Arabidopsis leaf hydraulic conductance is regulated by xylem sap pH,controlled, in turn,by a P-type H+-ATPase of vascular bundle sheath cells

The leaf vascular bundle sheath cells (BSCs) that tightly envelop the leaf veins, are a selective and dynamic barrier to xylem sap water and solutes radially entering the mesophyll cells. Under normal conditions, xylem sap pH below 6 is presumably important for driving and regulating the transmembranal solute transport. Having discovered recently a differentially high expression of a BSC proton pump, AHA2, we now test the hypothesis that it regulates the xylem sap pH and leaf radial water fluxes. We monitored the xylem sap pH in the veins of detached leaves of wild-type Arabidopsis, AHA mutants and aha2 mutants complemented with AHA2 gene solely in BSCs. We tested an AHA inhibitor (vanadate) and stimulator (fusicoccin), and different pH buffers. We monitored their impact on the xylem sap pH and the leaf hydraulic conductance (K_leaf), and the effect of pH on the water osmotic permeability (P_f) of isolated BSCs protoplasts. We found that AHA2 is necessary for xylem sap acidification, and in turn, for elevating K_leaf. Conversely, AHA2 knockdown, which alkalinized the xylem sap, or, buffering its pH to 7.5, reduced K_leaf, and elevating external pH to 7.5 decreased the BSCs P_f. All these showed a causative link between AHA2 activity in BSCs and leaf radial hydraulic water conductance.     

Stomatal Behaviours of Aspen (<Emphasis Type="Italic">Populus tremuloides</Emphasis>) Plants in Response to Low Root Temperature in Hydroponics

Hydroponic-grown seedlings of aspen (Populus tremuloides Michx.) were used to investigate how low root temperatures (5°C) affect stomatal conductance and water relations. An isohydric manner of the stomatal behaviour was found with the seedlings when their roots were subjected to the low temperature. Stomatal conductance rapidly and dramatically reduced in response to the low root temperature, while the xylem water potential did not significantly alter. Under the low root temperature, pH value of the xylem sap increased from 6.15 to 6.72 within the initial 4 h, while abscisic acid (ABA) concentration increased by the eighth hour of treatment. K⁺ concentration of the xylem sap significantly decreased within the 8th h and then reversed by the 24th h. The ion change was accompanied by a decrease and then an increase in the electrical conductivity, and an increase and then a decrease in the osmotic potential. The tempo of physiological responses to the low root temperature suggests that the rapid pH change of the xylem sap was the initial factor which triggered stomatal closure in low temperature-treated seedlings, and that the role of the more slowly accumulating ABA was likely to reinforce the stomatal closure. Xylem sap from the seedlings subjected low root temperature affected stomatal aperture on leaf discs when they were floated on the sap solution. The stomatal aperture correlated (P = 0.006) with the changed pattern of [K⁺] in the sap while the range of pH or ABA found in the xylem sap did not influence stomatal aperture of leaf discs in solution. The effect of xylem sap on stomatal aperture on leaf discs was different from on stomatal conductance in the intact seedlings. Comparison was made with previous study with the soil-grown seedlings.     

Changes in leaf hydraulic conductance correlate with leaf vein embolism in<Emphasis Type="Italic"> Cercis siliquastrum</Emphasis> L.

The impact of xylem cavitation and embolism on leaf (K _leaf) and stem (K _stem) hydraulic conductance was measured in current-year shoots of Cercis siliquastrum L. (Judas tree) using the vacuum chamber technique. K _stem decreased at leaf water potentials (Ψ_L) lower than ?1.0 MPa, while K _leaf started to decrease only at Ψ_L L K _leaf changes. Field measurements of leaf conductance to water vapour (g _L) and Ψ_L showed that stomata closed when Ψ_L decreased below the Ψ_L threshold inducing loss of hydraulic conductance in the leaf. The partitioning of hydraulic resistances within shoots and leaves was measured using the high-pressure flow meter method. The ratio of leaf to shoot hydraulic resistance was about 0.8, suggesting that stem cavitation had a limited impact on whole shoot hydraulic conductance. We suggest that stomatal aperture may be regulated by the cavitation-induced reduction of hydraulic conductance of the soil-to-leaf water pathway which, in turn, strongly depends on the hydraulic architecture of the plant and, in particular, on leaf hydraulics.     

