ALBERT

Description: Water deficit tolerance during post-germination stages is critical for seedling recruitment. In this work, we studied the effect of water deficit on morphological and biochemical responses in different organs of newly germinated mahogany ( Swietenia macrophylla King) seedlings, a woody species that occurs in the Amazon rainforest. The root : shoot ratio increased under water deficit. The leaf number and water potential were not altered, although reductions in leaf area and stomatal conductance were observed. Osmotic potential became more negative in leaves of seedlings under severe stress. Water deficit increased fructose, glucose, sucrose and myo-inositol levels in leaves. Stems accumulated fructose, glucose and l -proline. Nitric oxide (NO) levels increased in the vascular cylinder of roots under severe stress while superoxide anion levels decreased due to augmented superoxide dismutase activity in this organ. Water deficit induced glutathione reductase activity in both roots and stems. Upon moderate or severe stress, catalase activity decreased in leaves and remained unaffected in the other seedling organs, allowing for an increase of hydrogen peroxide (H 2 O 2 ) levels in leaves. Overall, the increase of signaling molecules in distinct organs—NO in roots, l -proline in stems and H 2 O 2 and myo-inositol in leaves—contributed to the response of mahogany seedlings to water deficit by triggering biochemical processes that resulted in the attenuation of oxidative stress and the establishment of osmotic adjustment. Therefore, this body of evidence reveals that the development of newly germinated mahogany seedlings may occur in both natural habitats and crop fields even when water availability is greatly limited.

Description: Uptake of nitrogen (N) by sequential root regions in six tree species was measured in roots of 16- to 26-month-old seedlings at 50 and 1500 µM NH 4 NO 3 concentration, at the cell level using oscillating microelectrodes and at the root region level using enriched 15 N application. Our objective was to determine the root regions making the greatest contribution to total N uptake in each species as measured by the two contrasting techniques. White and condensed tannin zones were the regions with the smallest surface area in all species, but these zones often had the highest net flux of NH 4 + and NO 3 – . For most species, little variation was found among root regions in N flux calculated using a 15 N mass balance approach, but where significant differences existed, high N flux was observed in white, cork or woody zones. When N fluxes measured by each of the two methods were multiplied by the estimated surface area or biomass of each root region, the effect of root region size had the greatest influence on regional N uptake. Root regions of greatest overall N uptake were the cork and woody zones, on average. Total N uptake may thus be greatest in older regions of tree seedling roots, despite low rates of uptake per unit area.

Description: Climate change is expected to increase both pest insect damage and the occurrence of severe drought. There is therefore a need to better understand the combined effects of biotic and abiotic damage on tree growth in order to predict the multi-factorial effect of climate change on forest ecosystem productivity. Indeed, the effect of stress interactions on tree growth is an increasingly important topic that greatly lacks experiments and data, and it is unlikely that the impact of combined stresses can be extrapolated from the outcomes of studies that focused on a single stress. We developed an original manipulative study under real field conditions where we applied artificial defoliation and induced water stress on 10-year-old (~10 m high) maritime pine trees ( Pinus pinaster Ait.). Tree response to combined stresses was quantitatively assessed following tree secondary growth and carbohydrate pools. Such a design allowed us to address the crucial question of combined stresses on trees under stand conditions, sharing soil supplies with neighboring trees. Our initial hypotheses were that (i) moderate defoliation can limit the impact of water stress on tree growth through reduced transpiration demand by a tree canopy partly defoliated and that (ii) defoliation results in reduced non-structural carbohydrate (NSC) pools, affecting tree tolerance to drought. Our results showed additive effects of defoliation and water stress on tree growth and contradict our initial hypothesis. Indeed, under stand conditions, we found that partial defoliation does not limit the impact of water stress through reduced transpiration. Our study also highlighted that, even if NSC in all organs were affected by defoliation, tree response to water stress was not triggered. We found that stem NSC were maintained or increased during the entire growing season, supporting literature-based hypotheses such as an active maintenance of the hydraulic system or another limiting resource for tree growth under defoliation. We also observed a significant decrease in root carbohydrates, which suggests a shift in the root carbon balance under defoliation. The decrease in carbohydrate supply under defoliation may not counterbalance the carbon use for mineral and water uptakes or a translocation to other tissues.

Description: In resprouting species, fire-induced topkill causes a reduction in height and leaf area without a comparable reduction in the size of the root system, which should lead to an increase in the efficiency of water transport after fire. However, large plants undergo a greater relative reduction in size, compared with small plants, so we hypothesized that this enhancement in hydraulic efficiency would be greatest among large growth forms. In the ecotone between long-leaf pine ( Pinus palustris Mill.) savannas and wetlands, we measured stomatal conductance ( g s ), mid-day leaf water potential ( leaf ), leaf-specific whole-plant hydraulic conductance ( K L.p ), leaf area and height of 10 species covering a range of growth forms in burned and unburned sites. As predicted, K L.p was higher in post-fire resprouts than in unburned plants, and the post-fire increase in K L.p was positively related to plant size. Specifically, large-statured species tended to undergo the greatest relative reductions in leaf area and height, and correspondingly experienced the greatest increases in K L.p . The post-fire increase in K L.p was smaller than expected, however, due to a decrease in absolute root hydraulic conductance (i.e., not scaled to leaf area). The higher K L.p in burned sites was manifested as an increase in g s rather than an increase in leaf . Post-fire increases in g s should promote high rates of photosynthesis for recovery of carbohydrate reserves and aboveground biomass, which is particularly important for large-statured species that require more time to recover their pre-fire size.

Description: Previous studies have shown that there are significant sexual differences in the morphological and physiological responses of Populus cathayana Rehder under stressful conditions. However, little is known about sex-specific differences in responses to nutrient deficiencies. In this study, the effects of nitrogen (N) and phosphorus (P) deficiencies on the morphological, physiological and chloroplast ultrastructural traits of P. cathayana males and females were investigated. The results showed that N and P deficiencies significantly decreased plant growth, foliar N and P contents, chlorophyll content, photosynthesis, and instantaneous photosynthetic N- and P-use efficiencies (PNUE and PPUE) in both sexes. Males had higher photosynthesis, higher PNUE and PPUE rates, and a lower accumulation of plastoglobules in chloroplasts than did females when exposed to N- and P-deficiency conditions. Nitrogen-deficient males had higher glutamate dehydrogenase and peroxidase activities, and a more intact chloroplast ultrastructure, but less starch accumulation than did N-deficient females. Phosphorus-deficient males had higher nitrate reductase, glutamine synthetase and acid phosphatase activities, but a lower foliar N : P ratio and less PSII damage than did P-deficient females. These results suggest that N and P deficiencies cause greater negative effects on females than on males, and that the different sexes of P. cathayana may employ different strategies to cope with N and P deficiencies.

Description: Root diameter, a critical indicator of root physiological function, varies greatly among tree species, but the underlying mechanism of this high variability is unclear. Here, we sampled 50 tree species across tropical and temperate zones in China, and measured root morphological and anatomical traits along the first five branch orders in each species. Our objectives were (i) to reveal the relationships between root diameter, cortical thickness and stele diameter among tree species in tropical and temperate forests, and (ii) to investigate the relationship of both root morphological and anatomical traits with divergence time during species radiation. The results showed that root diameter was strongly affected by cortical thickness but less by stele diameter in both tropical and temperate species. Changes in cortical thickness explained over 90% of variation in root diameter for the first order, and ~74–87% for the second and third orders. Thicker roots displayed greater cortical thickness and more cortical cell layers than thinner roots. Phylogenetic analysis demonstrated that root diameter, cortical thickness and number of cortical cell layers significantly correlated with divergence time at the family level, showing similar variation trends in geological time. The results also suggested that trees tend to decrease their root cortical thickness rather than stele diameter during species radiation. The close linkage of variations in root morphology and anatomy to phylogeny as demonstrated by the data from the 50 tree species should provide some insights into the mechanism of root diameter variability among tree species.

Description: Spring phenology of temperate forest trees is optimized to maximize the length of the growing season while minimizing the risk of freezing damage. The release from winter dormancy is environmentally mediated by species-specific responses to temperature and photoperiod. We investigated the response of early spring phenology to temperature and photoperiod at different stages of dormancy release in cuttings from four temperate tree species in controlled environments. By tracking bud development, we were able to identify the onset of bud swelling and bud growth in Acer pseudoplatanus L., Fagus sylvatica L., Quercus petraea (Mattuschka) Liebl. and Picea abies (L.) H. Karst. At a given early stage of dormancy release, the onset and duration of the bud swelling prior to bud burst are driven by concurrent temperature and photoperiod, while the maximum growth rate is temperature dependent only, except for Fagus , where long photoperiods also increased bud growth rates. Similarly, the later bud burst was controlled by temperature and photoperiod (in the photoperiod sensitive species Fagus, Quercus and Picea ). We conclude that photoperiod is involved in the release of dormancy during the ecodormancy phase and may influence bud burst in trees that have experienced sufficient chilling. This study explored and documented the early bud swelling period that precedes and defines later phenological stages such as canopy greening in conventional phenological works. It is the early bud growth resumption that needs to be understood in order to arrive at a causal interpretation and modelling of tree phenology at a large scale. Classic spring phenology events mark visible endpoints of a cascade of processes as evidenced here.

Description: In tropical dry forests, uppermost-canopy leaves of evergreen trees possess the ability to use water more conservatively compared with drought-deciduous trees, which may result from significant differences in the photoprotective mechanisms between functional types. We examined the seasonal variations in leaf gas exchange, chlorophyll fluorescence and the amounts of photosynthetic pigments within lamina of the uppermost-canopy leaves of three drought-deciduous trees ( Vitex peduncularis Wall., Xylia xylocarpa (Roxb.) W. Theob., Shorea siamensis Miq.), a semi-deciduous tree ( Irvingia malayana Miq.) and two evergreen trees ( Hopea ferrea Lanessan and Syzygium cumini (L.) Skeels) in Thailand. Area-based maximum carbon assimilation rates ( A max ) decreased during the dry season, except in S. siamensis . The electron transport rate (ETR) remained unchanged in deciduous trees, but decreased during the dry season in evergreen and semi-deciduous trees. In the principal component analysis, the first axis (Axis 1) accounted for 44.3% of the total variation and distinguished deciduous from evergreen trees. Along Axis 1, evergreen trees were characterized by a high Stern–Volmer non-photochemical quenching coefficient (NPQ), high xanthophyll cycle pigments/chlorophyll and a high de-epoxidation state of the xanthophyll cycle, whereas the deciduous trees were characterized by a high ETR, a high quantum yield of PSII ( PSII = ( F m ' – F )/ F m ' ) and a high mass-based A max under high-light conditions. These findings indicate that drought-deciduous trees showing less conservative water use tend to dissipate a large proportion of electron flow through photosynthesis or alternative pathways. In contrast, the evergreens showed more conservative water use, reduced A max and ETR and enhanced NPQ and xanthophyll cycle pigments/chlorophyll during the dry season, indicating that down-regulated photosynthesis with enhanced thermal dissipation of excess light energy played an important role in photoprotection. Trees with different water uses and leaf lifespans appear to employ different photoprotective mechanisms to overcome the unfavorable dry-season drought. Our data may suggest that future changes in precipitation will strongly impinge on forest structure and functions.

Description: Plant species use different strategies for maximizing growth and fitness under changing environmental conditions. At the ecosystem level, seedlings in particular compete with other vegetation components for light and nitrogen (N), which often constitute growth-limiting resources. In this study, we investigated the effect of light availability on the competition for N between seedlings of European beech and sycamore maple and analysed the consequences of this competition for the composition of N metabolites in fine roots. Our results show different strategies in N acquisition between beech and sycamore maple. Both species responded to reduced light availability by adapting their morphological and physiological traits with a decrease in biomass and net assimilation rate and an increase in specific leaf area and leaf area ratio. For beech seedlings, competition with sycamore maple led to a reduction in organic N uptake capacity. Reduced light availability led to a decrease in ammonium, but an increase in glutamine-N uptake capacity in sycamore maple. However, this response was stronger compared with that of beech and was accompanied by reduced growth. Thus, our results suggest better adaptation of N acquisition to reduced light availability in beech compared with sycamore maple seedlings.

Description: Shrubs of the Great Basin desert in Utah are subjected to a prolonged summer drought with the potential consequence of reduced water transport capability of the xylem due to drought-induced cavitation. Hydraulic redistribution (HR) is the passive movement of water from deep to shallow soil through plant roots. Hydraulic redistribution can increase water availability in shallow soil and ameliorate drought stress, providing better soil and root water status, which could affect shallow root conductivity ( K s ) and native root embolism. We tested this hypothesis in an Artemisia tridentata Nutt. mono-specific stand grown in a common garden in Utah. We enhanced HR artificially by applying a once a week deep-irrigation treatment increasing the water potential gradient between deep and shallow soil layers. Plants that were deep-watered had less negative water potentials and greater stomatal conductance and transpiration rates than non-watered control plants. After irrigation with labeled water ( D ), xylem water in stems and shallow roots of watered shrubs was enriched with respect to control shrubs, a clear indication of deep water uptake and HR. Shallow root conductivity was threefold greater and shrubs experienced lower native embolism when deep-watered. We found clear evidence of water transfer between deep and shallow roots through internal HR that delayed depletion of shallow soil water content, maintained K s and prevented root embolism. Overall, our results show a positive effect of HR on root water transport capacity in otherwise dry soil, with important implications for plant water status.

Description: Physiological changes in host plants in response to the broad spectrum of fungal modes of infection are still not well understood. The current study was conducted to better understand the infection of in vitro cultures of Pinus sylvestris L. seedlings by three trophically diverse fungal species, Fusarium oxysporum E. F. Sm. & Swingle, Trichoderma harzianum Rifai and Hebeloma crustuliniforme (Bull.) Quél. Biochemical methods and microscopy were utilized to determine (i) which factors (apoplastic and cellular pH, reactive oxygen species, glutathione and cell death) play a role in the establishment of pathogenic, saprotrophic and mycorrhizal fungi, and (ii) whether cell death is a common response of conifer seedling tissues when they are exposed to trophically diverse fungi. Establishment of the pathogen, F. oxysporum , was observed more frequently in the meristematic region of root tips than in the elongation zone, which was in contrast to T. harzianum and H. crustuliniforme . Ectomycorrhizal (ECM) hyphae, however, were occasionally observed in the studied root zone and caused small changes in the studied factors. Colonization of the meristematic zone occurred due to host cell death. Independently of the zone, changes in cellular pH resulting in an acidic cytoplasm conditioned the establishment of F. oxysporum . Additionally, cell death was negatively correlated with hydrogen peroxide (H 2 O 2 ) in roots challenged by a pathogenic fungus. Cell death was the only factor uniquely associated with the colonization of host roots by a saprotrophic fungus. The mechanism may differ, however, between the zones since apoplastic pH was negatively correlated with cell death in the elongation zone, whereas in the meristematic zone, none of the studied factors explained cell death. Colonization by the ECM fungus, H. crustuliniforme , was associated with a decreasing number of cells with acidic apoplast and by production of H 2 O 2 in the elongation zone resulting in cell death. Saprotrophic and ECM fungi had a greater effect on cell acidification in the meristematic zone than the pathogenic fungus.

