Dynamics of initial carbon allocation after drought release in mature Norway spruce-Increased belowground allocation of current photoassimilates covers only half of the carbon used for fine-root growth.

After drought events, tree recovery depends on sufficient carbon (C) allocation to the sink organs. The present study aimed to elucidate dynamics of tree-level C sink activity and allocation of recent photoassimilates (Cnew ) and stored C in c. 70-year-old Norway spruce (Picea abies) trees during a 4-week period after drought release. We conducted a continuous, whole-tree 13 C labeling in parallel with controlled watering after 5 years of experimental summer drought. The fate of Cnew to growth and CO2 efflux was tracked along branches, stems, coarse- and fine roots, ectomycorrhizae and root exudates to soil CO2 efflux after drought release. Compared with control trees, drought recovering trees showed an overall 6% lower C sink activity and 19% less allocation of Cnew to aboveground sinks, indicating a low priority for aboveground sinks during recovery. In contrast, fine-root growth in recovering trees was seven times greater than that of controls. However, only half of the C used for new fine-root growth was comprised of Cnew while the other half was supplied by stored C. For drought recovery of mature spruce trees, in addition to Cnew , stored C appears to be critical for the regeneration of the fine-root system and the associated water uptake capacity.

Additive or synergistic? Early ectomycorrhizal fungal community response to mixed tree plantings in boreal forest reclamation.

Ectomycorrhizal fungi are an important component to ecosystem function in the boreal forest. Underlying factors influencing fungal community composition and richness, such as host identity and soil type have been studied, but interactions between these factors have been less explored. Furthermore, mixed-species stands may have additive or synergistic effects on ectomycorrhizal fungi species richness, but this effect is challenging to test on natural sites due to difficulty in finding monospecific and mixed-species stands with similar site conditions and history. Forest reclamation areas can provide an opportunity to explore some of these fundamental questions, as site conditions and history are often known and managed, with the added benefit that knowledge emerging from these studies can be used to evaluate the recovery of degraded forest landscapes. Here, we compared the richness and composition of ectomycorrhizal fungi in young single- and mixed-species stands established on a reclamation area designed to inform strategies to restore upland boreal forests disturbed by oil sands mining. Seedlings of three host tree species (Populus tremuloides, Pinus banksiana, Picea glauca) were planted in single- and mixed-species stands on three different salvaged soils (forest floor material, peat, subsoil). After four growing seasons, there was no difference in total ectomycorrhizal fungi species richness and composition in mixed- versus combined single-species stands indicating that an additive effect of host tree species prevailed early in development. However, there were compositional shifts in fungal communities across both the host tree species and the salvaged soil type, with soil type being the strongest driver.

Living on next to nothing: tree seedlings can survive weeks with very low carbohydrate concentrations.

The usage of nonstructural carbohydrates (NSCs) to indicate carbon (C) limitation in trees requires knowledge of the minimum tissue NSC concentrations at lethal C starvation, and the NSC dynamics during and after severe C limitation. We completely darkened and subsequently released seedlings of two deciduous and two evergreen temperate tree species for varying periods. NSCs were measured in all major organs, allowing assessment of whole-seedling NSC balances. NSCs decreased fast in darkness, but seedlings survived species-specific whole-seedling starch concentrations as low as 0.4-0.8% per dry matter (DM), and sugar (sucrose, glucose and fructose) concentrations as low as 0.5-2.0% DM. After re-illumination, the refilling of NSC pools began within 3 wk, while the resumption of growth was delayed or restricted. All seedlings had died after 12 wk of darkness, and starch and sugar concentrations in most tissues were lower than 1% DM. We conclude that under the applied conditions, tree seedlings can survive several weeks with very low NSC reserves probably also using alternative C sources like lipids, proteins or hemicelluloses; lethal C starvation cannot be assumed, if NSC concentrations are higher than the minimum concentrations found in surviving seedlings; and NSC reformation after re-illumination occurs preferentially over growth.

Dying piece by piece: carbohydrate dynamics in aspen (Populus tremuloides) seedlings under severe carbon stress.