Functional coordination between branch hydraulic properties and leaf functional traits in miombo woodlands: implications for water stress management and species habitat preference

We investigated functional coordination between branch hydraulic properties and leaf functional traits among nine miombo woodlands canopy tree species differing in habitat preference and phenology. Specifically, we were seeking to answer the question: are branch hydraulic properties coordinated with leaf functional traits linked to plant drought tolerance in seasonally dry tropical forests and what are the implications for species habitat preference? The hydraulic properties investigated in this study were stem area specific hydraulic conductivity (K _S), Huber value (H _v), and xylem cavitation vulnerability (??₅₀). The leaf functional traits measured were specific leaf area (SLA), leaf dry matter content (LDMC), and mean leaf area (MLA). Generalists displayed significantly (P?<?0.05) higher cavitation resistance (??₅₀) and SLA, but lower sapwood specific hydraulic conductivity (K _S), leaf specific conductivity (K _L), MLA, and LDMC than mesic specialists. Although MLA was uncorrelated with ??₅₀, we found significant (P?<?0.05) positive and negative correlations between plant hydraulic properties and leaf functional traits linked to plant drought tolerance ability, indicating that the interactions between branch hydraulics and leaf functional traits related to plant drought tolerance ability may influence tree species habitat preference in water-limited ecosystems.     

Root hypoxia reduces leaf growth : role of factors in the transpiration stream

This study examined the potential role of restricted phloem export, or import of substances from the roots in the leaf growth response to root hypoxia. In addition, the effects of root hypoxia on abscisic acid (ABA) and zeatin riboside (ZR) levels were measured and their effects on in vitro growth determined. Imposition of root hypoxia in the dark when transpirational water flux was minimal delayed the reduction in leaf growth until the following light period. Restriction of phloem transport by stem girdling did not eliminate the hypoxia-induced reduction in leaf growth. In vitro growth of leaf discs was inhibited in the presence of xylem sap collected from hypoxic roots, and also by millimolar ABA. Disc growth was promoted by sap from aerated roots and by 0.1 micromolar ZR. The flux of both ABA and ZR was reduced in xylem sap from hypoxic roots. Leaf ABA transiently increased twofold after 24 hours of hypoxia exposure but there were no changes in leaf cytokinin levels.     

Diurnal and seasonal changes of leaf lamina hydraulic conductance in bur oak (Quercus macrocarpa) and trembling aspen (Populus tremuloides)

The aim of this study was to examine the diurnal and seasonal variations in the sensitivity of leaf lamina (K _lam) hydraulic conductance to irradiance in bur oak (Quercus macrocarpa Michx.) and trembling aspen (Populus tremuloides Michx.), which vary in their responses of K _lam to irradiance. K _lam was determined using the high-pressure method and the measurements were carried out in June, August and September. The irradiance response of K _lam in bur oak was present throughout the day and declined in senescing leaves. In trembling aspen, K _lam declined from morning to late afternoon and drastically decreased before the onset of leaf senescence, but it was not sensitive to irradiance. In both tree species, the capacity of the petioles to supply water to leaf lamina changed during the day in accordance with the ability of the leaf lamina to transport water. Petiole hydraulic conductivity (K _pet) declined during the season in bur oak leaves, while it tended to increase in trembling aspen leaves. There was no correlation between the K _lam values and air temperature or light intensity at the time of leaf collection. For trembling aspen, K _pet was negatively correlated with the air temperature suggesting sensitivity to drought. We conclude that the water transport properties of petioles and leaf lamina in the two studied tree species reflect their ecological adaptations. Trembling aspen leaves have high hydraulic conductivity and high stomatal conductance regardless of the irradiance level, consistent with the rapid growth and high demand for water. In contrast, the increased lamina hydraulic conductivity and stomatal conductance under high irradiance in bur oak trees reflect a water conservation strategy.     