Description: Drought reduces the carbon (C) assimilation of trees and decouples aboveground from belowground carbon fluxes, but little is known about the response of drought-stressed trees to rewetting. This study aims to assess dynamics and patterns of C allocation in beech saplings under dry and rewetted soil conditions. In October 2010, 5-year-old beech saplings from a forest site were transplanted into 20 l pots. In 2011, the saplings were subjected to different levels of soil drought ranging from non-limiting water supply (control) to severe water limitation with soil water potentials of less than –1.5 MPa. As a physiologically relevant measure of drought, the cumulated soil water potential (i.e., drought stress dose (DSD)) was calculated for the growing season. In late August, the saplings were transferred into a climate chamber and pulse-labeled with 13 C-depleted CO 2 ( 13 C of –47). Isotopic signatures in leaf and soil respiration were repeatedly measured. Five days after soil rewetting, a second label was applied using 99 atom% 13 CO 2 . After another 12 days, the fate of assimilated C in each sapling was assessed by calculating the 13 C mass balance. Photosynthesis decreased by 60% in saplings under severe drought. The mean residence time (MRT) of recent assimilates in leaf respiration was more than three times longer than under non-limited conditions and was positively correlated to DSD. Also, the appearance of the label in soil respiration was delayed. Within 5 days after rewetting, photosynthesis, MRT of recent assimilates in leaf respiration and appearance of the label in soil respiration recovered fully. Despite the fast recovery, less label was recovered in the biomass of the previously drought-stressed plants, which also allocated less C to the root compartment (45 vs 64% in the control). We conclude that beech saplings quickly recover from extreme soil drought, although transitional after-effects prevail in C allocation, possibly due to repair-driven respiratory processes.

Description: Despite the critical role that phloem plays in a number of plant functional processes and the potential impact of water stress on phloem structural and phloem sap compositional characteristics, little research has been done to examine how water stress influences phloem transport. The objectives of this study were to develop a more accurate understanding of how water stress affects phloem transport in trees, both in terms of the short-term impacts of water stress on phloem sap composition and the longer-term impacts on sieve cell anatomical characteristics. Phloem sieve cell conductivity ( k p ) was evaluated along a gradient of tree height and xylem water potential in Douglas-fir ( Pseudotsuga menziesii (Mirb.) Franco) trees in order to evaluate the influence of water stress on phloem transport capacity. The Hagen–Poiseuille equation was used with measurements of sieve cell anatomical characteristics, water content of phloem sap, non-structural carbohydrate content of phloem sap and shoot water potential ( l ) to evaluate impacts of water stress on k p . Based on regression analysis, for each 1 MPa decrease in mean midday l , sieve cell lumen radius decreased by 2.63 µm MPa –1 . Although there was no significant trend in sucrose content with decreasing l , glucose and fructose content increased significantly with water stress and sieve cell relative water content decreased by 13.5% MPa –1 , leading to a significant increase in sugar molar concentration of 0.46 mol l –1  MPa –1 and a significant increase in viscosity of 0.27 mPa s MPa –1 . Modeled k p was significantly influenced both by trends in viscosity as well as by water stress-related trends in sieve cell anatomy.

Description: In deciduous trees, shoot development in early spring is assumed to be achieved mainly at the expense of nitrogen (N) stores. Indeed, the possible compensation for poor autumn N storage by early spring N uptake has been little studied. We therefore determined the dynamics of spring N uptake in relation to spring N supply, carbon and N storage and shoot development. Young peach trees ( Prunus persica L. Batsch, cv. ‘GF305 ’ ) were raised outdoors in a hydroponic set-up during the spring and summer, with an excessive N supply. During the autumn, half of the trees were then N limited. The following spring, the N supply remained either high or low, or changed from high to low or low to high. Between 6 March and 13 May, N uptake was measured automatically on an hourly basis, while shoot growth was monitored once a week. These in situ measurements were completed by three destructive harvests to assess organ composition in N and total non-structural carbohydrates (TNC). Until the end of April, N uptake was dependent on the autumn N treatment, being higher in trees that had been N limited in the autumn. Total non-structural carbohydrate mobilization was also higher in those trees that had lost at least 17 g TNC by 24 April, while TNC levels in non-limited trees remained stable or even rose. Shoot development, estimated by the number of elongated axes and leaves per axis, was also slightly delayed by an N limitation in autumn. After 24 April, N uptake rates increased notably under all treatments and was determined by the spring N supply. In trees receiving a high N supply in the spring, the uptake rates also displayed marked short-term variations. That reduced the differences between treatments and by 13 May no differences could be evidenced between the trees in terms of organ biomass and TNC and N contents, whatever the treatment. We concluded that in the early spring, N uptake may compensate for a deficit of N storage insofar as large quantities of TNC can be mobilized for that purpose.

Description: This manuscript aims to evaluate the precision and accuracy of current methodology for estimating xylem phenology and tracheid production in trees. Through a simple approach, sampling at two positions on the stem of co-dominant black spruce trees in two sites of the boreal forest of Quebec, we were able to quantify variability among sites, between trees and within a tree for different variables. We demonstrated that current methodology is accurate for the estimation of the onset of xylogenesis, while the accuracy for the evaluation of the ending of xylogenesis may be improved by sampling at multiple positions on the stem. The pattern of variability in different phenological variables and cell production allowed us to advance a novel hypothesis on the shift in the importance of various drivers of xylogenesis, from factors mainly varying at the level of site (e.g., climate) at the beginning of the growing season to factors varying at the level of individual trees (e.g., possibly genetic variability) at the end of the growing season.

Description: Somatic embryogenesis is an important biotechnological tool in the large-scale propagation of elite genotypes and ex situ conservation of conifer species. Protocols for the induction and proliferation of embryogenic cultures (ECs) of Brazilian pine ( Araucaria angustifolia (Bert.) O. Ktze) are well established, although the proper formation of mature somatic embryos (SEs) is still problematic. Thus, the identification of molecular markers for the screening of ECs able to respond to maturation conditions (abscisic acid and osmotic agents) is highly desirable. To develop molecular markers for the early detection of ECs able to develop well-formed SEs under maturation conditions, we analyzed the proteins found during the proliferation phase of A. angustifolia cell lines with different embryogenic capabilities, with one cell line being responsive to maturation conditions (R cell line), and one cell line that presented blocked development of SEs (B cell line). In addition, based on the peptides identified, polyamine levels (free and conjugate), ethylene production and reactive oxygen species (ROS) emission were analyzed using both EC lines (R and B cell lines). A marked difference in the biochemistry of ECs between these two cell lines was observed. Eleven proteins that were differentially expressed in the cell lines were identified by the combination of two-dimensional electrophoresis (2-DE) and MALDI-TOF/TOF mass spectrometry. Among these, S -adenosylmethionine synthase, the enzyme associated with polyamines and ethylene biosynthesis, was observed exclusively in the R cell line, while a protein linked to the oxidative stress subunit F of NADH dehydrogenase was observed exclusively in the B cell lines. Additionally, B cell lines showed higher levels of diamine putrescine and lower levels of ethylene. Higher values of ethylene and ROS were observed for the cell line that showed normal development of SEs. Altogether, our results open new perspectives in the optimization of culture conditions for A. angustifolia somatic embryogenesis, as well as establishing biochemical markers for the early selection of ECs during maturation trials.

Description: Insights into how the simultaneous presence of organic and inorganic nitrogen (N) forms influences root absorption will help elucidate the relative importance of these N forms for plant nutrition in the field as well as for nursery cultivation of seedlings. Uptake of the individual N forms arginine, ammonium (NH 4 + ) and nitrate (NO 3 – ) was studied in Scots pine ( Pinus sylvestris (L.)) seedlings supplied as single N sources and additionally in mixtures of NO 3 – and NH 4 + or NO 3 – and arginine. Scots pine seedlings displayed a strong preference for NH 4 + -N and arginine-N as compared with NO 3 – -N. Thus, NO 3 – uptake was generally low and decreased in the presence of NH 4 + in the high-concentration range (500 µM N), but not in the presence of arginine. Moreover, uptake of NO 3 – and NH 4 + was lower in seedlings displaying a high internal N status as a result of high N pre-treatment, while arginine uptake was high in seedlings with a high internal N status when previously exposed to organic N. These findings may have practical implications for commercial cultivation of conifers.

Description: Oak species are well suited to water-limited conditions by either avoiding water stress through deep rooting or tolerating water stress through tight stomatal control. In co-occurring species where resources are limited, species may either partition resources in space and/or time or exhibit differing efficiencies in the use of limited resources. Therefore, this study seeks to determine whether two co-occurring oak species ( Quercus prinus L. and Quercus velutina Lam.) differ in physiological parameters including photosynthesis, stomatal conductance, water-use (WUE) and nitrogen-use efficiency (NUE), as well as to characterize transpiration and average canopy stomatal responses to climatic variables in a sandy, well-drained and nutrient-limited ecosystem. The study was conducted in the New Jersey Pinelands and we measured sap flux over a 3-year period, as well as leaf gas exchange, leaf nitrogen and carbon isotope concentrations. Both oak species showed relatively steep increases in leaf-specific transpiration at low vapor pressure deficit (VPD) values before maximum transpiration rates were achieved, which were sustained over a broad range in VPD. This suggests tight stomatal control over transpiration in both species, although Q. velutina showed significantly higher leaf-level and canopy-level stomatal conductance than Q. prinus . Average daytime stomatal conductance was positively correlated with soil moisture and both oak species maintained at least 75% of their maximum canopy stomatal conductance at soil moistures in the upper soil layer (0–0.3 m) as low as 0.03 m 3 m 3 –3 . Quercus velutina had significantly higher photosynthetic rates, maximum Rubisco-limited and electron-transport-limited carboxylation rates, dark respiration rates and nitrogen concentration per unit leaf area than Q. prinus . However, both species exhibited similar WUEs and NUEs. Therefore, Q. prinus has a more conservative resource-use strategy, while Q. velutina may need to exploit niches that are locally higher in nutrients and water. Likewise, both species appear to tap deep, stable water sources, highlighting the importance of rooting depth in modeling transpiration and stomatal conductance in many oak ecosystems.

Description: Leaf mass per area (LMA, g m –2 ) is an essential trait for modeling canopy function due to its strong association with photosynthesis, respiration and leaf nitrogen. Leaf mass per area, which is influenced by both leaf thickness and density (LMA = thickness  x  density), generally increases from the bottom to the top of tree canopies, yet the mechanisms behind this universal pattern are not yet resolved. For decades, the light environment was assumed to be the most influential driver of within-canopy variation in LMA, yet recent evidence has shown hydrostatic gradients to be more important in upper canopy positions, especially in tall evergreen trees in temperate and tropical forests. The aim of this study was to disentangle the importance of various environmental drivers on vertical LMA gradients in a mature sugar maple ( Acer saccharum Marshall) forest. We compared LMA, leaf density and leaf thickness relationships with height, light and predawn leaf water potential ( Pre ) within a closed and an exposed canopy to assess leaf morphological traits at similar heights but different light conditions. Contrary to our expectations and recent findings in the literature, we found strong evidence that light was the primary driver of vertical gradients in leaf morphology. At similar heights (13–23 m), LMA was greater within the exposed canopy than the closed canopy, and light had a stronger influence over LMA compared with Pre . Light also had a stronger influence over both leaf thickness and density compared with Pre ; however, the increase in LMA within both canopy types was primarily due to increasing leaf thickness with increasing light availability. This study provides strong evidence that canopy structure and crown exposure, in addition to height, should be considered as a parameter for determining vertical patterns in LMA and modeling canopy function.

Description: We investigated the role of glycolysis and sucrolysis in the difference in tolerance to root hypoxia between two Myrtaceae tree species, Melaleuca cajuputi (which shows superior tolerance to root hypoxia) and Eucalyptus camaldulensis (which does not). Analysis of the adenylate energy charge (AEC) in roots subjected to a 4-day hypoxic treatment (HT) in hydroponic culture revealed that the interspecies difference in tolerance corresponds to the ability to maintain energy status under root hypoxia: AEC was reduced by HT in E. camaldulensis , but not in M. cajuputi . The energy status in HT roots of E. camaldulensis was restored by feeding of glucose (Glc) but not sucrose (Suc). These data provide evidence that low substrate availability for glycolysis resulting from an impairment of sucrolysis suppresses ATP production under hypoxic conditions in this species. Measurements of the rates of O 2 consumption and CO 2 production in roots indicated that E. camaldulensis , but not M. cajuputi , failed to activate fermentation in HT roots. These results cannot be attributed to enzymatic dysfunction, because no inhibition of main glycolytic and fermentative enzymes was observed in both species, and Glc feeding had a beneficial effect on AEC of HT roots of E. camaldulensis . The impairment of sucrolysis was demonstrated by inhibited soluble acid invertase activity in HT roots of E. camaldulensis . In contrast, there was no inhibition in all sucrolytic enzymes tested in HT roots of M. cajuputi , suggesting that steady Suc degradation is essential for maintaining high energy status under root hypoxia. We conclude that root sucrolysis is one of the essential factors that determines the extent of tolerance to root hypoxia.

Description: The photosynthesis in evergreen trees living in Mediterranean ecosystems is subjected to multiple climatic stresses due to water shortage and high temperatures during the summer and to low temperatures during the winter. Mediterranean perennials deploy different photoprotective mechanisms to prevent damage to the photosynthetic system. Wax accumulation in leaves is a primary response which by enhancing light scattering in the leaf surface reduces incident radiation in the mesophyll. The existence of high variability in wax accumulation levels between coexisting individuals of a species has a visual effect on colour that provides distinguishable green and glaucous phenotypes. We explored this variability in a Mediterranean evergreen tree Juniperus thurifera (L.) to evaluate the impact of epicuticular wax on optical and ecophysiological properties and on the abundance of photoprotective pigments throughout an annual cycle. Because of light attenuation by waxes, we expected that glaucous phenotypes would lower the need for photoprotective pigments. We evaluated the effect of phenotype and season on reflectance, defoliation levels, photochemical efficiency and photoprotective pigment contents in 20 green and 20 glaucous junipers. Contrary to our expectations, the results showed that glaucous trees suffered from a diminution in photochemical efficiency, but there was no reduction in photoprotective pigments. Differences between glaucous and green phenotypes were greater in winter, which is the most stressful season for this species. Glaucous individuals also showed the highest levels of leaf defoliation. The lower photochemical efficiency of glaucous trees, together with higher defoliation rates and equal or greater number of physiological photoprotective mechanisms, suggests that in spite of wax accumulation, glaucous trees suffer from more severe stress than green ones. This result suggests that changes in colouration in Mediterranean evergreens may be a decline indicator.

Description: Low water potentials in xylem can result in damaging levels of cavitation, yet little is understood about which hydraulic traits have most influence in delaying the onset of hydraulic dysfunction during periods of drought. We examined three traits contributing to longer desiccation times in excised shoots of 11 species from two sites of contrasting aridity: (i) the amount of water released from plant tissues per decrease in xylem water potential ( W ); (ii) the minimum xylem water potential preceding acute water stress (defined as P 50L ; water potential at 50% loss of leaf conductance); and (iii) the integrated transpiration rate between the points of full hydration and P 50L ( W time ). The time required for species to reach P 50L varied markedly, ranging from 1.3 h to nearly 3 days. W , P 50L and W time all contributed significantly to longer desiccation times, explaining 28, 22 and 50% of the variance in the time required to reach P 50L . Interestingly, these three traits were nearly orthogonal to one another, suggesting that they do not represent alternative hydraulic strategies, but likely trade off with other ecological strategies not evaluated in this study. The majority of water lost during desiccation (60–91%) originated from leaves, suggesting an important role for leaf capacitance in small plants when xylem water potentials decrease below –2 MPa.

Description: Ozone is an air pollutant that causes oxidative stress by generation of reactive oxygen species (ROS) within the leaf. The capacity to detoxify ROS and repair ROS-induced damage may contribute to ozone tolerance. Ascorbate and glutathione are known to be key players in detoxification. Ozone effects on their biosynthesis and on amino acid metabolism were investigated in three Euramerican poplar genotypes ( Populus deltoides Bartr. x Populus nigra L.) differing in ozone sensitivity. Total ascorbate and glutathione contents were increased in response to ozone in all genotypes, with the most resistant genotype (Carpaccio) showing an increase of up to 70%. Reduced ascorbate (ASA) concentration at least doubled in the two most resistant genotypes (Carpaccio and Cima), whereas the most sensitive genotype (Robusta) seemed unable to regenerate ASA from oxidized ascorbate (DHA), leading to an increase of 80% of the oxidized form. Increased ascorbate (ASA + DHA) content correlated with the increase in gene expression in its biosynthetic pathway, especially the putative gene of GDP- l -galactose phosphorylase VTC2 . Increased cysteine availability combined with increased expression of -glutamylcysteine synthetase ( GSH1 ) and glutathione synthetase ( GSH2 ) genes allows higher glutathione biosynthesis in response to ozone, particularly in Carpaccio. In addition, ozone caused a remobilization of amino acids with a decreased pool of total amino acids and an increase of Cys and putrescine, especially in Carpaccio. In addition, the expression of genes encoding threonine aldolase was strongly induced only in the most tolerant genotype, Carpaccio. Reduced ascorbate levels could partly explain the sensitivity to ozone for Robusta but not for Cima. Reduced ascorbate level alone is not sufficient to account for ozone tolerance in poplar, and it is necessary to consider several other factors including glutathione content.

