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Tree Physiology

◐ Oxford University Press (OUP)

Preprints posted in the last 90 days, ranked by how well they match Tree Physiology's content profile, based on 24 papers previously published here. The average preprint has a 0.03% match score for this journal, so anything above that is already an above-average fit.

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Interconduit pit membranes of temperate angiosperms undergo changes in pit membrane thickness and electron density within the final growth ring

Schepers, J. R.; de Melo Silva, L.; Carmesin, C.; Huppenberger, A.; Kaack, L.; Korayem, N.; Trabi, C. L.; Jansen, S.

2026-08-03 plant biology 10.64898/2026.07.31.741446 medRxiv
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O_LIPit membranes play a crucial role in water transport between neighbouring xylem conduits by providing hydraulic safety and flow resistance. While effects of pit membranes thickness on embolism resistance and temporal changes in the ultrastructure of pit membranes have been documented between sapwood and heartwood, these changes are largely unknown within conduits of the current-year. C_LIO_LIWe studied interconduit pit membranes of branches of eight angiosperm species by sampling wood from a temperate forest over four consecutive seasons, focusing on the latest growth ring. We quantified the pit membrane thickness and greyscale intensity (as a proxy for electron density) using transmission electron microscopy and image analysis. C_LIO_LIOur observations showed considerable interspecific variation in changes to pit membrane thickness and electron density. Several species exhibited the thinnest pit membranes and highest electron density at the end of a growing season, while others showed minimal variation over time. Intra-tree variation showed that changes in pit membrane shrinkage and electron density were associated with conduit diameter: pit membranes in wide conduits showed larger modification over time than narrow ones. C_LIO_LIOur results indicate seasonal changes in the structure and chemistry of angiosperm pit membranes, even within the latest growth ring. While their shrinkage might increase resistance to flow, the occurrence of darker pit membranes indicates coating and penetration by polar lipids, affecting the behaviour of gas-liquid interfaces. We speculate that the degree of modification that pit membranes undergo is mechanistically driven by the sap flow rate, and possibly conduit dimensions. C_LI

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Stomatal and xylem plasticity, not growth rate, determines white spruce resilience to warmer and drier climates

Murphy, B. K.; Perkins, N.; Nagi, F.; Wang, S.; Muchos, T.; Boyle, J. A.; Isabel, N.; Ensminger, I.

2026-09-03 plant biology 10.64898/2026.09.02.748918 medRxiv
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In a warmer and drier climate, forest productivity will depend on trees' ability to maintain carbon uptake and hydraulic function. Whether fast-growing genotypes of boreal conifers are more vulnerable to combined climatic stress remains uncertain. Using a full-factorial field experiment, we investigated how progressive soil drying combined with extended warming affects growth, xylem development, and photosynthesis in two Picea glauca families with contrasting growth strategies. Rainout structures first reduced soil moisture from 25% to 18%, followed by a +5{degrees}C warming treatment applied using infrared heaters. During the warmest and driest period in August, air temperature reached 34.5{degrees}C in the warmed plots, while soil moisture declined to a low of 15% in the combined rainout and warming treatment. Contrary to expectations, both fast- and slow-growing white spruce families exhibited similar resilience to concurrent warming and soil drying. This finding challenges the prevailing theory that faster growth increases vulnerability to climatic stress. Despite an approximately 50% reduction in rainfall, pre-dawn water potential remained above -0.5 MPa across treatments, reflecting that seedlings were able to avoid hydraulic stress. Although the fast-growing family maintained greater height and diameter growth compared to the slow-growing family, both exhibited similar physiological and anatomical responses to warming. Warming decreased stomatal conductance, which increased intrinsic water-use efficiency. Latewood xylem traits related to hydraulic efficiency were also reduced under warming. Together, these coordinated stomatal and xylem adjustments decreased water loss and protected hydraulic function, enabling both families to maintain high photosynthesis and growth under simulated climate conditions. Overall, white spruce exhibits strong phenotypic plasticity, supporting intraspecific resilience to moderate warming and soil drying representative of projected 21st-century summer conditions for central and eastern Canada.

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Impact of secondary pests on carbon allocation in declining beech trees after the 2018-2020 drought episode

Gaertner, P.-A.; Breda, N.; Gerard, B.; Levillain, J.; Schmuck, H.; Larousse, T.; Badeau, V.; Saintonge, F.-X.; Massonnet, C.

