Residual water losses mediate the trade-off between growth and drought-survival across saplings of 12 tropical rainforest tree species with contrasting hydraulic strategies.

Knowledge of the physiological mechanisms underlying species vulnerability to drought is critical to better understand patterns of tree mortality. Investigating plant adaptive strategies to drought should thus help to fill this knowledge gap, especially in tropical rainforests exhibiting high functional diversity. In a semi-controlled drought experiment on 12 rainforest tree species, we investigated the diversity in hydraulic strategies and whether they determined the ability of saplings to use stored non-structural carbohydrates during an extreme imposed drought. We further explored the importance of water- and carbon-use strategies in relation to drought-survival through a modelling approach. Hydraulic strategies varied considerably across species with a continuum between dehydration- tolerance and -avoidance. During dehydration leading to hydraulic failure and irrespective of hydraulic strategies, species showed strong declines in whole-plant starch concentrations and a maintenance or even an increase in soluble sugar concentrations potentially favouring osmotic adjustments. Residual water losses mediated the trade-off between time to hydraulic failure and growth, suggesting that it is linked to the 'fast-slow' continuum of plant performances and that dehydration avoidance is an effective drought-survival strategy at the sapling stage. Further investigations on residual water losses may be key to understanding the response of tropical rainforest tree communities to climate change.

Functional trade-offs are driven by coordinated changes among cell types in the wood of angiosperm trees from different climates.

Wood performs several functions to ensure tree survival and carbon allocation to a finite stem volume leads to trade-offs among cell types. It is not known to what extent these trade-offs modify functional trade-offs and if they are consistent across climates and evolutionary lineages. Twelve wood traits were measured in stems and coarse roots across 60 adult angiosperm tree species from temperate, Mediterranean and tropical climates. Regardless of climate, clear trade-offs occurred among cellular fractions, but did not translate into specific functional trade-offs. Wood density was negatively related to hydraulic conductivity (Kth ) in stems and roots, but was not linked to nonstructural carbohydrates (NSC), implying a functional trade-off between mechanical integrity and transport but not with storage. NSC storage capacity was positively associated with Kth in stems and negatively in roots, reflecting a potential role for NSC in the maintenance of hydraulic integrity in stems but not in roots. Results of phylogenetic analyses suggest that evolutionary histories cannot explain covariations among traits. Trade-offs occur among cellular fractions, without necessarily modifying trade-offs in function. However, functional trade-offs are driven by coordinated changes among xylem cell types depending on the dominant role of each cell type in stems and roots.

Parenchyma fractions drive the storage capacity of nonstructural carbohydrates across a broad range of tree species.

PREMISE: Nonstructural carbohydrates (NSCs) play a key role in tree performance and functioning and are stored in radial and axial parenchyma (RAP) cells. Whether this relationship is altered among species and climates or is linked to functional traits describing xylem structure (wood density) and tree stature is not known. METHODS: In a systematic review, we collated data for NSC content and the proportion of RAP in stems for 68 tree species. To examine the relationships of NSCs and RAP with climatic factors and other functional traits, we also collected climatic data at each tree's location, as well as wood density and maximum height. A phylogenetic tree was constructed to examine the influence of species' evolutionary relationships on the associations among NSCs, RAP, and functional traits. RESULTS: Across all 68 tree species, NSCs were positively correlated with RAP and mean annual temperature, but relationships were only weakly significant in temperate species and angiosperms. When separating RAP into radial parenchyma (RP) and axial parenchyma (AP), both NSCs and wood density were positively correlated with RP but not with AP. Wood in taller trees was less dense and had lower RAP than in shorter trees, but height was not related to NSCs. CONCLUSIONS: In trees, NSCs are stored mostly in the RP fraction, which has a larger surface area in warmer climates. Additionally, NSCs were only weakly linked to wood density and tree height. Our analysis of evolutionary relationships demonstrated that RAP fractions and NSC content were always closely related across all 68 tree species, suggesting that RAP can act as a reliable proxy for potential NSC storage capacity in tree stems.

Drought effects on resource partition and conservation among leaf ontogenetic stages in epiphytic tank bromeliads.