Description: The rapid and widespread expansion of rubber plantations in Southeast Asia necessitates a greater understanding of tree physiology and the impacts of water consumption on local hydrology. Sap flow measurements were used to study the intra- and inter-annual variations in transpiration rate ( E t ) in a rubber stand in the low-elevation plain of central Cambodia. Mean stand sap flux density ( J S ) indicates that rubber trees actively transpire in the rainy season, but become inactive in the dry season. A sharp, brief drop in J S occurred simultaneously with leaf shedding in the middle of the dry season in January. Although the annual maxima of J S were approximately the same in the two study years, the maximum daily stand E t of ~2.0 mm day –1 in 2010 increased to ~2.4 mm day –1 in 2011. Canopy-level stomatal response was well explained by changes in solar radiation, vapor pressure deficit, soil moisture availability, leaf area, and stem diameter. Rubber trees had a relatively small potential to transpire at the beginning of the study period, compared with average diffuse-porous species. After 2 years of growth in stem diameter, transpiration potential was comparable to other species. The sensitivity of canopy conductance ( g c ) to atmospheric drought indicates isohydric behavior of rubber trees. Modeling also predicted a relatively small sensitivity of g c to the soil moisture deficit and a rapid decrease in g c under extreme drought conditions. However, annual observations suggest the possibility of a change in leaf characteristics with tree maturity and/or initiation of latex tapping. The estimated annual stand E t was 469 mm year –1 in 2010, increasing to 658 mm year –1 in 2011. Diagnostic analysis using the derived g c model showed that inter-annual change in stand E t in the rapidly growing young rubber stand was determined mainly by tree growth rate, not by differences in air and soil variables in the surrounding environment. Future research should focus on the potentially broad applicability of the relationship between E t and tree size as well as environmental factors at stands different in terms of clonal type and age.

Description: Temperature is the most important factor driving the cold edge distribution limit of temperate trees. Here, we identified the minimum temperatures for root growth in seven broad-leaved tree species, compared them with the species' natural elevational limits and identified morphological changes in roots produced near their physiological cold limit. Seedlings were exposed to a vertical soil-temperature gradient from 20 to 2 °C along the rooting zone for 18 weeks. In all species, the bulk of roots was produced at temperatures above 5 °C. However, the absolute minimum temperatures for root growth differed among species between 2.3 and 4.2 °C, with those species that reach their natural distribution limits at higher elevations also tending to have lower thermal limits for root tissue formation. In all investigated species, the roots produced at temperatures close to the thermal limit were pale, thick, unbranched and of reduced mechanical strength. Across species, the specific root length (m g –1 root) was reduced by, on average, 60% at temperatures below 7 °C. A significant correlation of minimum temperatures for root growth with the natural high elevation limits of the investigated species indicates species-specific thermal requirements for basic physiological processes. Although these limits are not necessarily directly causative for the upper distribution limit of a species, they seem to belong to a syndrome of adaptive processes for life at low temperatures. The anatomical changes at the cold limit likely hint at the mechanisms impeding meristematic activity at low temperatures.

Description: Stored non-structural carbohydrates (NSCs) could play an important role in tree survival in the face of a changing climate and associated stress-related mortality. We explored the effects of the stomata-blocking and defoliating fungal disease called Swiss needle cast on Douglas-fir carbohydrate reserves and growth to evaluate the extent to which NSCs can be mobilized under natural conditions of low water stress and restricted carbon supply in relation to potential demands for growth. We analyzed the concentrations of starch, sucrose, glucose and fructose in foliage, twig wood and trunk sapwood of 15 co-occurring Douglas-fir trees expressing a gradient of Swiss needle cast symptom severity quantified as previous-year functional foliage mass. Growth (mean basal area increment, BAI) decreased by ~80% and trunk NSC concentration decreased by 60% with decreasing functional foliage mass. The ratio of relative changes in NSC concentration and BAI, an index of the relative priority of storage versus growth, more than doubled with increasing disease severity. In contrast, twig and foliage NSC concentrations remained nearly constant with decreasing functional foliage mass. These results suggest that under disease-induced reductions in carbon supply, Douglas-fir trees retain NSCs (either actively or due to sequestration) at the expense of trunk radial growth. The crown retains the highest concentrations of NSC, presumably to maintain foliage growth and shoot extension in the spring, partially compensating for rapid foliage loss in the summer and fall.

Description: There is a need to incorporate the effects of herbivore damage into future models of plant volatile organic compound (VOC) emissions at leaf or canopy levels. Short-term (a few seconds to 48 h) changes in shoot VOC emissions of silver birch ( Betula pendula Roth) in response to feeding by geometrid moths ( Erannis defoliaria Hübner) were monitored online by proton transfer reaction time-of-flight mass spectrometry (PTR-TOF-MS). In addition, two separate field experiments were established to study the effects of long-term foliage herbivory (FH, 30–32 days of feeding by geometrids Agriopis aurantiaria (Clerck) and E. defoliaria in two consecutive years) and bark herbivory (BH, 21 days of feeding by the pine weevil ( Hylobius abietis L.) in the first year) on shoot and rhizosphere VOC emissions of three silver birch genotypes (gt14, gt15 and Hausjärvi provenance). Online monitoring of VOCs emitted from foliage damaged by geometrid larvae showed rapid bursts of green leaf volatiles (GLVs) immediately after feeding activity, whereas terpenoid emissions had a tendency to gradually increase during the monitoring period. Long-term FH caused transient increases in total monoterpene (MT) emissions from gt14 and sesquiterpene (SQT) emissions from Hausjärvi provenance, mainly in the last experimental season. In the BH experiment, genotype effects were detected, with gt14 trees having significantly higher total MT emissions compared with other genotypes. Only MTs were detected in the rhizosphere samples of both field experiments, but their emission rates were unaffected by genotype or herbivory. The results suggest that silver birch shows a rapid VOC emission response to short-term foliage herbivory, whereas the response to long-term foliage herbivory and bark herbivory is less pronounced and variable at different time points.

Description: Redwood branches provide all the carbohydrates for the most carbon-heavy forests on Earth, and recent whole-tree measurements have quantified trunk growth rates associated with complete branch inventories. Providing all of a tree's photosynthetic capacity, branches represent an increasing proportion of total aboveground wood production as trees enlarge. To examine branch development and its effects on wood volume growth, we dissected 31 branches from eight Sequoia sempervirens (D. Don) Endl. and seven Sequoiadendron giganteum Lindl. trees. The cambium-area-to-leaf-area ratio was maintained with size and age but increased with light availability, whereas the heartwood-deposition-area-to-leaf-area ratio increased with size and age but was insensitive to light availability. The proportion of foliage mass arrayed in 〈1-cm-diameter epicormic shoots increased with decreasing light and was higher in Sequoia (20–60%) than in Sequoiadendron (3–16%). Well-illuminated branches concentrated leaves higher and distally, while shaded branches distributed leaves lower and proximally. In similar light environments, older branches distributed leaves lower and more proximally than younger branches. Branch size, light, species, heartwood area, a heartwood-area–species interaction, and ovulate cone mass predicted 87.5% of the variability in wood volume growth of branches. After accounting for the positive effects of size and light, wood volume growth declined with heartwood area and age. The effect of age was trivial compared to the effect of heartwood area, suggesting that heartwood expansion caused the age-related decline in wood volume growth. Additionally, Sequoiadendron branches of similar size and light environment with more ovulate cones produced less wood, even though these cones were long-lived and photosynthetic, reflecting the energetic cost of seed production. These results contributed to a conceptual model of branch development in which light availability, injury, heartwood content, gravity, and time interact to produce the high degree of branch structural variation evident within redwood crowns.

Description: Understory plants in tropical forests often experience a low-light environment combined with high CO 2 concentration. We hypothesized that the high CO 2 concentration may compensate for leaf carbon loss caused by the low light, through increasing light-use efficiency of both steady-state and dynamic photosynthetic properties. To test the hypothesis, we examined CO 2 gas exchange in response to an artificial lightfleck in Dipterocarpus sublamellatus Foxw. seedlings under contrasting CO 2 conditions: 350 and 700 μmol CO 2  mol –1 air in a tropical rain forest, Pasoh, Malaysia. Total photosynthetic carbon gain from the lightfleck was about double when subjected to the high CO 2 when compared with the low CO 2 concentration. The increase of light-use efficiency in dynamic photosynthesis contributed 7% of the increased carbon gain, most of which was due to reduction of photosynthetic induction to light increase under the high CO 2 . The light compensation point of photosynthesis decreased by 58% and the apparent quantum yield increased by 26% at the high CO 2 compared with those at the low CO 2 . The study suggests that high CO 2 increases photosynthetic light-use efficiency under both steady-state and fluctuating light conditions, which should be considered in assessing the leaf carbon gain of understory plants in low-light environments.

Description: In trees exposed to prolonged drought, both carbon uptake (C source) and growth (C sink) typically decrease. This correlation raises two important questions: (i) to what degree is tree growth limited by C availability; and (ii) is growth limited by concurrent C storage (e.g., as nonstructural carbohydrates, NSC)? To test the relationships between drought, growth and C reserves, we monitored the changes in NSC levels and constructed stem growth chronologies of mature Pinus halepensis Miller trees of three drought stress levels growing in Yatir forest, Israel, at the dry distribution limit of forests. Moderately stressed and stressed trees showed 34 and 14% of the stem growth, 71 and 31% of the sap flux density, and 79 and 66% of the final needle length of healthy trees in 2012. In spite of these large reductions in growth and sap flow, both starch and soluble sugar concentrations in the branches of these trees were similar in all trees throughout the dry season (2–4% dry mass). At the same time, the root starch concentrations of moderately stressed and stressed trees were 47 and 58% of those of healthy trees, but never 〈2% dry mass. Our results show that all the studied trees maintain a fairly good coordination between C supply and demand, and even during prolonged drought there is more than one way for a tree to maintain a positive C balance.

Description: Long-lived tree species are subject to attack by various pests and pathogens during their lifetime. This problem is exacerbated by climate change, which may increase the host range for pathogens and extend the period of infestation by pests. Plant defences may involve preformed barriers or induced resistance mechanisms based on recognition of the invader, complex signalling cascades, hormone signalling, activation of transcription factors and production of pathogenesis-related (PR) proteins with direct antimicrobial or anti-insect activity. Trees have evolved some unique defence mechanisms compared with well-studied model plants, which are mostly herbaceous annuals. The genome sequence of Eucalyptus grandis W. Hill ex Maiden has recently become available and provides a resource to extend our understanding of defence in large woody perennials. This review synthesizes existing knowledge of defence mechanisms in model plants and tree species and features mechanisms that may be important for defence in Eucalyptus , such as anatomical variants and the role of chemicals and proteins. Based on the E. grandis genome sequence, we have identified putative PR proteins based on sequence identity to the previously described plant PR proteins. Putative orthologues for PR-1, PR-2, PR-4, PR-5, PR-6, PR-7, PR-8, PR-9, PR-10, PR-12, PR-14, PR-15 and PR-17 have been identified and compared with their orthologues in Populus trichocarpa Torr. & A. Gray ex Hook and Arabidopsis thaliana (L.) Heynh. The survey of PR genes in Eucalyptus provides a first step in identifying defence gene targets that may be employed for protection of the species in future. Genomic resources available for Eucalyptus are discussed and approaches for improving resistance in these hardwood trees, earmarked as a bioenergy source in future, are considered.

Description: Understanding the interactions between the hemiparasite Santalum album L. and its hosts has theoretical and practical significance in sandalwood plantations. In a pot study, we tested the effects of two non-N 2 -fixing ( Bischofia polycarpa (Levl.) Airy Shaw and Dracontomelon duperreranum Pierre) and two N 2 -fixing hosts ( Acacia confusa Merr. and Dalbergia odorifera T. Chen) on the growth characteristics and nitrogen (N) nutrition of S. album . Biomass production of shoot, root and haustoria, N and total amino acid were significantly greater in S. album grown with the two N 2 -fixing hosts. Foliage and root 15 N values of S. album were significantly lower when grown with N 2 -fixing than with non-N 2 -fixing hosts. Significantly higher photosynthetic rates and ABA (abscisic acid) concentrations were seen in S. album grown with D. odorifera . Similarity in the proportional amounts of amino acid of root xylem sap between S. album and its host D. odorifera was also evident, suggesting major access to nitrogenous solutes from D. odorifera to S. album . Irrespective of host species, S. album clearly appeared to optimize xylem sap extraction from its hosts by higher transpiration and lower water-use efficiency than its host. The growth of two non-N 2 -fixing hosts parasitized by S. album was significantly greater than the equivalent values for unparasitized treatments, and lower growth and photosynthesis were observed for parasitized A. confusa , and significant decreases in root N, photosynthesis and transpiration for parasitized D. odorifera compared with unparasitized treatments. Furthermore, foliage ABA concentrations were significantly higher in all hosts parasitized by S. album than in their unparasitized counterparts. Our study is probably the first to report on host dependence and preference in the hemiparasite S. album , and the generated results may have important implications for understanding of the physiological interactions between host species and parasitic plants, and for successfully mixing plantations of S. album with D. odorifera .

Description: Root systems serve important roles in carbon (C) storage and resource acquisition required for the increased photosynthesis expected in CO 2 -enriched atmospheres. For these reasons, understanding the changes in size, distribution and tissue chemistry of roots is central to predicting the ability of forests to capture anthropogenic CO 2 . We sampled 8000 cm 3 soil monoliths in a pine forest exposed to 14 years of free-air-CO 2 -enrichment and 6 years of nitrogen (N) fertilization to determine changes in root length, biomass, tissue C : N and mycorrhizal colonization. CO 2 fumigation led to greater root length (98%) in unfertilized plots, but root biomass increases under elevated CO 2 were only found for roots 〈1 mm in diameter in unfertilized plots (59%). Neither fine root [C] nor [N] was significantly affected by increased CO 2 . There was significantly less root biomass in N-fertilized plots (19%), but fine root [N] and [C] both increased under N fertilization (29 and 2%, respectively). Mycorrhizal root tip biomass responded positively to CO 2 fumigation in unfertilized plots, but was unaffected by CO 2 under N fertilization. Changes in fine root [N] and [C] call for further study of the effects of N fertilization on fine root function. Here, we show that the stimulation of pine roots by elevated CO 2 persisted after 14 years of fumigation, and that trees did not rely exclusively on increased mycorrhizal associations to acquire greater amounts of required N in CO 2 -enriched plots. Stimulation of root systems by CO 2 enrichment was seen primarily for fine root length rather than biomass. This observation indicates that studies measuring only biomass might overlook shifts in root systems that better reflect treatment effects on the potential for soil resource uptake. These results suggest an increase in fine root exploration as a primary means for acquiring additional soil resources under elevated CO 2 .