2026-07-24 ecology 10.64898/2026.07.22.739995 medRxiv
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O_LIDuring the extreme drought period of 2018-2020, secondary pest attacks were observed in some declining beech trees (Fagus sylvatica). The main species identified are Agrilus viridis and Taphrorychus bicolor, which are known to attack beech trees more frequently after periods of intense drought. The aim of this study is to investigate if and how these two cambium-feeding insects in interaction with drought contribute to beech decline. We assessed the modification of the carbon reserves stock and carbon allocation to growth and fruiting in declining beech trees. C_LIO_LIPast radial growth, non-structural carbohydrates (NSC) concentrations in several organs and the quantity of beechnuts were measured in 38 beech trees with contrasting levels of biotic attack in 2020. Total amount of carbon reserves was calculated on the basis of NSC concentrations, and the amount of carbon allocated to growth and fruiting at the tree level was assessed using allometric relationships. C_LIO_LIBeech trees most severely attacked by secondary pests showed lower radial growth for last ten years than trees that have not been attacked. NSC concentrations and the amount of carbon reserve in the coarse roots and stems of trees were also lower in beech trees with intense biotic symptoms compared to unaffected trees. However, the amount of carbon allocated to growth and fruiting during the 2020 growing season did not differ significantly among biotic attack classes. C_LIO_LISynthesis: Regardless of biotic attack intensity, declining beech trees exhibited abnormally low carbon reserves. Trees most severely affected by insects in 2020 appeared to be the most vulnerable, as evidenced by reduced radial growth over the past decade. Future experiments are needed to explore further whether insects impacted directly carbon reserves, or whether low carbon reserves in trees is a vulnerability factor promoting insects attacks. Finally, declining beech trees allocated a similar amount of carbon to fruiting as reported in healthy stands, suggesting that fruiting is a prioritised carbon sink, even under severe drought and biotic attack. C_LI

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Beyond air-seeding: Dynamic, multiphase interactionsreveal a two-step mechanism of embolism propagation inangiosperm xylem

Jansen, S.; Kaack, L.; Ahmed, M. A.; Beikircher, B.; Bittencourt, P.; Chen, R.; Flexas, J.; Gleason, S. M.; Guha, A.; Heuret, P.; Hoeltae, T.; Ingram, S.; Jiang, X.; Jotan, P.; Jupa, R.; Kanduc, M.; Korhonen, O.; Kotowska, M. M.; Kreinert, S.; Lauren, A.; Lens, F.; Levionnois, S.; Link, R.; Lintunen, A.; Mayr, S.; McAdam, S. A. M.; Mehltreter, K.; Michaud, J.; Miranda T., M.; Mocko, K.; Mondal, P. K.; Morris, H.; Nardini, A.; Ott, J.; Paligi, S. S.; Pires, G. S.; Plavcova, L.; Ribeiro V., R.; Rimer, I.; Rosner, S.; Rowland, L.; Sack, L.; Salmon, Y.; Scheire, A.; Schepers, J.; Schneck, E.; Schu

2026-08-06 plant biology 10.64898/2026.08.05.743004 medRxiv
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BackgroundThe mechanism underlying drought-induced embolism in angiosperm xylem has been attributed to air-seeding. This concept describes the bulk flow of gas from embolised to neighbouring conduits through the penetration of gas-liquid menisci across pores in interconduit pit membranes. While there is compelling evidence for the spatial propagation of embolism, air-seeding rests on various simplifying assumptions. Among others, air-seeding presumes that xylem sap has physical properties comparable to pure water, that pit membranes can be approximated as structures with simple pores, and that embolism occurs whenever a gas-liquid interface crosses a pit membrane. ScopeRecent experimental and theoretical work demonstrates that the biophysical conditions and processes governing gas-liquid interactions at interconduit pit membranes are fundamentally more dynamic and complex than assumed by air-seeding. These phenomena include: (1) gas movement through constriction pore networks, (2) insoluble, polar lipids at conduit surfaces and interfaces, (3) dynamic surface tension of xylem sap that depends on the local packing density of interfacial lipids, (4) bubble snap-off dynamics within pit membranes, (5) surfactant-stabilized nanobubbles in sap that is oversaturated with dissolved gas, and (6) electrostatic interactions between charged interfaces. Importantly, embolism propagation involves bubble generation and embolism formation as distinct, temporarily and spatially separated processes. Embolism formation occurs when nanobubbles become unstable, whereas nanobubbles below critical stability thresholds can remain stable in sap-filled conduits. ConclusionsTogether, these findings reconfirm that pit membranes function as safety valves enabling water transport according to the cohesion-tension theory, and provide mechanistic insights into embolism propagation. They address the question why plants do not suffer constant embolism formation despite negative xylem pressures. We conclude that a revised framework explicitly accounting for the 3D structure of pit membranes, and multiphase, dynamic processes operating within them are required to explain the biophysics underlying water transport and embolism resistance in angiosperm xylem.

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Plant Water-Carbon Relations In Huanglongbing-Affected Citrus Trees And The Impact Of Hlb Exposure Duration, Fruit Developmental Stage, And Source-Sink Manipulation On These Relations

Alade, D. O.; Vashisth, T.; Vincent, C.