Studying the response to drought stress of keystone epiphytes such as tank bromeliads is essential to better understand their resistance capacity to future climate change. The objective was to test whether there is any variation in the carbon, water and nutrient status among different leaf ontogenetic stages in a bromeliad rosette subjected to a gradient of drought stress. We used a semi-controlled experiment consisting in a gradient of water shortage in Aechmea aquilega and Lutheria splendens. For each bromeliad and drought treatment, three leaves were collected based on their position in the rosette and several functional traits related to water and nutrient status, and carbon metabolism were measured. We found that water status traits (relative water content, leaf succulence, osmotic and midday water potentials) and carbon metabolism traits (carbon assimilation, maximum quantum yield of photosystem II, chlorophyll and starch contents) decreased with increasing drought stress, while leaf soluble sugars and carbon, nitrogen and phosphorus contents remained unchanged. The different leaf ontogenetic stages showed only marginal variations when subjected to a gradient of drought. Resources were not reallocated between different leaf ontogenetic stages but we found a reallocation of soluble sugars from leaf starch reserves to the root system. Both species were capable of metabolic and physiological adjustments in response to drought. Overall, this study advances our understanding of the resistance of bromeliads faced with increasing drought stress and paves the way for in-depth reflection on their strategies to cope with water shortage.

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.

Fast growth involves high dependence on stored resources in seedlings of Mediterranean evergreen trees.

BACKGROUND AND AIMS: The carbon (C) and nitrogen (N) needed for plant growth can come either from soil N and current photosynthesis or through remobilization of stored resources. The contribution of remobilization to new organ growth on a whole-plant basis is quite well known in deciduous woody plants and evergreen conifers, but this information is very limited in broadleaf evergreen trees. This study compares the contribution of remobilized C and N to the construction of new organs in spring, and assesses the importance of different organs as C and N sources in 1-year-old potted seedlings of four ecologically distinct evergreen Mediterranean trees, namely Quercus ilex, Q. coccifera, Olea europaea and Pinus hapelensis. METHODS: Dual (13)C and (15)N isotope labelling was used to unravel the contribution of currently taken up and stored C and N to new growth. Stored C was labelled under simulated winter conditions. Soil N was labelled with the fertilization during the spring growth. KEY RESULTS: Oaks allocated most C assimilated under simulated winter conditions to coarse roots, while O. europaea and P. halepensis allocated it to the leaves. Remobilization was the main N source (>74 %) for new fine-root growth in early spring, but by mid-spring soil supplied most of the N required for new growth (>64 %). Current photosynthesis supplied >60 % of the C in new fine roots by mid-spring in most species. Across species, the proportion of remobilized C and N in new shoots increased with the relative growth rate. Quercus species, the slowest growing trees, primarily used currently acquired resources, while P. halepensis, the fastest growing species, mainly used reserves. Increases in the amount of stored N increased N remobilization, which fostered absolute growth both within and across species. Old leaves were major sources of remobilized C and N, but stems and roots also supplied considerable amounts of both in all species except in P. halepensis, which mainly relied on foliage formed in the previous growing season to supply stored resources. CONCLUSIONS: Seedlings of Mediterranean evergreen trees have distinct C and N storage physiologies, with relative growth rate driving the contribution of remobilized resources to new growth. These differences may reduce competition and facilitate species coexistence.

Does long-term drought or repeated defoliation affect seasonal leaf N cycling in young beech trees?

Forest trees adopt effective strategies to optimize nitrogen (N) use through internal N recycling. In the context of more recurrent environmental stresses due to climate change, the question remains of whether increased frequency of drought or defoliation threatens this internal N recycling strategy. We submitted 8-year-old beech trees to 2 years of either severe drought (Dro) or manual defoliation (Def) to create a state of N starvation. At the end of the second year before leaf senescence, we labeled the foliage of the Dro and Def trees, as well as that of control (Co) trees, with 15N-urea. Leaf N resorption, winter tree N storage (total N, 15N, amino acids, soluble proteins) and N remobilization in spring were evaluated for the three treatments. Defoliation and drought did not significantly impact foliar N resorption or N concentrations in organs in winter. Total N amounts in Def tree remained close to those in Co tree, but winter N was stored more in the branches than in the trunk and roots. Total N amount in Dro trees was drastically reduced (-55%), especially at the trunk level, but soluble protein concentrations increased in the trunk and fine roots compared with Co trees. During spring, 15N was mobilized from the trunk, branches and twigs of both Co and Def trees to support leaf growth. It was only provided through twig 15N remobilization in the Dro trees, thus resulting in extremely reduced Dro leaf N amounts. Our results suggest that stress-induced changes occur in N metabolism but with varying severity depending on the constraints: within-tree 15N transport and storage strategy changed in response to defoliation, whereas a soil water deficit induced a drastic reduction of the N amounts in all the tree organs. Consequently, N dysfunction could be involved in drought-induced beech tree mortality under the future climate.

Elevated CO2 and/or ozone modify lignification in the wood of poplars (Populus tremula x alba).