Description: The rising atmospheric CO 2 concentration ( C a ) has increased tree growth and intrinsic water-use efficiency (iWUE). However, the magnitude of this effect on long-term iWUE and whether this increase could stimulate the growth of riparian forests in extremely arid regions remain poorly understood. We investigated the relationship between growth [ring width; basal area increment (BAI)] and iWUE in a riparian Populus euphratica Oliv. forest to test whether growth was enhanced by increasing CO 2 and whether this compensated for environmental stresses in the lower reaches of the inland Heihe River, northwestern China. We accomplished this using dendrochronological methods and carbon ( 13 C) and oxygen ( 18 O) isotopic analysis. We found an increase in BAI before 1958, followed by a decrease from 1958 to 1977 and an increase to a peak around 2000. Tree-ring carbon discrimination () and 18 O indicated significant negative overall trends from 1920 to 2012. However, the relationship shifted in strength and direction around 1977 from significantly negative to a weak connection. The seasonal minimum temperature in April to July showed strong influence on , and 18 O was controlled by relative humidity (negatively correlated) and temperature (positively correlated) in June and July. The patterns of internal to atmospheric CO 2 ( C i /C a ) suggest a specific adaptation of tree physiology to increasing CO 2 . Intrinsic water-use efficiency increased significantly (by 36.4%) during the study period. The increased iWUE explained 19.8 and 39.1% of the observed yearly and high-frequency (first-order difference) variations in BAI, respectively, after 1977. Our results suggest significant CO 2 stimulation of riparian tree growth, which compensated for the negative influences of reductions in river streamflow and a drying climate during the study period.

Description: Plants protect themselves from pathogens and herbivores through fine-tuned resource allocation, including trade-offs among resource investments to support constitutive and inducible defences. However, empirical research, especially concerning conifers growing under natural conditions, is still scarce. We investigated the complexity of constitutive and induced defences in a natural Scots pine ( Pinus sylvestris L.) stand under growth-limiting conditions typical of alpine environments. Phenotypic trade-offs at three hierarchical levels were tested by investigating the behaviour of phenolic compounds and terpenoids of outer bark and phloem. We tested resource-derived phenotypic correlations between (i) constitutive and inducible defences vs tree ring growth, (ii) different constitutive defence metabolites and (iii) constitutive concentration and inducible variation of individual metabolites. Tree ring growth was positively correlated only with constitutive concentration of total terpenoids, and no overall phenotypic trade-offs between different constitutive defensive metabolites were found. At the lowest hierarchical level tested, i.e., at the level of relationship between constitutive and inducible variation of individual metabolites, we found that different compounds displayed different behaviours; we identified five different defensive metabolite response types, based on direction and strength of the response, regardless of tree age and growth rate. Therefore, under growth-limiting field conditions, Scots pine appears to utilize varied and complex outer bark and phloem defence chemistry, in which only part of the constitutive specialized metabolism is influenced by tree growth, and individual components do not appear to be expressed in a mutually exclusive manner in either constitutive or inducible metabolism.

Description: Phloem fibres in Mallotus japonicus Müll. Arg. were found to have a multi-layered structure that is S 1 + S 2 + n (G + L), where a non-lignified gelatinous layer (G) and a lignified layer (L) are formed alternately and n indicates the number of repetitions of these two layers. The aim of this study was to determine the process of xylan deposition and lignification in the multi-layered cell walls of phloem fibres. The formation process of the multi-layered structure of secondary phloem fibres was examined by light microscopy, ultraviolet microscopy and transmission electron microscopy. The distribution of glucuronoxylan was examined by immunoelectron microscopy. The activity of peroxidase was also determined using metal-enhanced diaminobenzidine substrates. Immunolabelling of glucuronoxylan occurred in lignified cell wall layers, except in the compound middle lamella (CML), i.e., the S 1 , S 2 and L layers but not the G layers. Change in immunolabelling density suggests that xylan deposition in these lignified layers occurs appositionally, i.e., xylan is deposited into the lignified layers directly and not by a penetrative mechanism, and deposition does not occur after the layers are fully deposited. Peroxidase activity was found in CML including cell corners during S 2 layer formation, then in developing G layers during G layer formation. Peroxidase activity was also found in the thin L layers that formed recently and was not found in the L layers already present. Xylan labelling was not found in the thin L layers that formed recently but did occur in L layers that developed earlier. Lignification of the S 1 and S 2 layers continued during the formation of the G layers, whereas in the L layers it finished just after deposition of the L layer.

Description: The mechanistic understanding of isotope fractionation processes is increasing but we still lack detailed knowledge of the processes that determine the isotopic composition of the tree-ring archive over the long term. Especially with regard to the path from leaf photosynthate production to wood formation, post-assimilation fractionations/processes might cause at least a partial decoupling between the leaf isotope signals that record processes such as stomatal conductance, transpiration and photosynthesis, and the wood or cellulose signals that are stored in the paleophysiological record. In this review, we start from the rather well understood processes at the leaf level such as photosynthetic carbon isotope fractionation, leaf water evaporative isotope enrichment and the issue of the isotopic composition of inorganic sources (CO 2 and H 2 O), though we focus on the less explored ‘downstream’ processes related to metabolism and transport. We further summarize the roles of cellulose and lignin as important chemical constituents of wood, and the processes that determine the transfer of photosynthate (sucrose) and associated isotopic signals to wood production. We cover the broad topics of post-carboxylation carbon isotope fractionation and of the exchange of organic oxygen with water within the tree. In two case studies, we assess the transfer of carbon and oxygen isotopic signals from leaves to tree rings. Finally we address the issue of different temporal scales and link isotope fractionation at the shorter time scale for processes in the leaf to the isotopic ratio as recorded across longer time scales of the tree-ring archive.

Description: Xylem hydraulic conductance varies in response to changes in sap solute content, and in particular of potassium (K + ) ion concentration. This phenomenon, known as the ‘ionic effect’, is enhanced in embolized stems, where it can compensate for cavitation-induced loss of hydraulic conductance. Previous studies have shown that in well-watered laurel plants ( Laurus nobilis L.), potassium concentration of the xylem sap and plant hydraulic conductance increased 24 h after fertilization with KCl. The aim of this work was to test whether water-stressed laurel plants, grown under low potassium availability, could recover earlier from stress when irrigated with a KCl solution instead of potassium-free water. Two-year-old potted laurel seedlings were subjected to water stress by suspending irrigation until leaf conductance to water vapour ( g L ) dropped to ~30% of its initial value and leaf water potential ( L ) reached the turgor loss point ( TLP ). Plants were then irrigated either with water or with 25 mM KCl and monitored for water status, gas exchange and plant hydraulics recovery at 3, 6 and 24 h after irrigation. No significant differences were found between the two experimental groups in terms of L , g L , plant transpiration, plant hydraulic conductance or leaf-specific shoot hydraulic conductivity. Analysis of xylem sap potassium concentration showed that there were no significant differences between treatments, and potassium levels were similar to those of potassium-starved but well-watered plants. In conclusion, potassium uptake from the soil solution and/or potassium release to the xylem appeared to be impaired in water-stressed plants, at least up to 24 h after relief from water stress, so that fertilization after the onset of stress did not result in any short-term advantage for recovery from drought.

Description: Wood represents one of the most important renewable commodities for humanity and plays a crucial role in terrestrial ecosystem carbon-cycling. Wood formation is the result of a multitude of events that require the concerted action of endogenous and exogenous factors under the influence of photoperiod, for instance genes and plant growth regulators. Beyond providing mechanical support and being responsible for the increase in stem radial diameter, woody tissues constitute the vascular system of trees and are capable of reacting to environmental stimuli, and as such are therefore quite plastic and responsive. Despite the ecological and economic importance of wood, not all aspects of its formation have been unveiled. Many gaps in our knowledge are still present, which hinder the maximal exploitation of this precious bioresource. This review aims at surveying the current knowledge of wood formation and the available molecular data addressing the relationship between wood production and environmental factors, which have crucial influences on the rhythmic regulation of cambial activity and exert profound effects on tree stem growth, wood yield and properties. We will here go beyond wood sensu stricto, i.e., secondary xylem, and extend our survey to other tissues, namely vascular cambium, phloem and fibres. The purpose is to provide the reader with an overview of the complexity of the topic and to highlight the importance of progressing in the future towards an integrated knowledge on the subject.

Description: Identifying how physiological responses are structured across environmental gradients is critical to understanding in what manner ecological factors determine tree performance. Here, we investigated the spatiotemporal patterns of signal strength of carbon isotope discrimination ( 13 C) and oxygen isotope composition ( 18 O) for three deciduous oaks ( Quercus faginea (Lam.), Q. humilis Mill. and Q. petraea (Matt.) Liebl.) and one evergreen oak ( Q. ilex L.) co-occurring in Mediterranean forests along an aridity gradient. We hypothesized that contrasting strategies in response to drought would lead to differential climate sensitivities between functional groups. Such differential sensitivities could result in a contrasting imprint on stable isotopes, depending on whether the spatial or temporal organization of tree-ring signals was analysed. To test these hypotheses, we proposed a mixed modelling framework to group isotopic records into potentially homogeneous subsets according to taxonomic or geographical criteria. To this end, carbon and oxygen isotopes were modelled through different variance–covariance structures for the variability among years (at the temporal level) or sites (at the spatial level). Signal-strength parameters were estimated from the outcome of selected models. We found striking differences between deciduous and evergreen oaks in the organization of their temporal and spatial signals. Therefore, the relationships with climate were examined independently for each functional group. While Q. ilex exhibited a large spatial dependence of isotopic signals on the temperature regime, deciduous oaks showed a greater dependence on precipitation, confirming their higher susceptibility to drought. Such contrasting responses to drought among oak types were also observed at the temporal level (interannual variability), with stronger associations with growing-season water availability in deciduous oaks. Thus, our results indicate that Mediterranean deciduous and evergreen oaks constitute two clearly differentiated functional groups in terms of their carbon and water economies, despite co-existing in a wide range of environments. In contrast, deciduous oaks form a rather homogeneous group in terms of climate sensitivity.

Description: To investigate the biological mechanism by which trees control the changes in microfibril (MF) orientation among secondary cell wall layers of conifer tracheids, we studied seasonal variation in the orientation of newly deposited MFs during tracheid cell wall development in Japanese cedar ( Cryptomeria japonica D. Don) trees growing in Central Japan (36°36'N, 140°39'E). Sample blocks were repeatedly collected from four 16-year-old clones of different origins during the growing season of 2010 to investigate the hypotheses that changes in cellulose MF orientation between wall layers exhibited seasonal and clonal differences. The progressive change in the orientation of newly deposited MFs on the primary and secondary cell wall layers of tracheids was detected by field-emission-scanning electron microscopy. Tracheid production and differentiation was studied by light microscopy. We observed a decreasing trend in the orientation of deposited MFs from earlywood to latewood in the S 2 and S 1 layers, where MFs appeared in a Z-helix. In contrast, no seasonal pattern in the orientation of the MFs in the S-helix was observed. Minor clonal variation was observed in the phenology of tracheid production and differentiation. We concluded that a seasonal decreasing trend in the orientation of the MFs in the Z-helix in S 1 and S 2 was present, whereas the MFs in other layers exhibited minor random variations. Thus, the orientation of the MFs in S 2 was affected by seasonal factors, whereas the MFs in other layers were more intrinsically controlled. The within-ring variations in the MF orientation and thus the resulting average MF angle might also be related to genotypic differences in the tracheid production and differentiation rate. However, our results do not exclude other intrinsic and environmental regulations in the change in MF orientation, which remains a topic for future studies.

Description: Planting provenances originating from southern to northern locations has been discussed as a strategy to speed up species migration and mitigate negative effects of climate change on forest stability and productivity. Especially for drought-susceptible species such as European beech ( Fagus sylvatica L.), the introduction of drought-tolerant provenances from the south could be an option. Yet, beech has been found to respond plastically to environmental conditions, suggesting that the climate on the plantation site might be more important for tree growth than the genetic predisposition of potentially drought-adapted provenances. In this study, we compared the radial growth, wood-anatomical traits and leaf phenology of four beech provenances originating from southern (Bulgaria, France) and northern locations (Sweden, the Netherlands) and planted in a provenance trial in the Netherlands. The distribution of precipitation largely differs between the sites of origin. The northern provenances experience a maximum and the southern provenances experience a minimum of rainfall in summer. We compared tree productivity and the anatomy of the water-conducting system for the period from 2000 to 2010, including the drought year 2003. In addition, tree mortality and the timing of leaf unfolding in spring were analysed for the years 2001, 2007 and 2012. Comparison of these traits in the four beech provenances indicates the influence of genetic predisposition and local environmental factors on the performance of these provenances under moderate site conditions. Variation in radial growth was controlled by environment, although the growth level slightly differed due to genetic background. The Bulgarian provenance had an efficient water-conducting system which was moreover unaffected by the drought in 2003, pointing to a high ability of this provenance to cope well with dry conditions. In addition, the Bulgarian provenance showed up as most productive in terms of height and radial growth. Altogether, we conclude that the similarity in ring-width variation among provenances points to environmental control of this trait, whereas the differences encountered in wood-anatomical traits between the well-performing Bulgarian provenance and the other three provenances, as well as the consistent differences in flushing pattern over 3 years under various environmental conditions, support the hypothesis of genetic control of these features.

Description: Despite its major economic importance and the vulnerability of Picea abies (L.) H. Karst. to climate change, how its radial growth at intra-annual resolution is influenced by weather conditions in forest stands with a high production capacity has scarcely been explored. Between 2009 and 2011, phenological variation in seasonal cambial cell production (CP) was analysed in adult P. abies trees from three contrasting sites, differing in altitude and latitude. The results indicate that the timing of cambial CP is a highly synchronic process within populations since in all cases the cambium simultaneously started and stopped producing xylem and phloem cells. Our results also demonstrate that the phenology of cambial CP is highly variable and plastic between years, depending on seasonal temperature and precipitation variation. Differences among sites, however, are only partially explained by different environmental (elevation and altitude) and climatic conditions, suggesting that local adaptation may also play a decisive role in the strategy of P. abies for adapting wood and phloem increments to function optimally under local conditions.

Description: Three methods are in widespread use to build vulnerability curves (VCs) to cavitation. The bench drying (BD) method is considered as a reference because embolism and xylem pressure are measured on large branches dehydrating in the air, in conditions similar to what happens in nature. Two other methods of embolism induction have been increasingly used. While the Cavitron (CA) uses centrifugal force to induce embolism, in the air injection (AI) method embolism is induced by forcing pressurized air to enter a stem segment. Recent studies have suggested that the AI and CA methods are inappropriate in long-vesselled species because they produce a very high-threshold xylem pressure for embolism (e.g., P 50 ) compared with what is expected from (i) their ecophysiology in the field (native embolism, water potential and stomatal response to xylem pressure) and (ii) the P 50 obtained with the BD method. However, other authors have argued that the CA and AI methods may be valid because they produce VCs similar to the BD method. In order to clarify this issue, we assessed VCs with the three above-mentioned methods on the long-vesselled Quercus ilex L. We showed that the BD VC yielded threshold xylem pressure for embolism consistent with in situ measurements of native embolism, minimal water potential and stomatal conductance. We therefore concluded that the BD method provides a reliable estimate of the VC for this species. The CA method produced a very high P 50 (i.e., less negative) compared with the BD method, which is consistent with an artifact related to the vessel length. The VCs obtained with the AI method were highly variable, producing P 50 ranging from –2 to –8.2 MPa. This wide variability was more related to differences in base diameter among samples than to differences in the length of samples. We concluded that this method is probably subject to an artifact linked to the distribution of vessel lengths within the sample. Overall, our results indicate that the CA and the AI should be used with extreme caution on long-vesselled species. Our results also highlight that several criteria may be helpful to assess the validity of a VC.

Description: Tissues of Scots pine ( Pinus sylvestris L.) contain several endophytic microorganisms of which Methylobacterium extorquens DSM13060 is a dominant species throughout the year. Similar to other endophytic bacteria, M. extorquens is able to colonize host plant tissues without causing any symptoms of disease. In addition to endophytic bacteria, plants associate simultaneously with a diverse set of microorganisms. Furthermore, plant-colonizing microorganisms interact with each other in a species- or strain-specific manner. Several studies on beneficial microorganisms interacting with plants have been carried out, but few deal with interactions between different symbiotic organisms and specifically, how these interactions affect the growth and development of the host plant. Our aim was to study how the pine endophyte M. extorquens DSM13060 affects pine seedlings and how the co-inoculation with ectomycorrhizal (ECM) fungi [ Suillus variegatus (SV) or Pisolithus tinctorius (PT)] alters the response of Scots pine. We determined the growth, polyamine and nutrient contents of inoculated and non-inoculated Scots pine seedlings in vitro. Our results show that M. extorquens is able to improve the growth of seedlings at the same level as the ECM fungi SV and PT do. The effect of co-inoculation using different symbiotic organisms was seen in terms of changes in growth and nutrient uptake. Inoculation using M. extorquens together with ECM fungi improved the growth of the host plant even more than single ECM inoculation. Symbiotic organisms also had a strong effect on the potassium content of the seedling. The results indicate that interaction between endophyte and ECM fungus is species dependent, leading to increased or decreased nutrient content and growth of pine seedlings.