2026-10-06 plant biology 10.64898/2026.10.01.755948 medRxiv
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Bulk phloem flow is driven by pressure gradients generated by water drawn from the xylem, coupling source-sink transport with water status. Huanglongbing (HLB), a phloem-affecting bacterial disease of citrus, disrupts phloem transport, and causes canopy water deficit. Although improved water management can enhance HLB-affected tree health, the effects of HLB on water-driven source-sink relations are unknown. Fruits function as both carbon sink and water reservoir, but the role of fruit in regulating whole-tree water status remain poorly understood. We examined leaf-fruit water relations in HLB-affected citrus and evaluated how HLB exposure duration, fruit developmental stage, and source-sink manipulation shape these dynamics. Source-sink relations were manipulated in branches of symptomatic trees with long-term (four years) and recent (six months) HLB exposure. We girdled branches with a single fruit as the sink and defoliated at (0%, 50%, or 75%), and an ungirdled, non-defoliated branch served as control. We measured stomatal conductance, leaf hydraulic traits, leaf and fruit soluble sugars, diurnal fruit shrinkage (to estimate xylem backflow) and fruit water loss. Source-sink manipulation did not affect shoot total soluble sugar. HLB-exposure resulted in less negative osmotic potential, more negative turgor potential and reduced modulus of elasticity and relative water content at turgor loss point. HLB exposure also increased diurnal fruit shrinkage, with shrinkage responses to source-sink manipulation varying across fruit development stages. Collectively, these findings suggest that long-term HLB exposure promotes leaf acclimation to water deficits, however, high sink demand may diminish the leaf water status regulation at Stage II, when fruit sink carbon demand is highest.

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13CO2 pulse labelling reveals species-specific alterations in carbon allocation and volatile organic compound emissions under heat stress

Dumberger, S.; Stock, C.; Meischner, M.; Wannenmacher, M.; Vogt, H.; Lua-Mellmann, P.; Kuehnhammer, K.; Kreuzwieser, J.; Werner, C.; Haberstroh, S.

2026-08-25 plant biology 10.64898/2026.08.24.746709 medRxiv
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Temperate forests increasingly face extreme air temperature, but plant physiological responses, particularly alterations in carbon allocation or protection via volatile organic compound (VOC) emissions, remain poorly understood. We pulse-labelled well-watered saplings of Fagus sylvatica and Pseudotsuga menziesii in a controlled heat stress experiment with 13CO2 to quantify heat-induced shifts in CO2, VOC and C pool exchange, specifically analyzing compound-specific {delta}13C of terpenoids, water-soluble organic matter (WSOM) and dark respiration. Under heat stress, up to 50% of fresh assimilates were directed to maintenance respiration and 1-2% to VOC emissions, while net assimilation and water use efficiency decreased by 50-75% in both species. Heat directly affected metabolic processes and reduced turnover rates of fresh assimilates in F. sylvatica, but accelerated them in P. menziesii. Strong 13C labelling of some compounds, particularly acyclic ones, suggested increased de novo synthesis of specific terpenoids for heat stress protection. By tracing the fate of recently assimilated 13CO2 we demonstrate that heat stress reduces net carbon uptake and water use efficiency, disrupts turnover of C pools and increases carbon loss via respiration and de novo synthesis of specific VOCs, potentially diminishing net carbon uptake of forests under future heat extremes.

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T50 as a proxy of PS II heat tolerance: time dependence, differences in calculation methods and implications for its suitability to assess the heat tolerance of tree species

Hauck, M.; Dulamsuren, C.

2026-07-24 ecology 10.64898/2026.07.24.740464 medRxiv
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O_LIThe heat tolerance of tree species has recently received increased attention, as it is critical for the response of forest ecosystems to climate change. Most studies rely on laboratory experiments with detached leaves, where the thermostability of photosystem II (PS II) is analyzed using chlorophyll fluorescence analysis. The temperature at which the maximum quantum yield (Fv/Fm) of PS II is reduced by 50% (T50) is often used as key measure of heat tolerance despite of weak mechanistic corroboration and inconsistent definitions. We propose a new metric consisting of a critical temperature (TIP) and a corresponding fluorescence value (FIP). C_LIO_LIWe analyzed the effect of incubation time and temperature on different versions of T50. C_LIO_LIT50 was strongly dependent on the duration of heat exposure, decreasing exponentially with incubation time. T50 and TIP values derived from short-term treatments of different incubation times <1 h are not comparable, but stabilize after longer intervals of heat exposure. C_LIO_LIT50 of Fv/Fm specified without considering incubation time is a meaningless metric. Specifications derived from short-term heat treatments should be avoided, because the result is highly influenced by the experimental setup. Different calculation methods for T50 influence the result and elucidate different aspects of the heat response. C_LI

8
CRISPR/Cas9 knockout of PtaAGL16 improves post-drought recovery and shoot vitality in hybrid poplar

Fendel, A.; Wodtke, S.; Fladung, M.; Bruegmann, T.

