Axial conduit widening, tree height, and height growth rate set the hydraulic transition of sapwood into heartwood.

The size-related xylem adjustments required to maintain a constant leaf-specific sapwood conductance (KLEAF) with increasing height (H) are still under discussion. Alternative hypotheses are that: (i) the conduit hydraulic diameter (Dh) at any position in the stem and/or (ii) the number of sapwood rings at stem base (NSWr) increase with H. In addition, (iii) reduced stem elongation (ΔH) increases the tip-to-base conductance through inner xylem rings, thus possibly the NSWr contributing to KLEAF. A detailed stem analysis showed that Dh increased with the distance from the ring apex (DCA) in all rings of a Picea abies and a Fagus sylvatica tree. Net of DCA effect, Dh did not increase with H. Using sapwood traits from a global dataset, NSWr increased with H, decreased with ΔH, and the mean sapwood ring width (SWrw) increased with ΔH. A numerical model based on anatomical patterns predicted the effects of H and ΔH on the conductance of inner xylem rings. Our results suggest that the sapwood/heartwood transition depends on both H and ΔH, and is set when the carbon allocation to maintenance respiration of living cells in inner sapwood rings produces a lower gain in total conductance than investing the same carbon in new vascular conduits.

No xylem phenotypic plasticity in mature Picea abies and Fagus sylvatica trees after 5 years of throughfall precipitation exclusion.

Forest trees are experiencing increasing frequency and intensity of drought events with climate change. We investigated xylem and phloem traits from mature Fagus sylvatica and Picea abies trees after 5 years of complete exclusion of throughfall precipitation during the growing season. Xylem and phloem anatomy, leaf and branch biomass were analysed along top branches of ~1.5 m lenght in 5 throughfall precipitation excluded (TE) and 5 control (CO) trees of both beech and spruce. Xylem traits were analysed on wood cores extracted from the stem at breast height. In the top branches of both species, the lumen diameter (or area) of xylem and phloem conduits did not differ between TE and CO trees. At breast height, TE trees of both species produced narrower xylem rings and conduits. While allocation to branch (BM) and needle biomass (LM) did not change between TE and CO in P. abies, TE F. sylvatica trees allocated proportionally more biomass to leaves (LM) than BM compared with CO. Despite artificial drought increased the mortality in the TE plots, our results revealed no changes in both xylem and phloem anatomies, undermining the hypothesis that successful acclimation to drought would primarily involve increased resistance against air embolism.

Plant respiration: Controlled by photosynthesis or biomass?

Two simplifying hypotheses have been proposed for whole-plant respiration. One links respiration to photosynthesis; the other to biomass. Using a first-principles carbon balance model with a prescribed live woody biomass turnover, applied at a forest research site where multidecadal measurements are available for comparison, we show that if turnover is fast the accumulation of respiring biomass is low and respiration depends primarily on photosynthesis; while if turnover is slow the accumulation of respiring biomass is high and respiration depends primarily on biomass. But the first scenario is inconsistent with evidence for substantial carry-over of fixed carbon between years, while the second implies far too great an increase in respiration during stand development-leading to depleted carbohydrate reserves and an unrealistically high mortality risk. These two mutually incompatible hypotheses are thus both incorrect. Respiration is not linearly related either to photosynthesis or to biomass, but it is more strongly controlled by recent photosynthates (and reserve availability) than by total biomass.

The total path length hydraulic resistance according to known anatomical patterns: What is the shape of the root-to-leaf tension gradient along the plant longitudinal axis?