Description: Predicting the large-scale consequences of drought in contrasting environments requires that we understand how drought effects differ among species originating from those environments. A previous meta-analysis of published experiments suggested that the effects of drought on both stomatal and non-stomatal limitations to photosynthesis may vary consistently among species from different hydroclimates. Here, we explicitly tested this hypothesis with two short-term water stress experiments on congeneric mesic and xeric species. One experiment was run in Australia using Eucalyptus species and the second was run in Spain using Quercus species as well as two more mesic species. In each experiment, plants were grown under moist conditions in a glasshouse, then deprived of water, and gas exchange was monitored. The stomatal response was analysed with a recently developed stomatal model, whose single parameter g 1 represents the slope of the relationship between stomatal conductance and photosynthesis. The non-stomatal response was partitioned into effects on mesophyll conductance ( g m ), the maximum Rubisco activity ( V cmax ) and the maximum electron transport rate ( J max ). We found consistency among the drought responses of g 1 , g m , V cmax and J max , suggesting that drought imposes limitations on Rubisco activity and RuBP regeneration capacity concurrently with declines in stomatal and mesophyll conductance. Within each experiment, the more xeric species showed relatively high g 1 under moist conditions, low drought sensitivity of g 1 , g m , V cmax and J max , and more negative values of the critical pre-dawn water potential at which V cmax declines most steeply, compared with the more mesic species. These results indicate adaptive interspecific differences in drought responses that allow xeric tree species to continue transpiration and photosynthesis for longer during periods without rain.

Description: The frequency of extreme weather has been rising in recent years. A 3-year study of street trees was undertaken in Tokyo to determine whether: (i) street trees suffer from severe water stress in unusually hot summer; (ii) species respond differently to such climatic fluctuations; and (iii) street trees are also affected by nitrogen (N) deficiency, photoinhibition and aerosol pollution. During the study period (2010–12), midsummers of 2010 and 2012 were unusually hot (2.4–2.8 °C higher maximum temperature than the long-term mean) and dry (6–56% precipitation of the mean). In all species, street trees exhibited substantially decreased photosynthetic rate in the extremely hot summer in 2012 compared with the average summer in 2011. However, because of a more conservative stomatal regulation (stomatal closure at higher leaf water potential) in the hot summer, apparent symptoms of hydraulic failure were not observed in street trees even in 2012. Compared with Prunus  x  yedoensis and Zelkova serrata , Ginkgo biloba , a gymnosperm, was high in stomatal conductance and midday leaf water potential even under street conditions in the unusually hot summer, suggesting that the species had higher drought resistance than the other species and was less susceptible to urban street conditions. This lower susceptibility might be ascribed to the combination of higher soil-to-leaf hydraulic conductance and more conservative water use. Aside from meteorological conditions, N deficiency affected street trees significantly, whereas photoinhibition and aerosol pollution had little effect. The internal CO 2 and 13 C suggested that both water and N limited the net photosynthetic rate of street trees simultaneously, but water was more limiting. From these results, we concluded that the potential risk of hydraulic failure caused by climatic extremes could be low in urban street trees in temperate regions. However, the size of the safety margin might be different between species.

Description: The lowland tropical rainforests in Xishuangbanna, Southwest (SW) China, mark the northern limit of Asian tropics. Fog has been hypothesized to play a role in maintaining rainforests and tropical crop production in this region, but the physiological mechanism has not been studied. The goals of this study were to characterize the seasonal dynamics in photosynthesis and to assess the potential for fog to mitigate chilling-induced photodamage for tropical trees and crops in Xishuangbanna. We measured seasonal dynamics in light-saturated net photosynthetic rate ( A a ), stomatal conductance ( g s ), intercellular CO 2 concentration, quantum yield of Photosystem II ( F v / F m ) and maximum P700 changes ( P m ; indicates the amount of active PSI complex), as well as chilling resistance and fog (light/shading) effects on low temperature-induced decline in F v / F m and P m for native tree and introduced lower latitude tree or woody shrub species grown in a tropical botanical garden. Despite significant decreases in A a , g s , P m and F v / F m , most species maintained considerably high A a during the cool season (2.51–14.6 μmol m –2  s –1 ). Shaded leaves exposed to seasonal low temperatures had higher F v / F m than sun-exposed leaves in the cool season. All species could tolerate 1.4 °C in the dark, whereas a combined treatment of low temperature and high light caused a distinctly faster decline in P m and F v / F m compared with low temperature treatment alone. Because fog persistence avoids or shortens the duration of high light condition in the morning when the temperatures are still low, our results provide support for the hypothesis that fog reduces chilling damage to tropical plants in this region and thus plays a role in maintaining tropical rainforests and agriculture in SW China.

Description: Trees growing in floodplains develop mechanisms by which to overcome anoxic conditions. Prioria copaifera Griseb. grows on the floodplains of the Atrato River, Colombia, and monodominant communities of this species remain flooded for at least 6 months a year. The aims of this study were as follows: (i) to compare variations in tree-ring structure with varying river water levels; and (ii) to reconstruct variations in water levels from the chronology of variations in the porosity of the tree rings. Discs were taken from 12 trees, and the number of vessels along 3-mm-wide radial transects was counted. Standard dendrochronological techniques were used to determine the mean number of vessels over 130 years, between 1877 and 2006; the signal-to-noise ratio was 13.3 and the expressed population signal 0.93. Furthermore, this series of vessel numbers was calibrated against variations in the water levels between 1977 and 2000; positive correlations were found with the mean for both the annual river water level and the level from June to August. The transfer function between the principal components of the mean annual water level and those of chronology allowed us to reconstruct the river levels over 130 years. Our conclusions are as follows: (i) the number of vessels per ring is an appropriate proxy for determining variations in water levels; and (ii) P. copaifera grows thicker and produces more vessels when water levels rise. The probable ecophysiological causes of this interesting behaviour are discussed.

Description: WRKY proteins are involved in various physiological processes in plants, especially in coping with diverse biotic and abiotic stresses. However, limited information is available on the roles of specific WRKY transcription factors in poplar defense. In this study, we reported the characterization of PtoWRKY60 , a Group IIa WRKY member, from Populus tomentosa Carr. The gene expression profile of PtoWRKY60 in various tissues showed that it significantly accumulated in old leaves. Phylogenetic analyses revealed that PtoWRKY60 had a close relationship with AtWRKY18, AtWRKY40 and AtWRKY60. PtoWRKY60 was induced mainly by salicylic acid (SA) and slightly by Dothiorella gregaria Sacc., jasmonic acid, wounding treatment, low temperature and salinity stresses. Overexpression of PtoWRKY60 in poplar resulted in increased resistance to D. gregaria . The defense-associated genes, such as PR5.1 , PR5.2 , PR5.4 , PR5.5 and C PR5 , were markedly up-regulated in transgenic plants overexpressing PtoWRKY60 . These results indicate that PtoWRKY60 might be partly involved in the signal transduction pathway initiated by SA in Populus .

Description: Total daily water use is a key factor influencing the growth of many terrestrial plants, and reflects both day-time and nocturnal water fluxes. However, while nocturnal sap flow ( E n ) and stomatal conductance ( g s,n ) have been reported across a range of species, ecosystems and microclimatic conditions, the regulation of these fluxes remains poorly understood. Here, we present a framework describing the role of abiotic and biotic factors in regulating E n and g s,n highlighting recent developments in this field. Across ecosystems, E n and g s,n generally increased with increasing soil water content and vapor pressure deficit, but the interactive effects of these factors and the potential roles of wind speed and other abiotic factors remain unclear. On average, g s,n and E n are higher in broad-leaved compared with needle-leaved plants, in C 3 compared with C 4 plants, and in tropical compared with temperate species. We discuss the impacts of leaf age, elevated [CO 2 ] and refilling of capacitance on night-time water loss, and how nocturnal g s,n may be included in vegetation models. Younger leaves may have higher g s,n than older leaves. Embolism refilling and recharge of capacitance may affect sap flow such that total plant water loss at night may be less than estimated solely from E n measurements. Our estimates of g s,n for typical plant functional types, based on the published literature, suggest that nocturnal water loss may be a significant fraction (10–25%) of total daily water loss. Counter-intuitively, elevated [CO 2 ] may increase nocturnal water loss. Assumptions in process-based ecophysiological models and dynamic global vegetation models that g s is zero when solar radiation is zero are likely to be incorrect. Consequently, failure to adequately consider nocturnal water loss may lead to substantial under-estimation of total plant water use and inaccurate estimation of ecosystem level water balance.

Description: Potassium (K) is a macro-nutrient understood to play a role in the physiological performance of plants under drought. In some plant species, sodium (Na) can partially substitute K. Although a beneficial role of Na is well established, information regarding its nutritional role in trees is scant and its function under conditions of drought is not fully understood. The objective of the present study was to evaluate the role of K and its possible replacement by Na in olive's ( Olea europaea L.) response to drought. Young and bearing olive trees were grown in soilless culture and exposed to gradual drought. In the presence of Na, trees were tolerant of extremely low K concentrations. Depletion of K and Na resulted in ~50% reduction in CO 2 assimilation rate when compared with sufficiently fertilized control plants. Sodium was able to replace K and recover the assimilation rate to nearly optimum level. The inhibitory effect of K deficiency on photosynthesis was more pronounced under high stomatal conductance. Potassium was not found to facilitate drought tolerance mechanisms in olives. Moreover, stomatal control machinery was not significantly impaired by K deficiency, regardless of water availability. Under drought, leaf water potential was affected by K and Na. High environmental K and Na increased leaf starch content and affected the soluble carbohydrate profile in a similar manner. These results identify olive as a species capable of partly replacing K by Na. The nutritional effect of K and Na was shown to be independent of plant water status. The beneficial effect of Na on photosynthesis and carbohydrates under insufficient K indicates a positive role of Na in metabolism and photosynthetic reactions.

Description: Stem injection techniques can be used to introduce 15 N into trees to overcome a low variation in natural abundance and label biomass with a distinct 15 N signature, but have tended to target small and young trees, of a variety of species, with little replication. We injected 98 atom% 15 N ammonium nitrate (NH 4 NO 3 ) solution into 13 mature, 9- to 13-m tall edge-profile Sitka spruce trees in order to produce a large quantity of labelled litter, examining the distribution of the isotope throughout the canopy after felling in terms of both total abundance of 15 N and relative distribution of the isotope throughout individual trees. Using a simple mass balance of the canopy alone, based on observed total needle biomass and modelled branch biomass, all of the isotope injected was accounted for, evenly split between needles and branches, but with a high degree of variability both within individual trees, and among trees. Both 15 N abundance and relative within-canopy distribution were biased towards the upper and middle crown in foliage. Recovery of the label in branches was much more variable than in needles, possibly due to differences in nitrogen allocation for both growth and storage, which differ seasonally between foliage and woody biomass.

Description: Global climate change causes an increase in ambient air temperature, a major environmental factor influencing plant physiology and growth that already has been perceived at the regional scale and is expected to become even more severe in the future. In the present study, we investigated the effect of elevated ambient air temperature on the nitrogen metabolism of two interior provenances of Douglas fir ( Pseudotsuga menziesii var. glauca ) originating from contrasting habitats, namely the provenances Monte Creek (MC) from a drier environment and Pend Oreille (PO) from a more humid environment. Three- to four-year-old seedlings of the two provenances were grown for 3 months in controlled environments under either control temperature (day 20 °C, night 15 °C) or high temperature (HT, 30/25 °C) conditions. Total nitrogen (N), soluble protein, chlorophyll and total amino acid (TAA) contents as well as individual amino acid concentrations were determined in both current-year and previous-year needles. Our results show that the foliar total N contents of the two provenances were unaffected by HT. Arginine, lysine, proline, glutamate and glutamine were the most abundant amino acids, which together contributed ~88% to the TAA pool of current- and previous-year needles. High temperature decreased the contents of most amino acids of the glutamate family (i.e., arginine, proline, ornithine and glutamine) in current-year needles. However, HT did not affect the concentrations of metabolites related to the photorespiratory pathway, such as $$N{H}_{4}^{+}$$ , glycine and serine. In general, current-year needles were considerably more sensitive to HT than previous-year needles. Moreover, provenance PO originating from a mesic environment showed stronger responses to HT than provenance MC. Our results indicate provenance-specific plasticity in the response of Douglas fir to growth temperature. Provenance-specific effects of elevated temperature on N-use efficiency suggest that origin might determine the sensitivity and growth potential of Douglas fir trees in a future warmer climate.

Description: Partial canopy cover promotes regeneration of many temperate forest trees, but the consequences of shading on seedling drought resistance are unclear. Reintroduction of blight-resistant American chestnut ( Castanea dentata (Marsh.) Borkh.) into eastern North American forests will often occur on water-limited sites and under partial canopy cover. We measured leaf pre-dawn water potential ( pd ), leaf gas exchange, and growth and biomass allocation of backcross hybrid American chestnut seedlings from three orchard sources grown under different light intensities (76, 26 and 8% full photosynthetically active radiation (PAR)) and subjected to well-watered or mid-season water-stressed conditions. Seedlings in the water-stress treatment were returned to well-watered conditions after wilting to examine recovery. Seedlings growing under medium- and high-light conditions wilted at lower leaf pd than low-light seedlings. Recovery of net photosynthesis ( A net ) and stomatal conductance ( g s ) was greater in low and medium light than in high light. Seed source did not affect the response to water stress or light level in most cases. Between 26 and 8% full PAR, light became limiting to the extent that the effects of water stress had no impact on some growth and morphological traits. We conclude that positive and negative aspects of shading on seedling drought tolerance and recovery are not mutually exclusive. Partial shade may help American chestnut tolerate drought during early establishment through effects on physiological conditioning.

Description: Tocochromanols are the most abundant lipid-soluble antioxidants in plants. Among them, α-tocopherol (α-Toc) shows a particularly high sensitivity to environmental stressors and its content is used as a stress biomarker even in non-photosynthetic tissues. Nevertheless, the presence of tocochromanols has not been described yet in the xylem of woody plants, even when their functions regarding cell membrane protection and the transport of photoassimilates may be crucial in this tissue and despite its potential utility in dendrometabolomics. Considering all these, we aimed to determine the presence and distribution of tocochromanols in the xylem of woody plants, to examine their responsiveness to high temperature and to evaluate their potential as environmental bioindicators. The analysis of 29 phyllogenetically diverse species showed that α-Toc is the most abundant and frequent tocochromanol in the xylem and is ubiquitously present in all the studied species, with a concentration ranging from 0.5 to 39.3 μg g –1 of dry weight. α-Tocopherol appeared to be mainly located in the parenchyma rays and was found in both the sapwood and the heartwood, suggesting that it is present even in dead parenchyma cells. The levels of α-Toc in the xylem did not change in response to locally induced xylem heating, but responded positively to the 3-year moving average of annual precipitation. The present findings suggest that α-Toc may be linked to changes in climatic stress. This should enhance further research on the environmental controls of α-Toc variation in the xylem as a first step towards a deeper understanding of dendrometabolomics.