Trees will have to cope with increasing levels of CO(2) and ozone in the atmosphere. The purpose of this work was to assess whether the lignification process could be altered in the wood of poplars under elevated CO(2) and/or ozone. Young poplars were exposed either to charcoal-filtered air (control), to elevated CO(2) (800 µl l(-1)), to ozone (200 nl l(-1)) or to a combination of elevated CO(2) and ozone in controlled chambers. Lignification was analysed at different levels: biosynthesis pathway activities (enzyme and transcript), lignin content, and capacity to incorporate new assimilates by using (13)C labelling. Elevated CO(2) and ozone had opposite effects on many parameters (growth, biomass, cambial activity, wood cell wall thickness) except on lignin content which was increased by elevated CO(2) and/or ozone. However, this increased lignification was due to different response mechanisms. Under elevated CO(2), carbon supply to the stem and effective lignin synthesis were enhanced, leading to increased lignin content, although there was a reduction in the level of some enzyme and transcript involved in the lignin pathway. Ozone treatment induced a reduction in carbon supply and effective lignin synthesis as well as transcripts from all steps of the lignin pathway and some corresponding enzyme activities. However, lignin content was increased under ozone probably due to variations in other major components of the cell wall. Both mechanisms seemed to coexist under combined treatment and resulted in a high increase in lignin content.

Non-structural carbohydrates and morphological traits of leaves, stems and roots from tree species in different climates.

OBJECTIVES: Carbon fixed during photosynthesis is exported from leaves towards sink organs as non-structural carbohydrates (NSC), that are a key energy source for metabolic processes in trees. In xylem, NSC are mostly stored as soluble sugars and starch in radial and axial parenchyma. The multi-functional nature of xylem means that cells possess several functions, including water transport, storage and mechanical support. Little is known about how NSC impacts xylem multi-functionality, nor how NSC vary among species and climates. We collected leaves, stem and root xylem from tree species growing in three climates and estimated NSC in each organ. We also measured xylem traits linked to hydraulic and mechanical functioning. DATA DESCRIPTION: The paper describes functional traits in leaves, stems and roots, including NSC, carbon, nitrogen, specific leaf area, stem and root wood density and xylem traits. Data are provided for up to 90 angiosperm species from temperate, Mediterranean and tropical climates. These data are useful for understanding the trade-offs in resource allocation from a whole-plant perspective, and to better quantify xylem structure and function related to water transportation, mechanical support and storage. Data will also give researchers keys to understanding the ability of trees to adjust to a changing climate.

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.

Inflorescence of grapevine (Vitis vinifera L.): a high ability to distribute its own assimilates.

The distribution of carbon (C) into whole grapevine fruiting cuttings was investigated during flower development to determine the relative contribution of inflorescence and leaf photoassimilates in the total C balance and to investigate their partitioning towards other plant organs. A (13)C labelling procedure was used to label C photoassimilates by leaves and inflorescences in grapevine. Investigations were carried out at various stages of flower/berry development, from separated cluster to fruit set, using grapevine fruiting cuttings with four leaves (Vitis vinifera L. cv. Chardonnay). This is the first study reporting that, during its development, (i) the carbon needs of the inflorescence were met by both leaf and inflorescence photosynthesis, and (ii) the inflorescence amazingly participated significantly to the total C balance of grapevine cuttings by redistributing an important part of its own assimilates to other plant organs. With regard to flowering, 29% of C assimilated by the inflorescence remained in the inflorescence, while partitioning towards the stem reached 42% and, as a lower proportion, 15% in leaves, and 14% in roots.

Tracing of recently assimilated carbon in respiration at high temporal resolution in the field with a tuneable diode laser absorption spectrometer after in situ 13CO2 pulse labelling of 20-year-old beech trees.

The study of the fate of assimilated carbon in respiratory fluxes in the field is needed to resolve the residence and transfer times of carbon in the atmosphere-plant-soil system in forest ecosystems, but it requires high frequency measurements of the isotopic composition of evolved CO2. We developed a closed transparent chamber to label the whole crown of a tree and a labelling system capable of delivering a 3-h pulse of 99% 13CO2 in the field. The isotopic compositions of trunk and soil CO2 effluxes were recorded continuously on two labelled and one control trees by a tuneable diode laser absorption spectrometer during a 2-month chase period following the late summer labelling. The lag times for trunk CO2 effluxes are consistent with a phloem sap velocity of about 1 m h(-1). The isotopic composition (delta13C) of CO2 efflux from the trunk was maximal 2-3 days after labelling and declined thereafter following two exponential decays with a half-life of 2-8 days for the first and a half-life of 15-16 days for the second. The isotopic composition of the soil CO2 efflux was maximal 3-4 days after labelling and the decline was also well fitted with a sum of two exponential functions with a half-life of 3-5 days for the first exponential and a half-life of 16-18 days for the second. The amount of label recovered in CO2 efflux was around 10-15% of the assimilated 13CO2 for soil and 5-13% for trunks. As labelling occurred late in the growing season, substantial allocation to storage is expected.