Xylem conduit diameter widens from leaf tip to stem base and how this widening affects the total hydraulic resistance (RTOT) and the gradient of water potential (Ψxyl) has never been thoroughly investigated. Data of conduit diameter of Acer pseudoplatanus,Fagus sylvatica and Picea abies were used to model the axial variation of RTOT and Ψxyl. The majority of RTOT (from 79 to 98%) was predicted to be confined within the leaf/needle. This means that the xylem conduits of stem and roots, accounting for nearly the total length of the hydraulic path, theoretically provide a nearly negligible contribution to RTOT. Consequently, a steep gradient of water potentials was predicted to develop within the leaf/needle base, whereas lower in the stem water potentials approximate those of rootlets. Our results would suggest that the strong partitioning of RTOT between leaves/needles coupled with basal conduit widening is of key importance for both hydraulic safety against drought-induced embolism formation and efficiency, as it minimizes the exposure of stem xylem to high tensions and makes the total plant's conductance substantially independent of body size.

Tree differences in primary and secondary growth drive convergent scaling in leaf area to sapwood area across Europe.

Trees scale leaf (AL ) and xylem (AX ) areas to couple leaf transpiration and carbon gain with xylem water transport. Some species are known to acclimate in AL  : AX balance in response to climate conditions, but whether trees of different species acclimate in AL  : AX in similar ways over their entire (continental) distributions is unknown. We analyzed the species and climate effects on the scaling of AL vs AX in branches of conifers (Pinus sylvestris, Picea abies) and broadleaved (Betula pendula, Populus tremula) sampled across a continental wide transect in Europe. Along the branch axis, AL and AX change in equal proportion (isometric scaling: b Ëœ 1) as for trees. Branches of similar length converged in the scaling of AL vs AX with an exponent of b = 0.58 across European climates irrespective of species. Branches of slow-growing trees from Northern and Southern regions preferentially allocated into new leaf rather than xylem area, with older xylem rings contributing to maintaining total xylem conductivity. In conclusion, trees in contrasting climates adjust their functional balance between water transport and leaf transpiration by maintaining biomass allocation to leaves, and adjusting their growth rate and xylem production to maintain xylem conductance.

X-ray microtomography observations of xylem embolism in stems of Laurus nobilis are consistent with hydraulic measurements of percentage loss of conductance.

Drought-induced xylem embolism is a serious threat to plant survival under future climate scenarios. Hence, accurate quantification of species-specific vulnerability to xylem embolism is a key to predict the impact of climate change on vegetation. Low-cost hydraulic measurements of embolism rate have been suggested to be prone to artefacts, thus requiring validation by direct visualization of the functional status of xylem conduits using nondestructive imaging techniques, such as X-ray microtomography (microCT). We measured the percentage loss of conductance (PLC) of excised stems of Laurus nobilis (laurel) dehydrated to different xylem pressures, and compared results with direct observation of gas-filled vs water-filled conduits at a synchrotron-based microCT facility using a phase contrast imaging modality. Theoretical PLC calculated on the basis of microCT observations in stems of laurel dehydrated to different xylem pressures overall were in agreement with hydraulic measurements, revealing that this species suffers a 50% loss of xylem hydraulic conductance at xylem pressures averaging -3.5 MPa. Our data support the validity of estimates of xylem vulnerability to embolism based on classical hydraulic techniques. We discuss possible causes of discrepancies between data gathered in this study and those of recent independent reports on laurel hydraulics.

Universal hydraulics of the flowering plants: vessel diameter scales with stem length across angiosperm lineages, habits and climates.

Angiosperm hydraulic performance is crucially affected by the diameters of vessels, the water conducting conduits in the wood. Hydraulic optimality models suggest that vessels should widen predictably from stem tip to base, buffering hydrodynamic resistance accruing as stems, and therefore conductive path, increase in length. Data from 257 species (609 samples) show that vessels widen as predicted with distance from the stem apex across angiosperm orders, habits and habitats. Standardising for stem length, vessels are only slightly wider in warm/moist climates and in lianas, showing that, rather than climate or habit, plant size is by far the main driver of global variation in mean vessel diameter. Terminal twig vessels become wider as plant height increases, while vessel density decreases slightly less than expected tip to base. These patterns lead to testable predictions regarding evolutionary strategies allowing plants to minimise carbon costs per unit leaf area even as height increases.