Description: Human-induced gene movement via afforestation and restoration programs is a widespread phenomenon throughout the world. However, its effects on the genetic composition of native populations have received relatively little attention, particularly in forest trees. Here, we examine to what extent gene flow from allochthonous plantations of Pinus pinaster Aiton impacts offspring performance in a neighboring relict natural population and discuss the potential consequences for the long-term genetic composition of the latter. Specifically, we conducted a greenhouse experiment involving two contrasting watering treatments to test for differences in a set of functional traits and mortality rates between P. pinaster progenies from three different parental origins: (i) local native parents, (ii) exotic parents and (iii) intercrosses between local mothers and exotic fathers (intraspecific hybrids). Our results showed differences among crosses in cumulative mortality over time: seedlings of exotic parents exhibited the lowest mortality rates and seedlings of local origin the highest, while intraspecific hybrids exhibited an intermediate response. Linear regressions showed that seedlings with higher water-use efficiency (WUE, 13 C) were more likely to survive under drought stress, consistent with previous findings suggesting that WUE has an important role under dry conditions in this species. However, differences in mortality among crosses were only partially explained by WUE. Other non-measured traits and factors such as inbreeding depression in the relict population are more likely to explain the lower performance of native progenies. Overall, our results indicated that intraspecific hybrids and exotic individuals are more likely to survive under stressful conditions than local native individuals, at least during the first year of development. Since summer drought is the most important demographic and selective filter affecting tree establishment in Mediterranean ecosystems, a potential early selective advantage of exotic and hybrid genotypes would enhance initial steps of introgression of non-native genes into the study relict population of P. pinaster .

Description: Climatic change causes gradual deforestation, partly through forest fires. However, fire has not been seen as an oxidative stressor on surviving forest trees. In addition, discrimination of stress-induced responses from acclimation steps cannot be examined under prolonged stress. Thus, four young Brutian pine ( Pinus brutia Ten.) trees, a fire-related species, were subjected to a simulation of a crown-fire event to evaluate its impact on the availability of soluble carbon (C) and nitrogen (N) and the redox status near fire-afflicted tissue. Total soluble sugars, amino acids and non-structural (NS) proteins in needles and phloem, the antioxidant ascorbic acid (AsA) and reactive oxygen species (ROS) in needles were investigated together with the phloem transport velocity. To examine the temporal progress of these parameters, samples were obtained prior to fire (pre-fire), 2 h after fire, the following day (Day 1) and the following week (Week 1). Findings were categorized into shock reactions (2 h) and acclimation steps. Phloem transport accelerated 2 h postfire by almost 30% and correlated negatively to phloem sugars. At the same time the phloem ratio of sugars/amino acids correlated negatively to needle ROS. The trees' main response at 2 h and particularly on Day 1 was a massive increase in phloem NS proteins. The acclimation process involved also significant increases in needle NS proteins and AsA, as well as significant depletion of phloem amino acids by 65% by Week 1. The highest availability of soluble C and N was recorded on Day 1 in the phloem. Regression models explained significantly the variability of most soluble compounds postfire. Our findings suggest sink control over the source and an advanced role of phloem transport in defense processes.

Description: Seasonal drought, typical of temperate and Mediterranean environments, creates problems in establishing plantations and affects development and yield, and it has been widely studied in numerous species. Forestry fast-growing species such as Eucalyptus spp. are an important resource in such environments, selected clones being generally used for production purposes in plantations in these areas. However, use of mono-specific plantations increases risk of plant loss due to abiotic stresses, making it essential to understand differences in an individual clone's physiological responses to drought stress. In order to study clonal differences in drought responses, nine Eucalyptus globulus (Labill.) clones (C14, C46, C97, C120, C222, C371, C405, C491 and C601) were gradually subjected to severe drought stress (〈14% of field capacity). A total of 31 parameters, physiological (e.g., photosynthesis, gas exchange), biochemical (e.g., chlorophyll content) and hormonal (abscisic acid [ABA] content), were analysed by classic and multivariate techniques. Relationships between parameters were established, allowing related measurements to be grouped into functional units (pigment, growth, water and ABA). Differences in these units showed that there were two distinct groups of E. globulus clones on the basis of their different strategies when faced with drought stress. The C14 group (C14, C120, C405, C491 and C601) clones behave as water savers, maintaining high water content and showing high stomatal adjustment, and reducing their aerial growth to a great extent. The C46 group (C46, C97, C222 and C371) clones behave as water spenders, reducing their water content drastically and presenting osmotic adjustment. The latter maintains the highest growth rate under the conditions tested. The method presented here can be used to identify appropriate E. globulus clones for drought environments, facilitating the selection of material for production and repopulation environments.

Description: Clearcutting a forest ecosystem can result in a drastic reduction of stand productivity. Despite the severity of this disturbance type, past studies have found that the productivity of young regenerating stands can quickly rebound, approaching that of mature undisturbed stands within a few years. One of the obvious reasons is increased leaf area (LA) with each year of recovery. However, a less obvious reason may be the variability in species composition and distribution during the natural regeneration process. The purpose of this study was to investigate to what extent the increase in gross ecosystem productivity (GEP), observed during the first 4 years of recovery in a naturally regenerating clearcut stand, was due to (i) an overall expansion of leaf area and (ii) an increase in the canopy's photosynthetic capacity stemming from either species compositional shifts or drift in physiological traits within species. We found that the multi-year rise in GEP following harvest was clearly attributed to the expansion of LA rather than a change in vegetation composition. Sizeable changes in the relative abundance of species were masked by remarkably similar leaf physiological attributes for a range of vegetation types present in this early-successional environment. Comparison of upscaled leaf-chamber estimates with eddy-covariance-based estimates of light-response curves revealed a broad consistency in both maximum photosynthetic capacity and quantum yield efficiency. The approaches presented here illustrate how chamber- and ecosystem-scale measurements of gas exchange can be blended with species-level LA data to draw conclusive inferences about changes in ecosystem processes over time in a highly dynamic environment.

Description: Forest productivity declines with tree age. This decline may be due to changes in metabolic functions, resource availability and/or changes in resource allocation (between growth, reproduction and storage) with tree age. Carbon and nitrogen remobilization/storage processes are key to tree growth and survival. However, studies of the effects of tree age on these processes are scarce and have not yet considered seasonal carbon and nitrogen variations in situ. This study was carried out in a chronosequence of sessile oak ( Quercus petraea Liebl.) for 1 year to survey the effects of tree age on the seasonal changes of carbon and nitrogen compounds in several tree compartments, focusing on key phenological stages. Our results highlight a general pattern of carbon and nitrogen function at all tree ages, with carbon reserve remobilization at budburst for growth, followed by carbon reserve formation during the leafy season and carbon reserve use during winter for maintenance. The variation in concentrations of nitrogen compounds shows less amplitude than that of carbon compounds. Storage as proteins occurs later, and mainly depends on leaf nitrogen remobilization and root uptake in autumn. We highlight several differences between tree age groups, in particular the loss of carbon storage function of fine and medium-sized roots with tree ageing. Moreover, the pattern of carbon compound accumulation in branches supports the hypothesis of a preferential allocation of carbon towards growth until the end of wood formation in juvenile trees, at the expense of the replenishment of carbon stores, while mature trees start allocating carbon to storage right after budburst. Our results demonstrate that at key phenological stages, physiological and developmental functions differ with tree age, and together with environmental conditions, influence the carbon and nitrogen concentration variations in sessile oaks.

Description: Generally ‘shade tolerance’ refers to the capacity of a plant to exist at low light levels but characteristics of shade can vary and must be taken into account in defining the term. We studied Abies amabilis Dougl. ex J.Forbes and Tsuga heterophylla (Raf.) Sarg. under a forest canopy in the northwest of the Olympic Peninsula, USA, which has low annual sunshine hours and frequent overcast days. Using BF3 sunshine sensors, we surveyed diffuse and total light received by saplings growing under a range of canopy openness up to 30%. We measured variation in photosynthetic capacity over the growing season and within days and estimated photosynthesis induction in relation to ambient light. Three components of shade tolerance are associated with variation in light climate: (i) Total light on the floor of an 88-year stand of naturally regenerated T. heterophylla was greater on overcast than clear days. Light on overcast days varied throughout the day sometimes with a cyclical pattern. (ii) Photosynthetic capacity, A max , varied both through the growing season and within days. A max was generally greater in the latter part of the growing season, being limited by temperature and stomatal conductance, g s , at times during the early part. Saplings in more shaded areas had lower A max and in the latter part of the growing season A max was found to decline from mid-afternoon. (iii) Two patterns of photosynthesis induction to increased light were found. In a mean ambient light of 139 μmol m –2  s –1 , induction had a curvilinear response to a step increase in light with a mean time constant, , of 112.3 s. In a mean ambient light of 74 μmol m –2  s –1 , induction had a two-part increase: one with 1 of 11.3 s and the other with 2 of 184.0 s. These are the smallest published values of to date. (iv) Both variation in photosynthetic capacity and induction are components of shade tolerance where light varies over time. A max acclimates to seasonal and diurnal changes in light and varies between microenvironments. The rapid induction processes can cause a rapid response of photosynthesis to changes in diffuse or direct light.

Description: Nocturnal sap flow ( Q n ) has been found to occur across many taxa, seasons and biomes. There is no general understanding as to how much Q n occurs and whether it is a significant contribution to total daily sap flow ( Q ). A synthesis of the literature and unpublished data was made to determine how significant is Q n , as a proportion of Q (% Q n ), across seasons, biomes, phylogenetic groups and different thermometric sap flow methods. A total of 98 species were analysed to find that % Q n , on average, was 12.03% with the highest average dataset of 69.00%. There was significantly less % Q n in winter than in other temperate seasons, and significantly less % Q n in the wet season than in the dry season. The equatorial and tropical biomes had significantly higher % Q n than the warm temperate and nemoral biomes. The heat ratio method (HRM) and the thermal dissipation (TDP) method had significantly higher % Q n than the heat balance method. Additional analysis between HRM and TDP found HRM to have significantly higher % Q n in winter, wet season and various biomes. In all but one out of 246 cases Q n occurred, demonstrating that Q n is significant and needs to be carefully considered in sap flow and related studies.

Description: The photosynthesis of various species or even a single plant varies dramatically in time and space, creating great spatial heterogeneity within a plant canopy. Continuous and spatially explicit monitoring is, therefore, required to assess the dynamic response of plant photosynthesis to the changing environment. This is a very challenging task when using the existing portable field instrumentation. This paper reports on the application of a technique, laser-induced fluorescence transient (LIFT), developed for ground remote measurement of photosynthetic efficiency at a distance of up to 50 m. The LIFT technique was used to monitor the seasonal dynamics of selected leaf groups within inaccessible canopies of deciduous and evergreen tree species. Electron transport rates computed from LIFT measurements varied over the growth period between the different species studied. The LIFT canopy data and light-use efficiency measured under field conditions correlated reasonably well with the single-leaf pulse amplitude-modulated measurements of broadleaf species, but differed significantly in the case of conifer tree species. The LIFT method has proven to be applicable for a remote sensing assessment of photosynthetic parameters on a diurnal and seasonal scale; further investigation is, however, needed to evaluate the influence of complex heterogeneous canopy structures on LIFT-measured chlorophyll fluorescence parameters.

Description: Photosynthetic carbon assimilation and transpirational water loss play an important role in the yield and the carbon sequestration potential of bioenergy-devoted cultures of fast-growing trees. For six poplar ( Populus ) genotypes in a short-rotation plantation, we observed significant seasonal and genotypic variation in photosynthetic parameters, intrinsic water-use efficiency (WUE i ) and leaf stable isotope composition ( 13 C and 18 O). The poplars maintained high photosynthetic rates (between 17.8 and 26.9 μmol m –2  s –1 depending on genotypes) until late in the season, in line with their fast-growth habit. Seasonal fluctuations were mainly explained by variations in soil water availability and by stomatal limitation upon photosynthesis. Stomatal rather than biochemical limitation was confirmed by the constant intrinsic photosynthetic capacity ( V cmax ) during the growing season, closely related to leaf nitrogen (N) content. Intrinsic water-use efficiency scaled negatively with carbon isotope discrimination ( 13 C bl ) and positively with the ratio between mesophyll diffusion conductance ( g m ) and stomatal conductance. The WUE i  –  13 C bl relationship was partly influenced by g m . There was a trade-off between WUE i and photosynthetic N-use efficiency, but only when soil water availability was limiting. Our results suggest that seasonal fluctuations in relation to soil water availability should be accounted for in future modelling studies assessing the carbon sequestration potential and the water-use efficiency of woody energy crops.

Description: Trees face the dilemma that achieving high plant productivity is accompanied by a risk of drought-induced hydraulic failure due to a trade-off in the trees' vascular system between hydraulic efficiency and safety. By investigating the xylem anatomy of branches and coarse roots, and measuring branch axial hydraulic conductivity and vulnerability to cavitation in 4-year-old field-grown aspen plants of five demes ( Populus tremula L. and Populus tremuloides Michx.) differing in growth rate, we tested the hypotheses that (i) demes differ in wood anatomical and hydraulic properties, (ii) hydraulic efficiency and safety are related to xylem anatomical traits, and (iii) aboveground productivity and hydraulic efficiency are negatively correlated to cavitation resistance. Significant deme differences existed in seven of the nine investigated branch-related anatomical and hydraulic traits but only in one of the four coarse-root-related anatomical traits; this likely is a consequence of high intra-plant variation in root morphology and the occurrence of a few ‘high-conductivity roots’. Growth rate was positively related to branch hydraulic efficiency (xylem-specific conductivity) but not to cavitation resistance; this indicates that no marked trade-off exists between cavitation resistance and growth. Both branch hydraulic safety and hydraulic efficiency significantly depended on vessel size and were related to the genetic distance between the demes, while the xylem pressure causing 88% loss of hydraulic conductivity ( P 88 value) was more closely related to hydraulic efficiency than the commonly used P 50 value . Deme-specific variation in the pit membrane structure may explain why vessel size was not directly linked to growth rate. We conclude that branch hydraulic efficiency is an important growth-influencing trait in aspen, while the assumed trade-off between productivity and hydraulic safety is weak.

Description: The goal of this study was to determine the effects of atmospheric demand on both plant water relations and daily whole-tree water balance across the upper limit of a cloud forest at the mean base height of the trade wind inversion in the tropical trade wind belt. We measured the microclimate and water relations (sap flow, water potential, stomatal conductance, pressure–volume relations) of Metrosideros polymorpha Gaudich. var. polymorpha in three habitats bracketing the cloud forest's upper limit in Hawai‘i to understand the role of water relations in determining ecotone position. The subalpine shrubland site, located 100 m above the cloud forest boundary, had the highest vapor pressure deficit, the least amount of rainfall and the highest levels of nighttime transpiration ( E N ) of all three sites. In the shrubland site, on average, 29% of daily whole-tree transpiration occurred at night, while on the driest day of the study 50% of total daily transpiration occurred at night. While E N occurred in the cloud forest habitat, the proportion of total daily transpiration that occurred at night was much lower (4%). The average leaf water potential ( leaf ) was above the water potential at the turgor loss point ( TLP ) on both sides of the ecotone due to strong stomatal regulation. While stomatal closure maintained a high leaf , the minimum leaf water potential ( leafmin ) was close to TLP , indicating that drier conditions may cause drought stress in these habitats and may be an important driver of current landscape patterns in stand density.

Description: The most interesting factors associated with seed and cone production of Aleppo pine were largely reviewed to identify broad patterns and potential effectiveness of reforestation efforts and planning. Aleppo pine cone production and seed yields are relatively variable, with differences between spatial and temporal influences. These differences are considered, mainly between (i) year, (ii) stand characteristics and (iii) individual tree measurements. Annual variability among populations was recorded for cone production per tree, based on influencing factors such as genetic characteristics, wetness, nutrient availability, insect pests and disease. In addition, some factors may affect Aleppo pine tree growth directly but may be affecting seed and cone production indirectly. Therefore, reduced stand density results in less competition among Aleppo pine trees and accompanying understory flora, which subsequently increases the stem diameter and other tree dimensions, including seed production. This review suggests that reforestation planning, particularly thinning, will result in improved tree morphology that will increase Aleppo pine seed and cone crops. Wildfire intensity and stand conditions such as light and soil nutrient status are also examined.

