ALBERT

Description: The impact of stored water on estimates of transpiration from scaled sap flux measurements was assessed in mature Pinus taeda (L.) at the Duke Free-Air CO 2 Enrichment (FACE) site. We used a simple hydraulic model with measurements of sap flux ( J ) at breast height and the base of the live crown for 26 trees over 6 months to examine the effects of elevated CO 2 (eCO 2 ) and fertilization (N F ) treatments, as well as temporal variation in soil moisture ( M ( t ) ), on estimates of the hydraulic time constant (). At low M ( t ) , there was little (〈12%) difference in of different treatments. At high M ( t ) , differences were much greater, with reductions of 27, 52 and 34% in eCO 2 , N F and eCO 2 x N F respective to the control. Incorporating with these effects into the analysis of a larger data set of previous J measurements at this site (1998–2008) improved agreement between modeled and measured values in 92% of cases. However, a simplified calibration of that neglected treatment and soil moisture effects performed more dependably, improving agreement in 98% of cases. Incorporating had the effect of increasing estimates of reference stomatal conductance at 1 kPa vapor pressure deficit (VPD) and saturating photosynthetic active radiation (PAR) an average of 12–14%, while increasing estimated sensitivities to VPD and PAR. A computationally efficient hydraulic model, such as the one presented here, incorporated into a hierarchical model of stomatal conductance presents a novel approach to including hydraulic time constants in estimates of stomatal responses from long-term sap flux data sets.

Description: Inferences on climate change effects are reliable only if they are based on a causal relationship rather than simple statistical predictive capacity. To assess for causal links between climate and mature black spruce ( Picea mariana (Mills.) BSP) radial growth, we combined the use of wood anatomy, cambium phenology, climate and soil measurements (air temperature and humidity, precipitations, soil temperature and water content, photosynthetically active radiation), and a model selection approach proceeding backwards from a full model. Results show that the number of tracheids is responsible for 88% of the variation in ring width whereas mean tracheid diameter accounts for the remaining 12%. The number of tracheids produced depends on factors related to photosynthesis during tracheid production, i.e., daily light intensity and maximum temperature between the day of initiation and the day of cessation of tracheid production, plus soil temperature during August of the previous year which is an important period for determining the number of new needles produced. It is also important to consider duration of the period for tracheid production. These results imply that short-term climate change should increase black spruce radial growth. They also suggest that the typical use of post-growth ring width sampling individually linked to air temperature and precipitations is not sufficient to infer climate change effects accurately on radial growth where there is no strong single climatic limitation but multiple limitations instead.

Description: It is generally assumed that the largest vessels are occurring in the roots and that vessel diameters and the related hydraulic conductance in the xylem are decreasing acropetally from roots to leaves. With this study in five tree species of a perhumid tropical rainforest in Sulawesi (Indonesia), we searched for patterns in hydraulic architecture and axial conductivity along the flow path from small-diameter roots through strong roots and the trunk to distal sun-canopy twigs. Wood density differed by not more than 10% across the different flow path positions in a species, and branch and stem wood density were closely related in three of the five species. Other than wood density, the wood anatomical and xylem hydraulic traits varied in dependence on the position along the flow path, but were unrelated to wood density within a tree. In contrast to reports from conifers and certain dicotyledonous species, we found a hump-shaped variation in vessel diameter and sapwood area-­specific conductivity along the flow path in all five species with a maximum in the trunk and strong roots and minima in both small roots and twigs; the vessel size depended on the diameter of the organ. This pattern might be an adaptation to the perhumid climate with a low risk of hydraulic failure. Despite a similar mean vessel diameter in small roots and twigs, the two distal organs, hydraulically weighted mean vessel diameters were on average 30% larger in small roots, resulting in ~ 85% higher empirical and theoretical specific conductivities. Relative vessel lumen area in percent of sapwood area decreased linearly by 70% from roots to twigs, reflecting the increase in sclerenchymatic tissue and tracheids in acropetal direction in the xylem. Vessel size was more closely related to the organ diameter than to the distance along the root-to-shoot flow path. We conclude that (i) the five co-occurring tree species show convergent patterns in their hydraulic architecture despite different growth strategies, and (ii) the paradigm assuming continuous acropetal vessel tapering and decrease in specific conductance from fine roots towards distal twigs needs reconsideration.

Description: In this study, we employ a network of thermal dissipation probes (TDPs) monitoring sap flux density to estimate leaf-specific transpiration ( E L ) and stomatal conductance ( G S ) in Pinus taeda (L.) and Liquidambar styraciflua L. exposed to +200 ppm atmospheric CO 2 levels (eCO 2 ) and nitrogen fertilization. Scaling half-hourly measurements from hundreds of sensors over 11 years, we found that P. taeda in eCO 2 intermittently (49% of monthly values) decreased stomatal conductance ( G S ) relative to the control, with a mean reduction of 13% in both total E L and mean daytime G S . This intermittent response was related to changes in a hydraulic allometry index ( A H ), defined as sapwood area per unit leaf area per unit canopy height, which decreased a mean of 15% with eCO 2 over the course of the study, due mostly to a mean 19% increase in leaf area ( A L ). In contrast, L. styraciflua showed a consistent (76% of monthly values) reduction in G S with eCO 2 with a total reduction of 32% E L , 31% G S and 23% A H (due to increased A L per sapwood area). For L. styraciflua , like P. taeda , the relationship between A H and G S at reference conditions suggested a decrease in G S across the range of A H . Our findings suggest an indirect structural effect of eCO 2 on G S in P. taeda and a direct leaf level effect in L. styraciflua . In the initial year of fertilization, P. taeda in both CO 2 treatments, as well as L. styraciflua in eCO 2 , exhibited higher G S with N F than expected from shifts in A H , suggesting a transient direct effect on G S . Whether treatment effects on mean leaf-specific G S are direct or indirect, this paper highlights that long-term treatment effects on G S are generally reflected in A H as well.

Description: Estimating sapwood area is one of the main sources of error when upscaling point scale sap flow measurements to whole-tree water use. In this study, the potential use of electrical resistivity tomography (ERT) to determine the sapwood–heartwood (SW–HW) boundary is investigated for Pinus elliottii Engelm var. elliottii x Pinus caribaea Morelet var. hondurensis growing in a subtropical climate. Specifically, this study investigates: (i) how electrical resistivity is correlated to either wood moisture content, or electrolyte concentration, or both, and (ii) how the SW–HW boundary is defined in terms of electrical resistivity. Tree cross-sections at breast height are analysed using ERT before being felled and the cross-section surface sampled for analysis of major electrolyte concentrations, wood moisture content and density. Electrical resistivity tomography results show patterns with high resistivities occurring in the inner part of the cross-section, with much lower values towards the outside. The high-resistivity areas were generally smaller than the low-resistivity areas. A comparison between ERT and actual SW area measured after felling shows a slope of the linear regression close to unity (=0.96) with a large spread of values ( R 2 = 0.56) mostly due to uncertainties in ERT. Electrolyte concentrations along sampled radial transects (cardinal directions) generally showed no trend from the centre of the tree to the bark. Wood moisture content and density show comparable trends that could explain the resistivity patterns. While this study indicates the potential for application of ERT for estimating SW area, it shows that there remains a need for refinement in locating the SW–HW boundary (e.g., by improvement of the inversion method, or perhaps electrode density) in order to increase the robustness of the method.

Description: Palms are an important component of tropical ecosystems, living alongside dicotyledonous trees, even though they have a very different growth pattern and vascular system. As monocots, vessels in palms are located within vascular bundles and, without a vascular cambium that many dicotyledonous trees possess, palms cannot add additional vessels to their vascular system as they get older and taller. This means that hydraulic architecture in palms is more predetermined, which may require a highly efficient hydraulic system. This preset nature, along with the decoupling of hydraulic and mechanical functioning to different cell types, may allow palms to have a more efficient hydraulic system than dicotyledonous trees. Therefore, this study seeks to determine the efficiency of the hydraulic system in the palm Iriartea deltoidea (Ruiz & Pav.) and compare this efficiency with other tree forms. We measured cross-sectional areas of roots, stems and fronds as well as leaf areas of I. deltoidea saplings. Likewise, cross-sections were made and vessel diameters and frequencies measured. This allowed for the calculation of theoretical specific conductivity ( K S,calc ), theoretical leaf-specific conductivity ( K L,calc ), and vessel diameter and vessel number ratios between distal and proximal locations in the palms. Iriartea deltoidea palms were found to have the largest, least frequent vessels that diverged most from the square packing limit (maximum number of vessels that fit into a given area) compared with other major tree forms, and they therefore invested the least space and carbon into water transport structures. Likewise, conduits tapered by ~1/3 between ranks (root, bole and petiole), which represents an efficient ratio with regard to the trade-offs between safety and efficiency of the conducting system. Conduits also exhibited a high conservation of the sum of the conduit radii cubed ( r 3 ) across ranks, thereby approximating Murray's law patterning. Therefore, our results indicate that the palm I. deltoidea has a very efficient hydraulic system in terms of maintaining a large conducting capacity with a minimal vascular investment. This efficiency may allow palms to compete well with dicotyledonous trees in tropical and subtropical climates but other developmental factors largely restrict palms from regions that experience prolonged freezing temperatures.

Description: Functional attributes determine the survival and growth of planted seedlings in reforestation projects. Nitrogen (N) and water are important resources in the cultivation of forest species, which have a strong effect on plant functional traits. We analyzed the influence of N nutrition on drought acclimation of Pinus pinea L. seedlings. Specifically, we addressed if high N fertilization reduces drought and frost tolerance of seedlings and whether drought hardening reverses the effect of high N fertilization on stress tolerance. Seedlings were grown under two N fertilization regimes (6 and 100 mg N per plant) and subjected to three drought-hardening levels (well-watered, moderate and strong hardening). Water relations, gas exchange, frost damage, N concentration and growth at the end of the drought-hardening period, and survival and growth of seedlings under controlled xeric and mesic outplanting conditions were measured. Relative to low-N plants, high-N plants were larger, had higher stomatal conductance (27%), residual transpiration (11%) and new root growth capacity and closed stomata at higher water potential. However, high N fertilization also increased frost damage (24%) and decreased plasmalemma stability to dehydration (9%). Drought hardening reversed to a great extent the reduction in stress tolerance caused by high N fertilization as it decreased frost damage, stomatal conductance and residual transpiration by 21, 31 and 24%, respectively, and increased plasmalemma stability to dehydration (8%). Drought hardening increased tissue non-structural carbohydrates and N concentration, especially in high-fertilized plants. Frost damage was positively related to the stability of plasmalemma to dehydration ( r = 0.92) and both traits were negatively related to the concentration of reducing soluble sugars. No differences existed between moderate and strong drought-hardening treatments. Neither N nutrition nor drought hardening had any clear effect on seedling performance under xeric outplanting conditions. However, fertilization increased growth under mesic conditions, whereas drought hardening decreased growth. We conclude that drought hardening and N fertilization applied under typical container nursery operational conditions exert opposite effects on the physiological stress tolerance of P. pinea seedlings. While drought hardening increases overall stress tolerance, N nutrition reduces it and yet has no effect on the drought acclimation capacity of seedlings.

Description: We investigated Scots pine adaptive responses to the light spectra by measuring hypocotyl length in seeds sampled from three natural Scots pine ecotypes across a latitudinal cline ranging from 63° to 68° N in Sweden where the adaptive cline is known to be steeper. Seeds were germinated under dark (D) and three monochromatic continuous light wavelengths: blue (B), red (R) and far-red (FR). Analysis of variance revealed a northward decrease in the inhibitory effect of FR with respect to D, the so-called far red high irradiance response. Ecotypic variation for hypocotyl development was observed under the FR and D treatments, while the trends for the B and R treatments were not statistically significant. Under FR the ecotypic variation showed an increase in hypocotyl length northwards, in contrast to the treatment under D which showed a decrease in the hypocotyl length northwards. These results could be interpreted in view of the previously reported northward increase in FR requirement to maintain growth in Norway spruce and Scots pine. Prior to the performance of the main light experiment, the maternal effect on progeny performance was investigated, which showed the absence of maternal environment effect on the performance of the seedlings.

Description: Drought stress can induce closure of stomata, thus leading to photoinhibition. The effects of prolonged severe drought under natural growing conditions on photosystem I (PSI), photosystem II (PSII) and cyclic electron flow (CEF) in drought-tolerant tree species are unclear. In spring 2010, southwestern China confronted severe drought that lasted several months. Using three dominant evergreen species, Cleistanthus sumatranus (Miq.) Muell. Arg. (Euphorbiaceae), Celtis philippensis Bl. (Ulmaceae) and Pistacia weinmannifolia J. Poisson ex Franch. (Anacardiaceae) that are native to a tropical limestone forest, we investigated the influence of this stress on PSI and PSII activities as well as light energy distribution in the PSII and P700 redox state. By the end of the drought period, predawn leaf water potential ( pd ) largely declined in each species, especially in C. sumatranus . Photosystem I activity strongly decreased in the three species, especially in C. sumatranus which showed a decrease of 65%. The maximum quantum yield of PSII after dark adaptation remained stable in P. weinmannifolia and C. philippensis but significantly decreased in C. sumatranus . Light response curves indicated that both linear electron flow and non-photochemical quenching were severely inhibited in C. sumatranus along with disappearance of CEF, resulting in deleterious excess light energy in PSII. We conclude that PSI is more sensitive than PSII to prolonged severe drought in these three drought-tolerant species, and CEF is essential for photoprotection in them.

Description: We investigated changes in chlorophyll a fluorescence from alternate leaf surfaces to assess the intraleaf light acclimation patterns in combination with natural variations in radiation, leaf angles, leaf mass per area (LMA), chlorophyll content (Chl) and leaf optical parameters. Measurements were conducted on bottom- and top-layer leaves of Tilia cordata Mill. (a shade-tolerant sub-canopy species, sampled at heights of 11 and 16 m) and Populus tremula L. (a light-demanding upper canopy species, sampled at canopy heights of 19 and 26 m). The upper canopy species P. tremula had a six times higher PSII quantum yield ( II ) and ratio of open reaction centres (qP), and a two times higher LMA than T. cordata . These species-specific differences were also present when the leaves of both species were in similar light conditions. Leaf adaxial/abaxial fluorescence ratio was significantly larger in the case of more horizontal leaves. Populus tremula (more vertical leaves), had smaller differences in fluorescence parameters between alternate leaf sides compared with T. cordata (more horizontal leaves). However, optical properties on alternate leaf sides showed a larger difference for P. tremula . Intraspecifically, the measured optical parameters were better correlated with LMA than with leaf Chl. Species-specific differences in leaf anatomy appear to enhance the photosynthetic potential of leaf biochemistry by decreasing the interception of excess light in P. tremula and increasing the light absorptance in T. cordata . Our results indicate that intraleaf light absorption gradient, described here as leaf adaxial/abaxial side ratio of chlorophyll a fluorescence, varies significantly with changes in leaf light environment in a multi-layer multi-species tree canopy. However, this variation cannot be described merely as a simple function of radiation, leaf angle, Chl or LMA, and species-specific differences in light acclimation strategies should also be considered.

Description: Norway maple ( Acer platanoides L), which is among the most invasive tree species in forests of eastern North America, is associated with reduced regeneration of the related native species, sugar maple ( Acer saccharum Marsh) and other native flora. To identify traits conferring an advantage to Norway maple, we grew both species through an entire growing season under simulated light regimes mimicking a closed forest understorey vs. a canopy disturbance (gap). Dynamic shade-houses providing a succession of high-intensity direct-light events between longer periods of low, diffuse light were used to simulate the light regimes. We assessed seedling height growth three times in the season, as well as stem diameter, maximum photosynthetic capacity, biomass allocation above- and below-ground, seasonal phenology and phenotypic plasticity. Given the north European provenance of Norway maple, we also investigated the possibility that its growth in North America might be increased by delayed fall senescence. We found that Norway maple had significantly greater photosynthetic capacity in both light regimes and grew larger in stem diameter than sugar maple. The differences in below- and above-ground biomass, stem diameter, height and maximum photosynthesis were especially important in the simulated gap where Norway maple continued extension growth during the late fall. In the gap regime sugar maple had a significantly higher root : shoot ratio that could confer an advantage in the deepest shade of closed understorey and under water stress or browsing pressure. Norway maple is especially invasive following canopy disturbance where the opposite (low root : shoot ratio) could confer a competitive advantage. Considering the effects of global change in extending the potential growing season, we anticipate that the invasiveness of Norway maple will increase in the future.