Short-term nitrogen dynamics are impacted by defoliation and drought in Fagus sylvatica L. branches.

The predicted recurrence of adverse climatic events such as droughts, which disrupt nutrient accessibility for trees, could jeopardize the nitrogen (N) metabolism in forest trees. Internal tree N cycling capacities are crucial to ensuring tree survival but how the N metabolism of forest trees responds to intense, repeated environmental stress is not well known. For 2 years, we submitted 9-year-old beech (Fagus sylvatica L.) trees to either a moderate or a severe prolonged drought or a yearly removal of 75% of the foliage to induce internal N cycling changes. During the second year of stress, in spring and summer, we sprayed 15N-urea on the leaves (one branch per tree). Then, for 14 days, we traced the 15N dynamics through the leaves, into foliar proteins and into the branch compartments (leaves and stems segments), as well as its long-distance transfer from the labeled branches to the tree apical twigs. Defoliation caused a short- and mid-term N increase in the leaves, which remained the main sink for N. Whatever the treatment and the date, most of the leaf 15N stayed in the leaves and was invested in soluble proteins (60-68% of total leaf N). 15N stayed more in the proximal part of the branch in response to drought compared with other treatments. The long-distance transport of N was maintained even under harsh drought, highlighting efficient internal N recycling in beech trees. Under extreme constraints creating an N and water imbalance, compensation mechanisms operated at the branch level in beech trees and allowed them (i) to maintain leaf N metabolism and protein synthesis and (ii) to ensure the seasonal short- and long-distance transfer of recycled leaf N even under drastic water shortage conditions.

Do elevation of CO(2) concentration and nitrogen fertilization alter storage and remobilization of carbon and nitrogen in pedunculate oak saplings?

Soil nitrogen can alter storage and remobilization of carbon and nitrogen in forest trees and affect growth responses to elevated carbon dioxide concentration ([CO(2)]). We investigated these effects in oak saplings (Quercus robur L.) exposed for two years to ambient or twice ambient [CO(2)] in combination with low- (LN, 0.6 mmol N l(-1)) or high-nitrogen (HN, 6.1 mmol N l(-1)) fertilization. Autumn N retranslocation efficiency from senescing leaves was less in HN saplings than in LN saplings, but about 15% of sapling N was lost to the litter. During the dormant season, nonstructural carbohydrates made up 20 to 30% of the dry mass of perennial organs. Starch was stored mainly in large roots where it represented 35-46% of dry mass. Accumulation of starch increased in large roots in response to LN but was unaffected by elevated [CO(2)]. The HN treatment resulted in high concentrations of N-soluble compounds, and this effect was reduced by elevated [CO(2)], which decreased soluble protein N (-17%) and amino acid N (-37%) concentrations in the HN saplings. Carbon and N reserves were labeled with (13)C and (15)N, respectively, at the end of the first year. In the second year, about 20% of labeled C and 50% of labeled N was remobilized for spring growth in all treatments. At the end of leaf expansion, 50-60% of C in HN saplings originated from assimilation versus only 10-20% in LN saplings. In HN saplings only, N uptake occurred, and some newly assimilated N was allocated to new shoots. Through effects on the C and N content of perennial organs, elevated [CO(2)] and HN increased remobilization capacity, thereby supporting multiple shoot flushes, which increased leaf area and subsequent C acquisition in a positive feedback loop.

Non-structural carbohydrates in woody plants compared among laboratories.