Description: The effects of elevated ozone (1.4 x ambient) and temperature (ambient +1.3 °C) alone and in combination were studied on the needle cell structure of soil-grown Norway spruce seedlings in the late growing season and winter. Temperature treatment continued over winter and lengthened the snow-free period. Elevated temperature caused microscopic changes related to photosynthesis (decreased chloroplast size and increased number), carbon storage (reduced starch and increased cytoplasmic lipids) and defence (decreased mitochondrial size and proportion per cytoplasm, increased peroxisomes and plastoglobuli, altered appearance of tannins). The results suggest increased oxidative stress by elevated temperature and altered allocation of limited carbon reserve to defence. The number of peroxisomes and plastoglobuli remained high in the outer cells of needles of ozone-exposed seedlings but decreased in the inner cells. This may indicate defence allocation to cells close to the stomata and surface, which are experiencing more oxidative stress. Ozone reduced winter hardiness based on seasonal changes in chloroplast shape and location in the cells. The effects of ozone became evident at the end of the growing season, indicating the effect of cumulative ozone dose or that the seedlings were vulnerable to ozone at the later phases of winter hardening. Elevated temperature increased cellular damage in early winter and visible damage in spring, and the damage was enhanced by ozone. In conclusion, the study suggests that modest air temperature elevation increases stress at the cell structural level in spruce seedlings and is enhanced by low ozone elevation. Future climatic conditions where snow cover is formed later or is lacking but temperatures are low can increase the risk of severe seedling damage, and current and future predicted ozone concentrations increase this risk.

Description: Gas exchange, growth, water transport and carbon (C) metabolism diminish during drought according to their respective sensitivities to declining water status. The timing of this sequence of declining physiological functions may determine how water and C relations compromise plant survival. In this paper, we test the hypothesis that the degree of asynchrony between declining C supply (photosynthesis) and C demand (growth and respiration) determines the rate and magnitude of changes in whole-plant non-structural carbohydrates (NSC) during drought. Two complementary experiments using two tree species ( Eucalyptus globulus Labill. and Pinus radiata D. Don) with contrasting drought response strategies were performed to (i) assess changes in radial stem growth, transpiration, leaf water potential and gas exchange in response to chronic drought, and (ii) evaluate the concomitant impacts of these drought responses on the temporal patterns of NSC during terminal drought. The three distinct phases of water stress were delineated by thresholds of growth cessation and stomatal closure that defined the ‘carbon safety margin’ (i.e., the difference between leaf water potential when growth is zero and leaf water potential when net photosynthesis is zero). A wider C safety margin in E. globulus was defined by an earlier cessation of growth relative to photosynthesis that reduced the demand for NSC while maintaining C acquisition. By contrast, the narrower C safety margin in P. radiata was characterized by a synchronous decline in growth and photosynthesis, whereby growth continued under a declining supply of NSC from photosynthesis. The narrower C safety margin in P. radiata was associated with declines in starch concentrations after ~90 days of chronic drought and significant depletion of starch in all organs at mortality. The observed divergence in the sensitivity of drought responses is indicative of a potential trade-off between maintaining hydraulic safety and adequate C availability.

Description: Plant species may show a preference for uptake of particular nitrogen (N) forms, but little is known about how N form preference is influenced by soil temperature. Potential future changes in soil N form availability and plant N form preference in warmer soils might shift competitive interactions among forest tree species. We compared the N uptake and growth of three conifer species from contrasting environments grown at rhizosphere temperatures of 10, 16 or 20 °C and supplied with ammonium (NH 4 + ) or nitrate (NO 3 – ) or a mix of arginine and alanine. Short-term N uptake was assessed using ion-selective microelectrodes and application of 15 N, and long-term uptake was assessed by plant N status. Species exhibited preferences for particular N forms, and these preferences related to the N form most available in native soils. Specifically, Douglas-fir ( Pseudotsuga menziesii (Mirb.) Franco) showed a preference for nitrate (a N form commonly found in warmer areas), Sitka spruce ( Picea sitchensis (Bong.) Carr.) preferred ammonium (a N form abundant in cold soils) and Engelmann spruce ( Picea engelmannii Parry ex Engelm.) showed a preference for ammonium and organic N (organic N is often abundant in cold soils). Relative N form preference, as indicated by plant growth, changed with temperature in some species, indicating that these species could acclimate to changing rhizosphere temperatures. Understanding how conifers utilize available soil nutrients at different temperatures can help to predict species' future performance as soil temperatures rise.

Description: In cloud forests, foliar uptake (FU) of water has been reported for numerous species, possibly acting to relieve daily water and carbon stress. While the prevalence of FU seems common, how daily variation in fog timing may affect this process has not been studied. We examined the quantity of FU, water potentials, gas exchange and abiotic variation at the beginning and end of a 9-day exposure to fog in a glasshouse setting. Saplings of Abies fraseri (Pursh) Poir. and Picea rubens Sarg. were exposed to morning (MF), afternoon (AF) or evening fog (EF) regimes to assess the ability to utilize fog water at different times of day and after sustained exposure to simulated fog. The greatest amount of FU occurred during MF (up to 50%), followed by AF (up to 23%) and then EF, which surprisingly had no FU. There was also a positive relationship between leaf conductance and FU, suggesting a role of stomata in FU. Moreover, MF and AF lead to the greatest improvements in daily water balance and carbon gain, respectively. Foliar uptake was important for improving plant ecophysiology but was influenced by diurnal variation in fog. With climate change scenarios predicting changes to cloud patterns and frequency that will likely alter diurnal patterns, cloud forests that rely on this water subsidy could be affected.

Description: Difficult access to 40-m-tall emergent trees in tropical rainforests has resulted in a lack of data related to vertical variations in wood CO 2 efflux, even though significant variations in wood CO 2 efflux are an important source of errors when estimating whole-tree total wood CO 2 efflux. This study aimed to clarify vertical variations in wood CO 2 efflux for emergent trees and to document the impact of the variations on the whole-tree estimates of stem and branch CO 2 efflux. First, we measured wood CO 2 efflux and factors related to tree morphology and environment for seven live emergent trees of two dipterocarp species at four to seven heights of up to ~40 m for each tree using ladders and a crane. No systematic tendencies in vertical variations were observed for all the trees. Wood CO 2 efflux was not affected by stem and air temperature, stem diameter, stem height or stem growth. The ratios of wood CO 2 efflux at the treetop to that at breast height were larger in emergent trees with relatively smaller diameters at breast height. Second, we compared whole-tree stem CO 2 efflux estimates using vertical measurements with those based on solely breast height measurements. We found similar whole-tree stem CO 2 efflux estimates regardless of the patterns of vertical variations in CO 2 efflux because the surface area in the canopy, where wood CO 2 efflux often differed from that at breast height, was very small compared with that at low stem heights, resulting in little effect of the vertical variations on the estimate. Additionally, whole-tree branch CO 2 efflux estimates using measured wood CO 2 efflux in the canopy were considerably different from those measured using only breast height measurements. Uncertainties in wood CO 2 efflux in the canopy did not cause any bias in stem CO 2 efflux scaling, but affected branch CO 2 efflux.

Description: The endophytic bacterial communities of six Prunus avium L. genotypes differing in their growth patterns during in vitro propagation were identified by culture-dependent and culture-independent methods. Five morphologically distinct isolates from tissue culture material were identified by 16S rDNA sequence analysis. To detect and analyze the uncultivable fraction of endophytic bacteria, a clone library was established from the amplified 16S rDNA of total plant extract. Bacterial diversity within the clone libraries was analyzed by amplified ribosomal rDNA restriction analysis and by sequencing a clone for each identified operational taxonomic unit. The most abundant bacterial group was Mycobacterium sp., which was identified in the clone libraries of all analyzed Prunus genotypes. Other dominant bacterial genera identified in the easy-to-propagate genotypes were Rhodopseudomonas sp. and Microbacterium sp. Thus, the community structures in the easy- and difficult-to-propagate cherry genotypes differed significantly. The bacterial genera, which were previously reported to have plant growth-promoting effects, were detected only in genotypes with high propagation success, indicating a possible positive impact of these bacteria on in vitro propagation of P. avium , which was proven in an inoculation experiment.

Description: Stem CO 2 efflux is known to vary seasonally and vertically along tree stems. However, annual tree- and stand-scale efflux estimates are commonly based on measurements made only a few times a year, during daytime and at breast height. In this study, the effect of these simplifying assumptions on annual efflux estimates and their influence on the estimates of the importance of stems in stand-scale carbon cycling are evaluated. In order to assess the strength of seasonal, diurnal and along-stem variability in CO 2 efflux, half-hourly measurements were carried out at three heights on three mature Norway spruce ( Picea abies (L . ) Karst.) trees over a period of 3 years. Making the common assumption of breast height efflux rates being representative of the entire stem was found to result in underestimations of 10–17% in the annual tree-scale CO 2 efflux. Upscaling using only daytime measurements from breast height increased the underestimation to 15–20%. Furthermore, the results show that the strength of the vertical gradient varies seasonally, being strongest in the early summer and non-existent during the cool months. The observed seasonality in the vertical CO 2 efflux gradient could not be explained by variation in stem temperature, temperature response of the CO 2 efflux ( Q 10 ), outer-bark permeability, CO 2 transport in the xylem or CO 2 release from the phloem. However, the estimated CO 2 concentration immediately beneath the bark was considerably higher in the upper stem during the main period of diameter growth, coinciding with the strongest vertical efflux gradient. These results suggest that higher growth rates in the upper stem are the main cause for the observed vertical variation in the stem CO 2 effluxes. Furthermore, the results indicate that accounting for the vertical efflux variation is essential for assessments of the importance of stems in stand-scale carbon cycling.

Description: Plant defence theories have recently evolved in such a way that not only the quantity but also the quality of mineral nutrients is expected to influence plant constitutive defence. Recently, an extended prediction derived from the protein competition model (PCM) suggested that nitrogen (N) limitation is more important for the production of phenolic compounds than phosphorus (P). We aimed at studying sexual differences in the patterns of carbon allocation to growth and constitutive defence in relation to N and P availability in Populus tremula L. seedlings. We compared the gender responses in photosynthesis, growth and whole-plant allocation to phenolic compounds at different combination levels of N and P, and studied how they are explained by the main plant defence theories. We found no sexual differences in phenolic concentrations, but interestingly, slow-growing females had higher leaf N concentration than did males, and genders differed in their allocation priority. There was a trade-off between growth and the production of flavonoid-derived phenylpropanoids on one hand, and between the production of salicylates and flavonoid-derived phenylpropanoids on the other. Under limited nutrient conditions, females prioritized mineral nutrient acquisition, flavonoid and condensed tannin (CT) production, while males invested more in above-ground biomass. Salicylate accumulation followed the growth differentiation balance hypothesis as low N mainly decreased the production of leaf and stem salicylate content while the combination of both low N and low P increased the amount of flavonoids and CTs allocated to leaves and to a lesser extent stems, which agrees with the PCM. We suggest that such a discrepancy in the responses of salicylates and flavonoid-derived CTs is linked to their clearly distinct biosynthetic origins and/or their metabolic costs.

Description: To investigate the effects of light quality (wavelength) on shoot elongation and flower-bud formation in Japanese pear ( Pyrus pyrifolia (Burm. f.) Nakai), we treated 1-year-old trees with the following: (i) 8 h sunlight + 16 h dark (SD); (ii) 8 h sunlight + 16 h red light (LD(SD + R)); or (iii) 8 h sunlight + 16 h far-red (FR) light (LD(SD + FR)) daily for 4 months from early April (before the spring flush) until early August in 2009 and 2010. In both years, shoot elongation stopped earlier in the LD(SD + FR) treatment than in the SD and LD(SD + R) treatments. After 4 months of treatments, 21% (2009) or 40% (2010) of LD(SD + FR)-treated trees formed flower buds in the shoot apices, whereas all the shoot apices from SD or LD(SD + R)-treated plants remained vegetative. With an additional experiment conducted in 2012, we confirmed that FR light at 730 nm was the most efficacious wavelength to induce flower-bud formation. Reverse transcription–quantitative polymerase chain reaction revealed that the expression of two floral meristem identity gene orthologues, LEAFY ( PpLFY2a ) and APETALA1 ( PpMADS2-1a ), were up-regulated in the shoot apex of LD(SD + FR). In contrast, the expression of a flowering repressor gene, TERMINAL FLOWER 1 ( PpTFL1-1a , PpTFL1-2a ), was down-regulated. In addition, expression of an orthologue of the flower-promoting gene FLOWERING LOCUS T ( PpFT1a ) was positively correlated with flower-bud formation, although the expression of another orthologue, PpFT2a , was negatively correlated with shoot growth. Biologically active cytokinin and gibberellic acid concentrations in shoot apices were reduced with LD(SD + FR) treatment. Taken together, our results indicate that pear plants are able to regulate flowering in response to the R : FR ratio. Furthermore, LD(SD + FR) treatment terminated shoot elongation and subsequent flower-bud formation in the shoot apex at an earlier time, possibly by influencing the expression of flowering-related genes and modifying plant hormone concentrations.

Description: The timing of budburst of temperate trees is known to be controlled by complicated interactions of temperature and photoperiod. To improve the phenological models of budburst, better knowledge of the internal bud development preceding budburst in relation to environmental cues is needed. We studied the effect of accumulated chilling and forcing temperatures on the internal development of vegetative buds preceding budburst in Norway spruce [ Picea abies (L.) Karst.]. Branches from 17-year-old trees of southern Finnish origin were transferred eight times at 1- to 2-week intervals from October to December 2007 from the field at Punkaharju (61°48'N, 29°20'E) to the greenhouse with forcing conditions (day length 12 h, +20 °C). After seven different durations of forcing, the developmental phase and primordial shoot growth of the buds were analysed at the stereomicroscopic level. Air temperature was recorded hourly throughout the study period. The accumulated chilling unit sum had a significant effect on the temperature sum that was required to attain a certain developmental phase; a higher amount of chilling required a lower amount of forcing. The variation in the rate of development of different buds within each sample branch in relation to the chilling unit and forcing temperature sum was low. Regarding primordial shoot growth, there was also an inverse relation between accumulated chilling and forcing, i.e., a higher accumulated chilling unit sum before forcing required a lower temperature sum to initiate primordial shoot growth and resulted in a stronger effect of accumulated forcing. A second-order regression model with an interaction of chilling and forcing explained the variation of primordial shoot growth with high precision ( R 2 = 0.88). However, further studies are required to determine the final parameter values to be used in phenological modelling.

Description: Photosynthetically active radiation (PAR, 400–700 nm) is one of the primary controls of forest carbon and water relations. In complex terrain, PAR has high spatial variability. Given the high cost of commercial datalogging equipment, spatially distributed measurements of PAR have been typically modeled using geographic coordinates and terrain indices. Here, we present a design for a low-cost, field-deployable device for measuring and recording PAR built around an Arduino microcontroller—named PARduino. PARduino provides for widely distributed sensor arrays and tests the feasibility of using open-source, hobbyist-grade electronics for collecting scientific data. PARduino components include a quantum sensor, an EME Systems signal converter/amplifier and an Arduino Pro Mini microcontroller. Additional components include a real-time clock, a microSD Flash memory card and a custom printed circuit board. The components were selected for ease of assembly. We found strong agreement between the PARduino datalogger system and National Institute of Standards and Technology traceable sensors logged by an industry standard datalogger (slope = 0.99, SE 〈 0.01, P 〈 0.01; intercept = – 14.84, SE = 0.78, P 〈 0.01). The average difference between the two systems was 22.0 µmol m –2  s –1 with PARduino typically underestimating PAR. The average percentage difference between systems was 3.49%. On average, PARduino performed within the factory absolute calibration of the PAR sensor; however, larger errors occurred at low PAR levels. Using open-source technologies such as this can make it possible to develop a spatially distributed sensor network within the constraints of a typical research budget.