The Role of Water Channel Proteins in Facilitating Recovery of Leaf Hydraulic Conductance from Water Stress in Populus trichocarpa

Gas exchange is constrained by the whole-plant hydraulic conductance (K _plant). Leaves account for an important fraction of K _plant and may therefore represent a major determinant of plant productivity. Leaf hydraulic conductance (K _leaf) decreases with increasing water stress, which is due to xylem embolism in leaf veins and/or the properties of the extra-xylary pathway. Water flow through living tissues is facilitated and regulated by water channel proteins called aquaporins (AQPs). Here we assessed changes in the hydraulic conductance of Populus trichocarpa leaves during a dehydration-rewatering episode. While leaves were highly sensitive to drought, K _leaf recovered only 2 hours after plants were rewatered. Recovery of K _leaf was absent when excised leaves were bench-dried and subsequently xylem-perfused with a solution containing AQP inhibitors. We examined the expression patterns of 12 highly expressed AQP genes during a dehydration-rehydration episode to identify isoforms that may be involved in leaf hydraulic adjustments. Among the AQPs tested, several genes encoding tonoplast intrinsic proteins (TIPs) showed large increases in expression in rehydrated leaves, suggesting that TIPs contribute to reversing drought-induced reductions in K _leaf. TIPs were localized in xylem parenchyma, consistent with a role in facilitating water exchange between xylem vessels and adjacent living cells. Dye uptake experiments suggested that reversible embolism formation in minor leaf veins contributed to the observed changes in K _leaf.     

Xylem and Phloem transport and the functional economy of carbon and nitrogen of a legume leaf

Exchanges of CO₂ and changes in content of C and N were studied over the life of a leaf of Lupinus albus L. These data were combined with measurements of C:N weight ratios of xylem (upper stem tracheal) and phloem (petiole) sap to determine net fluxes of C and N between leaf and plant. Phase 1 of leaf development (first 11 days, leaf to one-third area) showed increasing net import of C and N, with phloem contributing 61% of the imported C and 18% of the N. ¹⁴C feeding studies suggested the potential for simultaneous import and export through phloem over the period 9 to 12 days. Phase 2 (11-20 days, leaf attaining maximum area and net photosynthesis rate) exhibited net import through xylem and increasing export through phloem. Eighty-two% of xylem-delivered N was consumed in leaf growth, the remainder exported in phloem. Phase 3 (20-38 days) showed high but declining rates of photosynthesis, translocation, and net export of N. Phase 4 (38-66 days) exhibited substantial losses of N and declining photosynthesis and translocation of C. C:N ratio of xylem sap remained constant (2.3-2.6) during leaf life; petiole phloem sap C:N ratio varied from 25 to 135 over leaf development. The relationships between net photosynthesis and N import in xylem were: phase 1, 4.8 milligrams C per milligram N; phase 2, 24.7 milligrams C per milligram N; phase 3, 91.9 milligrams C per milligram N; and phase 4, 47.7 milligrams C per milligram N.     

How the roots contribute to the ability of Phaseolus vulgaris L. to cope with chilling-induced water stress

Intact plants and stem-girdled plants of Phaseolus vulgaris grown hydroponically were exposed to 5 degrees C for up to 4 d; stem girdling was used to inhibit the phloem transport from the leaves to the roots. After initial water stress, stomatal closure and an amelioration of root water transport properties allowed the plants to rehydrate and regain turgor. Chilling augmented the concentration of abscisic acid (ABA) content in leaves, roots and xylem sap. In intact plants stomatal closure and leaf ABA accumulation were preceded by a slight alkalinization of xylem sap, but they occurred earlier than any increase in xylem ABA concentration could be detected. Stem girdling did not affect the influence of chilling on plant water relations and leaf ABA content, but it reduced slightly the alkalinization of xylem sap and, principally, prevented the massive ABA accumulation in root tissues and the associated transport in the xylem that was observed in non-girdled plants. When the plants were defoliated just prior to chilling or after 10 h at 5 degrees C, root and xylem sap ABA concentration remained unchanged throughout the whole stress period. When the plants were chilled under conditions preventing the occurrence of leaf water deficit (i.e. at 100% relative humidity), there were no significant variations in endogenous ABA levels. The increase in root hydraulic conductance in chilled plants was a response neither to root ABA accretion, nor to some leaf-borne chemical signal transported downwards in the phloem, nor to low temperature per se, as indicated by the results of the experiments with defoliated or girdled plants and with plants chilled at 100% relative humidity. It was concluded that the root system contributed substantially to the bean's ability to cope with chilling-induced water stress, but not in an ABA-dependent manner.     