Description: Fungal colonization can significantly affect the secondary metabolism of the host plants. We tested the impact of a common below-ground symbiosis, i.e., ectomycorrhiza formation, on poplar leaf chemical components that are involved in the defence against a common disease, i.e., rust fungi, in N-deficient soil. A rust-susceptible poplar clone ( Populus trichocarpa x deltoides ‘Beaupré’) was (a) non-associated with ectomycorrhizal fungus (EM) Hebeloma mesophaeum (Pers.) Quélet MÜN and non-infected with rust fungus Melampsora larici-populina Kleb. (isolate 98AG31), (b) associated with EM, (c) inoculated with rust fungus and (d) associated with EM and inoculated with rust fungus. Poplar leaves were analysed by photometric and mass spectrometric techniques (liquid chromatography–tandem mass spectrometry (LC-MS/MS), pyrolysis–field ionization mass spectrometry (Py-FIMS)). Both rust infection and mycorrhiza formation led to increased proportions of condensed tannins in relation to total phenolics (13% in the control, 18–19% in the fungal treatments). In contrast, salicylic acid concentration (6.8 µg g –1 in the control) was higher only in the rust treatments (17.9 and 25.4 µg g –1 with rust infection). The Py-FIMS analysis revealed that the rust-infected treatments were significantly separated from the non-rust-infected treatments on the basis of six flavonoids and one lipid. The relative abundance of these components, which have known functions in plant defence, was decreased after rust infection of non-mycorrhizal plants, but not in mycorrhizal plants. The results indicate that the ectomycorrhizal formation compensated the rust infection by a decrease in the flavonoid syntheses. The study provides new evidence for an interactive response of mycorrhizal colonization and infection with rust fungi in the metabolism of poplar.

Description: Apple proliferation (AP) represents a serious threat to several fruit-growing areas and is responsible for great economic losses. Several studies have highlighted the key role played by the cell wall in response to pathogen attack. The existence of a cell wall integrity signaling pathway which senses perturbations in the cell wall architecture upon abiotic/biotic stresses and activates specific defence responses has been widely demonstrated in plants. More recently a role played by cell wall-related genes has also been reported in plants infected by phytoplasmas. With the aim of shedding light on the cell wall response to AP disease in the economically relevant fruit-tree Malus x domestica Borkh., we investigated the expression of the cellulose ( CesA ) and callose synthase ( CalS ) genes in different organs (i.e., leaves, roots and branch phloem) of healthy and infected symptomatic outdoor-grown trees, sampled over the course of two time points (i.e., spring and autumn 2011), as well as in in vitro micropropagated control and infected plantlets. A strong up-regulation in the expression of cell wall biosynthetic genes was recorded in roots from infected trees. Secondary cell wall CesA s showed up-regulation in the phloem tissue from branches of infected plants, while either a down-regulation of some genes or no major changes were observed in the leaves. Micropropagated plantlets also showed an increase in cell wall-related genes and constitute a useful system for a general assessment of gene expression analysis upon phytoplasma infection. Finally, we also report the presence of several ‘knot’-like structures along the roots of infected apple trees and discuss the occurrence of this interesting phenotype in relation to the gene expression results and the modalities of phytoplasma diffusion.

Description: The availability of a system for direct transfer of antifungal candidate genes into European chestnut ( Castanea sativa Mill.) would offer an alternative approach to conventional breeding for production of chestnut trees tolerant to ink disease caused by Phytophthora spp. For the first time, a chestnut thaumatin-like protein gene ( CsTL1 ), isolated from chestnut cotyledons, has been overexpressed in three chestnut somatic embryogenic lines. Transformation experiments have been performed using an Agrobacterium tumefaciens Smith and Townsend vector harboring the neomycin phosphotransferase ( NPTII ) selectable and the green fluorescent protein ( EGFP ) reporter genes. The transformation efficiency, determined on the basis of the fluorescence of surviving explants, was clearly genotype dependent and ranged from 32.5% in the CI-9 line to 7.1% in the CI-3 line. A total of 126 independent transformed lines were obtained. The presence and integration of chestnut CsTL1 in genomic DNA was confirmed by polymerase chain reaction (PCR) and Southern blot analyses. Quantitative real-time PCR revealed that CsTL1 expression was up to 13.5-fold higher in a transgenic line compared with its corresponding untransformed line. In only one of the 11 transformed lines tested, expression of the CsTL1 was lower than the control. The remaining 115 transformed lines were successfully subjected to cryopreservation. Embryo proliferation was achieved in all of the transgenic lines regenerated and the transformed lines showed a higher mean number of cotyledonary stage embryos and total number of embryos per embryo clump than their corresponding untransformed lines. Transgenic plants were regenerated after maturation and germination of transformed somatic embryos. Furthermore, due to the low plantlet conversion achieved, axillary shoot proliferation cultures were established from partially germinated embryos (only shoot development), which were multiplied and rooted according to procedures already established. Transgenic plants were acclimatized and grown in a greenhouse. No phenotypic differences were found with control plants, suggesting no potential cytotoxic effects of the green fluorescent protein. The results reported in the present work could be considered as a first step toward the production of fungal-disease tolerant cisgenic chestnut plants.

Description: Studying the dynamics of stem water content (   ) in living trees has an outstanding physiological interest but all the available techniques to measure exhibit major drawbacks. In this work, we present a new methodology to estimate variations in along with sap velocity using the compensated heat pulse (CHP) technique. One lab experiment was performed on several wooden blocks obtained from three different tree species. Samples were slowly dried and their moisture loss was monitored by both gravimetric approaches and time-domain reflectometry (TDR) or CHP probes in order to contrast the validity of our methodology (volumetric specific heat (VSH)-CHP) over a range of water contents. In addition, a field experiment was conducted to monitor fluctuations in standing olive trees ( Olea europaea L. cv. ‘Arbequina’) growing under three different irrigation regimes. In the lab test, the actual values deduced gravimetrically differed from the estimates yielded by the VSH-CHP method. However, it could successfully track relative changes in the water stored for the range of expected in living wood. Furthermore, the field experiment showed a seasonal change in , which was similar in shape and magnitude to those reported in the literature for olive and other Mediterranean tree species. On the other hand, differences in the seasonal patterns of between irrigation treatments strongly corresponded with those of sap flow and some leaf water potential measurements. The results of this work suggest that the CHP technique could be employed to monitor the dynamics of both and sap flow simultaneously in standing trees and evidence that seasonal changes in might be used as a long-term water status indicator.

Description: The emerald ash borer ( Agrilus planipennis Fairmaire, EAB) is an alien, invasive wood-boring insect that is responsible for killing millions of ash trees since its discovery in North America in 2002. All North American ash species ( Fraxinus spp.) that EAB has encountered have shown various degrees of susceptibility, while Manchurian ash ( Fraxinus mandshurica Ruprecht), which shares a co-evolutionary history with this insect, is resistant. Recent studies have looked into constitutive resistance mechanisms in Manchurian ash, concentrating on the secondary phloem, which is the feeding substrate for the insect. In addition to specialized metabolism and defense-related components, primary metabolites and nutritional summaries can also be important to understand the feeding behavior of insect herbivores. Here, we have compared the nutritional characteristics (water content, total protein, free amino acids, total soluble sugars and starch, percent carbon and nitrogen, and macro- and micronutrients) of outer bark and phloem from black, green, white and Manchurian ash to determine their relevance to resistance or susceptibility to EAB. Water content and concentrations of Al, Ba, Cu, Fe, K, Li, tryptophan and an unknown compound were found to separate black and Manchurian ash from green and white ash in a principal component analysis (PCA), confirming their phylogenetic placements into two distinct clades. The traits that distinguished Manchurian ash from black ash in the PCA were water content and concentrations of total soluble sugars, histidine, lysine, methionine, ornithine, proline, sarcosine, tyramine, tyrosol, Al, Fe, K, Na, V and an unknown compound. However, only proline, tyramine and tyrosol were significantly different, and higher, in Manchurian ash than in black ash.

Description: According to well-known biochemical and biophysical mechanisms, the stimulation of C 3 photosynthesis by elevated atmospheric CO 2 concentration ([CO 2 ]) is strongly modified by changes in temperature and radiation. In order to investigate whether a static parameterization of the commonly used Farquhar et al. model of photosynthesis (i.e., without CO 2 -induced seasonal or thermal acclimation of photosynthetic capacity) can accurately predict these interactions in mature boreal Norway spruce ( Picea abies (L.) Karst.) during the frost-free part of the growing season, shoot gas exchange was continuously measured on trees during their second/third year of exposure to ambient or doubled [CO 2 ] inside whole-tree chambers. The relative CO 2 -induced enhancement of net photosynthesis ( A n ) at a given temperature remained stable over the study period, but increased strongly with temperature and radiation, in agreement with predictions by the model. Light-saturated A n (+67% at 20 °C), dark respiration (+36%) and intercellular to ambient [CO 2 ] ratio ( c i / c a ; +27%) were significantly increased by CO 2 treatment. Stomatal conductance ( g s ) was not significantly affected. Our results demonstrate that the Farquhar et al. model of photosynthesis has the capability to predict interactions between [CO 2 ] and seasonal weather variability on A n in Norway spruce during the non-frost growing season without accounting for CO 2 -induced seasonal and/or thermal photosynthetic acclimation. However, stomatal model assumptions of reduced g s and constant c i / c a under rising atmospheric [CO 2 ] did not hold.

Description: Radial stem growth and the isotopic composition of growth rings are commonly used to quantify the effects of droughts on trees. However, often these parameters are quantified only at one stem height, e.g., 1.3 m, and it is not known how representative this is for the whole stem. This study investigated radial growth at four stem heights (1.3, 5.5, 9.8 and 14 m) of 21, and wood 13 C and 18 O at two heights (1.3 and 14 m) of 10 (co-)dominant Norway spruce trees from heavily (HT) and moderately thinned (MT) stands to assess whether different thinning intensities influenced the drought response of stems at different tree heights. Annual basal area increments (BAIs) and stable isotopes in earlywood and latewood were compared between thinning treatments and among the different stem heights. For BAIs, linear correlations with climate were analysed as well. The response of radial growth and isotopic composition to drought was similar at different stem heights in HT trees, but varied with height in MT trees, which were also more sensitive to climatic variations. Recovery of radial growth after drought was more rapid in trees from HT compared with MT stands, except for the topmost height. Basal area increments at breast height (1.3 m) provided good estimates of the volume growth response to drought for the whole stem, but not for its recovery. The faster recovery of radial growth at 1.3 m height of HT compared with MT trees after the 2003 drought was not accompanied by differences in recovery of isotopic composition. However, this is likely to be related to differences between treatments in remobilization of stored C and in tree structure (leaf area, root systems).

Description: Crown architecture is an important determinant of biomass production and yield of any bio-energy plantation since it determines leaf area display and hence light interception. Four Populus genotypes—of different species and hybrids and with contrasting productivity and leaf area—were examined in terms of their branch characteristics in relation to crown architecture during the first and second growing seasons after plantation establishment. The trees were planted at high density (8000 ha –1 ) on two different former land use types, cropland and pasture. We documented significant differences in branch architecture among the genotypes and for the first year among the former land use types. Land use effects only affected factors not related to canopy closure and wood production, and decreased after the first growing season. This suggested that both former land use types were equally suited for the establishment success of a poplar bio-energy plantation. Tree height and branch dimensions—branch diameter and branch length—were the most important determinants of wood production and maximum leaf area index. Despite the secondary importance of the number of sylleptic branches, these branches contributed significantly to the total leaf area in three out of the four studied genotypes. This indicated that enhanced syllepsis accelerates leaf area development and hence carbon assimilation, especially in the early stages of a high-density plantation with poplar.

Description: Polyamine (PA) metabolism was studied in liquid cultures of Scots pine ( Pinus sylvestris L.) embryogenic cells. The focus of the study was on the metabolic changes at the interphase between the initial lag phase and the exponential growth phase. PA concentrations fluctuated in the liquid cultures as follows. Putrescine (Put) concentrations increased, whereas spermidine (Spd) concentrations decreased in both free and soluble conjugated PA fractions. The concentrations of free and soluble conjugated spermine (Spm) remained low, and small amounts of excreted PAs were also found in the culture medium. The minor production of secondary metabolites reflected the undifferentiated stage of the embryogenic cell culture. Put was produced via the arginine decarboxylase (ADC) pathway. Futhermore, the gene expression data suggested that the accumulation of Put was caused neither by an increase in Put biosynthesis nor by a decrease in Put catabolism, but resulted mainly from the decrease in the biosynthesis of Spd and Spm. Put seemed to play an important role in cell proliferation in Scots pine embryogenic cells, but the low pH of the culture medium could also, at least partially, be the reason for the accumulation of endogenous Put. High Spd concentrations at the initiation of the culture, when cells were exposed to stress and cell death, suggested that Spd may act not only as a protector against stress but also as a growth suppressor, when proliferative growth is not promoted. All in all, Scots pine embryogenic cell culture was proved to be a favourable experimental platform to study PA metabolism and, furthermore, the developed system may also be beneficial in experiments where, e.g., the effect of specific stressors on PA metabolism is addressed.

Description: Several heat-based sap flow methods, such as the heat field deformation method and the heat ratio method, include the thermal diffusivity D of the sapwood as a crucial parameter. Despite its importance, little attention has been paid to determine D in a plant physiological context. Therefore, D is mostly set as a constant, calculated during zero flow conditions or from a method of mixtures, taking into account wood density and moisture content. In this latter method, however, the meaning of the moisture content is misinterpreted, making it theoretically incorrect for D calculations in sapwood. A correction to this method, which includes the correct application of the moisture content, is proposed. This correction was tested for European and American beech and Eucalyptus caliginosa Blakely & McKie. Depending on the dry wood density and moisture content, the original approach over- or underestimates D and, hence, sap flux density by 10% and more.

Description: The evergreen holm oak Quercus ilex L. is the most representative tree in Mediterranean forests. Accurate estimation of the limiting factors of photosynthesis for Q. ilex and the prediction of ecosystem water-use efficiency by mechanistic models can be achieved only by establishing whether this species shows heterogenic stomatal aperture, and, if so, the circumstances in which this occurs. Here, we collected gas-exchange and chlorophyll fluorescence data in Q. ilex leaves from a nursery to measure the effects of stomatal oscillations on PSII quantum yield ( PSII ) under water stress. Stomatal conductance ( g s ) was used as an integrative indicator of the degree of water stress. Images of chlorophyll fluorescence showed heterogeneous PSII when g s was 〈50 mmol H 2 O m –2 s –1 , representative of severe drought and corresponding to a container capacity 〈45%. Stomatal patchiness was related to a coefficient of variation (CV) of PSII values 〉2.5%. A parallel study in the forest confirmed heterogeneous PSII values in leaves in response to declining water availability. Three kinds of Q. ilex individuals were distinguished: those resprouting after a clear-cut (resprouts, R); intact individuals growing in the same clear-cut area as resprouts (controls, C); and intact individuals in a nearby, undisturbed area (forest controls, CF). Patchiness increased in C and CF in response to increasing drought from early May to late July, whereas in R, PSII values were maintained as a result of their improved water relations since the pre-existing roots were associated with a smaller aerial biomass. Patchiness was related to a % CV of PSII values 〉4 and associated in the summer with mean g s values of 30 mmol H 2 O m –2 s –1 . Under milder drought in spring, PSII patchiness was less strictly related to g s variations, pointing to biochemical limitants of photosynthesis. The occurrence of heterogenic photosynthesis caused by patchy stomatal closure in Q. ilex during severe drought should be taken into account in ecosystem modelling in which harsher water stress conditions associated with climate change are predicted.