Description: The growth rate of triploid European aspen ( Populus tremula L.) and hybrid aspen ( P. tremula   x   Populus tremuloides Michx.) significantly exceeds that of diploid aspen, but the underlying physiological controls of the superior growth rates of these genotypes are not known. We tested the hypothesis that the superior growth rate of triploid and hybrid aspen reflects their greater net photosynthesis rate. Micropropagated clonal plants varying in age from 2.5 to 19 months were used to investigate the ploidy and plant age interaction. The quantum yield of net CO 2 fixation ( ) in leaves of young 2.5-month-old hybrid aspen was lower than that of diploid and triploid trees. However, in 19-month-old hybrid aspen was equal to that in triploid aspen and higher than that in diploid aspen. and the rate of light-saturated net photosynthesis ( A NS ) increased with plant age, largely due to higher leaf dry mass per unit area in older plants. A NS in leaves of 19-month-old trees was highest in hybrid, medium in triploid and lowest in diploid aspen. Light-saturated photosynthesis had a broad temperature optimum between 20 and 35 °C. Rate of respiration in the dark ( R DS ) did not vary among the genotypes in 2.5-month-old plants, and the shape of the temperature response was also similar. R DS increased with plant age, but R DS was still not significantly different among the leaves of 19-month-old diploid and triploid aspen, but it was significantly lower in leaves of 19-month-old hybrid plants. The initial differences in the growth of plants with different ploidy were minor up to the age of 19 months, but during the next 2 years, the growth rate of hybrid aspen exceeded that of triploid plants by 2.7 times and of diploid plants by five times, in line with differences in A NS of 19-month-old plants of these species. It is suggested that differences in photosynthesis and growth became more pronounced with tree aging, indicating that ontogeny plays a key role in the expression of superior traits determining the productivity of given genotypes.

Description: Sugars play an important role in various physiological processes during plant growth and development; however, the developmental roles and regulatory functions of hexoses other than glucose are still largely unclear. Recent studies suggest that blocked embryo development in Norway spruce ( Picea abies (L.) Karst) is associated with accumulation of fructose. In the present study, the potential biochemical regulatory mechanism of glucose and fructose was studied during development of somatic embryos of Norway spruce from pro-embryogenic masses to mature embryos. The changes in protein fluorescence, a marker of the Maillard reaction, were monitored in two cell lines of Norway spruce that were grown on media containing sucrose (control), glucose or fructose. Manual time-lapse photography showed that growth of embryogenic cultures on medium containing sucrose was characterized by normal development of mature embryos whereas the embryogenic cultures that were grown on media containing glucose or fructose did not develop mature embryos. The biochemical analyses of embryogenic samples collected during embryo development showed that: (i) the content of glucose and fructose in the embryogenic cultures increased significantly during growth on each medium, respectively; (ii) the accumulation of Maillard products in the embryogenic cultures was highly correlated with the endogenous content of fructose but not glucose; and (iii) the embryogenic cultures grown on fructose displayed the highest protein carbonyl content and DNA damage whereas the highest content of glutathione was recorded in the embryogenic cultures that had grown on sucrose. Our data suggest that blocked development of embryos in the presence of fructose may be associated with the Maillard reaction.

Description: We explored the impact of canopy position on leaf respiration ( R ) and associated traits in tree and shrub species growing in a lowland tropical rainforest in Far North Queensland, Australia. The range of traits quantified included: leaf R in darkness ( R D ) and in the light ( R L ; estimated using the Kok method); the temperature ( T )-sensitivity of R D ; light-saturated photosynthesis ( A sat ); leaf dry mass per unit area (LMA); and concentrations of leaf nitrogen (N), phosphorus (P), soluble sugars and starch. We found that LMA, and area-based N, P, sugars and starch concentrations were all higher in sun-exposed/upper canopy leaves, compared with their shaded/lower canopy and deep-shade/understory counterparts; similarly, area-based rates of R D , R L and A sat (at 28 °C) were all higher in the upper canopy leaves, indicating higher metabolic capacity in the upper canopy. The extent to which light inhibited R did not differ significantly between upper and lower canopy leaves, with the overall average inhibition being 32% across both canopy levels. Log–log R D – A sat relationships differed between upper and lower canopy leaves, with upper canopy leaves exhibiting higher rates of R D for a given A sat (both on an area and mass basis), as well as higher mass-based rates of R D for a given [N] and [P]. Over the 25–45 °C range, the T- sensitivity of R D was similar in upper and lower canopy leaves, with both canopy positions exhibiting Q 10 values near 2.0 (i.e., doubling for every 10 °C rise in T ) and T max values near 60 °C (i.e., T where R D reached maximal values). Thus, while rates of R D at 28 °C decreased with increasing depth in the canopy, the T -dependence of R D remained constant; these findings have important implications for vegetation-climate models that seek to predict carbon fluxes between tropical lowland rainforests and the atmosphere.

Description: Boron (B) deficiency is a widespread problem in many crops, including Citrus . The effects of B-deficiency on gas exchange, carbohydrates, organic acids, amino acids, total soluble proteins and phenolics, and the activities of key enzymes involved in organic acid and amino acid metabolism in ‘Xuegan’ [ Citrus sinensis (L.) Osbeck] leaves and roots were investigated. Boron-deficient leaves displayed excessive accumulation of nonstructural carbohydrates and much lower CO 2 assimilation, demonstrating feedback inhibition of photosynthesis. Dark respiration, concentrations of most organic acids [i.e., malate, citrate, oxaloacetate (OAA), pyruvate and phosphoenolpyruvate] and activities of enzymes [i.e., phosphoenolpyruvate carboxylase (PEPC), NAD-malate dehydrogenase, NAD-malic enzyme (NAD-ME), NADP-ME, pyruvate kinase (PK), phosphoenolpyruvate phosphatase (PEPP), citrate synthase (CS), aconitase (ACO), NADP-isocitrate dehydrogenase (NADP-IDH) and hexokinase] involved in glycolysis, the tricarboxylic acid (TCA) cycle and the anapleurotic reaction were higher in B-deficient leaves than in controls. Also, total free amino acid (TFAA) concentration and related enzyme [i.e., NADH-dependent glutamate 2-oxoglutarate aminotransferase (NADH-GOGAT) and glutamate OAA transaminase (GOT)] activities were enhanced in B-deficient leaves. By contrast, respiration, concentrations of nonstructural carbohydrates and three organic acids (malate, citrate and pyruvate), and activities of most enzymes [i.e., PEPC, NADP-ME, PK, PEPP, CS, ACO, NAD-isocitrate dehydrogenase, NADP-IDH and hexokinase] involved in glycolysis, the TCA cycle and the anapleurotic reaction, as well as concentration of TFAA and activities of related enzymes (i.e., nitrate reductase, NADH-GOGAT, glutamate pyruvate transaminase and glutamine synthetase) were lower in B-deficient roots than in controls. Interestingly, leaf and root concentration of total phenolics increased, whereas that of total soluble protein decreased, in response to B-deficiency. In conclusion, respiration, organic acid (i.e., glycolysis and the TCA cycle) metabolism, the anapleurotic pathway and amino acid biosynthesis were upregulated in B-deficient leaves with excessive accumulation of carbohydrates to ‘consume’ the excessive carbon available, but downregulated in B-deficient roots with less accumulation of carbohydrates to maintain the net carbon balance.

Description: Dwarf mistletoes, obligate, parasitic plants with diminutive aerial shoots, have long-term effects on host tree water relations, hydraulic architecture and photosynthetic gas exchange and can eventually induce tree death. To investigate the long-term (1886–2010) impacts of dwarf mistletoe on the growth and gas exchange characteristics of host western hemlock, we compared the diameter growth and tree-ring cellulose stable carbon (C) and oxygen (O) isotope ratios ( 13 C cell , 18 O cell ) of heavily infected and uninfected trees. The relative basal area growth of infected trees was significantly greater than that of uninfected trees in 1886–90, but declined more rapidly in infected than uninfected trees through time and became significantly lower in infected than uninfected trees in 2006–10. Infected trees had significantly lower 13 C cell and 18 O cell than uninfected trees. Differences in 18 O cell between infected and uninfected trees were unexpected given that stomatal conductance and environmental variables that were expected to influence the 18 O values of leaf water were similar for both groups. However, estimates of mesophyll conductance ( g m ) were significantly lower and estimates of effective path length for water movement ( L ) were significantly higher in leaves of infected trees, consistent with their lower values of 18 O cell . This study reconstructs the long-term physiological responses of western hemlock to dwarf mistletoe infection. The long-term diameter growth and 13 C cell trajectories suggested that infected trees were growing faster than uninfected trees prior to becoming infected and subsequently declined in growth and leaf-level photosynthetic capacity compared with uninfected trees as the dwarf mistletoe infection became severe. This study further points to limitations of the dual-isotope approach for identifying sources of variation in 13 C cell and indicates that changes in leaf internal properties such as g m and L that affect 18 O cell must be considered.

Description: Branch/tree-level measurements of carbon (C)-acquisition provide an integration of the physical and biological processes driving the C gain of all individual leaves. Most research dealing with the interacting effects of high-irradiance environments and soil-induced water stress on the C-gain of fruit tree species has focused on leaf-level measurements. The C-gain of both sun-exposed leaves and branches of adult almond trees growing in a semi-arid climate was investigated to determine the respective costs of structural and biochemical/physiological protective mechanisms involved in the behaviour at branch scale. Measurements were performed on well-watered (fully irrigated, FI) and drought-stressed (deficit irrigated, DI) trees. Leaf-to-branch scaling for net CO 2 assimilation was quantified by a global scaling factor ( f g ), defined as the product of two specific scaling factors: (i) a structural scaling factor ( f s ), determined under well-watered conditions, mainly involving leaf mutual shading; and (ii) a water stress scaling factor ( f ws,b ) involving the limitations in C-acquisition due to soil water deficit. The contribution of structural mechanisms to limiting branch net C-gain was high (mean f s ~0.33) and close to the projected-to-total leaf area ratio of almond branches (  = 0.31), while the contribution of water stress mechanisms was moderate (mean f ws,b ~0.85), thus supplying an f g ranging between 0.25 and 0.33 with slightly higher values for FI trees with respect to DI trees. These results suggest that the almond tree (a drought-tolerant species) has acquired mechanisms of defensive strategy (survival) mainly based on a specific branch architectural design. This strategy allows the potential for C-gain to be preserved at branch scale under a large range of soil water deficits. In other words, almond tree branches exhibit an architecture that is suboptimal for C-acquisition under well-watered conditions, but remarkably efficient to counteract the impact of DI and drought events.

Description: Physiological characteristics of saplings can be considered one of the most basic constraints on species distribution. The shade-tolerant arborescent palm Euterpe edulis Mart. is endemic to the Atlantic Forest of Argentina, Brazil and Paraguay. At a local scale, saplings of this species growing in native forests are absent in gaps. We tested the hypothesis whether sensitivity to photoinhibition or hydraulic architecture constrains the distribution of E. edulis saplings in sun-exposed forest environments. Using shade houses and field studies, we evaluated growth, survival, hydraulic traits and the susceptibility of Photosystem II to photoinhibition in E. edulis saplings under different growth irradiances. Survival rates in exposed sites in the field were very low (a median of 7%). All saplings exhibited photoinhibition when exposed to high radiation levels, but acclimation to a high radiation environment increased the rate of recovery. Petiole hydraulic conductivity was similar across treatments regardless of whether it was expressed per petiole cross-sectional area or per leaf area. At the plant level, investment in conductive tissues relative to leaf area (Huber values) increased with increasing irradiance. Under high irradiance conditions, plants experienced leaf water potentials close to the turgor-loss point, and leaf hydraulic conductance decreased by 79% relative to its maximum value. Euterpe edulis saplings were able to adjust their photosynthetic traits to different irradiance conditions, whereas hydraulic characteristics at the leaf level did not change across irradiance treatments. Our results indicate that uncoupling between water demand and supply to leaves apparently associated with high resistances to water flow at leaf insertion points, in addition to small stems with low water storage capacity, weak stomatal control and high vulnerability of leaves to hydraulic dysfunction, are the main ecophysiological constraints that prevent the growth and survival of E. edulis saplings in gaps in the native forest where native lianas and bamboos show aggressive growth.

Description: Metal pollution has been widely reported in mangrove wetlands; however, the mechanisms involved in metal detoxification by mangroves are still poorly understood. This study aimed to investigate the possible function of root anatomy and lignification/suberization on metal uptake and tolerance in seedlings of six species of mangroves. The results revealed that the three rhizophoraceous species ( Bruguiera gymnorrhiza (L.) Poir, Kandelia obovata Sheue, Liu & Yong and Rhizophora stylosa Griff) consistently exhibited higher metal tolerances than the three pioneer species ( Aegiceras corniculatum (Linn.) Blanco, Acanthus ilicifolius L. and Avicennia marina (Forsk.) Viern.). Moreover, metal-tolerant species often exhibited a thick exodermis with high lignification and suberization. The tolerance indices of the mangroves were found to be positively correlated with the amounts of lignin and suberin deposition within the exodermal cell walls. The observed metal uptake by the excised roots further illustrated that a lignified/suberized exodermis directly delayed the entry of metals into the roots, and thereby contributed to a higher tolerance to heavy metals. In summary, the present study proposes a barrier property of the lignified/suberized exodermis in dealing with the stresses of heavy metals, such that the mangroves which possessed more extensive lignification/suberization within the exodermis appeared to exhibit higher metal tolerance.

Description: Dicotyledons have evolved a strategy to compensate for the increase in hydraulic resistance to water transport with height growth by widening xylem conduits downwards. In monocots, the accumulation of hydraulic resistance with height should be similar, but the absence of secondary growth represents a strong limitation for the maintenance of xylem hydraulic efficiency during ontogeny. The hydraulic architecture of monocots has been studied but it is unclear how monocots arrange their axial vascular structure during ontogeny to compensate for increases in height. We measured the vessel lumina and estimated the hydraulic diameter ( D h ) at different heights along the stem of two arborescent monocots, Bactris gasipaes (Kunth) and Guadua angustifolia (Kunth). For the former, we also estimated the variation in D h along the leaf rachis. Hydraulic diameter increased basally from the stem apex to the base with a scaling exponent ( b ) in the range of those reported for dicot trees ( b = 0.22 in B. gasipaes ; b = 0.31 and 0.23 in G. angustifolia ). In B. gasipaes , vessels decrease in D h from the stem's centre towards the periphery, an opposite pattern compared with dicot trees. Along the leaf rachis, a pattern of increasing D h basally was also found ( b = 0.13). The hydraulic design of the monocots studied revealed an axial pattern of xylem conduits similar to those evolved by dicots to compensate and minimize the negative effect of root-to-leaf length on hydrodynamic resistance to water flow.

Description: For obvious practical reasons, tree phenological data obtained in warming and photoperiod experiments are generally conducted on juvenile trees (saplings and seedlings) or on watered or rooted cuttings collected from adult trees. As juvenile trees differ from adult trees in their phenological response to environmental conditions, they represent inappropriate plant material to experimentally assess the phenological responses of forests to seasonality. Cuttings are physiologically closer to adult trees, but cutting itself and the disruption of hormonal signals may create artefacts. This study aimed to investigate the potential deviation between phenological responses of cuttings vs donor trees. We hypothesized that, once dormant, buds may respond autonomously to environmental influences such as chilling, photoperiod and warming, and, thus, cuttings may exhibit similar phenological responses to mature trees. We compared bud development of seedlings, saplings and mature trees of three deciduous tree species with bud development of cuttings that were excised from both saplings and adults and positioned in situ in the vicinity of adult trees within a mature mixed forest in the foothills of the Swiss Jura Mountains. No significant difference was detected in the timing of bud burst between cuttings and donor trees for the three studied tree species when the vertical thermal profile was accounted for. However, a significant difference in the timing of flushing was found between seedlings, saplings and adults, with earlier flushing during the juvenile stage. At least for the three studied species, this study clearly demonstrates that cuttings are better surrogates than juvenile trees to assess potential phenological responses of temperate forests to climate change in warming and photoperiod experiments.