Consistent decrease in conifer embolism resistance from the stem apex to base resulting from axial trends in tracheid and pit traits.

Introduction: Drought-induced embolism formation in conifers is associated with several tracheid and pit traits, which vary in parallel from stem apex to base. We tested whether this axial anatomical variability is associated with a progressive variation in embolism vulnerability along the stem from apex to base. Methods: We assessed the tracheid hydraulic diameter (Dh), mean pit membrane area (PMA) and the xylem pressure at 50% loss of conductivity (P50) on longitudinal stem segments extracted at different distances from the stem apex (DFA) in a Picea abies and an Abies alba tree. Results: In both trees, Dh and PMA scaled with DFA 0.2. P50 varied for more than 3 MPa from the treetop to the stem base, according to a scaling of -P50 with DFA-0.2 . The largest Dh, PMA and P50 variation occurred for DFA<1.5 m. PMA and Dh scaled more than isometrically (exponent b=1.2). Pit traits vary proportionally with tracheid lumen diameter. Discussion and conclusions: Apex-to-base trends in tracheid and pit traits, along with variations in P50, suggest a strong structure-function relationship that is influenced by DFA. Although the effect of DFA on P50 has not been extensively explored previously, we propose that analyzing the relationship between P50 and DFA could be crucial for a comprehensive assessment of embolism vulnerability at the individual level.

Perfluoroalkyl substances exposure alters stomatal opening and xylem hydraulics in willow plants.

Climate change and pollution are increasingly important stress factors for life on Earth. Dispersal of poly- and perfluoroalkyl substances (PFAS) are causing worldwide contamination of soils and water tables. PFAS are partially hydrophobic and can easily bioaccumulate in living organisms, causing metabolic alterations. Different plant species can uptake large amounts of PFAS, but little is known about its consequences for the plant water relation and other physiological processes, especially in woody plants. In this study, we investigated the fractionation of PFAS bioaccumulation from roots to leaves and its effects on the conductive elements of willow plants. Additionally, we focused on the stomal opening and the phytohormonal content. For this purpose, willow cuttings were exposed to a mixture of 11 PFAS compounds and the uptake was evaluated by LC-MS/MS. Stomatal conductance was measured and the xylem vulnerability to air embolism was tested and further, the abscisic acid and salicylic acid contents were quantified using LC-MS/MS. PFAS accumulated from roots to leaves based on their chemical structure. PFAS-exposed plants showed reduced stomatal conductance, while no differences were observed in abscisic acid and salicylic acid contents. Interestingly, PFAS exposure caused a higher vulnerability to drought-induced xylem embolism in treated plants. Our study provides novel information about the PFAS effects on the xylem hydraulics, suggesting that the plant water balance may be affected by PFAS exposure. In this perspective, drought events may be more stressful for PFAS-exposed plants, thus reducing their potential for phytoremediation.

Widening of xylem conduits in a conifer tree depends on the longer time of cell expansion downwards along the stem.

The diameter of vascular conduits increases towards the stem base. It has been suggested that this profile is an efficient anatomical feature for reducing the hydraulic resistance when trees grow taller. However, the mechanism that controls the cell diameter along the plant is not fully understood. The timing of cell differentiation along the stem was investigated. Cambial activity and cell differentiation were investigated in a Picea abies tree (11.5 m in height) collecting microsamples at nine different heights (from 1 to 9 m) along the stem with a 4 d time interval. Wood sections (8-12 µm thick) were stained and observed under a light microscope with polarized light to differentiate the developing xylem cells. Cell wall lignification was detected using cresyl violet acetate. The first enlarging cells appeared almost simultaneously along the tree axis indicating that cambium activation is not height-dependent. A significant increase in the duration of the cell expansion phase was observed towards the tree base: at 9 m from the ground, xylem cells expanded for 7 d, at 6 m for 14 d, and at 3 m for 19 d. The duration of the expansion phase is positively correlated with the lumen area of the tracheids (r(2)=0.68, P < 0.01) at the same height. By contrast, thickness of the cell wall of the earlywood did not show any trend with height. The lumen area of the conduits down the stem appeared linearly dependent on time during which differentiating cells remained in the expansion phase. However, the inductive signal of such long-distance patterned differentiation remains to be identified.