Description: The freezing pattern and frost killing temperatures of apple ( Malus domestica Borkh.) xylem were determined by differential thermal analysis and infrared differential thermal analysis (IDTA). Results from detached or attached twigs in controlled freezing experiments and during natural field freezing of trees were compared. Non-lethal freezing of apoplastic water in apple xylem as monitored during natural winter frosts in the field occurred at –1.9 ± 0.4 °C and did not change seasonally. The pattern of whole tree freezing was variable and specific to the environmental conditions. On detached twigs high-temperature freezing exotherms (HTEs) occurred 2.8 K below the temperature observed under natural frosts in the field with a seasonal mean of –4.7 ± 0.5 °C. Microporous apple xylem showed freezing without a specific pattern within a few seconds in IDTA images during HTEs, which is in contrast to macroporous xylem where a 2D freezing pattern mirrors anatomical structures. The pith tissue always remained unfrozen. Increasing twig length increased ice nucleation temperature; for increased twig diameter the effect was not significant. In attached twigs frozen in field portable freezing chambers, HTEs were recorded at a similar mean temperature (–4.6 ± 1.0 °C) to those for detached twigs. Upon lethal intracellular freezing of apple xylem parenchyma cells (XPCs) low-temperature freezing exotherms (LTEs) can be recorded. Low-temperature freezing exotherms determined on detached twigs varied significantly between a winter minimum of –36.9 °C and a summer maximum –12.7 °C. Within the temperature range wherein LTEs were recorded by IDTA in summer (–12.7 ± 0.5 to –20.3 ± 1.1 °C) various tiny clearly separated discontinuous freezing events could be detected similar to that in other species with contrasting XPC anatomy. These freezing events appeared to be initially located in the primary and only later in the secondary xylem. During the LTE no freezing events in the bark and central pith tissue were recorded. Attached twigs were exposed to various freezing temperatures at which LTEs occur. Even if 60% of XPCs were frost-damaged twigs were able to recuperate and showed full re-growth indicating a high regeneration capacity even after severe frost damage to XPCs.

Description: Nitrogen (N) retranslocation within tree canopies has been intensively studied and assumed to function as a one-way process (e.g., from older to newer leaves). However, recent studies have found that both N output and input occur in individual leaves, suggesting that ‘gross’ N retranslocation exists behind ‘net’ N retranslocation. In the present study, the amount and direction of gross N retranslocation within a canopy of deciduous oak Quercus serrata Thunb. ex. Murray saplings were investigated. Labeling was conducted with leaves of Q. serrata saplings cultivated under conditions of low-N (LN) or high-N (HN) fertility. Subsequently, N movement within the canopy was traced. Leaves at two different positions in the canopy (top and lateral) were labeled to determine the direction of gross N retranslocation. To detect seasonal differences, the leaf-labeling experiment was conducted twice during the early and late phases of the growing season. In addition, to compare the quantitative importance of gross N retranslocation and root N uptake, the latter was determined by labeling Q. serrata roots. The N-labeling experiment revealed gross N retranslocation among leaves, i.e., from top to lateral, lateral to top and lateral to lateral positions. Gross N retranslocation was quantitatively more important than root uptake, especially for plants cultivated at LN fertility. Season also affected the amount of gross N retranslocation, and these effects differed between LN and HN fertilities. These findings suggest that N allocation within a canopy is controlled dynamically by both gross N output and input. The mechanisms controlling gross N output and input likely function as key determinants of N allocation within a tree canopy.

Description: Natural temperature gradient (NTG) can be a significant problem in thermal sap flow measurements, particularly in dry environments with sparse vegetation. To resolve this problem, we propose a novel correction method called cyclic heat dissipation (CHD) in its thermal dissipation probe (TDP) application. The CHD method is based on cyclic, switching ON/OFF power schema measurements and a three-exponential model, extrapolating measured signal to steady state thermal equilibrium. The extrapolated signal OFF represents NTG, whereas the extrapolated signal ON represents standard TDP signal, biased by NTG. Therefore, subtraction of the OFF signal from the ON signal allows defining the unbiased TDP signal, finally processed according to standard Granier calibration. The in vivo Kalahari measurements were carried out in three steps on four different tree species, first as NTG, then as standard TDP and finally in CHD mode, each step for ~1–2 days. Afterwards, each tree was separated from its stem following modified Roberts' (1977) procedure, and CHD verification was applied. The typical NTG varying from ~0.5 °C during night-time to –1 °C during day-time, after CHD correction, resulted in significant reduction of sap flux densities ( J p ) as compared with the standard TDP, particularly distinct for low J p . The verification of the CHD method indicated ~20% agreement with the reference method, largely dependent on the sapwood area estimate. The proposed CHD method offers the following advantages: (i) in contrast to any other NTG correction method, it removes NTG bias from the measured signal by using in situ, extrapolated to thermal equilibrium signal; (ii) it does not need any specific calibration making use of the standard Granier calibration; (iii) it provides a physical background to the proposed NTG correction; (iv) it allows for power savings; (v) it is not tied to TDP, and so can be adapted to other thermal methods. In its current state, the CHD data processing is not yet fully automated.

Description: Patchy stomatal closure occurs in plants with heterobaric leaves, in which vertical extensions of bundle sheath cells delimit the mesophyll and restrict the diffusion of CO 2 . The scale of patchy stomatal behavior was investigated in this study. The distribution of PSII quantum yield ( II ) obtained from chlorophyll fluorescence images was used to evaluate the scale of stomatal patchiness and its relationship with leaf photosynthesis in the sun leaves of 2-year-old saplings of Quercus crispula Blume. Fluorescent patches were observed only during the day with low stomatal conductance. Comparison of numerical simulation of leaf gas exchange and chlorophyll fluorescence images showed that heterogeneous distribution of electron transport rate through PSII ( J ) was observed following stomatal closure with a bimodal manner under both natural and saturated photosynthetic photon flux densities. Thus, fluorescence patterns can be interpreted in terms of patchy stomatal closure. The mapping of J from chlorophyll fluorescence images showed that the scale of stomatal patchiness was approximately 2.5-fold larger than that of anatomical patches (lamina areas bounded by bundle sheath extensions within lamina). Our results suggest the spatial scale of stomatal patches in Q. crispula leaves.

Description: Predicted future changes in air temperature and atmospheric CO 2 concentration ([CO 2 ]), coupled with altered precipitation, are expected to substantially affect tree growth. Effects on growth may vary considerably across a species range, as temperatures vary from sub-optimal to supra-optimal for growth. We performed an experiment simultaneously at two locations in the current range of loblolly pine, a cool site and a warm site, to examine the effect of future climate conditions on growth of loblolly pine seedlings in contrasting regions of the species range. At both sites 1-year-old loblolly pine seedlings were grown in current (local ambient temperature and [CO 2 ]) and predicted future atmospheric conditions (ambient +2 °C temperature and 700 mol mol –1 [CO 2 ]). Additionally, high and low soil moisture treatments were applied within each atmospheric treatment at each site by altering the amount of water provided to the seedlings. Averaged across water treatments, photosynthesis ( A net ) was 31% greater at the cool site and 34% greater at the warm site in elevated temperature and [CO 2 ] compared with ambient temperature. Biomass accumulation was also stimulated by 38% at the cool site and by 24% at the warm site in that treatment. These results suggest that a temperature increase of 2 °C coupled with an increase in [CO 2 ] (predicted future climate) will create conditions favorable for growth of this species. Reduced soil moisture decreased growth in both current and predicted atmospheric conditions. Biomass accumulation and A net were reduced by ~39 and 17%, respectively, in the low water treatment. These results suggest that any benefit of future atmospheric conditions may be negated if soil moisture is reduced by altered precipitation patterns.

Description: Conifer bark beetles are often associated with fungal complexes whose components have different ecological roles. Some associated species are nutritionally obligate fungi, serving as nourishment to the larvae, whereas others are pathogenic blue-stain fungi known to be involved in the interaction with host defenses. In this study we characterized the local and systemic defense responses of Scots pine ( Pinus sylvestris L.) against Ophiostoma brunneo-ciliatum Math. (a blue-stain pathogen) and Hyalorhinocladiella macrospora (Franke-Grosm.) Harr. (a nutritional fungus). These fungi are the principal associates of the pine engraver beetle, Ips acuminatus (Gyll.). Host responses were studied following inoculation with the fungi, singly and as a fungal complex, and by identifying and quantifying terpenoids, phenolic compounds and lignin. Although the length of the necrotic lesions differed between control (wound) and fungal treatments, only two compounds (pinosylvin monomethyl ether and (+)-α-pinene) were significantly affected by the presence of the fungi, indicating that Scots pine has a generic, rather than specific, induced response. The fact that both nutritional and blue-stain fungi triggered comparable induced defense responses suggests that even a non-pathogenic fungus may participate in exhausting host plant defenses, indirectly assisting in the beetle establishment process. Our findings contribute to the further development of current theory on the role of associated fungal complexes in bark beetle ecology.

Description: Hydraulic traits were studied for six Nothofagus species from South America (Argentina and Chile), and for three of these species two populations were studied. The main goal was to determine if properties of the water conductive pathway in stems and leaves are functionally coordinated and to assess if leaves are more vulnerable to cavitation than stems, consistent with the theory of hydraulic segmentation along the vascular system of trees in ecosystems subject to seasonal drought. Vulnerability to cavitation, hydraulic conductivity of stems and leaves, leaf water potential, wood density and leaf water relations were examined. Large variations in vulnerability to cavitation of stems and leaves were observed across populations and species, but leaves were consistently more vulnerable than stems. Water potential at 50% loss of maximum hydraulic efficiency ( P 50 ) ranged from –0.94 to –2.44 MPa in leaves and from –2.6 to –5.3 MPa in stems across species and populations. Populations in the driest sites had sapwood and leaves more vulnerable to cavitation than those grown in the wettest sites. Stronger diurnal down-regulation in leaf hydraulic conductance compared with stem hydraulic conductivity apparently has the function to slow down potential water loss in stems and protect stem hydraulics from cavitation. Species-specific differences in wood density and leaf hydraulic conductance ( K Leaf ) were observed. Both traits were functionally related: species with higher wood density had lower K Leaf . Other stem and leaf hydraulic traits were functionally coordinated, resulting in Nothofagus species with an efficient delivery of water to the leaves. The integrity of the more expensive woody portion of the water transport pathway can thus be maintained at the expense of the replaceable portion (leaves) of the stem–leaf continuum under prolonged drought. Compensatory adjustments between hydraulic traits may help to decrease the rate of embolism formation in the trees more vulnerable to cavitation.

Description: Sap flow measurements conducted with thermal dissipation probes (TDPs) are vulnerable to natural temperature gradient (NTG) bias. Few studies, however, attempted to explain the dynamics underlying the NTG formation and its influence on the sensors' signal. This study focused on understanding how the TDP signals are affected by negative and positive temperature influences from NTG and tested the novel cyclic heat dissipation (CHD) method to filter out the NTG bias. A series of three experiments were performed in which gravity-driven water flow was enforced on freshly cut stem segments of Fagus sylvatica L., while an artificial temperature gradient (ATG) was induced. The first experiment sought to confirm the incidence of the ATG on sensors. The second experiment established the mis-estimations caused by the biasing effect of the ATG on standard TDP measurements. The third experiment tested the accuracy of the CHD method to account for the ATG biasing effect, as compared with other cyclic correction methods. During experiments, sap flow measured by TDP was assessed against gravimetric measurements. The results show that negative and positive ATGs were comparable in pattern but substantially larger than field NTGs. Second, the ATG bias caused an overestimation of the standard TDP sap flux density of ~17 cm 3 cm –2 h –1 by 76%, and the sap flux density of ~2 cm 3 cm –2 h –1 by over 800%. Finally, the proposed CHD method successfully reduced the max. ATG bias to 25% at ~11 cm 3 cm –2 h –1 and to 40% at ~1 cm 3 cm –2 h –1 . We concluded that: (i) the TDP method is susceptible to NTG especially at low flows; (ii) the CHD method successfully corrected the TDP signal and resulted in generally more accurate sap flux density estimates (mean absolute percentage error ranging between 11 and 21%) than standard constant power TDP method and other cyclic power methods; and (iii) the ATG enforcing system is a suitable way of re-creating NTG for future tests.

Description: Two mature clones of Norway spruce ( Picea abies (L.) Karst.) that have previously been shown to have differential degrees of resistance towards the necrotrophic pathogen Heterobasidion parviporum (Niemelä & Korhonen) were compared with respect to the primed defense expression of transcripts related to biosynthesis of lignin, stilbenes and other phenolic compounds from one year to the next. The host's response to physical wounding and pathogen inoculation was examined in the initial year, whereas indications of heightened basal defense level or primed response, and responses to re-wounding, were examined the following year. The responses of the two clones to wounding and pathogen inoculation, examined in the initial year, differed; the increases in lignin and phenolics were more distinct in response to the pathogen than to wounding alone. The more resistant clone 589 had higher initial lignin concentrations in the cell walls when compared with clone 409, and these remained higher in clone 589 over both years and increased after the treatments. Both clones responded at the transcriptional and chemical levels to wounding; changes were evident both in the initial wounds and when re-wounded the following year. There were distinct differences in the basal transcript levels of the lignin pathway-related genes, phenolics and total lignin levels in healthy tissue from the initial year to the following year indicative of a primed host response or at least altered constitutive level of defense expression.

Description: In a world of diminishing water reservoirs and a rising demand for food, the practice and development of water stress indicators and sensors are in rapid progress. The heat dissipation method, originally established by Granier, is herein applied and modified to enable sap flow measurements in date palm trees in the southern Arava desert of Israel. A long and tough sensor was constructed to withstand insertion into the date palm's hard exterior stem. This stem is wide and fibrous, surrounded by an even tougher external non-conducting layer of dead leaf bases. Furthermore, being a monocot species, water flow does not necessarily occur through the outer part of the palm's stem, as in most trees. Therefore, it is highly important to investigate the variations of the sap flux densities and determine the preferable location for sap flow sensing within the stem. Once installed into fully grown date palm trees stationed on weighing lysimeters, sap flow as measured by the modified sensors was compared with the actual transpiration. Sap flow was found to be well correlated with transpiration, especially when using a recent calibration equation rather than the original Granier equation. Furthermore, inducing the axial variability of the sap flux densities was found to be highly important for accurate assessments of transpiration by sap flow measurements. The sensors indicated no transpiration at night, a high increase of transpiration from 06:00 to 09:00, maximum transpiration at 12:00, followed by a moderate reduction until 08:00; when transpiration ceased. These results were reinforced by the lysimeters' output. Reduced sap flux densities were detected at the stem's mantle when compared with its center. These results were reinforced by mechanistic measurements of the stem's specific hydraulic conductivity. Variance on the vertical axis was also observed, indicating an accelerated flow towards the upper parts of the tree and raising a hypothesis concerning dehydrating mechanisms of the date palm tree. Finally, the sensors indicated reduction in flow almost immediately after irrigation of field-grown trees was withheld, at a time when no climatic or phenological conditions could have led to reduction in transpiration.

Description: Northern forests are currently experiencing increasing mean temperatures, especially during autumn and spring. Consequently, alterations in carbon sequestration, leaf biochemical quality and freezing tolerance (FT) are likely to occur. The interactive effects of elevated temperature and ozone (O 3 ), the most harmful phytotoxic air pollutant, on Norway spruce ( Picea abies (L.) Karst.) seedlings were studied by analysing phenology, metabolite concentrations in the needles, FT and gas exchange. Sampling was performed in September and May. The seedlings were exposed to a year-round elevated temperature (+1.3 °C), and to 1.4 x ambient O 3 concentration during the growing season in the field. Elevated temperature increased the concentrations of amino acids, organic acids of the citric acid cycle and some carbohydrates, and reduced the concentrations of phenolic compounds, some organic acids of the shikimic acid pathway, sucrose, cyclitols and steroids, depending on the timing of the sampling. Although growth onset occurred earlier at elevated temperature, the temperature of 50% lethality (LT 50 ) was similar in the treatments. Photosynthesis and the ratio of photosynthesis to dark respiration were reduced by elevated temperature. Elevated concentrations of O 3 reduced the total concentration of soluble sugars, and tended to reduce LT 50 of the needles in September. These results show that alterations in needle chemical quality can be expected at elevated temperatures, but the seedlings' sensitivity to autumn and spring frosts is not altered. Elevated O 3 has the potential to disturb cold hardening of Norway spruce seedlings in autumn, and to alter the water balance of the seedling through changes in stomatal conductance ( g s ), while elevated temperature is likely to reduce g s and consequently reduce the O 3 -flux inside the leaves.