Non-structural carbohydrates (NSC) in plant tissue are frequently quantified to make inferences about plant responses to environmental conditions. Laboratories publishing estimates of NSC of woody plants use many different methods to evaluate NSC. We asked whether NSC estimates in the recent literature could be quantitatively compared among studies. We also asked whether any differences among laboratories were related to the extraction and quantification methods used to determine starch and sugar concentrations. These questions were addressed by sending sub-samples collected from five woody plant tissues, which varied in NSC content and chemical composition, to 29 laboratories. Each laboratory analyzed the samples with their laboratory-specific protocols, based on recent publications, to determine concentrations of soluble sugars, starch and their sum, total NSC. Laboratory estimates differed substantially for all samples. For example, estimates for Eucalyptus globulus leaves (EGL) varied from 23 to 116 (mean = 56) mg g(-1) for soluble sugars, 6-533 (mean = 94) mg g(-1) for starch and 53-649 (mean = 153) mg g(-1) for total NSC. Mixed model analysis of variance showed that much of the variability among laboratories was unrelated to the categories we used for extraction and quantification methods (method category R(2) = 0.05-0.12 for soluble sugars, 0.10-0.33 for starch and 0.01-0.09 for total NSC). For EGL, the difference between the highest and lowest least squares means for categories in the mixed model analysis was 33 mg g(-1) for total NSC, compared with the range of laboratory estimates of 596 mg g(-1). Laboratories were reasonably consistent in their ranks of estimates among tissues for starch (r = 0.41-0.91), but less so for total NSC (r = 0.45-0.84) and soluble sugars (r = 0.11-0.83). Our results show that NSC estimates for woody plant tissues cannot be compared among laboratories. The relative changes in NSC between treatments measured within a laboratory may be comparable within and between laboratories, especially for starch. To obtain comparable NSC estimates, we suggest that users can either adopt the reference method given in this publication, or report estimates for a portion of samples using the reference method, and report estimates for a standard reference material. Researchers interested in NSC estimates should work to identify and adopt standard methods.

Pulse-labelling trees to study carbon allocation dynamics: a review of methods, current knowledge and future prospects.

Pulse-labelling of trees with stable or radioactive carbon (C) isotopes offers the unique opportunity to trace the fate of labelled CO(2) into the tree and its release to the soil and the atmosphere. Thus, pulse-labelling enables the quantification of C partitioning in forests and the assessment of the role of partitioning in tree growth, resource acquisition and C sequestration. However, this is associated with challenges as regards the choice of a tracer, the methods of tracing labelled C in tree and soil compartments and the quantitative analysis of C dynamics. Based on data from 47 studies, the rate of transfer differs between broadleaved and coniferous species and decreases as temperature and soil water content decrease. Labelled C is rapidly transferred belowground-within a few days or less-and this transfer is slowed down by drought. Half-lives of labelled C in phloem sap (transfer pool) and in mature leaves (source organs) are short, while those of sink organs (growing tissues, seasonal storage) are longer. (13)C measurements in respiratory efflux at high temporal resolution provide the best estimate of the mean residence times of C in respiratory substrate pools, and the best basis for compartmental modelling. Seasonal C dynamics and allocation patterns indicate that sink strength variations are important drivers for C fluxes. We propose a conceptual model for temperate and boreal trees, which considers the use of recently assimilated C versus stored C. We recommend best practices for designing and analysing pulse-labelling experiments, and identify several topics which we consider of prime importance for future research on C allocation in trees: (i) whole-tree C source-sink relations, (ii) C allocation to secondary metabolism, (iii) responses to environmental change, (iv) effects of seasonality versus phenology in and across biomes, and (v) carbon-nitrogen interactions. Substantial progress is expected from emerging technologies, but the largest challenge remains to carry out in situ whole-tree labelling experiments on mature trees to improve our understanding of the environmental and physiological controls on C allocation.

Carbon and nitrogen mobilization in Larix x eurolepis leafy stem cuttings assessed by dual (13)C and (15)N labeling: relationships with rooting.

Changes in use of both stored and newly synthesized sources of carbon (C) and nitrogen (N) were investigated during rooting of leafy cuttings of Larix x eurolepis A. Henry. We used dual (13)C and (15)N long-term labeling of reserves of stock plants and followed isotope dilution of the labels in the cuttings to determine the respective proportions of C and N derived from stock plant reserves (Q(C,old), Q(N,old)) and from newly synthesized sources (Q(C,new), Q(N,new)). We also assessed their partitioning into the growing needles and roots. Because rooting development occurred over several months, destructive harvests were made during three periods (t(0): April 9; t(1): May 15 to June 5; t(2): June 12 to June 26). Total C content (Q(C)) of cuttings did not change with time, whereas total N content (Q(N)) decreased significantly between t(0) and t(2). In rooted cuttings, unlike unrooted cuttings, total N concentration decreased less markedly in needles and in the lower stem component between t(1) and t(2) than between t(0) and t(2). The lower stem of rooted cuttings contained more Q(N,old) than that of unrooted cuttings and showed lower C:N and (13)C:(15)N ratios. The increase in Q(N,new) in the lower stem was positively correlated with rooting status. At t(1), new roots appeared and grew mainly at the expense of Q(C,new) and Q(N,old). By t(2), root growth was mainly dependent on new assimilates. We conclude that in leafy cuttings the initial amount of N reserves, rather than C reserves, may constitute a limiting factor for rooting.