Hydraulic constraints limit height growth in trees at high altitude.

• Low temperatures limit the fixation of photosynthates and xylogenesis. Here, we hypothesized that reduced longitudinal growth in trees at high altitude is related to the lower hydraulic efficiency of the transport system. • Apical buds of Norway spruce (Picea abies) trees at high and low elevation were heated during 2006 and 2007. At the end of the experiment, trees were felled. Longitudinal increments and tracheid lumen areas were measured along the stem. Apical hydraulic conductivity (k) was estimated from anatomical data. • Before heating, high-altitude trees showed fewer (P = 0.002) and smaller (P = 0.008) apical conduits, and therefore reduced k (P = 0.016) and stem elongation (P < 0.0001), in comparison with trees at low elevation. After 2 yr of heating, k increased at both high (P = 0.014) and low (P = 0.047) elevation. Only high-altitude trees showed increased stem elongation, which reached the same magnitude as that of controls at low elevation (P = 0.735). Heating around the apical shoots did not appear to induce significant changes in conduit dimension along the rest of the stem. • The total number and size of xylem elements at the stem apex are strongly constrained by low temperatures. Trees at high altitude are therefore prevented from building up an efficient transport system, and their reduced longitudinal growth reflects strong hydraulic limitations.

Comment on "The blind men and the elephant: the impact of context and scale in evaluating conflicts between plant hydraulic safety and efficiency" by Meinzer et al. (2010).

A recent paper by Meinzer et al. (Oecologia, 2010, 164:287-296) has reviewed the topic of conflicts between hydraulic efficiency and safety in vascular plants. The authors raised the question that reliable interpretations of results on the mechanisms behind this trade-off requires an integrated approach that considers the variations in hydraulic traits throughout the entire plant and between different individuals, species and environments. Nonetheless, the literature review was only partial. Here, we discuss that an important feature of the whole tree xylem architecture, conduit tapering, is one of traits that must be considered when planning analyses of hydraulic properties of plants and interpreting the results.

Susceptibility to Xylella fastidiosa and functional xylem anatomy in Olea europaea: revisiting a tale of plant-pathogen interaction.

Xylella fastidiosa is a xylem-limited bacterium causing the Olive Quick Decline Syndrome, which is currently devastating the agricultural landscape of Southern Italy. The bacterium is injected into the xylem vessels of leaf petioles after the penetration of the insect vector's stylet. From here, it is supposed to colonize the xylem vasculature moving against water flow inside conductive vessels. Widespread vessel clogging following the bacterial infection and causing the failure of water transport seemed not to fully supported by the recent empirical xylem anatomical observations in infected olive trees. We tested the hypothesis that the higher susceptibility to the X. fastidiosa's infection in Cellina di Nardò compared with Leccino is associated to the higher vulnerability to air embolism of its larger vessels. Such hypothesis is motivated by the recognized ability of X. fastidiosa in degrading pit membranes and also because air embolism would possibly provide microenvironmental conditions more favourable to its more efficient aerobic metabolism. We revised the relevant literature on bacterium growth and xylem physiology, and carried out empirical field, mid-summer measurements of xylem anatomy and native embolism in olive cultivars with high (Cellina di Nardò) and low susceptibility (Leccino) to the infection by X. fastidiosa. Both cultivars had similar shoot mass traits and vessel length (~80 cm), but the highly susceptible one had larger vessels and a lower number of vessels supplying a given leaf mass. Native air embolism reduced mean xylem hydraulic conductance by ~58 % (Cellina di Nardò) and ~38 % (Leccino). The higher air-embolism vulnerability of the larger vessels in Cellina di Nardò possibly facilitates the X. fastidiosa's infection compared to Leccino. Some important characteristics of the vector-pathogen-plant interactions still require deep investigations acknowledging both the pathogen metabolic pathways and the biophysical principles of xylem hydraulics.