Description: Sunflecks are brief, intermittent periods of high photon flux density (PFD) that can significantly improve carbon gain in shaded forest understories and lower canopies of trees. In this review, we discuss the physiological basis of leaf-level responses to sunflecks and the mechanisms plants use to tolerate sudden changes in PFD and leaf temperature induced by sunflecks. We also examine the potential effects of climate change stresses (including elevated temperatures, rising CO 2 concentrations and drought) on the ability of tree species to use sunflecks, and advocate more research to improve our predictions of seedling and tree carbon gain in future climates. Lastly, while we have the ability to model realistic responses of photosynthesis to fluctuating PFD, dynamic responses of photosynthesis to sunflecks are not accounted for in current models of canopy carbon uptake, which can lead to substantial overestimates of forest carbon fixation. Since sunflecks are a critical component of seasonal carbon gain for shaded leaves, sunfleck regimes and physiological responses to sunflecks should be incorporated into models to more accurately capture forest carbon dynamics.

Description: In this study we investigated the role that ascorbate (AA) and glutathione (GSH) play in the plant pathogen interaction of susceptible Eucalyptus sieberi L. A. Johnson and resistant Eucalyptus sideroxylon Woolls with Phytophthora cinnamomi Rands root infection. In a glasshouse study, seedlings were grown in soil-free plant boxes to facilitate the inoculation of the root systems by a P. cinnamomi zoospore solution. Ascorbate and GSH concentrations were measured in infected roots and leaves, along with leaf gas exchange, chlorophyll fluorescence and carbohydrate concentrations over a time course up to 312 h (13 days) post-inoculation (pi). At the early stages of infection (from 24 h pi), significant decreases in AA and GSH concentrations were observed in the infected roots and leaves of the susceptible E. sieberi seedlings. At the later stage of infection (312 h pi), the earlier AA decreases in the leaves of infected plants had become significant increases. In contrast, late, significant AA increases in the absence of any GSH changes were observed in the infected roots of the resistant E. sideroxylon seedlings. In E. sideroxylon leaves, a significant GSH increase occurred at 24 h pi; however, by 312 h pi the earlier increase had become a significant decrease, while no changes occurred in AA. In E. sieberi , photosynthesis ( A ), stomatal conductance ( g s ) and PSII quantum efficiency ( PSII ) were reduced by ~60, 80 and 30%, respectively, in infected plants and remained significantly lower than uninfected controls for the duration of the experiment. Significant reductions in these parameters did not occur until later (120 h pi for g s and 312 h pi for A and PSII ), and to a lesser extent in the resistant species. Non-structural carbohydrate analysis of roots and leaves indicate that carbohydrate metabolism and resource flow between shoots and roots may have been altered at later infection stages. This study suggests that reduced antioxidant capacity, leaf physiological function and carbohydrate metabolism are associated with susceptibility in E. sieberi to P. cinnamomi infection, while AA increases and new root formation were associated with resistance in E . sideroxylon .

Description: Interspecific gene flow is common in oaks. In the Mediterranean, this process produced geographical differentiations and new species, which may have contributed to the diversification of the production of volatile terpenes in the oak species of this region. The endemic North African deciduous oak Quercus afares (Pomel) is considered to be a stabilized hybrid between the evergreen Quercus suber (L.) and the deciduous Quercus canariensis (Willd.), presumably being monoterpene and isoprene emitters, respectively. In a common garden experiment, we examined the terpene emission capacities, terpene synthase (TPS) activities and nuclear genetic markers in 52 trees of these three oak species. All but one of the Q. suber and Q. canariensis trees were found to be genetically pure, whereas most Q. afares trees possessed a mixed genotype with a predominance of Q. suber alleles. Analysis of the foliar terpene emissions and TPS activities revealed that all the Q. canariensis trees strongly produced isoprene while all the Q. suber trees were strong monoterpene producers. Quercus afares trees produced monoterpenes as well but at more variable and significantly lower rates, and with a monoterpene pattern different than that observed in Q. suber . Among 17 individuals tested, one Q. afares tree emitted only an insignificant amount of terpenes. No mixed isoprene/monoterpene emitter was detected. Our results suggest that the capacity and pattern of volatile terpene production in Algerian Q. afares populations have strongly diverged from those of its parental species and became quantitatively and qualitatively reduced, including the complete suppression of isoprene production.

Description: Low temperature plays a crucial role in seasonal development of woody plants and may directly impact crop production, more particularly in temperate fruit trees. Given its high genetic variability and adaptability to different climatic conditions, almond ( Prunus dulcis Mill.) is an interesting model to understand the mechanisms regulating low temperature sensing in fruit trees. In this paper, we report the cloning and characterization of two genes ( PdCBF1 and PdCBF2 ) belonging to the C-repeat-binding factor (CBF) family of transcription factors. Southern blotting analysis showed that this family is composed of at least five members. In almond shoots propagated in vitro, transcription of these genes was rapidly induced by low temperature, suggesting an involvement in cold acclimation. Transactivation assays showed that PdCBF1 and PdCBF2 could bind to dehydration responsive element/C-repeat containing sequences, as activators of gene expression. In addition, induction of both PdCBF s by cold was higher towards the end of the day, which agreed with the expression pattern of PdDehydrin1 , a predicted CBF target gene. Furthermore, PdCBF1 and PdCBF2 were also transiently induced by abscisic acid and drought treatments. Considering the bin mapping analysis that correlated PdCBF s and PdDHN1 (respectively in linkage groups 5 and 7) with two different quantitative trait locicontrolling blooming time, it is relevant to perform further association studies that may validate their effect on this trait.

Description: Leaves, the distal section of the soil–plant–atmosphere continuum, exhibit the lowest water potentials in a plant. In contrast to angiosperm leaves, knowledge of the hydraulic architecture of conifer needles is scant. We investigated the hydraulic efficiency and safety of Pinus pinaster needles, comparing different techniques. The xylem hydraulic conductivity ( k s ) and embolism vulnerability ( P 50 ) of both needle and stem were measured using the cavitron technique. The conductance and vulnerability of whole needles were measured via rehydration kinetics, and Cryo-SEM and 3D X-ray microtomographic observations were used as reference tools to validate physical measurements. The needle xylem of P. pinaster had lower hydraulic efficiency ( k s = 2.0 x 10 –4  m 2  MPa –1  s –1 ) and safety ( P 50 = – 1.5 MPa) than stem xylem ( k s = 7.7 x 10 –4  m 2  MPa –1  s –1 ; P 50 = – 3.6 to – 3.2 MPa). P 50 of whole needles (both extra-vascular and vascular pathways) was – 0.5 MPa, suggesting that non-vascular tissues were more vulnerable than the xylem. During dehydration to – 3.5 MPa, collapse and embolism in xylem tracheids, and gap formation in surrounding tissues were observed. However, a discrepancy in hydraulic and acoustic results appeared compared with visualizations, arguing for greater caution with these techniques when applied to needles. Our results indicate that the most distal parts of the water transport pathway are limiting for hydraulics of P. pinaster . Needle tissues exhibit a low hydraulic efficiency and low hydraulic safety, but may also act to buffer short-term water deficits, thus preventing xylem embolism.

Description: The lignification process in poplar tension wood lignified cell wall layers, specifically the S 1 and S 2 layers and the compound middle lamella (CML), was analysed using ultraviolet (UV) and transmission electron microscopy (TEM). Variations in the thickness of the gelatinous layer (G-layer) were also measured to clarify whether the lignified cell wall layers had completed their lignification before the deposition of G-layers, or, on the contrary, if lignification of these layers was still active during G-layer formation. Observations using UV microscopy and TEM indicated that both UV absorbance and the degree of potassium permanganate staining increased in the CML and S 1 and S 2 layers during G-layer formation, suggesting that the lignification of these lignified layers is still in progress during G-layer formation. In the context of the cell-autonomous monolignol synthesis hypothesis, our observations suggest that monolignols must go through the developing G-layer during the lignification of CML and the S 1 and S 2 layers. The alternative hypothesis of external synthesis (in the rays) does not require that monolignols go through the G-layer before being deposited in the CML, or the S 1 and S 2 layers. Interestingly, the previous observation of lignin in the poplar G-layer was not confirmed with the microscopy techniques used in the present study.

Description: Oak forests dominate much of the eastern USA, but their future is uncertain due to a number of threats and widespread failure of oak regeneration. A sudden loss of oaks ( Quercus spp . ) could be accompanied by major changes in forest nitrogen (N) cycles with important implications for plant nutrient uptake and tree species composition. In this study, we measured the changes in N use and growth rates of black birch trees ( Betula lenta L.) following oak girdling at the Black Rock Forest in southeastern New York, USA. Data were collected from nine experimental plots composed of three treatments: 100% oaks girdled (OG), 50% oaks girdled (O50) and control (C). Foliar N concentration and foliar 15 N abundance increased significantly in the oak-girdled plots relative to the control, indicating that the loss of oaks significantly altered N cycling dynamics. As mineralization and nitrification rates increase following oak loss, black birch trees increase N absorption as indicated by higher foliar N content and increased growth rates. Foliar N concentration increased by 15.5% in the O50 and 30.6% in the OG plots relative to the control, while O50 and OG plots were enriched in 15 N by 1.08 and 3.33, respectively ( P 〈 0.0001). A 641% increase in black birch growth rates in OG plots suggests that this species is able to respond to additional N availability and/or increased light availability. The loss of oaks and subsequent increase in black birch productivity may have a lasting impact on ecosystem form and function.

Description: Introducing nitrogen-fixing tree species in fast-growing eucalypt plantations has the potential to improve soil nitrogen availability compared with eucalypt monocultures. Whether or not the changes in soil nutrient status and stand structure will lead to mixtures that out-yield monocultures depends on the balance between positive interactions and the negative effects of interspecific competition, and on their effect on carbon (C) uptake and partitioning. We used a C budget approach to quantify growth, C uptake and C partitioning in monocultures of Eucalyptus grandis (W. Hill ex Maiden) and Acacia mangium (Willd.) (treatments E100 and A100, respectively), and in a mixture at the same stocking density with the two species at a proportion of 1 : 1 (treatment MS). Allometric relationships established over the whole rotation, and measurements of soil CO 2 efflux and aboveground litterfall for ages 4–6 years after planting were used to estimate aboveground net primary production (ANPP), total belowground carbon flux (TBCF) and gross primary production (GPP). We tested the hypotheses that (i) species differences for wood production between E. grandis and A. mangium monocultures were partly explained by different C partitioning strategies, and (ii) the observed lower wood production in the mixture compared with eucalypt monoculture was mostly explained by a lower partitioning aboveground. At the end of the rotation, total aboveground biomass was lowest in A100 (10.5 kg DM m –2 ), intermediate in MS (12.2 kg DM m –2 ) and highest in E100 (13.9 kg DM m –2 ). The results did not support our first hypothesis of contrasting C partitioning strategies between E. grandis and A. mangium monocultures: the 21% lower growth ( B w ) in A100 compared with E100 was almost entirely explained by a 23% lower GPP, with little or no species difference in ratios such as TBCF/GPP, ANPP/TBCF, B w /ANPP and B w /GPP. In contrast, the 28% lower B w in MS than in E100 was explained both by a 15% lower GPP and by a 15% lower fraction of GPP allocated to wood growth, thus partially supporting our second hypothesis: mixing the two species led to shifts in C allocations from above- to belowground, and from growth to litter production, for both species.

Description: Soil temperature is proposed to affect the photosynthetic rate and carbon allocation in boreal trees through sink limitation. The aim of this study was to investigate the effect of temperature on CO 2 exchange, biomass partitioning and ectomycorrhizal (ECM) fungi of boreal tree species. We measured carbon allocation, above- and below-ground CO 2 exchange and the species composition of associated ECM fungi in the rhizosphere of Scots pine ( Pinus sylvestris L.), Norway spruce ( Picea abies K.) and silver birch ( Betula pendula Roth) seedlings grown in soil maintained at 7–12, 12–15 and 16–22 °C. We found increased root biomass and photosynthetic rate at higher soil temperatures, but simultaneously with photosynthesis rate, higher temperature generally increased soil respiration as well as shoot, and root and rhizosphere respiration. The net CO 2 exchange and seedling biomass did not increase significantly with increasing temperature due to a concomitant increase in carbon assimilation and respiration rates. The 2-month-long growth period in different soil temperatures did not alter the ECM fungi species composition and the below-ground carbon sink strength did not seem to be directly related to ECM biomass and species composition in any of the tree species. Ectomycorrhizal species composition and number of mycorrhiza did not explain the CO 2 exchange results at different temperatures.

Description: Wood production represents a large but variable fraction of gross primary production (GPP) in highly productive Eucalyptus plantations. Assessing patterns of carbon (C) partitioning (C flux as a fraction of GPP) between above- and belowground components is essential to understand mechanisms driving the C budget of these plantations. Better knowledge of fluxes and partitioning to woody and non-woody tissues in response to site characteristics and resource availability could provide opportunities to increase forest productivity. Our study aimed at investigating how C allocation varied within one apparently homogeneous 90 ha stand of Eucalyptus grandis (W. Hill ex Maiden) in Southeastern Brazil. We assessed annual above­ground net primary production (ANPP: stem, leaf, and branch production) and total belowground C flux (TBCF: the sum of root production and respiration and mycorrhizal production and respiration), GPP (computed as the sum of ANPP, TBCF and estimated aboveground respiration) on 12 plots representing the gradient of productivity found within the stand. The spatial heterogeneity of topography and associated soil attributes across the stand likely explained this fertility gradient. Component fluxes of GPP and C partitioning were found to vary among plots. Stem NPP ranged from 554 g C m –2 year –1 on the plot with lowest GPP to 923 g C m –2 year –1 on the plot with highest GPP. Total belowground carbon flux ranged from 497 to 1235 g C m –2 year –1 and showed no relationship with ANPP or GPP. Carbon partitioning to stem NPP increased from 0.19 to 0.23, showing a positive trend of increase with GPP ( R 2 = 0.29, P = 0.07). Variations in stem wood production across the gradient of productivity observed at our experimental site were a result of the variability in C partitioning to different forest system components.

Description: We review approaches to predicting carbon and nitrogen allocation in forest models in terms of their underlying assumptions and their resulting strengths and limitations. Empirical and allometric methods are easily developed and computationally efficient, but lack the power of evolution-based approaches to explain and predict multifaceted effects of environmental variability and climate change. In evolution-based methods, allocation is usually determined by maximization of a fitness proxy, either in a fixed environment, which we call optimal response (OR) models, or including the feedback of an individual's strategy on its environment (game-theoretical optimization, GTO). Optimal response models can predict allocation in single trees and stands when there is significant competition only for one resource. Game-theoretical optimization can be used to account for additional dimensions of competition, e.g., when strong root competition boosts root allocation at the expense of wood production. However, we demonstrate that an OR model predicts similar allocation to a GTO model under the root-competitive conditions reported in free-air carbon dioxide enrichment (FACE) experiments. The most evolutionarily realistic approach is adaptive dynamics (AD) where the allocation strategy arises from eco-evolutionary dynamics of populations instead of a fitness proxy. We also discuss emerging entropy-based approaches that offer an alternative thermodynamic perspective on allocation, in which fitness proxies are replaced by entropy or entropy production. To help develop allocation models further, the value of wide-ranging datasets, such as FLUXNET, could be greatly enhanced by ancillary measurements of driving variables, such as water and soil nitrogen availability.