Carbon starvation as a mechanism of tree mortality is poorly understood. We exposed seedlings of aspen (Populus tremuloides) to complete darkness at 20 or 28 °C to identify minimum non-structural carbohydrate (NSC) concentrations at which trees die and to see if these levels vary between organs or with environmental conditions. We also first grew seedlings under different shade levels to determine if size affects survival time under darkness due to changes in initial NSC concentration and pool size and/or respiration rates. Darkness treatments caused a gradual dieback of tissues. Even after half the stem had died, substantial starch reserves were still present in the roots (1.3-3% dry weight), indicating limitations to carbohydrate remobilization and/or transport during starvation in the absence of water stress. Survival time decreased with increased temperature and with increasing initial shade level, which was associated with smaller biomass, higher respiration rates, and initially smaller NSC pool size. Dead tissues generally contained no starch, but sugar concentrations were substantially above zero and differed between organs (~2% in stems up to ~7.5% in leaves) and, at times, between temperature treatments and initial, pre-darkness shade treatments. Minimum root NSC concentrations were difficult to determine because dead roots quickly began to decompose, but we identify 5-6% sugar as a potential threshold for living roots. This variability may complicate efforts to identify critical NSC thresholds below which trees starve.

Nonstructural carbohydrate dynamics of lodgepole pine dying from mountain pine beetle attack.

Bark beetle outbreaks are an important cause of tree death, but the process by which trees die remains poorly understood. The effect of beetle attack on whole-tree nonstructural carbohydrate (NSC) dynamics is particularly unclear, despite the potential role of carbohydrates in plant defense and survival. We monitored NSC dynamics of all organs in attacked and protected lodgepole pines (Pinus contorta) during a mountain pine beetle (Dendroctonus ponderosae) outbreak in British Columbia, starting before beetle flight in June 2011 through October 2012, when most attacked trees had died. Following attack, NSC concentrations were first reduced in the attacked region of the bole. The first NSC reduction in a distant organ appeared in the needles at the end of 2011, while branch and root NSC did not decline until much later in 2012. Attacked trees that were still alive in October 2012 had less beetle damage, which was negatively correlated with initial bark sugar concentrations in the attack region. The NSC dynamics of dying trees indicate that trees were killed by a loss of water conduction and not girdling. Further, our results identify locally reduced carbohydrate availability as an important mechanism by which stressors like drought may increase tree susceptibility to biotic attack.

Reserves accumulated in non-photosynthetic organs during the previous growing season drive plant defenses and growth in aspen in the subsequent growing season.

Plants store non-structural carbohydrates (NSC), nitrogen (N), as well as other macro and micronutrients, in their stems and roots; the role of these stored reserves in plant growth and defense under herbivory pressure is poorly understood, particularly in trees. Trembling aspen (Populus tremuloides) seedlings with different NSC and N reserves accumulated during the previous growing season were generated in the greenhouse. Based on NSC and N contents, seedlings were assigned to one of three reserve statuses: Low N-Low NSC, High N-Medium NSC, or High N-High NSC. In the subsequent growing season, half of the seedlings in each reserve status was subjected to defoliation by forest tent caterpillar (Malacosoma disstria) while the other half was left untreated. Following defoliation, the effect of reserves was measured on foliar chemistry (N, NSC) and caterpillar performance (larval development). Due to their importance in herbivore feeding, we also quantified concentrations of phenolic glycoside compounds in foliage. Seedlings in Low N-Low NSC reserve status contained higher amounts of induced phenolic glycosides, grew little, and supported fewer caterpillars. In contrast, aspen seedlings in High N-Medium or High NSC reserve statuses contained lower amounts of induced phenolic glycosides, grew faster, and some of the caterpillars which fed on these seedlings developed up to their fourth instar. Furthermore, multiple regression analysis indicated that foliar phenolic glycoside concentration was related to reserve chemistry (NSC, N). Overall, these results demonstrate that reserves accumulated during the previous growing season can influence tree defense and growth in the subsequent growing season. Additionally, our study concluded that the NSC/N ratio of reserves in the previous growing season represents a better measure of resources available for use in defense and growth than the foliar NSC/N ratios.

Tracing carbon and nitrogen reserve remobilization during spring leaf flush and growth following defoliation.