The challenge of tree height in Eucalyptus regnans: when xylem tapering overcomes hydraulic resistance.

*Recent research suggests that increasing conduit tapering progressively reduces hydraulic constraints caused by tree height. Here, we tested this hypothesis using the tallest hardwood species, Eucalyptus regnans. *Vertical profiles of conduit dimensions and vessel density were measured for three mature trees of height 47, 51 and 63 m. *Mean hydraulic diameter (Dh) increased rapidly from the tree apex to the point of crown insertion, with the greatest degree of tapering yet reported (b > 0.33). Conduit tapering was such that most of the total resistance was found close to the apex (82-93% within the first 1 m of stem) and the path length effect was reduced by a factor of 2000. Vessel density (VD) declined from the apex to the base of each tree, with scaling parameters being similar for all trees (a = 4.6; b = -0.5). *Eucalyptus regnans has evolved a novel xylem design that ensures a high hydraulic efficiency. This feature enables the species to grow quickly to heights of 50-60 m, beyond the maximum height of most other hardwood trees.

Scots pine trees react to drought by increasing xylem and phloem conductivities.

Drought limits the long-distance transport of water in the xylem due to the reduced leaf-to-soil water potential difference and possible embolism-related losses of conductance and of sugars in the phloem due to the higher viscosity of the dehydrated sugary solution. This condition can have cascading effects in water and carbon (C) fluxes that may ultimately cause tree death. We hypothesize that the maintenance of xylem and phloem conductances is fundamental for survival also under reduced resource availability, when trees may produce effective and low C cost anatomical adjustments in the xylem and phloem close to the treetop where most of the hydraulic resistance is concentrated. We analyzed the treetop xylem and phloem anatomical characteristics in coexisting Scots pine trees, symptomatic and non-symptomatic of drought-induced dieback. We selected the topmost 55 cm of the main stem and selected several sampling positions at different distances from the stem apex to test for differences in the axial patterns between the two groups of trees. We measured the annual ring area, the tracheid hydraulic diameter (Dh) and cell wall thickness (CWT), the conductive phloem area and the average lumen diameter of the 20 largest phloem sieve cells (Dph). Declining trees grew less than the non-declining ones, and despite the similar axial scaling of anatomical traits, had larger Dh and lower CWT. Moreover, declining trees had wider Dph. Our results demonstrate that even under drought stress, maintenance of xylem and phloem efficiencies is of primary importance for survival, even if producing fewer larger tracheids may lead to a xylem more vulnerable to embolism formation.

Plant physiology in theory and practice: an analysis of the WBE model for vascular plants.

The theoretical model of West, Brown and Enquist (hereafter WBE) proposed the fractal geometry of the transport system as the origin of the allometric scaling laws observed in nature. The WBE model has either been criticized for some restrictive and biologically unrealistic constraints or its reliability debated on the evidence of empirical tests. In this work, we revised the structure of the WBE model for vascular plants, highlighting some critical assumptions and simplifications and discuss them with regard to empirical evidence from plant anatomy and physiology. We conclude that the WBE model had the distinct merit of shedding light on some important features such as conduit tapering. Nonetheless, it is over-simplistic and a revised model would be desirable with an ontogenetic perspective that takes some important phenomena into account, such as the transformation of the inner sapwood into heartwood and the effect of hydraulic constraints in limiting the growth in height.

Similarities and differences in the balances between leaf, xylem and phloem structures in Fraxinus ornus along an environmental gradient.

