowground carbon allocation with consequences for plant nitrogen uptake and soil organic carbon decomposition.

Grassland plants allocate photosynthetically fixed carbon (C) belowground to root biomass and rhizodeposition, but also to support arbuscular mycorrhizal fungi (AMF). These C allocation pathways could increase nutrient scavenging, but also mining of nutrients through enhanced organic matter decomposition. While important for grassland ecosystem functioning, methodological constraints have limited our ability to measure these processes under field conditions. We used 13CO2 and 15N pulse labelling methods to examine belowground C allocation to root biomass production, rhizodeposition and AMF colonisation during peak plant growth in a grassland field experiment after three years of N fertilisation (0 and 40 kg N ha-1 year-1) and defoliation frequency treatments ("low" and "high", with 3-4 and 6-8 simulated grazing events per year, mimicking moderate and intense grazing, respectively). Moreover, we quantified the consequences for plant nitrogen (N) uptake and decomposition of soil organic C (SOC). Nitrogen fertilisation increased rhizodeposition and AMF colonisation (by 63 % and 54 %), but reduced root biomass (by 25 %). With high defoliation frequency, AMF colonisation increased (by 60 %), but both root biomass and rhizodeposition declined (by 35 % and 58 %). Plant N uptake was highest without N fertilisation and low defoliation frequency, and positively related to root biomass and the number of root tips. Therefore, when N supply is low and the capacity to produce C through photosynthesis is high, belowground C allocation to root production and associated root tips was important to scavenge for N in the soil. In contrast, the strong positive relationship between the rate of rhizodeposition and SOC decomposition, suggests that rhizodeposition may help plants to mine for nutrients locked in SOC. Taken together, the results of this study suggest that belowground C allocation pathways affected by N fertilisation and defoliation frequency affect plant N scavenging and mining with important consequences for long-term grassland C dynamics.

In situ assessment of the velocity of carbon transfer by tracing 13 C in trunk CO2 efflux after pulse labelling: variations among tree species and seasons.

Phloem is the main pathway for transferring photosynthates belowground. In situ(13) C pulse labelling of trees 8-10 m tall was conducted in the field on 10 beech (Fagus sylvatica) trees, six sessile oak (Quercus petraea) trees and 10 maritime pine (Pinus pinaster) trees throughout the growing season. Respired (13) CO2 from trunks was tracked at different heights using tunable diode laser absorption spectrometry to determine time lags and the velocity of carbon transfer (V). The isotope composition of phloem extracts was measured on several occasions after labelling and used to estimate the rate constant of phloem sap outflux (kP ). Pulse labelling together with high-frequency measurement of the isotope composition of trunk CO2 efflux is a promising tool for studying phloem transport in the field. Seasonal variability in V was predicted in pine and oak by bivariate linear regressions with air temperature and soil water content. V differed among the three species consistently with known differences in phloem anatomy between broadleaf and coniferous trees. V increased with tree diameter in oak and beech, reflecting a nonlinear increase in volumetric flow with increasing bark cross-sectional area, which suggests changes in allocation pattern with tree diameter in broadleaf species. Discrepancies between V and kP indicate vertical changes in functional phloem properties.

The impact of prolonged drought on phloem anatomy and phloem transport in young beech trees.

Phloem failure has recently been recognized as one of the mechanisms causing tree mortality under drought, though direct evidence is still lacking. We combined 13C pulse-labelling of 8-year-old beech trees (Fagus sylvatica L.) growing outdoors in a nursery with an anatomical study of the phloem tissue in their stems to examine how drought alters carbon transport and phloem transport capacity. For the six trees under drought, predawn leaf water potential ranged from -0.7 to -2.4 MPa, compared with an average of -0.2 MPa in five control trees with no water stress. We also observed a longer residence time of excess 13C in the foliage and the phloem sap in trees under drought compared with controls. Compared with controls, excess 13C in trunk respiration peaked later in trees under moderate drought conditions and showed no decline even after 4 days under more severe drought conditions. We estimated higher phloem sap viscosity in trees under drought. We also observed much smaller sieve-tube radii in all drought-stressed trees, which led to lower sieve-tube conductivity and lower phloem conductance in the tree stem. We concluded that prolonged drought affected phloem transport capacity through a change in anatomy and that the slowdown of phloem transport under drought likely resulted from a reduced driving force due to lower hydrostatic pressure between the source and sink organs.

Short-term carbon dynamics in a temperate grassland and heathland ecosystem exposed to 104 days of drought followed by irrigation.

Temperate ecosystems are susceptible to drought events. The effect of a severe drought (104 days) followed by irrigation on the plant C uptake, its assimilation and input of C in soil were examined using a triple 13CO2 pulse-chase labelling experiment in model grassland and heathland ecosystems. First 13CO2 pulse at day 0 of the experiment revealed much higher 13C tracer uptake for shoots, roots and soil compared to the second pulse (day 44), where all plants showed significantly lower 13C tracer uptake. After the third 13CO2 pulse (day 70), very low 13C uptake in shoots led to a negligible allocation of 13C into roots and soil. During irrigation after the severe drought, the 13C tracer that was allocated in plant tissues during the second and third pulse labelling was re-allocated in roots and soil, as soon as the irrigation started. This re-allocation was higher and longer lasting in heathland compared to grassland ecosystems.