Woody plants rely on the remobilization of carbon (C) and nitrogen (N) reserves to support growth and survival when resource demand exceeds supply at seasonally predictable times like spring leaf flush and following unpredictable disturbances like defoliation. However, we have a poor understanding of how reserves are regulated and whether distance between source and sink tissues affects remobilization. This leads to uncertainty about which reserves-and how much-are available to support plant functions like leaf growth. To better understand the source of remobilized reserves and constraints on their allocation, we created aspen saplings with organ-specific labeled reserves by using stable isotopes (13C,15N) and grafting unlabeled or labeled stems to labeled or unlabeled root stocks. We first determined which organs had imported root or stem-derived C and N reserves after spring leaf flush. We then further tested spatial and temporal variation in reserve remobilization and import by comparing 1) upper and lower canopy leaves, 2) early and late leaves, and 3) early flush and re-flush leaves after defoliation. During spring flush, remobilized root C and N reserves were preferentially allocated to sinks closer to the reserve source (i.e., lower vs upper canopy leaves). However, the reduced import of 13C in late versus early leaves indicates reliance on C reserves declined over time. Following defoliation, re-flush leaves imported the same proportion of root N as spring flush leaves, but they imported a lower proportion of root C. This lower import of reserve C suggests that, after defoliation, leaf re-flush rely more heavily on current photosynthate, which may explain the reduced leaf mass recovery of re-flush canopies (31% of initial leaf mass). The reduced reliance on reserves occurred even though roots retained significant starch concentrations (~5% dry wt), suggesting aspen prioritizes the maintenance of root reserves at the expense of fast canopy recovery.

Variation in carbon availability, defense chemistry and susceptibility to fungal invasion along the stems of mature trees.

If carbon (C) sinks withdraw carbohydrates as they are transported along tree stems, carbohydrate availability may depend on local sink strength and distance from sources. Defenses, including monoterpenes--a major component of resin--limit the invasibility of pines. Since carbohydrate reserves fund monoterpene synthesis, we hypothesized that monoterpene concentrations in pine stems would decrease from the crown to the lower stem, and susceptibility to fungal infection would increase. Here, we measured carbohydrate and monoterpene concentrations along the stems of lodgepole pine trees (Pinus contorta var. latifolia) before inoculating with a blue-stain fungus at different heights. After 6 wk, we assessed tree responses to fungal infection based on lesion length and carbohydrate mobilization. Concentrations of carbohydrates and monoterpenes in the phloem before inoculation decreased with distance from the crown, whereas lesion lengths after inoculation increased. However, trees mobilized sugars in response to fungal infection such that carbohydrate reserves near lesions were similar at all heights. Despite C mobilization, the lower stem was more vulnerable than the upper stem. Consistent with predictions based on sink-source relationships, vulnerability occurred where carbohydrates were less available, and likely resulted from C withdrawal by sinks higher in the supply chain.

Seed release in serotinous lodgepole pine forests after mountain pine beetle outbreak.

There are concerns that large-scale stand mortality due to mountain pine beetle (MPB) could greatly reduce natural regeneration of serotinous Rocky Mountain (RM) lodgepole pine (Pinus contorta var. latifolia) because the closed cones are held in place without the fire cue for cone opening. We selected 20 stands (five stands each of live [control], 3 years since MPB [3-yr-MPB], 6 years since MPB [6-yr-MPB], and 9 years since MPB [9-yr-MPB] mortality) in north central British Columbia, Canada. The goal was to determine partial loss of serotiny due to fall of crown-stored cones via breakage of branches and in situ opening of canopy cones throughout the 2008 and 2009 growing seasons. We also quantified seed release by the opening of forest-floor cones, loss of seed from rodent predation, and cone burial. Trees killed by MPB three years earlier dropped approximately 3.5 times more cones via branch breakage compared to live stands. After six years, MPB-killed stands had released 45% of their canopy seed bank through cone opening, cone fall due to breakage, and squirrel predation. Further losses of canopy seed banks are expected with time since we found 9-yr-MPB stands had 38% more open canopy cones. This was countered by the development of a modest forest-floor seed bank (6% of the original canopy seed bank) from burial of cones; this seed bank may be ecologically important if a fire or anthropogenic disturbance reexposes these cones. If adequate levels of regeneration are to occur, disturbances to create seedbeds must occur shortly after tree mortality, before the seed banks are lost. Our findings also suggest that the sustained seed rain (over at least nine years) after MPB outbreak may be beneficial for population growth of ground-foraging vertebrates. Our study adds insight to the seed ecology of serotinous pines under a potentially continental-wide insect outbreak, threatening vast forests adapted to regeneration after fire. Key words: biotic disturbance; cone burial; cone opening; Dendroctonus ponderosae; ground-foraging vertebrates; mountain pine beetle; natural regeneration; Pinus contorta var. latifolia; Rocky Mountain lodgepole pine; seed banks; serotiny (canopy seed storage); Tamiasciurus hudsonicus.