The plant carbon balance depends on the coordination between photosynthesis and the long-distance transport of water and sugars. How plants modify the allocation to the different structures affecting this coordination under different environmental conditions has been poorly investigated. In this study, we evaluated the effect of soil water availability on the allocation to leaf, xylem and phloem structures in Fraxinus ornus L. We selected small individuals of F. ornus (height ~2 m) from sites contrasting in soil water availability (wet vs dry). We measured how the leaf (LM) and stem + branch biomass (SBM) are cumulated along the stem. Moreover, we assessed the axial variation in xylem (XA) and phloem tissue area (PA), and in lumen area of xylem vessels (CAxy) and phloem sieve elements (CAph). We found a higher ratio of LM:SBM in the trees growing under drier conditions. The long-distance transport tissues of xylem and phloem followed axial patterns with scaling exponents (b) independent of site conditions. PA scaled isometrically with XA (b ~ 1). While CAxy was only marginally higher at the wet sites, CAph was significantly higher at the drier sites. Our results showed that under reduced soil water availability, F. ornus trees allocate relatively more to the leaf biomass and produce more conductive phloem, which is likely to compensate for the drought-related hydraulic limitations to the leaf gas exchanges and the phloem sap viscosity.

The potential of Mid-Infrared spectroscopy for prediction of wood density and vulnerability to embolism in woody angiosperms.

Xylem resistance to embolism formation determines the species-specific drought tolerance and the survival prospects of plants under extreme climatic conditions. Fourier Transform-Infrared (FTIR) spectroscopy is a cost-effective and rapid analytical tool with potential beyond its current use in plant physiology. We tested the use of FTIR spectroscopy as a method for estimating wood density (WD) and xylem resistance to embolism formation (P50) in 24 angiosperm species. Higher WD was associated with more negative P50 (r2 = 0.41). Partial least squares regression was applied to establish models of FTIR spectra and the reference data. They showed a high predictive quality for WD (r2 = 0.73), whereas the prediction of P50 was weaker (r2 = 0.49). By including WD in the model as an additional factor influencing P50, its predictive power significantly increased (r2 = 0.59). The spectral range in the model elaboration has been also narrowed (bands of lignin, cellulose, hemicellulose), but this did not influence the model descriptors, suggesting that for P50 prediction broad spectral range is more informative than narrow band regions reflecting main wood constituents. In conclusion, FTIR spectroscopy associated with WD measurements has proven to be a promising alternative to traditional methods for screening of individual- or species-specific resistance to embolism in angiosperms.

Hydraulic recovery from xylem embolism in excised branches of twelve woody species: Relationships with parenchyma cells and non-structural carbohydrates.

Embolism repair ability has been documented in numerous species. Although the actual mechanism driving this phenomenon is still debated, experimental findings suggest that non-structural carbohydrates (NSC) stored in wood parenchyma would provide the osmotic forces to drive the refilling of embolized conduits. We selected 12 broadleaved species differing in vulnerability to xylem embolism (P50) and amount of wood parenchyma in order to check direct evidence about the possible link(s) between parenchyma cells abundance, NSC availability and species-specific capacity to reverse xylem embolism. Branches were dehydrated until ∼50% loss of hydraulic conductivity was recorded (PLC ∼50%). Hydraulic recovery (ΔPLC) and NSC content was, then, assessed after 1h of rehydration. Species showed a different ability to recover their hydraulic conductivity from PLC∼50%. Removing the bark in the species showing hydraulic recovery inhibited the embolism reversal. Strong correlations between the ΔPLC and: a) the amount of parenchyma cells (mainly driven by the pith area), b) the consumption of soluble NSC have been recorded. Our results support the hypothesis that refilling of embolized vessels is mediated by the mobilization of soluble NSC and it is mainly recorded in species with a higher percentage of parenchyma cells that may be important in the hydraulic recovery mechanism as a source of carbohydrates and/or as a source of water.