Description: Deep root water uptake and hydraulic redistribution (HR) have been shown to play a major role in forest ecosystems during drought, but little is known about the impact of climate change, fertilization and soil characteristics on HR and its consequences on water and carbon fluxes. Using data from three mid-rotation loblolly pine plantations, and simulations with the process-based model MuSICA, this study indicated that HR can mitigate the effects of soil drying and had important implications for carbon uptake potential and net ecosystem exchange (NEE), especially when N fertilization is considered. At the coastal site (C), characterized by deep organic soil, HR increased dry season tree transpiration ( T ) by up to 40%, and such an increase affected NEE through major changes in gross primary productivity (GPP). Deep-rooted trees did not necessarily translate into a large volume of HR unless soil texture allowed large water potential gradients to occur, as was the case at the sandy site (S). At the Piedmont site (P) characterized by a shallow clay-loam soil, HR was low but not negligible, representing up to 10% of T . In the absence of HR, it was predicted that at the C, S and P sites, annual GPP would have been diminished by 19, 7 and 9%, respectively. Under future climate conditions HR was predicted to be reduced by up to 25% at the C site, reducing the resilience of trees to precipitation deficits. The effect of HR on T and GPP was predicted to diminish under future conditions by 12 and 6% at the C and P sites, respectively. Under future conditions, T was predicted to stay the same at the P site, but to be marginally reduced at the C site and slightly increased at the S site. Future conditions and N fertilization would decrease T by 25% at the C site, by 15% at the P site and by 8% at the S site. At the C and S sites, GPP was estimated to increase by 18% and by 〉70% under future conditions, respectively, with little effect of N fertilization. At the P site, future conditions would stimulate GPP by only 12%, but future conditions plus N fertilization would increase GPP by 24%. As a consequence, in all sites, water use efficiency was predicted to improve dramatically with future conditions. Modeling the effect of reduced annual precipitation indicated that limited water availability would decrease all carbon fluxes, including NEE and respiration. Our simulations highlight the interactive effects of nutrients and elevated CO 2 , and showed that the effect of N fertilization would be greater under future climate conditions.

Description: Warmer climates induced by elevated atmospheric CO 2 (eCO 2 ) are expected to increase damaging bark beetle activity in pine forests, yet the effect of eCO 2 on resin production—the tree's primary defense against beetle attack—remains largely unknown. Following growth-differentiation balance theory, if extra carbohydrates produced under eCO 2 are not consumed by respiration or growth, resin production could increase. Here, the effect of eCO 2 on resin production of mature pines is assessed. As predicted, eCO 2 enhanced resin flow by an average of 140% ( P = 0.03) in canopy dominants growing in low-nitrogen soils, but did not affect resin flow in faster-growing fertilized canopy dominants or in carbohydrate-limited suppressed individuals. Thus, pine trees may become increasingly protected from bark beetle attacks in an eCO 2 climate, except where they are fertilized or are allowed to become overcrowded.

Description: Research on the degree to which carbon (C) availability limits growth in trees, as well as recent trends in climate change and concurrent increases in drought-related tree mortality, have led to a renewed focus on the physiological mechanisms associated with tree growth responses to current and future climate. This has led to some dispute over the role of stored non-structural C compounds as indicators of a tree's current demands for photosynthate. Much of the uncertainty surrounding this issue could be resolved by developing a better understanding of the potential functions of non-structural C stored within trees. In addition to functioning as a buffer to reconcile temporal asynchrony between C demand and supply, the storage of non-structural C compounds may be under greater regulation than commonly recognized. We propose that in the face of environmental stochasticity, large, long-lived trees may require larger C investments in storage pools as safety margins than previously recognized, and that an important function of these pools may be to maintain hydraulic transport, particularly during episodes of severe stress. If so, survival and long-term growth in trees remain a function of C availability. Given that drought, freeze–thaw events and increasing tree height all impose additional constraints on vascular transport, the common trend of an increase in non-structural carbohydrate concentrations with tree size, drought or cold is consistent with our hypothesis. If the regulated maintenance of relatively large constitutive stored C pools in trees serves to maintain hydraulic integrity, then the minimum thresholds are expected to vary depending on the specific tissues, species, environment, growth form and habit. Much research is needed to elucidate the extent to which allocation of C to storage in trees is a passive vs. an active process, the specific functions of stored C pools, and the factors that drive active C allocation to storage.

Description: In the present experiment, the single and combined effects of elevated temperature and ozone (O 3 ) on four silver birch genotypes (gt12, gt14, gt15 and gt25) were studied in an open-air field exposure design. Above- and below-ground biomass accumulation, stem growth and soil respiration were measured in 2008. In addition, a 13 C-labelling experiment was conducted with gt15 trees. After the second exposure season, elevated temperature increased silver birch above- and below-ground growth and soil respiration rates. However, some of these variables showed that the temperature effect was modified by tree genotype and prevailing O 3 level. For instance, in gt14 soil respiration was increased in elevated temperature alone (T) and in elevated O 3 and elevated temperature in combination (O 3 + T) treatments, but in other genotypes O 3 either partly (gt12) or totally nullified (gt25) temperature effects on soil respiration, or acted synergistically with temperature (gt15). Before leaf abscission, all genotypes had the largest leaf biomass in T and O 3 + T treatments, whereas at the end of the season temperature effects on leaf biomass depended on the prevailing O 3 level. Temperature increase thus delayed and O 3 accelerated leaf senescence, and in combination treatment O 3 reduced the temperature effect. Photosynthetic : non-photosynthetic tissue ratios (P : nP ratios) showed that elevated temperature increased foliage biomass relative to woody mass, particularly in gt14 and gt12, whereas O 3 and O 3 + T decreased it most clearly in gt25. O 3 -caused stem growth reductions were clearest in the fastest-growing gt14 and gt25, whereas mycorrhizal root growth and sporocarp production increased under O 3 in all genotypes. A labelling experiment showed that temperature increased tree total biomass and hence 13 C fixation in the foliage and roots and also label return was highest under elevated temperature. Ozone seemed to change tree 13 C allocation, as it decreased foliar 13 C excess amount, simultaneously increasing 13 C excess obtained from the soil. The present results suggest that warming has potential to increase silver birch growth and hence carbon (C) accumulation in tree biomass, but the final magnitude of this C sink strength is partly counteracted by temperature-induced increase in soil respiration rates and simultaneous O 3 stress. Silver birch populations' response to climate change will also largely depend on their genotype composition.

Description: The dynamics of rapid changes in carbon (C) partitioning within forest ecosystems are not well understood, which limits improvement of mechanistic models of C cycling. Our objective was to inform model processes by describing relationships between C partitioning and accessible environmental or physiological measurements, with a special emphasis on short-term C flux through a forest ecosystem. We exposed eight 7-year-old loblolly pine ( Pinus taeda L.) trees to air enriched with 13 CO 2 and then implemented adjacent light shade (LS) and heavy shade (HS) treatments in order to manipulate C uptake and flux. The impacts of shading on photosynthesis, plant water potential, sap flow, basal area growth, root growth and soil CO 2 efflux rate (CER) were assessed for each tree over a 3-week period. The progression of the 13 C label was concurrently tracked from the atmosphere through foliage, phloem, roots and surface soil CO 2 efflux. The HS treatment significantly reduced C uptake, sap flow, stem growth and fine root standing crop, and resulted in greater residual soil water content to 1 m depth. Soil CER was strongly correlated with sap flow on the previous day, but not the current day, with no apparent treatment effect on the relationship. Although there were apparent reductions in new C flux belowground, the HS treatment did not noticeably reduce the magnitude of belowground autotrophic and heterotrophic respiration based on surface soil CER, which was overwhelmingly driven by soil temperature and moisture. The 13 C label was immediately detected in foliage on label day (half-life = 0.5 day), progressed through phloem by Day 2 (half-life = 4.7 days), roots by Days 2–4, and subsequently was evident as respiratory release from soil which peaked between Days 3 and 6. The 13 C of soil CO 2 efflux was strongly correlated with phloem 13 C on the previous day, or 2 days earlier. While the 13 C label was readily tracked through the ecosystem, the fate of root C through respiratory, mycorrhizal or exudative release pathways was not assessed. These data detail the timing and relative magnitude of C flux through various components of a young pine stand in relation to environmental conditions.

Description: Understanding the underlying mechanisms that account for the impact of potassium (K) fertilization and its replacement by sodium (Na) on tree growth is key to improving the management of forest plantations that are expanding over weathered tropical soils with low amounts of exchangeable bases. A complete randomized block design was planted with Eucalyptus grandis (W. Hill ex Maiden) to quantify growth, carbon uptake and carbon partitioning using a carbon budget approach. A combination of approaches including the establishment of allometric relationships over the whole rotation and measurements of soil CO 2 efflux and aboveground litterfall at the end of the rotation were used to estimate aboveground net production (ANPP), total belowground carbon flux and gross primary production (GPP). The stable carbon isotope ( 13 C) of stem wood α-cellulose produced every year was used as a proxy for stomatal limitation of photosynthesis. Potassium fertilization increased GPP and decreased the fraction of carbon allocated belowground. Aboveground net production was strongly enhanced, and because leaf lifespan increased, leaf biomass was enhanced without any change in leaf production, and wood production (P W ) was dramatically increased. Sodium application decreased the fraction of carbon allocated belowground in a similar way, and enhanced GPP, ANPP and P W , but to a lesser extent compared with K fertilization. Neither K nor Na affected 13 C of stem wood α-cellulose, suggesting that water-use efficiency was the same among the treatments and that the inferred increase in leaf photosynthesis was not only related to a higher stomatal conductance. We concluded that the response to K fertilization and Na addition on P W resulted from drastic changes in carbon allocation.

Description: 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.

Description: Urbanization is accelerating across the globe, elevating the importance of studying urban ecology. Urban environments exhibit several factors affecting plant growth and function, including high temperatures (particularly at night), CO 2 concentrations and atmospheric nitrogen deposition. We investigated the effects of urban environments on growth in Quercus rubra L. seedlings. We grew seedlings from acorns for one season at four sites along an urban–rural transect from Central Park in New York City to the Catskill Mountains in upstate New York (difference in average maximum temperatures of 2.4 °C; difference in minimum temperatures of 4.6 °C). In addition, we grew Q. rubra seedlings in growth cabinets (GCs) mimicking the seasonal differential between the city and rural sites (based on a 5-year average). In the field experiment, we found an eightfold increase in biomass in urban-grown seedlings relative to those grown at rural sites. This difference was primarily related to changes in growth allocation. Urban-grown seedlings and seedlings grown at urban temperatures in the GCs exhibited a lower root: shoot ratio (urban ~0.8, rural/remote ~1.5), reducing below-ground carbon costs associated with construction and maintenance. These urban seedlings instead allocated more growth to leaves than did rural-grown seedlings, resulting in 10-fold greater photosynthetic area but no difference in photosynthetic capacity of foliage per unit area. Seedlings grown at urban temperatures in both the field and GC experiments had higher leaf nitrogen concentrations per unit area than those grown at cooler temperatures (increases of 23% in field, 32% in GC). Lastly, we measured threefold greater 13 C enrichment of respired CO 2 (relative to substrate) in urban-grown leaves than at other sites, which may suggest greater allocation of respiratory function to growth over maintenance. It also shows that lack of differences in total R flux in response to environmental conditions may mask dramatic shifts in respiratory functioning. Overall, our findings indicating greater seedling growth and establishment at a critical regeneration phase of forest development may have important implications for the ecology of urban forests as well as the predicted growth of the terrestrial biosphere in temperate regions in response to climate change.

Description: Establishing quantitative links between plant hydraulic properties and the response of transpiration to environmental factors such as atmospheric vapor pressure deficit ( D ) is essential for improving our ability to understand plant water relations across a wide range of species and environmental conditions. We studied stomatal responses to D in irrigated trees in the urban landscape of Los Angeles, California. We found a strong linear relationship between the sensitivity of tree-level transpiration estimated from sap flux ( m T ; slope of the relationship between tree transpiration and ln D ) and transpiration at D = 1 kPa ( E Tref ) that was similar to previous surveys of stomatal behavior in natural environments. In addition, m T was significantly related to vulnerability to cavitation of branches ( P 50 ). While m T did not appear to differ between ring- and diffuse-porous species, the relationship between m T and P 50 was distinct by wood anatomy. Therefore, our study confirms systematic differences in water relations in ring- versus diffuse-porous species, but these differences appear to be more strongly related to the relationship between stomatal sensitivity to D and vulnerability to cavitation rather than to stomatal sensitivity per se.

Description: Global change challenges forest adaptability at the distributional limit of species. We studied ring-porous Quercus canariensis Willd. xylem traits to analyze how they adjust to spatio-temporal variability in climate. Trees were sampled along altitudinal transects, and annual time series of radial growth (ring width (RW)) and several earlywood vessel (EV) traits were built to analyze their relationships with climate. The trees responded to increasing water constraints with decreasing altitude and changes in climate in the short term but the analyses showed that xylem did not acclimate in response to long-term temperature increase during the past 30 years. The plants' adjustment to climate variability was expressed in a different but complementary manner by the different xylem traits. At low elevations, trees exhibited higher correlations with water stress indices and trees acclimated to more xeric conditions at low elevations by reducing radial growth and hydraulic diameter ( D H ) but increasing the density of vessels (DV). Average potential conductivity ( K H ) was similar for trees at different altitudes. However, inter-tree differences in xylem traits were higher than those between altitudes, suggesting a strong influence of individual genetic features or micro-site conditions. Trees exhibited higher RW those years with larger D H and particularly the linear density of vessels (DV l ), but partly, climatic signals expressed in RW differed from those in EVs. Trees produced larger D H after cold winters and wet years. Ring width responded positively to wet and cool weather in fall and spring, whereas the response to climate of DV and K H was generally opposite to that of RW. These relationships likely expressed the negative impact of high respiration rates in winter on the carbon pools used to produce the EVs in the next spring and the overall positive influence of water availability for trees. Our results showed that trees at different sites were able to adjust their hydraulic architecture to climatic variability and temperature increase during recent decades coordinating several complementary traits. Nonetheless, it should be monitored whether they will succeed to acclimate to future climatic scenarios of increasing water stress.

Description: Despite considerable investigations of diurnal water use characteristics in different plant functional groups, the research on daily water use strategies of woody bamboo grasses remains lacking. We studied the daily water use and gas exchange of Sinarundinaria nitida (Mitford) Nakai, an abundant subtropical bamboo species in Southwest China. We found that the stem relative water content (RWC) and stem hydraulic conductivity ( K s ) of this bamboo species did not decrease significantly during the day, whereas the leaf RWC and leaf hydraulic conductance ( K leaf ) showed a distinct decrease at midday, compared with the predawn values. Diurnal loss of K leaf was coupled with a midday decline in stomatal conductance ( g s ) and CO 2 assimilation. The positive root pressures in the different habitats were of sufficient magnitude to refill the embolisms in leaves. We concluded that (i) the studied bamboo species does not use stem water storage for daily transpiration; (ii) diurnal down-regulation in K leaf and g s has the function to slow down potential water loss in stems and protect the stem hydraulic pathway from cavitation; (iii) since K leaf did not recover during late afternoon, refilling of embolism in bamboo leaves probably fully depends on nocturnal root pressure. The embolism refilling mechanism by root pressure could be helpful for the growth and persistence of this woody monocot species.

Description: Pinus radiata D. Don is one of the most abundant species in the north of Spain. Knowledge of drought response mechanisms is essential to guarantee plantation survival under reduced water supply as predicted in the future. Tolerance mechanisms are being studied in breeding programs, because information on such mechanisms can be used for genotype selection. In this paper, we analyze the changes of leaf water potential, hydraulic conductance ( K leaf ), stomatal conductance and phytohormones under drought in P. radiata breeds (O1, O2, O3, O4, O5 and O6) from different climatology areas, hypothesizing that they could show variable drought tolerance. As a primary signal, drought decreased cytokinin (zeatin and zeatin riboside—Z + ZR) levels in needles parallel to K leaf and gas exchange. When Z + ZR decreased by 65%, indole-3-acetic acid (IAA) and abscisic acid (ABA) accumulation started as a second signal and increments were higher for IAA than for ABA. When plants decreased by 80%, Z + ZR and K leaf doubled their ABA and IAA levels, the photosystem II yield decreased and the electrolyte leakage increased. At the end of the drought period, less tolerant breeds increased IAA over 10-fold compared with controls. External damage also induced jasmonic acid accumulation in all breeds except in O5 ( P. radiata var. radiata x var. cedrosensis ), which accumulated salicylic acid as a defense mechanism. After rewatering, only the most tolerant plants recovered their K leaf, perhaps due to an IAA decrease and 1-aminocyclopropane-1-carboxylic acid maintenance. From all phytohormones, IAA was the most representative ‘water deficit signal’ in P. radiata .