2026-10-06 plant biology 10.64898/2026.10.04.756531 medRxiv
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Drought stress severely limits tree growth and survival, a problem intensifying under climate change due to the rising frequency of extreme weather events. Identifying genetic variants that modulate drought stress responses and enhance resilience is therefore of great interest for developing climate-adapted trees. While the MADS-box transcription factor AGL16 has been implicated in drought responses and stomatal regulation in herbaceous species, the function of its homolog in woody perennials remains largely unexplored. Here, we investigate the role of PtaAGL16 in the poplar hybrid Populus tremula x P. alba. We generated CRISPR/Cas9 knockout lines of PtaAGL16 in the poplar clone INRA 717-1B4. Plants were subjected to a severe drought stress regime followed by a re-watering recovery phase. Morphological and physiological traits, including growth, stomatal conductance, shoot vitality, and stomatal density, together with stress-associated metabolites and drought-responsive gene expression, were analyzed. All agl16 knockout lines exhibited significantly reduced drought-induced damage compared to the wild-type. While the wild-type suffered extensive defoliation and persistent stem necrosis, the knockout lines retained higher shoot vitality and resumed growth after re-watering, including the formation of new leaves. Consequently, vitality in the knockout lines reached up to 86%, compared to only 34% in the wild-type. This improved resilience was accompanied by reduced persistent oxidative membrane damage, as indicated by lower MDA levels after recovery. These findings indicate that the disruption of PtaAGL16 is associated with enhanced drought resilience in a woody perennial species and highlight its potential as a genetic target for improving climate adaptation in fast-growing forest trees.

9
Constraints on the xylem safety-efficiency trade-off

Tezeh, F. R.; Link, R. M.; Petrik, P.; Richter, R.; Weigelt, A.; Wirth, C.; Schuldt, B.

2026-09-28 plant biology 10.64898/2026.09.25.754362 medRxiv
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The xylem safety-efficiency trade-off is a central paradigm in plant hydraulics, yet empirical support remains inconsistent. In this study, we highlight how this inconsistency may arise from variation in environmental and evolutionary constraints and from differences in observational scale. We analysed 38 diffuse-porous angiosperm and 24 conifer species grown in a common garden experiment to quantify the relationship between embolism resistance (P50) and specific hydraulic conductivity (Ks) across and within species, and its differences between three species groups differing in evolutionary ancestry and eco-hydrological strategy. When pooling over all species, we found evidence for a safety-efficiency trade-off both across and within species. However, this pattern was driven almost exclusively by the mesic broadleaved species. In contrast, the across-species pattern was suppressed in coniferous species due to the evolutionary constraints on efficiency posed by their tracheid-based wood anatomy, while there was no evidence for a trade-off on either level in hydrophilic broadleaved species, which evolved under conditions that pose reduced selective pressure on embolism resistance. Our results demonstrate that the safety-efficiency trade-off is not universal, but emerges only in specific ecological and evolutionary contexts, highlighting the importance of multi-scale analyses and constraint-based frameworks for interpreting plant hydraulic strategies.

10
Pectin is a critical contributor to the mechanical properties of Chara cell walls

He, Z.; Kim, J.; Chen, Z.; Svetkey, I. D.; Rockwell, F.; Suo, Z.; Holbrook, N. M.

2026-09-24 biophysics 10.64898/2026.09.16.751926 medRxiv
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Cellulose has long been viewed as the major load-bearing component in plant primary cell walls. Recent studies, however, suggest that pectin plays a significant role in cell wall mechanics despite a kPa-scaled modulus. Here we quantify the mechanical properties of centimeter-sized internodal cell walls of Chara corallina following treatments to remove calcium crosslinks or digest pectin. We also characterize the flow of water and gas through the cell wall as a function of pressure. We found that removal of calcium crosslinks did not affect wall strength and stress-relaxation behavior, yet it decreased the elastic modulus and increased permeability to water. In contrast, pectin removal had a dramatic impact on all measurements. Samples from which pectin had been removed exhibited lower strength and stiffness and had greater stress relaxation under constant strain. Pectin removal increased the permeability of the cell wall to water above that observed when calcium was removed and reduced the pressure at which gas "tunnels" through the cell wall ~ 2-fold. Our findings support the idea that pectin locally restricts the movement of cellulose microfibrils, with implications for understanding functional properties of primary cell walls in vascular plants and developing high-performance double-network materials.

11
Large differences in photorespiration and its temperature response among temperate trees

Tiwari, R.; David, P.; Muscarella, R.