Viability of forest floor and canopy seed banks in Pinus contorta var. latifolia (Pinaceae) forests after a mountain pine beetle outbreak.

PREMISE OF THE STUDY: Seed banks are important for the natural regeneration of many forest species. Most of the seed bank of serotinous lodgepole pine is found in the canopy, but after an outbreak of mountain pine beetle (MPB), a considerable forest-floor seed bank develops through the falling of canopy cones. After large-scale mortality of pine stands from MPB, however, the viability of seeds in both the canopy and the forest-floor cone bank is uncertain. METHODS: We sampled cones in five stands 3 yr after MPB (3y-MPB); five stands 6 yr after MPB (6y-MPB); and 10 stands 9 yr after MPB (9y-MPB), in central British Columbia, Canada. Seeds were extracted and viability tested using germination techniques. KEY RESULTS: Forest-floor cones had seed with high germination capacity (GC): 82% for embedded (partly buried) closed cones vs. 45% for buried partly open cones. For canopy cones, GC steeply declined about 15 yr after cone maturation and by 25 yr, GC was 50%, compared with 98% in the first year. In the 3y- and 6y-MPB stands, seeds from cones that were 7 to 9 yr old had similar GC on dead and living trees; however, seeds from the dead trees had lower vigor than seeds from living trees. CONCLUSIONS: We demonstrate for the first time that a serotinous pine can form a viable soil seed bank by cone burial, which may facilitate natural regeneration if a secondary disturbance occurs. Seeds contained in 15-yr-old cones showed a steep decline in viability, which could limit regeneration if there is a long delay before a secondary disturbance.

Splitting the Difference: Heterogeneous Soil Moisture Availability Affects Aboveground and Belowground Reserve and Mass Allocation in Trembling Aspen.

When exploring the impact of resource availability on perennial plants, artificial treatments often apply conditions homogeneously across space and time, even though this rarely reflects conditions in natural systems. To investigate the effects of spatially heterogeneous soil moisture on morphological and physiological responses, trembling aspen (Populus tremuloides) saplings were used in a split-pot experiment. Following the division of the root systems, saplings were established for a full year and then subjected to either heterogeneous (portion of the root system exposed to non-lethal drought) or homogeneous (whole root system exposed to non-lethal drought or well-watered) treatments. Above- and belowground growth and non-structural carbohydrate (NSC) reserves (soluble sugars and starch) were measured to determine how allocation of reserves and mass between and within organs changed in response to variation in soil moisture availability. In contrast to saplings in the homogeneous drought treatment, which experienced reduced shoot growth, leaf abscission and fine root loss, saplings exposed to the heterogeneous conditions maintained similar aboveground growth and increased root system allocation compared to well-watered saplings. Interestingly under heterogeneous soil moisture conditions, the portion of the root system that was resource limited had no root dieback and increased carbon reserve concentrations, while the portion of the root system that was not resource limited added new roots (30% increase). Overall, saplings subjected to the heterogeneous soil moisture regime over-compensated belowground, both in mass and NSC reserves. These results indicate that the differential allocation of mass or reserves between above- and belowground organs, but also within the root system can occur. While the mechanisms and processes involved in these patterns are not clear, these responses could be interpreted as adaptations and acclimations to preserve the integrity of the entire sapling and suggests that different portions of plant organs might respond autonomously to local conditions. This study provides further appreciation of the complexity of the mechanisms by which plants manage heterogeneous conditions and offers evidence that spatial and temporal variability of resource availability, particularly belowground, needs to be accounted for when extrapolating and modeling stress responses at larger temporal and spatial scales.

Choices on sampling, sequencing, and analyzing DNA influence the estimation of community composition of plant fungal symbionts.