Description: Understanding the response of leaf hydraulic conductance ( K leaf ) to light is a challenge in elucidating plant–water relationships. Recent data have shown that the effect of light on K leaf is not systematically related to aquaporin regulation, leading to conflicting conclusions. Here we investigated the relationship between light, K leaf , and aquaporin transcript levels in five tree species ( Juglans regia L., Fagus sylvatica L., Quercus robur L., Salix alba L. and Populus tremula L.) grown in the same environmental conditions, but differing in their K leaf responses to light. Moreover, the K leaf was measured by two independent methods (high-pressure flow metre (HPFM) and evaporative flux method (EFM)) in the most ( J. regia ) and least ( S. alba ) responsive species and the transcript levels of aquaporins were analyzed in perfused and unperfused leaves. Here, we found that the light-induced K leaf value was closely related to stronger expression of both the PIP1 and PIP2 aquaporin genes in walnut ( J. regia ), but to stimulation of PIP1 aquaporins alone in F. sylvatica and Q. robur . In walnut, all newly identified aquaporins were found to be upregulated in the light and downregulated in the dark, further supporting the relationship between the light-mediated induction of K leaf and aquaporin expression in walnut. We also demonstrated that the K leaf response to light was quality-dependent, K leaf being 60% lower in the absence of blue light. This decrease in K leaf was correlated with strong downregulation of three PIP2 aquaporins and of all the PIP1 aquaporins tested. These data support a relationship between light-mediated K leaf regulation and the abundance of aquaporin transcripts in the walnut tree.

Description: Photosynthetic acclimation to highly variable local irradiance within the tree crown plays a primary role in determining tree carbon uptake. This study explores the plasticity of leaf structural and physiological traits in response to the interactive effects of ontogeny, water stress and irradiance in adult almond trees that have been subjected to three water regimes (full irrigation, deficit irrigation and rain-fed) for a 3-year period (2006–08) in a semiarid climate. Leaf structural (dry mass per unit area, N and chlorophyll content) and photosynthetic (maximum net CO 2 assimilation, A max , maximum stomatal conductance, g s,max , and mesophyll conductance, g m ) traits and stem-to-leaf hydraulic conductance ( K s-l ) were determined throughout the 2008 growing season in leaves of outer south-facing (S-leaves) and inner northwest-facing (NW - leaves) shoots. Leaf plasticity was quantified by means of an exposure adjustment coefficient ( = 1- X NW / X S ) for each trait ( X ) of S- and NW-leaves. Photosynthetic traits and K s-l exhibited higher irradiance-elicited plasticity (higher ) than structural traits in all treatments, with the highest and lowest plasticity being observed in the fully irrigated and rain-fed trees, respectively. Our results suggest that water stress modulates the irradiance-elicited plasticity of almond leaves through changes in crown architecture. Such changes lead to a more even distribution of within-crown irradiance, and hence of the photosynthetic capacity, as water stress intensifies. Ontogeny drove seasonal changes only in the of area- and mass-based N content and mass-based chlorophyll content, while no leaf age-dependent effect was observed on as regards the physiological traits. Our results also indicate that the irradiance-elicited plasticity of A max is mainly driven by changes in leaf dry mass per unit area, in g m and, most likely, in the partitioning of the leaf N content.

Description: Seedlings of a conifer ( Pinus radiata D. Don) and a broad leaf angiosperm ( Eucalyptus globulus Labill.) were grown for 100 days in two growth cabinets at 45 or 65% relative humidity. The seedlings were exposed to treatments designed to modify carbon assimilation rates and capacities, stomatal conductance and transpiration to test conceptual models that attempt to clarify the interpretation of carbon isotope discrimination ( 13 C) by using oxygen isotope enrichment ( 18 O). Differences in relative humidity and within-cabinet treatments (including lower irradiance, lower nitrogen inputs, higher leaf temperature and lower moisture status than control seedlings) produced significant differences in assimilation rates, photosynthetic capacities, stomatal conductance, leaf transpiration rates and leaf evaporative enrichment. The dual-isotope approach accurately interpreted the cause of variation in wood cellulose 13 C for some of the treatments, but not for others. We also tested whether we could use 13 C variation to constrain the interpretation of 18 O variation. Carbon isotope discrimination appears to be influenced by transpiration (providing information on leaf evaporative enrichment), but the results did not provide a clear way to interpret such variation. The dual-isotope approach appears to be valid conceptually, but more work is needed to make it operational under different scenarios.

Description: Severe droughts may increase physiological stress on long-lived woody vegetation, occasionally leading to rapid defoliation and progressive increase in mortality of overstorey trees. Over the last few years, episodes of drought-induced tree dieback have been documented in a variety of woodlands and forests around the world. However, the factors determining tree survival and subsequent recovery are still poorly understood, especially in resprouter species. We have studied the effects of a single drought episode on crown condition in a holm oak ( Quercus ilex L.) forest located in NE Spain 7 years after the drought event. Generalized linear models were used to study the environmental correlates of forest crown condition 7 years after the drought event. Additionally, we evaluated the association between crown condition and the carbon and nutrient reserves stored in lignotubers 7 years after the drought. Our study reveals the multifactor nature of a drought-driven forest dieback in which soil depth and the characteristics of individual trees, particularly their number of stems, determined a complex spatial pattern of tree-level responses. This dieback was associated with a depletion of the carbon reserves in lignotubers 7 years after the episode, representing a reduction of up to 60% in highly drought-damaged trees. Interestingly, in the absence of new acute droughts, successive surveys in 2007–11 showed a direct association between carbon reserves depletion and further deterioration of crown condition. More frequent droughts, as predicted by climate change projections, may lead to a progressive depletion of carbon reserves and to a loss of resilience in Mediterranean resprouter species.

Description: Leaves of Mediterranean evergreens experience large variations in gas exchange rates over their life span due to aging and seasonally changing environmental conditions. Accounting for the changing respiratory physiology of leaves over time will help improve estimations of leaf and whole-plant carbon balances. Here we examined seasonal variations in light-saturated net CO 2 assimilation ( A max ), dark respiration ( R d ) and the proportional change in R d per 10 °C change in temperature ( Q 10 of R d ) in previous-year (PY) and current-year (CY) leaves of the broadleaved evergreen tree Quercus ilex L. A max and R d were lower in PY than in CY leaves. Differences in nitrogen between cohorts only partly explained such differences, and rates of A max and R d expressed per unit of leaf nitrogen were still significantly different between cohorts. The decline in A max in PY leaves did not result in the depletion of total non-structural carbohydrates, whose concentration was in fact higher in PY than CY leaves. Leaf-level carbon balance modeled from gas exchange data was positive at all ages. Q 10 of R d did not differ significantly between leaf cohorts; however, failure to account for distinct R d between cohorts misestimated canopy leaf respiration by 13% across dates when scaling up leaf measurements to the canopy. In conclusion, the decline in A max in old leaves that are close to or exceed their mean life span does not limit the availability of carbohydrates, which are probably needed to sustain new growth, as well as R d and nutrient resorption during senescence. Accounting for leaf age as a source of variation of R d improves the estimation of foliar respiratory carbon release at the stand scale.

Description: We investigated whether timing and rate of growth are related to the life strategies and fitness of three conifer species. Intra-annual dynamics of wood formation, shoot elongation and needle phenology were monitored over 3 years in five Norway spruces ( Picea abies (L.) Karst.), five Scots pines ( Pinus sylvestris L.) and five silver firs ( Abies alba Mill.) grown intermixed. For the three species, the growing season (delimited by cambial activity onset and cessation) lasted about 4 months, while the whole process of wood formation lasted 5–6 months. Needle unfolding and shoot elongation followed the onset of cambial activity and lasted only one-third of the season. Pines exhibited an ‘extensive strategy’ of cambial activity, with long durations but low growth rates, while firs and spruces adopted an ‘intensive strategy’ with shorter durations but higher growth rates. We estimated that about 75% of the annual radial increment variability was attributable to the rate of cell production, and only 25% to its duration. Cambial activity rates culminated at the same time for the three species, whereas shoot elongation reached its maximal rate earlier in pines. Results show that species-specific life strategies are recognizable through functional traits of intra-annual growth dynamics. The opposition between Scots pine extensive strategy and silver fir and Norway spruce intensive strategy supports the theory that pioneer species are greater resource expenders and develop riskier life strategies to capture resources, while shade-tolerant species utilize resources more efficiently and develop safer life strategies. Despite different strategies, synchronicity of the maximal rates of cambial activity suggests a strong functional convergence between co-existing conifer species, resulting in head-on competition for resources.

Description: Foliar anatomy and morphology are strongly related to physiological performance; therefore, phenotypic plasticity in leaves to variations in environmental conditions, such as irradiance and soil moisture availability, can be related to growth rate and survivorship, mainly during critical growth phases, such as establishment. The aim of this work was to analyze changes in the foliar internal anatomy (tissue proportions and cell dimensions) and external morphology (leaf length, width and area) of Nothofagus pumilio (Poepp. et Endl.) Krasser seedlings growing in a greenhouse under controlled irradiance (three levels) and soil moisture (two levels) during one growing season (measured three times), and to relate them to physiological traits. Three irradiance levels (4, 26 and 64% of the natural incident light) and two soil moisture levels (40 and 80% soil capacity) were evaluated during November, January and March. Internal foliar anatomy of seedlings was analyzed using digital photographs of histological cuttings, while leaf gross morphology was measured using digital calipers and image analysis software. Most internal anatomical variables presented significant differences under different irradiance levels during the growing season, but differences were not detected between soil moisture levels. Palisade parenchyma was the tissue most sensitive to irradiance levels, and high irradiance levels (64% natural incident light) produced greater values in most of the internal anatomical variables than lower irradiance levels (4–24% natural incident light). Complementarily, larger leaves were observed in medium and low irradiance levels, as well as under low soil moisture levels (40% soil capacity). The relationship of main results with some eco-physiological traits was discussed. Foliar internal anatomical and external morphological plasticity allows quick acclimation of seedlings to environmental changes (e.g., during harvesting). These results can be used to propose new forest practices that consider soil moisture and light availability changes to maintain high physiological performance of seedlings.

Description: The phenotypic responses of functional traits in natural populations are driven by genetic diversity and phenotypic plasticity. These two mechanisms enable trees to cope with rapid climate change. We studied two European temperate tree species (sessile oak and European beech), focusing on (i) in situ variations of leaf functional traits (morphological and physiological) along two altitudinal gradients and (ii) the extent to which these variations were under environmental and/or genetic control using a common garden experiment. For all traits, altitudinal trends tended to be highly consistent between species and transects. For both species, leaf mass per area displayed a positive linear correlation with altitude, whereas leaf size was negatively correlated with altitude. We also observed a significant increase in leaf physiological performance with increasing altitude: populations at high altitudes had higher maximum rates of assimilation, stomatal conductance and leaf nitrogen content than those at low altitudes. In the common garden experiment, genetic differentiation between populations accounted for 0–28% of total phenotypic variation. However, only two traits (leaf mass per area and nitrogen content) exhibited a significant cline. The combination of in situ and common garden experiments used here made it possible to demonstrate, for both species, a weaker effect of genetic variation than of variations in natural conditions, suggesting a strong effect of the environment on leaf functional traits. Finally, we demonstrated that intrapopulation variability was systematically higher than interpopulation variability, whatever the functional trait considered, indicating a high potential capacity to adapt to climate change.

Description: Xylem cavitation resistance is a key physiological trait correlated with species tolerance to extreme drought stresses. Little is known about the genetic variability and phenotypic plasticity of this trait in natural tree populations. Here we measured the cavitation resistance of 17 Fagus sylvatica populations representative of the full range of the species in Europe. The trees were grown in three field trials under contrasting climatic conditions. Our findings suggest that the genotypic variability of cavitation resistance is high between genotypes of a given population. By contrast, no significant differences were found for this trait across populations, the mean population cavitation resistance being remarkably constant in each trial. We found a significant site effect and a significant site x population interaction, suggesting that cavitation resistance has a high phenotypic plasticity and that this plasticity is under genetic control. The implications of our findings for beech forest management in a context of climate change are discussed.

Description: Future climate will alter the soil cover of mosses and snow depths in the boreal forests of eastern Canada. In field manipulation experiments, we assessed the effects of varying moss and snow depths on the physiology of black spruce ( Picea ­mariana (Mill.) B.S.P.) and trembling aspen ( Populus tremuloides Michx.) in the boreal black spruce forest of western Québec. For 1 year, naturally regenerated 10-year-old spruce and aspen were grown with one of the following treatments: additional N fertilization, addition of sphagnum moss cover, removal of mosses, delayed soil thawing through snow and hay addition, or accelerated soil thawing through springtime snow removal. Treatments that involved the addition of insulating moss or snow in the spring caused lower soil temperature, while removing moss and snow in the spring caused elevated soil temperature and thus had a warming effect. Soil warming treatments were associated with greater temperature variability. Additional soil cover, whether moss or snow, increased the rate of photosynthetic recovery in the spring. Moss and snow removal, on the other hand, had the opposite effect and lowered photosynthetic activity, especially in spruce. Maximal electron transport rate (ETR max ) was, for spruce, 39.5% lower after moss removal than with moss addition, and 16.3% lower with accelerated thawing than with delayed thawing. Impaired photosynthetic recovery in the absence of insulating moss or snow covers was associated with lower foliar N concentrations. Both species were affected in that way, but trembling aspen generally reacted less strongly to all treatments. Our results indicate that a clear negative response of black spruce to changes in root-zone temperature should be anticipated in a future climate. Reduced moss cover and snow depth could adversely affect the photosynthetic capacities of black spruce, while having only minor effects on trembling aspen.

Description: Winter frost resistance (WFR), midwinter frost hardening and frost dehardening potential of Pinus cembra L. were determined in situ by means of a novel low-temperature freezing system at the alpine timberline ecotone (1950 m a.s.l., Mt Patscherkofel, Innsbruck, Austria). In situ liquid nitrogen (LN 2 )-quenching experiments should check whether maximum WFR of P. cembra belonging to the frost hardiest conifer group, being classified in US Department of Agriculture climatic zone 1, suffices to survive dipping into LN 2 (–196 °C). Viability was assessed in a field re-growth test. Maximum in situ WFR (LT 50 ) of leaves was 〈– 75 °C and that of buds was less (–70.3 °C), matching the lowest water contents. In midwinter, in situ freezing exotherms of leaves, buds and the xylem were often not detectable. Ice formed in the xylem at a mean of –2.8 °C and in leaves at –3.3 °C. In situ WFR of P. cembra was higher than that obtained on detached twigs, as reported earlier. In situ LN 2 -quenching experiments were lethal in all cases even when twigs of P. cembra were exposed to an in situ frost hardening treatment (12 days at –20 °C followed by 3 days at –50 °C) to induce maximum WFR. Temperature treatments applied in the field significantly affected the actual WFR. In January a frost hardening treatment (21 days at –20 °C) led to a significant increase of WFR (buds: –62 °C to 〈– 70 °C; leaves: –59.6 °C to –65.2 °C), showing that P. cembra was not at its specific maximum WFR. In contrast, simulated warm spells in late winter led to premature frost dehardening (buds: –32.6 °C to –10.2 °C; leaves: –32.7 to –16.4 °C) followed by significantly earlier bud swelling and burst in late winter. Strikingly, both temperature treatments, either increased air temperature (+10.1 °C) or increased soil temperature (+6.5 °C), were similarly effective. This high readiness to frost harden and deharden in winter in the field must be considered to be of great significance for future winter survival of P. cembra . Determination of WFR in field re-growth tests appears to be a valuable tool for critically judging estimates of WFR obtained on detached twigs in an ecological context.