Land use in mountain grasslands alters drought response and recovery of carbon allocation and plant-microbial interactions.

Mountain grasslands have recently been exposed to substantial changes in land use and climate and in the near future will likely face an increased frequency of extreme droughts. To date, how the drought responses of carbon (C) allocation, a key process in the C cycle, are affected by land-use changes in mountain grassland is not known.We performed an experimental summer drought on an abandoned grassland and a traditionally managed hay meadow and traced the fate of recent assimilates through the plant-soil continuum. We applied two 13 CO 2 pulses, at peak drought and in the recovery phase shortly after rewetting.Drought decreased total C uptake in both grassland types and led to a loss of above-ground carbohydrate storage pools. The below-ground C allocation to root sucrose was enhanced by drought, especially in the meadow, which also held larger root carbohydrate storage pools.The microbial community of the abandoned grassland comprised more saprotrophic fungal and Gram(+) bacterial markers compared to the meadow. Drought increased the newly introduced AM and saprotrophic (A+S) fungi:bacteria ratio in both grassland types. At peak drought, the 13C transfer into AM and saprotrophic fungi, and Gram(-) bacteria was more strongly reduced in the meadow than in the abandoned grassland, which contrasted the patterns of the root carbohydrate pools.In both grassland types, the C allocation largely recovered after rewetting. Slowest recovery was found for AM fungi and their 13C uptake. In contrast, all bacterial markers quickly recovered C uptake. In the meadow, where plant nitrate uptake was enhanced after drought, C uptake was even higher than in control plots. Synthesis. Our results suggest that resistance and resilience (i.e. recovery) of plant C dynamics and plant-microbial interactions are negatively related, that is, high resistance is followed by slow recovery and vice versa. The abandoned grassland was more resistant to drought than the meadow and possibly had a stronger link to AM fungi that could have provided better access to water through the hyphal network. In contrast, meadow communities strongly reduced C allocation to storage and C transfer to the microbial community in the drought phase, but in the recovery phase invested C resources in the bacterial communities to gain more nutrients for regrowth. We conclude that the management of mountain grasslands increases their resilience to drought.

Split-root labelling to investigate 15N rhizodeposition by Pinus sylvestris and Picea abies.

We investigated the transfer of 15N into the soil via 15N uptake and release by tree roots, which involves the principles of the split-root technique. One half of the root system received an injection of (15NH4)2SO4 and the other half equivalent amounts of (NH4)2SO4 at 15N natural abundance level. 15N was transferred from one side of the root system (15N side) to the other side (14N side) and released into the soil. The method was conducted with Scots pine (Pinus sylvestris L.) and Norway spruce (Picea abies [L.] Karst). Two concentration levels of (NH4)2SO4 were used, corresponding with annual N deposition in the Netherlands (30 kg N ha-1) and a twelfth of that (2.5 kg N ha-1). Samples were taken 3 and 6 weeks after labelling and divided into needles + stem, roots, rhizosphere and bulk soil. Already 3 weeks after labelling, Scots pine took up 23.7 % of the low and 9.1 % of the high amounts of 15N, while Norway spruce took up 21.5 and 32.1 %, respectively. Both species transported proportions of 15N to the rhizosphere (0.1-0.2 %) and bulk soil (0.3-0.9 %). The method is a useful tool to investigate the fate of root-derived N in soils, for example, for the formation of stable forms of soil organic matter.

Drought history affects grassland plant and microbial carbon turnover during and after a subsequent drought event.

Drought periods are projected to become more severe and more frequent in many European regions. While effects of single strong droughts on plant and microbial carbon (C) dynamics have been studied in some detail, impacts of recurrent drought events are still little understood.We tested whether the legacy of extreme experimental drought affects responses of plant and microbial C and nitrogen (N) turnover to further drought and rewetting. In a mountain grassland, we conducted a 13C pulse-chase experiment during a naturally occurring drought and rewetting event in plots previously exposed to experimental droughts and in ambient controls (AC). After labelling, we traced 13C below-ground allocation and incorporation into soil microbes using phospholipid fatty acid biomarkers.Drought history (DH) had no effects on the standing shoot and fine root plant biomass. However, plants with experimental DH displayed decreased shoot N concentrations and increased fine root N concentrations relative to those in AC. During the natural drought, plants with DH assimilated and allocated less 13C below-ground; moreover, fine root respiration was reduced and not fuelled by fresh C compared to plants in AC.Regardless of DH, microbial biomass remained stable during natural drought and rewetting. Although microbial communities initially differed in their composition between soils with and without DH, they responded to the natural drought and rewetting in a similar way: gram-positive bacteria increased, while fungal and gram-negative bacteria remained stable. In soils with DH, a strongly reduced uptake of recent plant-derived 13C in microbial biomarkers was observed during the natural drought, pointing to a smaller fraction of active microbes or to a microbial community that is less dependent on plant C. Synthesis. Drought history can induce changes in above- vs. below-ground plant N concentrations and affect the response of plant C turnover to further droughts and rewetting by decreasing plant C uptake and below-ground allocation. DH does not affect the responses of the microbial community to further droughts and rewetting, but alters microbial functioning, particularly the turnover of recent plant-derived carbon, during and after further drought periods.