Plant root symbionts, namely mycorrhizal fungi, can be characterized using a variety of methods, but most of these rely on DNA. While Sanger sequencing still fulfills particular research objectives, next-generation sequencing currently dominates the field, thus understanding how the two methods differ is important for identifying both opportunities and limitations to characterizing fungal communities. In addition to testing sequencing methods, we also examined how roots and soils may yield different fungal communities and how disturbance may affect those differences. We sequenced DNA from ectomycorrhizal fungi colonizing roots of Pinus banksiana and found that operational taxonomic unit richness was higher, and compositional variance lower, for Illumina MiSeq-sequenced communities compared to Sanger-sequenced communities. We also found that fungal communities associated with roots were distinct in composition compared to those associated with soils and, moreover, that soil-associated fungi were more clustered in composition than those of roots. Finally, we found community dissimilarity between roots and soils was insensitive to disturbance; however, rarefying read counts had a sizeable influence on trends in fungal richness. Although interest in mycorrhizal communities is typically focused on the abiotic and biotic filters sorting fungal species, our study shows that the choice of methods to sample, sequence, and analyze DNA can also influence the estimation of community composition.

Hydraulic acclimation to shading in boreal conifers of varying shade tolerance.

The purpose of this study was to determine how shading affects the hydraulic and wood-anatomical characteristics of four boreal conifers (Pinus banksiana, Pinus contorta, Picea glauca and Picea mariana) that differ in shade tolerance. Plants were grown in an open field and under a deciduous-dominated overstory for 6 years. Sapwood- and leaf-area specific conductivity, vulnerability curves, and anatomical measurements (light and scanning electron microscopy) were made on leading shoots from six to nine trees of each treatment combination. There was no difference in sapwood-area specific conductivity between open-grown and understory conifers, although two of four species had larger tracheid diameters in the open. Shaded conifers appeared to compensate for small diameter tracheids by changes in pit membrane structure. Scanning electron microscopy revealed that understory conifers had thinner margo strands, greater maximum pore size in the margo, and more torus extensions. All of these trends may contribute to inadequate sealing of the torus. This is supported by the fact that all species showed increased vulnerability to cavitation when grown in the understory. Although evaporative demand in an understory environment is low, a rapid change into fully exposed conditions could be detrimental for shaded conifers.

Fire drives transcontinental variation in tree birch defense against browsing by snowshoe hares.

Fire has been the dominant disturbance in boreal America since the Pleistocene, resulting in a spatial mosaic in which the most fire occurs in the continental northwest. Spatial variation in snowshoe hare (Lepus americanus) density reflects the fire mosaic. Because fire initiates secondary forest succession, a fire mosaic creates variation in the abundance of early successional plants that snowshoe hares eat in winter, leading to geographic variation in hare density. We hypothesize that fire is the template for a geographic mosaic of natural selection: where fire is greatest and hares are most abundant, hare browsing has most strongly selected juvenile-phase woody plants for defense. We tested the hypothesis at multiple spatial scales using Alaska birch (Betula neoalaskana) and white birch (Betula papyrifera). We also examined five alternative hypotheses for geographic variation in antibrowsing defense. The fire-hare-defense hypothesis was supported at transcontinental, regional, and local scales; alternative hypotheses were rejected. Our results link transcontinental variation in species interactions to an abiotic environmental driver, fire. Intakes of defense toxins by Alaskan hares exceed those by Wisconsin hares, suggesting that the proposed selection mosaic may coincide with a geographic mosaic of coevolution.

Identifying the relevant carbohydrate storage pools available for remobilization in aspen roots.

Nonstructural carbohydrate (NSC) remobilization remains poorly understood in trees. In particular, it remains unclear (i) which tissues (e.g., living bark or xylem) and compounds (sugars or starch) in woody plants are the main sources of remobilized carbon, (ii) to what extent these NSC pools can be depleted and (iii) whether initial NSC mass or concentration is a better predictor of regrowth potential following disturbance. To address these questions, we collected root segments from a large mature trembling aspen stand; we then allowed them to resprout (sucker) in the dark and remobilize NSC until all sprouts had died. We found that initial starch mass, not concentration, was the best predictor of subsequent sprout mass. In total, more NSC mass (~4×) was remobilized from the living inner bark than the xylem of the roots. After resprouting, root starch was generally depleted to <0.6% w/w in both tissues. In contrast, a large portion of sugars appear unavailable for remobilization: sugar concentrations were only reduced to 12% w/w in the bark and 2% in the xylem. These findings suggest that in order to test whether plant processes like resprouting are limited by storage we need to (i) measure storage in the living bark, not just the xylem, (ii) consider storage pool size-not just concentration-and (iii) carefully determine which compounds are actually components of the storage pool.

Standardized protocols and procedures can precisely and accurately quantify non-structural carbohydrates.