Description: Genetic differences in 13 C (isotopic composition of dry matter carbon) have been evidenced among poplar genotypes at juvenile stages. To check whether such differences were maintained with age in trees growing in plantations, we investigated the time course of 13 C as recorded in annual tree rings from different genotypes growing at three sites in southwestern France and felled at ~15–17 years. Wood cores were cut from tree discs to record the time course of annual basal area increment (BAI). The isotopic ratio 13 C was recorded in bulk wood and in extracted cellulose from the annual rings corresponding to the period 1996–2005. Discrimination against 13 C between atmosphere and tissues ( 13 C) was computed by taking into account the inter-annual time course of 13 C in the atmosphere. Annual BAI increased steadily and stabilized at about 8 years. An offset in 13 C of ~1 was recorded between extracted cellulose and bulk wood. It was relatively stable among genotypes within sites but varied among sites and increased slightly with age. Site effects as well as genotype differences were detected in 13 C recorded from the cellulose fraction. Absolute values as well as the genotype ranking of 13 C remained stable with age in the three sites. Genotype means of 13 C were not correlated to annual BAI. We conclude that genotypic differences of 13 C occur in older poplar trees in plantations, and that the differences as well as the genotype ranking remain stable while trees age until harvest.

Description: This study examined the role of ectomycorrhizal associations in nitrogen assimilation of Populus tremuloides seedlings. Seedlings were inoculated with Hebeloma crustuliniforme and compared with non-inoculated plants. Nitrogen-metabolizing enzymatic properties were also determined in H. crustuliniforme grown in sterile culture. The seedlings and fungal cultures were subjected to nitrogen treatments (including NO 3 – , NH 4 + and a combination of NO 3 – + NH 4 + ) for 2 months to examine the effects on growth, nitrogen-assimilating enzyme activities and xylem sap concentrations of NH 4 + and NO 3 – . Seedlings were also provided for 3 days with 15 N-labeled NH 4 + and NO 3 – , and leaf and root 15 N content relative to total nitrogen was measured. Both NO 3 – and NH 4 + were effective in supporting seedling growth when either form was provided separately. When NO 3 – and NH 4 + were provided together, seedling growth decreased while enzymatic assimilation of NH 4 + increased. Additionally, nitrogen assimilation in inoculated seedlings was less affected by the form of nitrogen compared with non-inoculated plants. Fungal ability to enzymatically respond to and assimilate NH 4 + combined with aspen's enzymatic responsiveness to NO 3 – was likely the reason for efficient assimilation of both nitrogen forms by mycorrhizal plants.

Description: Boron (B) is an essential micronutrient whose deficiency is common both in agriculture and in silviculture. Boron deficiency impairs the growth of plants and affects many metabolic processes like carbohydrate metabolism. Boron deficiency and also excess B may decrease the sink demand by decreasing the growth and sugar transport which may lead to the accumulation of carbohydrates and down-regulation of photosynthesis. In this study, we investigated the effects of B nutrition on the soluble and storage carbohydrate concentrations of summer leaves and autumn buds in a deciduous tree species, Betula pendula Roth. In addition, we investigated the changes in the pools of condensed tannins between summer and autumn harvests. One-year-old birch seedlings were fertilized with a complete nutrient solution containing three different levels of B: 0, 30 and 100% of the standard level for complete nutrient solution. Half of the seedlings were harvested after summer period and another half when leaves abscised. The highest B fertilization level (B100) caused an accumulation of starch and a decrease in the concentrations of hexoses (glucose and fructose) in summer leaves, whereas in the B0 seedlings, hexoses (mainly glucose) accumulated and starch decreased. These changes in carbohydrate concentrations might be related to the changes in the sink demand since the autumn growth was the smallest for the B100 seedlings and largest for the B30 seedlings that did not accumulate carbohydrates. The autumn buds of B30 seedlings contained the lowest levels of glucose, glycerol, raffinose and total polyols, which was probably due to the dilution effect of the deposition of other substances like phenols. Condensed tannins accumulated in high amounts in the birch stems during the hardening of seedlings and the largest accumulation was detected in the B30 treatment. Our results suggest that B nutrition of birch seedlings affects the carbohydrate and phenol metabolism and may play an important role in the hardening process of the seedlings.

Description: Norway spruce [ Picea abies (L.) Karst.] is one of the economically most important conifer species in Europe. The major pathogen on Norway spruce is Heterobasidion parviporum (Fr.) Niemelä & Korhonen. To achieve a better understanding of Norway spruce's defence mechanisms, transcriptional responses in bark to H. parviporum infection were compared with the response to wounding using cDNA-amplified fragment length polymorphism. The majority of the recovered transcript-derived fragments (TDFs) showed a similar expression pattern for infection and wounding treatment, although inoculated samples showed an enhanced reaction. Genes related to systemic acquired resistance, e.g., PR1 , accumulated after H. parviporum infection. Simultaneously, several transcripts involved in various aspects of jasmonic acid (JA)- and ethylene (ET)-mediated signalling accumulated. Genes involved in the ubiquitin/proteasome system were also regulated. Expression patterns have been confirmed by quantitative polymerase chain reaction. The expression patterns of the isolated TDFs suggest that infection with H. parviporum in Norway spruce induces a broad defence, with many similarities to non-specific defence responses in angiosperms. The parallel induction of salicylic acid- and JA/ET-mediated pathways implies spatially separated responses in different cell layers, with and without hyphal contact. A set of TDFs were analysed in an independent experiment with unrelated material treated with wounding or with inoculation with H. parviporum or Phlebiopsis gigantea , verifying the original observations and underlining the non-specific defence responses. In addition, our data suggest that rerouting of carbon in secondary metabolism is an integral part of Norway spruce induced defence. We report the sequences of three 3-deoxy-d-arabino-heptulosonate 7-phosphate synthase genes ( PaDAHP1 , PaDAHP2 and PaDAHP3) and their relative expression in response to wounding and infection with H. parviporum and P. gigantea . The results clearly indicate differential regulation of the three DAHPs in the induced defence responses in Norway spruce. This study gives insights into the central mechanisms in the induced defences in Norway spruce.

Description: When stomata first evolved they initiated the greening of terrestrial earth, and now more than 400 million years later these simple bi-cellular valves in the leaf surface regulate global fluxes of water and carbon. Despite their importance and superficial simplicity, the behaviour of stomata remains a great challenge to understand. Different approaches to studying stomatal control have yielded rather disparate models for how stomata respond to environmental stimuli. Much of this discord arises from the diversity of mechanisms apparently involved in changing guard cell turgor and hence the aperture of the stomatal pore. On the one hand, the physical tension produced by dragging water from the soil through the xylem to the leaves directly influences leaf and guard cell turgor, while on the other hand, phytohormone levels (most importantly abscisic acid), light, photosynthesis and atmospheric gases induce active changes in guard cell turgor by triggering ionic pumping. Each stomatal control mechanism has its own champion and no model has ever successfully integrated all components. In such an environment there is great value in examining how different parts of the stomatal control network interact, particularly the competition between ‘hydraulic’ signals related to leaf water content and ‘metabolic’ signals related to ambient photosynthetic conditions.

Description: In woody plants, pioneer roots are the main roots used to expand the root system horizontally and vertically whereas fibrous ‘feeder’ roots are chiefly used in the absorption of water and nutrients. Because of their different roles, we expected newly emerged pioneer and fibrous roots to respond differently to restrictions in soil moisture. We hypothesized that fibrous roots would exhibit greater growth plasticity and greater physiological impairment from soil moisture deficits, especially under heterogeneous conditions. We compared the responses of fibrous and pioneer roots of olive seedlings ( Olea europaea ) to localized and uniform soil moisture deficits in transparent containers in the greenhouse. In comparison with uniformly wet conditions, uniformly dry conditions caused reduced shoot photosynthesis and reduced shoot growth, but no significant effect on root morphology, root respiration (measured in aerated buffer solution using excised roots) or electrolyte leakage as a function of root age. Under heterogeneous soil moisture conditions, root growth tended to preferentially occur in the moist sector, especially in the pioneer roots. In comparison with pioneer roots in the moist sector, pioneer roots in the dry sector had higher tissue density and higher suberin content, but no shift in root respiration, non-structural carbohydrates or electrolyte leakage. In contrast, fibrous roots in the dry sector exhibited evidence of impaired physiology in older (〉38 days) roots compared with similar age fibrous roots in the moist sector. While we anticipated that, compared with pioneer roots, fibrous roots would be more sensitive to soil moisture deficits as expressed by higher electrolyte leakage, we did not expect the strong growth plasticity of pioneer roots under heterogeneous soil moisture conditions. Differentiating the responses of these two very different root types can improve our understanding of how different portions of the root system of woody plants cope with soil moisture deficits.

Description: Knowledge about nocturnal transpiration ( E night ) of trees is increasing and its impact on regional water and carbon balance has been recognized. Most of this knowledge has been generated in temperate or equatorial regions. Yet, little is known about E night and tree water use ( Q ) in semi-arid regions. We investigated the influence of atmospheric conditions on daytime ( Q day ) and nighttime water transport ( Q night ) of Eucalyptus victrix L.A.S. Johnson & K.D. Hill growing over shallow groundwater (not 〉1.5 m in depth) in semi-arid tropical Australia. We recorded Q day and Q night at different tree heights in conjunction with measurements of stomatal conductance ( g s ) and partitioned E night from refilling processes. Q of average-sized trees (200–400 mm diameter) was 1000–3000 l month –1 , but increased exponentially with diameter such that large trees (〉500 mm diameter) used up to 8000 l month –1 . Q was remarkably stable across seasons. Water flux densities ( J s ) varied significantly at different tree heights during day and night. We show that g s remained significantly different from zero and E night was always greater than zero due to vapor pressure deficits ( D ) that remained 〉1.5 kPa at night throughout the year. Q night reached a maximum of 50% of Q day and was 〉0.03 mm h –1 averaged across seasons. Refilling began during afternoon hours and continued well into the night. Q night eventually stabilized and closely tracked D night . Coupling of Q night and D night was particularly strong during the wet season ( R 2 = 0.95). We suggest that these trees have developed the capacity to withstand a pronounced desiccation–rehydration cycle in a semi-arid environment. Such a cycle has important implications for local and regional hydrological budgets of semi-arid landscapes, as large nighttime water fluxes must be included in any accounting.

Description: In common gardens along an ~900 km latitudinal transect through Wisconsin and Illinois, USA, tree species typical of boreal and temperate forests were compared with respect to the nature and magnitude of leaf respiratory acclimation to contrasting climates. The boreal representatives were trembling aspen ( Populus tremuloides Michx.) and paper birch ( Betula papyrifera Marsh.), while the temperate species were eastern cottonwood ( Populus deltoides Bartr ex. Marsh var. deltoides ) and sweetgum ( Liquidambar styraciflua L.). Assessments were conducted on seedlings grown from seed sources collected near southern and northern range boundaries, respectively. Nighttime rates of leaf dark respiration ( R d ) at common temperatures, as well as R d 's short-term temperature sensitivity (energy of activation, E o ), were assessed for all species and gardens twice during a growing season. Little evidence of R d thermal acclimation was observed, despite a 12 °C range in average air temperature across gardens. Instead, R d variation at warm temperatures was linked most closely with prior leaf photosynthetic performance, while R d variation at cooler temperatures was most strongly related to leaf nitrogen concentration. Moreover, E o differences across species and gardens appeared to stem from the somewhat independent limitations on warm versus cool R d . Based on this construct, an empirical model relying on R d estimates from leaf photosynthesis and nitrogen concentration explained 55% of the observed E o variation.

Description: Nothofagus nitida (Phil.) Krasser (Nothofagaceae) regenerates under the canopy in microsites protected from high light. Nonetheless, it is common to find older saplings in clear areas and adults as emergent trees of the Chilean evergreen forest. We hypothesized that this shade to sun transition in N. nitida is supported by an increase in photochemical and non-photochemical energy dissipation capacities of both photosystems in parallel with the increase in plant size and light availability. To dissect the relative contribution of light environment and plant developmental stage to these physiological responses, the photosynthetic performance of both photosystems was studied from the morpho-anatomical to the biochemical level in current-year leaves of N. nitida plants of different heights (ranging from 0.1 to 7 m) growing under contrasting light environments (integrated quantum flux (IQF) 5–40 mol m –2  day –1 ). Tree height (TH) and light environment (IQF) independently increased the saturated electron transport rates of both photosystems, as well as leaf and palisade thickness, but non-photochemical energy flux, photoinhibition susceptibility, state transition capacity, and the contents of D1 and PsbS proteins were not affected by IQF and TH. Spongy mesophyll thickness and palisade cell diameter decreased with IQF and TH. A max , light compensation and saturation points, Rubisco and nitrogen content (area basis) only increased with light environment (IQF), whereas dark respiration ( R d ) decreased slightly and relative chlorophyll content was higher in taller trees. Overall, the independent effects of more illuminated environment and tree height mainly increased the photochemical instead of the non-photochemical energy flux. Regardless of the photochemical increase with TH, carbon assimilation only significantly improved with higher IQF. Therefore it seems that mainly acclimation to the light environment supports the phenotypic transition of N. nitida from shade to sun.

Description: Conifers and other trees are constantly adapting to changes in light conditions, water/nutrient supply and temperatures by physiological and morphological modifications of their foliage. However, the relationship between physiological processes and anatomical characteristics of foliage has been little explored in trees. In this study we evaluated needle structure and function in Norway spruce families exposed to different light conditions and transpiration regimes. We compared needle characteristics of sun-exposed and shaded current-year needles in a control plot and a thinned plot with 50% reduction in stand density. Whole-tree transpiration rates remained similar across plots, but increased transpiration of lower branches after thinning implies that sun-exposed needles in the thinned plot were subjected to higher water stress than sun-exposed needles in the control plot. In general, morphological and anatomical needle parameters increased with increasing tree height and light intensity. Needle width, needle cross-section area, needle stele area and needle flatness (the ratio of needle thickness to needle width) differed most between the upper and lower canopy. The parameters that were most sensitive to the altered needle water status of the upper canopy after thinning were needle thickness, needle flatness and percentage of stele area in needle area. These results show that studies comparing needle structure or function between tree species should consider not only tree height and light gradients, but also needle water status. Unaccounted for differences in needle water status may have contributed to the variable relationship between needle structure and irradiance that has been observed among conifers.

Description: Transport phloem is no longer associated with impermeable pipes, but is instead considered as a leaky system in which loss and retrieval mechanisms occur. Local stem chilling is often used to study these phenomena. In this study, 5-cm- lengths of stems of 3-year-old oak trees ( Quercus robur L.) were locally chilled for 1 week to investigate whether observations at stem and leaf level can be explained by the leakage-retrieval mechanism. The chilling experiment was repeated three times across the growing season. Measurements were made of leaf photosynthesis, carbohydrate concentrations in leaves and bark, stem growth and maximum daily stem shrinkage. Across the growing season, a feedback inhibition in leaf photosynthesis was observed, causing increased dark respiration and starch concentration. This inhibition was attributed to the total phloem resistance which locally increased due to the cold temperatures. It is hypothesized that this higher phloem resistance increased the phloem pressure above the cold block up to the source leaves, inducing feedback inhibition. In addition, an increase in radial stem growth and carbohydrate concentration was observed above the cold block, while the opposite occurred below the block. These observations indicate that net lateral leakage of carbohydrates from the phloem was enhanced above the cold block and that translocation towards regions below the block decreased. This behaviour is probably also attributable to the higher phloem resistance. The chilling effects on radial stem growth and carbohydrate concentration were significant in the middle of the growing season, while they were not at the beginning and near the end of the growing season. Furthermore, maximum daily shrinkages were larger above the cold block during all chilling experiments, indicating an increased resistance in the xylem vessels, also generated by low temperatures. In conclusion, localized stem chilling altered multiple carbon processes in the source leaves and the main stem by changing hydraulic resistances.