2026-08-09 plant biology 10.1101/2025.11.22.689893 medRxiv
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Photorespiration significantly influences terrestrial carbon fluxes, yet empirical measurements of its variability across tree species and temperature conditions remain limited, constraining predictions of vegetation and climate models. We quantified apparent photorespiratory CO2 loss (Lapp) and its temperature response for seven temperate broadleaf tree species in northern Europe, using in situ O2-shift measurements in Uppsala, Sweden during peak summer. Apparent loss was derived as the difference between net CO2 assimilation under ambient (Anet) and O2-free conditions at three leaf temperatures (25, 30, and 35 {degrees}C), spanning typical and heat-wave scenarios. Apparent photorespiratory CO2 loss showed pronounced interspecific variation and increased with temperature, while net photosynthesis remained relatively stable. The ratio of apparent loss to net photosynthesis ({phi} = Lapp/Anet) rose sharply with temperature, reaching species-mean values up to 0.94 at 35 {degrees}C, indicating that photorespiration can represent nearly the entirety of net carbon gain under heat stress even when leaves remain net CO2 sinks. Suppression of photorespiration under N2 and associated changes in leaf temperature systematically reallocated photosynthetic electron transport: the fraction of ambient electron transport rate (ETR) allocated to net CO2 assimilation declined with temperature, whereas the complementary fraction allocated to apparent photorespiratory loss and other O2-dependent sinks increased, with ETR-based apparent loss and its proportional expression rising steeply across the 25-35 {degrees}C range. Together, these in situ flux and partitioning measurements reveal high variability and strong temperature sensitivity in apparent photorespiration among temperate trees. Compared to crop-based parameterisations, the {phi} values we report for temperate trees are substantially higher and more temperature-dependent, providing species-specific constraints that can improve Farquhar-von Caemmerer-Berry-type vegetation model representations of photorespiration in forest ecosystems.

12
Studying the role of climate and sex in flowering dynamics of black poplar across time and space

Cazenave, X.; Marc, V.; Julien, D.; Veronique, J.; Durufle, H.

2026-10-06 plant biology 10.64898/2026.10.05.756621 medRxiv
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Global change has a major impact on rivers and their associated terrestrial compartment, the riparian forest. The black poplar (Populus nigra L.), the dominant species of the European riparian forest, is considered an indicator of the biodiversity dynamics of these ecosystems, but is threatened with extinction in several parts of its range. The species is dioecious and shows wide variability in many traits, enabling it to adapt to a broad range of environmental conditions. A detailed characterization of this natural variability is therefore a key issue both for the conservation of natural populations and for biological engineering aimed at restoring riverbanks. Phenology is both an indicator of climate change and an adaptation lever for populations, particularly in perennial species such as trees. In this study, we characterized the spatial and temporal variability in flowering phenology by studying 13 natural black poplar populations along the Loire River, France. Flowering date and duration showed substantial variability both within and between populations, and males generally flowered earlier than females. Despite this variability, the ranking of individual trees' flowering dates was highly consistent across years. Most male-female pairs shared several days of overlapping flowering each year, and this overlap was moderately stable across years. Flowering dates advanced by about five to six days per degree Celsius during early spring, with no detectable difference in sensitivity between sexes. This work contributes to the characterization of flowering variability in an economically important tree species and may inform both the conservation of genetic resources and breeding.

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Intercellular BVOC accumulation reflects sustainedantioxidant defenses without additional carbon loss underozone exposure in Eugenia uniflora

do Nascimento, A.; Anselmo-Moreira, F.; da Costa, B. R. .B.; Siqueira, M. H. P.; Furlan, C. M.; Souza, S. R.