Non-structural carbohydrates (NSCs), the stored products of photosynthesis, building blocks for growth and fuel for respiration, are central to plant metabolism, but their measurement is challenging. Differences in methods and procedures among laboratories can cause results to vary widely, limiting our ability to integrate and generalize patterns in plant carbon balance among studies. A recent assessment found that NSC concentrations measured for a common set of samples can vary by an order of magnitude, but sources for this variability were unclear. We measured a common set of nine plant material types, and two synthetic samples with known NSC concentrations, using a common protocol for sugar extraction and starch digestion, and three different sugar quantification methods (ion chromatography, enzyme, acid) in six laboratories. We also tested how sample handling, extraction solvent and centralizing parts of the procedure in one laboratory affected results. Non-structural carbohydrate concentrations measured for synthetic samples were within about 11.5% of known values for all three methods. However, differences among quantification methods were the largest source of variation in NSC measurements for natural plant samples because the three methods quantify different NSCs. The enzyme method quantified only glucose, fructose and sucrose, with ion chromatography we additionally quantified galactose, while the acid method quantified a large range of mono- and oligosaccharides. For some natural samples, sugars quantified with the acid method were two to five times higher than with other methods, demonstrating that trees allocate carbon to a range of sugar molecules. Sample handling had little effect on measurements, while ethanol sugar extraction improved accuracy over water extraction. Our results demonstrate that reasonable accuracy of NSC measurements can be achieved when different methods are used, as long as protocols are robust and standardized. Thus, we provide detailed protocols for the extraction, digestion and quantification of NSCs in plant samples, which should improve the comparability of NSC measurements among laboratories.

Carbon isotope discrimination and water stress in trembling aspen following variable retention harvesting.

Variable retention harvesting (VRH) has been proposed as a silvicultural practice to maintain biodiversity and ecosystem integrity. No previous study has examined tree carbon isotope discrimination to provide insights into water stress that could lead to dieback and mortality of trees following VRH. We measured and compared the carbon isotope ratios (delta(13)C) in stem wood of trembling aspen (Populus tremuloides Michx.) before and after VRH. Eight trees were sampled from isolated residual, edge and control (interior of unharvested stand) positions from each of seven plots in three regions (Calling Lake and Drayton Valley, Alberta and Lac Duparquet, Québec). After VRH, the general trend in mean delta(13)C was residual > edge > control trees. Although this trend is indicative of water stress in residual trees, it also suggests that edge trees received some sheltering effect, reducing their stress compared with that of residuals. A strong inverse relationship was found between the delta(13)C values and the mean annual precipitation in each region. The trend in mean delta(13)C signature was Calling Lake > Drayton Valley > Lac Duparquet trees. These results suggest that residual or edge trees in drier regions are more likely to suffer water stress following VRH. We also observed a trend of greater delta(13)C in stout trees compared with slender trees, both before and after VRH. The evidence of greater water stress in stout trees likely occurred because of a positive relationship between stem diameter and crown volume per basal area. Our results provide evidence that water stress could be the driving mechanism leading to dieback and mortality of residual trees shortly after VRH. Additionally, the results from edge trees indicate that leaving hardwood residuals in larger patches or more sheltered landscape positions could reduce the water stress to which these trees are subjected, thereby reducing dieback and mortality.

Stress differentially causes roots of tree seedlings to exude carbon.

How carbon (C) flows through plants into soils is poorly understood. Carbon exuded comes from a pool of non-structural carbohydrates (NSC) in roots. Simple models of diffusion across concentration gradients indicate that the more C in roots, the more C should be exuded from roots. However, the mechanisms underlying the accumulation and loss of C from roots may differ depending on the stress experienced by plants. Thus, stress type may influence exudation independent of NSC. We tested this hypothesis by examining the relationship between NSC in fine roots and exudation of organic C in aspen (Populus tremuloides Michx.) seedlings after exposure to shade, cold soils and drought in a controlled environment. Fine root concentrations of NSC varied by treatment. Mass-specific C exudation increased with increasing fine root sugar concentration in all treatments, but stress type affected exudation independently of sugar concentration. Seedlings exposed to cold soils exuded the most C on a per mass basis. Through 13C labeling, we also found that stressed seedlings allocated relatively more new C to exudates than roots compared with unstressed seedlings. Stress affects exudation of C via mechanisms other than changes in root carbohydrate availability.