Description: The Castanea sativa SCL1 gene ( CsSCL1 ) has previously been shown to be induced by auxin during adventitious root (AR) formation in rooting-competent microshoots. However, its expression has not previously been analyzed in rooting-incompetent shoots. This study focuses on the regulation of CsSCL1 during maturation and the role of the gene in the formation of AR. The expression of CsSCL1 in rooting-incompetent microshoots and other tissues was investigated by quantitative reverse transcriptase–­polymerase chain reaction. The analysis was complemented by in situ hybridization of the basal segments of rooting-competent and -­incompetent microshoots during AR induction, as well as in AR and lateral roots. It was found that CsSCL1 is upregulated by auxin in a cell-type- and phase-dependent manner during the induction of AR. In root-forming shoots, CsSCL1 mRNA was specifically located in the cambial zone and derivative cells, which are rooting-competent cells, whereas in rooting-incompetent shoots the hybridization signal was more diffuse and evenly distributed through the phloem and parenchyma. CsSCL1 expression was also detected in lateral roots and axillary buds. The different CsSCL1 expression patterns in rooting-competent and -incompetent microshoots, together with the specific location of transcripts in cell types involved in root meristem initiation and in the root primordia of AR and lateral roots, indicate an important role for the gene in determining whether certain cells will enter the root differentiation pathway and its involvement in meristem maintenance.

Description: Forest canopies exchange a large part of the mass and energy between the earth and the atmosphere. The processes that regulate these exchanges have been of interest to scientists from a diverse range of disciplines for a long time. The International Union of Forest Research Organizations (IUFRO) Canopy Processes Working Group provides a forum for these scientists to explore canopy processes at scales ranging from the leaf to the ecosystem. Given the changes in climate that are being experienced in response to rising [CO 2 ], there is a need to understand how forest canopy processes respond to altered environments. Globally, native and managed forests represent the largest terrestrial biome and, in wood and soils, the largest terrestrial stores of carbon. Changing climates have significant implications for carbon storage in forests, as well as their water use, species diversity and management. In order to address these issues, the Canopy Processes Working Group held a travelling workshop in south-east Australia during October 2010 to examine the impact of changing climates on forest canopies, highlighting knowledge gaps and developing new research directions.

Description: The ongoing global change could be an additional threat to the establishment and the long-term survival of Populus nigra L., an emblematic European riparian species. With the general aim of gaining insights into the adaptive potential of this species, we (i) quantified variations within and among three French P. nigra populations for key physiological attributes, i.e., water-use efficiency (assessed from bulk leaf carbon isotope discrimination, 13 C), growth performance and related leaf traits, (ii) examined genotype and population by environment interactions, and (iii) explored the relationship between 13 C and growth. Thirty genotypes were sampled in each of three naturally established populations and grown in two different sites, Orléans (ORL) and Guémené-Penfao (GMN). In ORL, two similar plots were established and different watering regimes were applied in order to test for the drought response. Significant variations were observed for all traits within and among populations irrespective of site and watering. Trait variation was larger within than among populations. The effect of drought was neither genotype- nor population-dependent, contrary to the effect of site. The population ranking was maintained in all sites and watering regimes for the two most complex traits: 13 C and growth. Moreover, these two traits were unrelated, which indicates that (i) water-use efficiency and growth are largely uncoupled in this species, and (ii) the environmental factors driving genetic structuration for 13 C and growth act independently. The large variations found within populations combined with the consistent differences among populations suggest a large adaptive potential for P. nigra .

Description: Abies alba and Abies pinsapo are two closely related fir species that occur in the Iberian Peninsula under very different environmental conditions. Abies alba proliferates in the humid European mountains, including the Spanish Pyrenees. In contrast, A. pinsapo is a relict species that occurs in some restricted areas of the Mediterranean mountain ranges in Spain and Morocco, which experience intense summer drought periods. To cope with the high atmospheric evaporative demand during summer, A. pinsapo may either have a high resistance to xylem cavitation or develop a very efficient conducting system to reduce the soil-to-leaf water potential gradient. To investigate such hypotheses, we measured (i) the xylem vulnerability to cavitation for different populations, and (ii) several anatomical and hydraulic parameters indicating xylem sufficiency for ­supplying water to the shoot in two contrasting populations of both species. Our results show that the resistance to cavitation was not different between species or populations. However, hydraulic conductivity ( K h ), specific hydraulic conductivity ( K s ), leaf-specific conductivity (LSC) and whole-shoot hydraulic conductance ( K shoot ) were higher in A. pinsapo , indicating a higher efficiency of water transport, which should contribute to maintaining its xylem tension below the threshold for rapidly increasing cavitation. The higher K s in A. pinsapo was largely a result of its wider tracheids, suggesting that this species may be much more vulnerable to freeze–thaw-induced cavitation than A. alba . This is consistent with the absence of A. pinsapo in northern mountain ranges with cooler winters. These physiological differences could partly explain the niche segregation and the geographical separation of these two firs.

Description: With their ratification of the Kyoto Protocol, many countries have established forests on previously non-forested land with the view of offsetting greenhouse gas emissions. While these forests indisputably result in increased carbon storage in above-ground biomass, consideration of other major implications is often neglected. Forest establishment results in changes in albedo and soil carbon storage, reduced runoff and downstream water supply, and effects on biodiversity. Such effects of forest establishment may be less desirable from environmental, economic and social perspectives. While there have been many studies of the impacts of forest establishment on individual aspects, policy makers need to be able to integrate the benefits and consequences to assist in making decisions on land management. Further, the relative magnitude of the effects of forestry needs to be considered in the context of elevated atmospheric carbon dioxide partial pressure and climate change resulting in increasing temperature and changes in the amount and distribution of rainfall. This introductory review highlights the major benefits and consequences of forest establishment and demonstrates progress in integrating across the services provided by forests. New modelling approaches are being developed that allow analysis of benefits, consequences and trade-offs to assist policy makers in decisions to manage the provision of multiple resources.

Description: The carbon ( 13 C) and oxygen ( 18 O) stable isotope composition is widely used to obtain information on the linkages between environmental drivers and tree physiology over various time scales. The tree-ring archive can especially be exploited to reconstruct inter- and intra-annual variation of both climate and physiology. There is, however, a lack of information on the processes potentially affecting 13 C and 18 O on their way from assimilation in the leaf to the tree ring. As a consequence, the aim of this study was to trace the isotope signals in European beech ( Fagus sylvatica L.) from leaf water ( 18 O) and leaf assimilates ( 13 C and 18 O) to tree-ring wood via phloem-transported compounds over a whole growing season. Phloem and leaf samples for 13 C and 18 O analyses as well as soil water, xylem water, leaf water and atmospheric water vapour samples for 18 O analysis were taken approximately every 2 weeks during the growing season of 2007. The 13 C and 18 O samples from the tree rings were dated intra-annually by monitoring the tree growth with dendrometers. 18 O in the phloem organic matter and tree-ring whole wood was not positively related to leaf water evaporative enrichment and 18 O of canopy organic matter pools. This finding implies a partial uncoupling of the tree-ring oxygen isotopic signal from canopy physiology. At the same time, internal carbon storage and remobilization physiology most likely prevented 13 C in tree-ring whole wood from being closely related to intra-annual variation in environmental drivers. Taking into account the post-photosynthetic isotope fractionation processes resulting in alterations of 13 C and 18 O not only in the tree ring but also in phloem carbohydrates, as well as the intra-annual timing of changes in the tree internal physiology, might help to better understand the meaning of the tree-ring isotope signal not only intra- but also inter-annually.

Description: The effects of management practices on energy, water and carbon exchanges were investigated in a young pine plantation in south-west France. In 2009–10, carbon dioxide (CO 2 ), H 2 O and heat fluxes were monitored using the eddy covariance and sap flow techniques in a control plot (C) with a developed gorse layer, and an adjacent plot that was mechanically weeded and thinned (W). Despite large differences in the total leaf area index and canopy structure, the annual net radiation absorbed was only 4% lower in plot W. We showed that higher albedo in this plot was offset by lower emitted long-wave radiation. Annual evapotranspiration (ET) from plot W was 15% lower, due to lower rainfall interception and transpiration by the tree canopy, partly counterbalanced by the larger evaporation from both soil and regrowing weedy vegetation. The drainage belowground from plot W was larger by 113 mm annually. The seasonal variability of ET was driven by the dynamics of the soil and weed layers, which was more severely affected by drought in plot C. Conversely, the temporal changes in pine transpiration and stem diameter growth were synchronous between sites despite higher soil water content in the weeded plot. At the annual scale, both plots were carbon sinks, but thinning and weeding reduced the carbon uptake by 73%: annual carbon uptake was 243 and 65 g C m –2 on plots C and W, respectively. Summer drought dramatically impacted the net ecosystem exchange: plot C became a carbon source as the gross primary production (GPP) severely decreased. However, plot W remained a carbon sink during drought, as a result of decreases in both GPP and ecosystem respiration ( R E ). In winter, both plots were carbon sources, plots C and W emitting 67.5 and 32.4 g C m –2 , respectively. Overall, this study highlighted the significant contribution of the gorse layer to mass and energy exchange in young pine plantations.

Description: Potassium (K) influences the photosynthesis process in a number of ways; however, the mechanisms underlying the photosynthetic response to differences in K supply are not well understood. Concurrent measurements of gas exchange and chlorophyll fluorescence were made to investigate the effect of K nutrition on photosynthetic efficiency and mesophyll conductance ( g m ) in hickory seedlings ( Carya cathayensis Sarg.) in a greenhouse. The results show that leaf K concentrations 〈0.7–0.8% appeared to limit the leaf net CO 2 assimilation rate ( A ), and that the relative limitation of photosynthesis due to g m and stomatal conductance ( g s ) decreased with increasing supplies of K. However, a sensitivity analysis indicated that A was most sensitive to the maximum carboxylation rate of Rubisco ( V c,max ) and the maximum rate of electron transport ( J max ). These results indicate that the photosynthetic rate is primarily limited by the biochemical processes of photosynthesis ( V c,max and J max ), rather than by g m and g s in K-deficient plants. Additionally, g m was closely correlated with g s and the leaf dry mass per unit area ( M A ) in hickory seedlings, which indicates that decreased g m and g s may be a consequence of leaf anatomical adaptation.

Description: The tropical conifer Callitris intratropica (Cupressaceae) produces clear annual growth rings, and has been shown to be potentially useful for understanding past climate variability in northern Australia. As climate patterns in this region become less predictable, an understanding of plant responses to different weather patterns is of importance. In this paper, we examine tree water relations using a parameter here called tree water deficit ( D ), determined from de-trended stem size variability in densely grown (‘grove’) and isolated trees. This parameter provides an integrated measure of the trees' response to water supply and demand under constantly changing environmental conditions. The work, conducted over 12 months, found that daily variation in tree water deficit was determined mainly by soil water availability, but temperature and relative humidity contributed more to the variability over some periods. Isolated and grove trees exhibited quite distinct patterns of D development during the year, but particularly during the transition between the dry and wet seasons. The results of this work suggest that the dendrochronological interpretation of tree rings in the context of strongly seasonal water availability should incorporate an understanding of the development of seasonal drought in isolated trees compared with trees experiencing strong intra-specific competition. Different responses based on the ecological situations of the trees will affect their patterns of stem growth, and ultimately the climatic information that is incorporated in ring width variability.

Description: Elevated atmospheric [CO 2 ] (e C a ) often decreases stomatal conductance, which may delay the start of drought, as well as alleviate the effect of dry soil on plant water use and carbon uptake. We studied the interaction between drought and e C a in a whole-tree chamber experiment with Eucalyptus saligna . Trees were grown for 18 months in their C a treatments before a 4-month dry-down. Trees grown in e C a were smaller than those grown in ambient C a (a C a ) due to an early growth setback that was maintained throughout the duration of the experiment. Pre-dawn leaf water potentials were not different between C a treatments, but were lower in the drought treatment than the irrigated control. Counter to expectations, the drought treatment caused a larger reduction in canopy-average transpiration rates for trees in the e C a treatment compared with a C a . Total tree transpiration over the dry-down was positively correlated with the decrease in soil water storage, measured in the top 1.5 m, over the drying cycle; however, we could not close the water budget especially for the larger trees, suggesting soil water uptake below 1.5 m depth. Using neutron probe soil water measurements, we estimated fractional water uptake to a depth of 4.5 m and found that larger trees were able to extract more water from deep soil layers. These results highlight the interaction between rooting depth and response of tree water use to drought. The responses of tree water use to e C a involve interactions between tree size, root distribution and soil moisture availability that may override the expected direct effects of e C a . It is essential that these interactions be considered when interpreting experimental results.

Description: This study describes the physiological response of two co-occurring tree species ( Eucalyptus marginata and Corymbia ­calophylla ) to seasonal drought at low- and high-quality restored bauxite mine sites in south-western Australia. Seasonal changes in photosynthesis ( A ), stomatal conductance ( g s ), leaf water potential (), leaf osmotic potential (), leaf relative water content (RWC) and pressure–volume analysis were captured over an 18-month field study to (i) determine the nature and severity of physiological stress in relation to site quality and (ii) identify any physiological differences between the two species. Root system restriction at the low-quality site reduced maximum rates of gas exchange ( g s and A ) and increased water stress ­(midday and daily RWC) in both species during drought. Both species showed high stomatal sensitivity ­during drought; however, E. marginata demonstrated a higher dehydration tolerance where and RWC fell to –3.2 MPa and 73% compared with –2.4 MPa and 80% for C. calophylla. Corymbia calophylla showed lower g s and higher and RWC during drought, indicating higher drought tolerance. Pressure–volume curves showed that cell-wall elasticity of E. marginata leaves increased in response to drought, while C. calophylla leaves showed lower osmotic potential at zero turgor in summer than in winter, indicating osmotic adjustment. Both species are clearly able to tolerate seasonal drought at hostile sites; however, by C. ­calophylla closing stomata earlier in the drought cycle, maintaining a higher water status during drought and having the additional mechanism of osmotic adjustment, it may have a greater capacity to survive extended periods of drought.

Description: Mesophyll conductance, g m , was estimated from measurements of stomatal conductance to carbon dioxide transfer, g s , photosynthesis, A , and chlorophyll fluorescence for Year 0 (current-year) and Year 1 (1-year-old) fully sunlit leaves from short (2 m tall, 10-year-old) and tall (15 m tall, 120-year-old) Nothofagus solandrii var. cliffortiodes trees growing in adjacent stands. Rates of photosynthesis at saturating irradiance and ambient CO 2 partial pressure, A satQ , were 25% lower and maximum rates of carboxylation, V cmax , were 44% lower in Year 1 leaves compared with Year 0 leaves across both tree sizes. Although g s and g m were not significantly different between Year 0 and Year 1 leaves and g s was not significantly different between tree heights, g m was significantly (19%) lower for leaves on tall trees compared with leaves on short trees. Overall, V cmax was 60% higher when expressed on the basis of CO 2 partial pressure at the chloroplasts, C c , compared with V cmax on the basis of intercellular CO 2 partial pressure, C i , but this varied with leaf age and tree size. To interpret the relative stomatal and mesophyll limitations to photosynthesis, we used a model of carbon isotopic composition for whole leaves incorporating g m effects to generate a surface of ‘operating values’ of A over the growing season for all leaf classes. Our analysis showed that A was slightly higher for leaves on short compared with tall trees, but lower g m apparently reduced actual A substantially compared with A satQ . Our findings showed that lower rates of photosynthesis in Year 1 leaves compared with Year 0 leaves were attributable more to increased biochemical limitation to photosynthesis in Year 1 leaves than differences in g m . However, lower A in leaves on tall trees compared with those on short trees could be attributed in part to lower g m and higher stomatal, L s , and mesophyll, L m , limitations to photosynthesis, consistent with steeper hydraulic gradients in tall trees.

Description: Trees adapted to mesic and xeric habits may differ in a suite of physiological responses that affect leaf-level carbon balance, including the relationship between photosynthesis ( A ) and respiration at night ( R n ). Understanding the factors that regulate physiological function in mesic and xeric species is critical for predicting changes in growth and distribution under changing climates. In this study, we examined the relationship between A and R n , and leaf traits that may regulate A and R n , in six Eucalyptus species native to mesic or xeric ecosystems, during two 24-h cycles in a common garden under high soil moisture. Peak A and R n generally were higher in xeric compared with mesic species. Across species, A and R n covaried, correlated with leaf mass per area, leaf N per unit area and daytime soluble sugar accumulation. A also covaried with g s , which accounted for 93% of the variation in A within species. These results suggest that A and R n in these six Eucalyptus species were linked through leaf N and carbohydrates. Further, the relationship between A and R n across species suggests that differences in this relationship between mesic and xeric Eucalyptus species in their native habitats may be largely driven by environmental factors rather than inter-specific genetic variation.