2026-08-11 plant biology 10.64898/2026.08.10.743946 medRxiv
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Tropospheric ozone (O) is a major atmospheric pollutant that affects plant carbon metabolism, redox homeostasis, and secondary metabolism, including the biosynthesis and emission of biogenic volatile organic compounds (BVOCs). However, the contribution of BVOCs to O3 tolerance, particularly in tropical woody species, remains poorly understood. Here, we investigated whether acute O exposure (cumulative AOT40 of 3497.82 ppb h) induces alterations in photosynthetic performance, redox homeostasis, and BVOC partitioning in Eugenia uniflora. We evaluated gas exchange, photosynthetic pigments, ascorbate and glutathione pools, emitted BVOCs, modeled intercellular BVOC concentrations, and the relative carbon cost associated with BVOC emissions. O exposure significantly increased net CO2 assimilation without affecting stomatal conductance, transpiration, leaf water status, or chlorophyll concentrations, indicating maintenance of photosynthetic performance. Carotenoid concentrations and total glutathione decreased, whereas glutathione redox status was maintained. O induced marked compound-specific changes in BVOC composition and partitioning. Several monoterpenes appeared exclusively under O exposure, {gamma}-elemene emission increased significantly, and the relative distribution of individual BVOCs between the modeled intercellular and emitted pools was altered. These findings show that the response of E. uniflora to acute O exposure was characterized by interplay among carbon assimilation, glutathione redox regulation, and BVOC partitioning rather than by increased total volatile emission. Enhanced carbon assimilation occurred without additional carbon loss through BVOC release, while changes in the modeled intercellular pool indicate that part of the volatile response remained within the leaf. Our findings highlight BVOC partitioning as an important dimension of the plant response to oxidative stress and demonstrate that emission measurements alone may not fully capture the fate and potential physiological role of volatile carbon under O exposure. O_FIG O_LINKSMALLFIG WIDTH=178 HEIGHT=200 SRC="FIGDIR/small/743946v1_ufig1.gif" ALT="Figure 1"> View larger version (52K): org.highwire.dtl.DTLVardef@653af1org.highwire.dtl.DTLVardef@ca5forg.highwire.dtl.DTLVardef@1e641bforg.highwire.dtl.DTLVardef@1e68fae_HPS_FORMAT_FIGEXP M_FIG C_FIG BVOC Partitioning Contributes to Oxidative Stress Defence Under Acute O Exposure

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Physiological signatures but no evidence for universal genomic predictors of extreme drought responses in European beech

Li, C.; de Witte, L. C.; Czyz, E. A.; Moradi, A.; Braun, S.; Hopf, S.-E.; Schmid, B.; Schaepman, M.; Schuman, M.

2026-09-22 ecology 10.64898/2026.09.20.752377 medRxiv
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Following a severe 2018 drought affecting much of Europe, European beech (Fagus sylvatica) exhibited patchy canopy dieback. To investigate the drivers of this mortality, we used a paired case-control design across Swiss forests, integrating phenotypic assessments and genomic analyses. Phenotypically, crown transparency and discoloration were used to classify tree damage state, while several other characteristics were measured to evaluate whether paired, neighboring damaged and undamaged trees were otherwise comparable, and to identify physiological stress signatures. Elevated leaf calcium content and red edge reflectance in dried leaves emerged as robust indicators of physiological stress. Genomic association models using reference-based SNP markers and reference-free k-mers were heavily confounded by micro-environmental variance and local population stratification even within our geographically limited study region. Our cross-validated genomic outliers showed no spatial overlap with predictive drought-associated loci previously identified in an independent German cohort. These findings indicate that drought survival in European beech is not governed by a universal set of major-effect alleles. Instead, tree performance appears highly context-dependent, likely shaped by micro-environmental heterogeneity and phenotypic condition. However, our results do not exclude polygenic drought responses involving multiple small-effect loci.

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The rhizosphere of Picea abies is a hotspot of terpenoid production

Meischner, M.; Steuerle, A.; Rinnan, R.; Werner, C.

2026-08-13 plant biology 10.64898/2026.08.12.744374 medRxiv
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Forest soils are an important source of volatile organic compounds (VOCs), yet little is known about how different tree species influence soil VOC emissions and the role of rhizosphere processes in mediating VOC release form roots. We analysed soil VOC emissions from the soil surface and bulk soil as well as from roots with intact rhizosphere and washed roots of Picea abies and Fagus sylvatica. Tree saplings were grown on natural forest soil, and VOC emissions and gas exchange of soils and roots were measured under controlled conditions using online gas analysers integrated into an automated system. To assess the contribution of rhizosphere soil and microbial communities to root VOC emissions, roots were analysed (a) without washing, preserving the rhizosphere, (b) water-washed, and (c) ethanol-washed (70 vol%) to minimize microbial contributions. Species-specific VOC emission patterns were observed in both soils and roots. P. abies showed higher total emission rates and a more diverse, terpenoid-rich VOC profile dominated by -pinene, {beta}-pinene, {beta}-myrcene, and -phellandrene than F. sylvatica. Notably, these differences were evident not only at the soil surface but also in root and litter free bulk soil. Root washing further revealed that the rhizosphere is a hotspot of terpenoid production in P. abies, with significantly higher monoterpenoid emissions from unwashed roots than from water or ethanol-washed roots. This study demonstrates how tree species shape net soil VOC emissions, potentially leading to cascading effects on atmospheric VOC concentrations, and highlights the importance of the rhizosphere in regulating belowground VOC production.

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Early warning indicators for heat-induced mortality in temperate tree saplings

Stock, C.; Dumberger, S.; Meischner, M.; Wannenmacher, M.; Kuehnhammer, K.; Kreuzwieser, J.; Haberstroh, S.; Werner, C.

2026-08-23 ecology 10.64898/2026.08.18.745401 medRxiv
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{middle dot} Globally, forest ecosystems face widespread mortality events. However, the independent impacts of distinct stressors, such as heat stress vs edaphic drought, remain poorly understood and physiological early warning indicators for tree mortality are urgently required. {middle dot} We exposed well-watered saplings of Fagus sylvatica, Pseudotsuga menziesii and Picea abies to summer heat waves and subsequent natural winter-desiccation. Physiological parameters (e.g. gas exchange, water uptake velocity via 2H labelling, and volatile organic compound emissions) were monitored throughout the growing season and survival was assessed regularly until subsequent spring to capture immediate and delayed mortality as a consequence of legacy effects. {middle dot} Heat exposure without soil water deficit, followed by winter desiccation, triggered species-specific mortality rates (51.8% F. sylvatica, 48.2% P. abies, 16.9% P. menziesii), with P. abies exhibiting significantly faster mortality response than the other species. Reduced water uptake, lower stomatal conductance, impaired photosynthetic efficiency, and altered VOC emissions distinguished non-surviving from surviving saplings months before visible damage in all three species. {middle dot} Heat stress drives mortality independent of edaphic drought, with sub-lethal physiological indicators detectable up to 10 months before visual signs. These early warning indicators could enable damage detection before lethal thresholds are crossed, offering new strategies for mitigating climate change-driven forest decline.

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Reconciling intrinsic and ecosystem water-use efficiency reveals physiological mechanisms underlying carbon-water coupling across biological scales

Puchi, P. F.; Dalmonech, D.; Castagneri, D.; Vangi, E.; Lehmann, M. H.; Saurer, M.; Klesse, S.; Hortnagl, L.; Collalti, A.

2026-10-08 ecology 10.64898/2026.10.07.757244 medRxiv
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Understanding how carbon-water coupling scales from leaves to ecosystems remains a major challenge because water-use efficiency (WUE) metrics integrate physiological, hydrological, and ecological processes. We integrated tree-ring width, triple stable isotopes ({delta}13C, {delta}18O and {delta}2H), eddy covariance (EC) observations, and a process-based model to investigate the drivers of intrinsic (iWUE) and ecosystem water-use efficiency (WUEeco) across biological scales in a mature Fagus sylvatica forest between 2004 and 2023. Modelled and isotope-derived iWUE were strongly correlated (r = 0.61, p < 0.05) and both indicated an approximately 20% long-term increase, consistent with increasing gross primary productivity. In contrast, WUEeco derived from EC was lower than model estimates and exhibited a positive trend not reproduced by the model. This divergence primarily reflected differences in biological representation and hydrological integration, as the single-species model underestimated ecosystem evapotranspiration by excluding co-occurring deciduous and evergreen species together with understory vegetation. Triple-isotope analyses showed that the long-term increase in iWUE was driven by enhanced photosynthetic assimilation rather than sustained stomatal closure, whereas {delta}18O and {delta}2H variability was mainly controlled by source-water dynamics. These findings provide a mechanistic framework for reconciling leaf- and ecosystem-scale WUE and improving predictions of forest carbon-water coupling under climate change.

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Recurring daytime and nighttime modes of VOC emissions in a cool-temperate oak forest

Sekimoto, K.; Suyama, Y.; Kita, Y.; Fukuyama, D.; Koss, A.; Matsukami, A.; Tomihira, S.; Yamagishi, H.; Shiojiri, K.; Saito, T.; Yazaki, K.

2026-07-24 physiology 10.64898/2026.07.20.739470 medRxiv
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Plants emit substantial amounts of biogenic volatile organic compounds (VOCs) that link plant physiological activity to ecological interactions and atmospheric chemistry. However, the processes regulating VOC emissions within the forest air and at the interface directly above the canopy remain poorly characterized. In this study, we investigated forest-scale VOC dynamics in a cool-temperate deciduous forest dominated by Quercus crispula using high-time- and high-mass-resolution proton-transfer-reaction time-of-flight mass spectrometry (PTR-ToF-MS) integrated with positive matrix factorization (PMF). This non-targeted, process-oriented framework was applied to forest-interior and canopy-top atmospheres during rain-free summer days in 2024 and 2025 to extract dominant modes of VOC emissions. The PMF consistently resolved two recurring modes characterized by the daytime and nighttime enhancement patterns. The daytime mode was dominated by isoprene and its oxidation products and demonstrated strong light- and temperature-dependence, whereas the nighttime mode was enriched in mono- and sesquiterpenes. The daytime contribution exhibited a pronounced morning-afternoon asymmetry, indicating non-linear physiological and canopy-scale controls. These patterns were reproducible across the years. This study demonstrates that combining PTR-ToF-MS with PMF enables robust, top-down identification of recurring modes of forest VOC variability within and above forest canopies, linking leaf-level physiology and ecosystem-scale atmospheric exchange. HighlightForest-scale VOC emissions were resolved into recurring daytime and nighttime modes using a non-targeted PTR-ToF-MS and PMF framework.

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Stomatal sensitivity to VPD across 38 tree species reflects past drought responses but is little explained by stomatal and leaf economic traits

Sachsenmaier, L.; Richter, R.; Ahner, C.; Kretz, L.; Kahl, A.; Schellenberger Costa, D.; Schnabel, F.; Staude, I.; Sabot, M.; Wirth, C.

2026-09-11 ecology 10.64898/2026.09.09.750341 medRxiv
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Climate change is exposing trees to rising atmospheric water demand. Because stomata regulate plant water loss, understanding stomatal behaviour is critical. However, few comparative studies examine species-specific stomatal responses to atmospheric demand. Here, we quantify stomatal behaviour in 38 temperate tree species (27 broadleaved, 11 conifers) in a German research arboretum during a warm summer without soil drought. We measured the diurnal stomatal conductance (gs) of 787 leaves and modelled gs species-specific responses to leaf-to-air vapour pressure deficit (VPDL). Four descriptors of the gs-VPDL relationship were derived: maximum stomatal conductance (gmax), the VPDL that maximises gs, the VPDL associated with a 50% reduction in gs, and the stomatal closure rate. We relate these metrics to drought-induced shifts of carbon isotope ratios and to leaf functional traits. Large variation in stomatal behaviour exists, ranging from early-closure to VPDL-tolerant strategies. Species maintaining gs at higher VPDL also show weaker physiological responses to past drought. gmax is partly explained by leaf economic traits; other gs-VPDL descriptors are not related to stomatal anatomy and leaf economy. Our interspecific comparison reveals substantial variation in stomatal responses to atmospheric demand that is relevant to drought responses, but cannot be readily inferred from commonly measured leaf traits.

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Switchgrass Root Cell Wall Composition and Anatomy Vary with Depth, Suggesting Approaches for Trait Enhancement

Panahabadi, R.; Jewell, J. B.; Biswal, A. K.; Engle, N. L.; Nonavinakere Chandrakanth, N.; Poisson, J.; Mohanty, S. S.; Tschaplinski, T. J.; Mohnen, D.; Harman-Ware, A. E.; Bartley, L. E.

2026-08-19 plant biology 10.64898/2026.08.14.744798 medRxiv
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Plant root cellular architecture and cell wall composition influence plant productivity, stress resilience, biotic interactions, and potentially soil carbon accumulation. This study establishes comprehensive compositional parameters for roots of a lowland switchgrass genotype, DVR3. Root traits were analyzed in 12.5 cm depth segments, from Zone 1 near the surface to Zone 4 down to 50 cm. Mean abundance ({micro}g/mg) for major cell wall components included cellulose 470 {+/-} 20, xylose 250 {+/-} 20, lignin 170 {+/-} 15, and total suberin 35 {+/-} 5. Composition and cellular anatomy varied with depth, in a partially coordinated manner. Cross sections showed extensive aerenchyma in mature root regions despite greater root mass density, corresponding to abundant lignin and cellulose. Deep roots were enriched for pectin-associated traits, including arabinogalactan II, homogalacturonan, and arabinose-associated linkages. Suberin content did not vary significantly, though Casparian strip formation, endoderm and exoderm thickening, and suberin surface staining progressed with development. Similar trends in root lignin and specific root length were observed for another lowland switchgrass genotype, AP13. These results suggest that it may be possible to genetically enhance native switchgrass root chemistry to promote soil penetration and below-ground carbon accumulation by reducing variability with development, potentially via cell-type specific adjustments. HighlightOlder, shallower switchgrass crown roots are enriched in lignin and cellulose, and deeper, younger roots are pectin-rich with juvenile cellular anatomy. A more uniform compositional distribution might enhance below-ground traits. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=120 SRC="FIGDIR/small/744798v1_ufig1.gif" ALT="Figure 1"> View larger version (47K): org.highwire.dtl.DTLVardef@159de98org.highwire.dtl.DTLVardef@124d714org.highwire.dtl.DTLVardef@1a49c14org.highwire.dtl.DTLVardef@2fa67_HPS_FORMAT_FIGEXP M_FIG C_FIG Schematic summary of switchgrass root anatomy and composition across four 12.5-cm depth zones of a 50-cm root system. Zone 1 represents older, shallow roots and Zone 4 includes younger roots and root tips. Representative cross-sections show greater aerenchyma development in older roots than in young root tips. The compositional heatmap shows higher cellulose, lignin, and xylose in Zone 1, higher pectin and nitrogen in Zone 4, and relatively little variation in suberin across zones.