Neither xylem collapse,cavitation, or changing leaf conductance drive stomatal closure in wheat

Identifying the drivers of stomatal closure and leaf damage during stress in grasses is a critical prerequisite for understanding crop resilience. Here, we investigated whether changes in stomatal conductance (g_s) during dehydration were associated with changes in leaf hydraulic conductance (K_leaf), xylem cavitation, xylem collapse, and leaf cell turgor in wheat (Triticum aestivum). During soil dehydration, the decline of g_s was concomitant with declining K_leaf under mild water stress. This early decline of leaf hydraulic conductance was not driven by cavitation, as the first cavitation events in leaf and stem were detected well after K_leaf had declined. Xylem vessel deformation could only account for <5% of the observed decline in leaf hydraulic conductance during dehydration. Thus, we concluded that changes in the hydraulic conductance of tissues outside the xylem were responsible for the majority of K_leaf decline during leaf dehydration in wheat. However, the contribution of leaf resistance to whole plant resistance was less than other tissues (<35% of whole plant resistance), and this proportion remained constant as plants dehydrated, indicating that K_leaf decline during water stress was not a major driver of stomatal closure.     

Spontaneous Phloem bleeding from cryopunctured fruits of a ureide-producing legume

The vasculature of the dorsal suture of cowpea (Vigna unguiculata [L.] Walp) fruits bled a sugar-rich exudate when punctured with a fine needle previously cooled in liquid N₂. Bleeding continued for many days at rates equivalent to 10% of the estimated current sugar intake of the fruit. A phloem origin for the exudate was suggested from its high levels (0.4-0.8 millimoles per milliliter) of sugar (98% of this as sucrose) and its high K⁺ content and high ratio of Mg²⁺ to Ca²⁺. Fruit cryopuncture sap became labeled with ¹⁴C following feeding of [¹⁴C]urea to leaves or adjacent walls of the fruit, of ¹⁴CO₂ to the pod gas space, and of [¹⁴C] asparagine or [¹⁴C]allantoin to leaflets or cut shoots through the xylem. Rates of translocation of ¹⁴C-assimilates from a fed leaf to the puncture site on a subtended fruit were 21 to 38 centimeters per hour. Analysis of ¹⁴C distribution in phloem sap suggested that [¹⁴C]allantoin was metabolized to a greater extent in its passage to the fruit than was [¹⁴C] asparagine. Amino acid:ureide:nitrate ratios (nitrogen weight basis) of NO₃-fed, non-nodulated plants were 20:2:78 in root bleeding xylem sap versus 90:10:0.1 for fruit phloem sap, suggesting that the shoot utilized NO₃-nitrogen to synthesize amino acids prior to phloem transfer of nitrogen to the fruit. Feeding of ¹⁵NO₃ to roots substantiated this conclusion. The amino acid:ureide ratio (nitrogen weight basis) of root xylem sap of symbiotic plants was 23:77 versus 89:11 for corresponding fruit phloem sap indicating intense metabolic transfer of ureide-nitrogen to amino acids by vegetative parts of the plant.     

Stem and whole-plant hydraulics in olive (Olea europaea) and kiwifruit (Actinidia deliciosa)

A field study and an experiment under controlled conditions using pressure-flux relationships were conducted to compare the stem and whole-plant conductance in olive (Olea europaea) and kiwifruit (Actinidia deliciosa) species. Anatomical observations were also made on one-year-old stem to determine the conductive area of vessels (A _ves) and the total xylem area (A _xyl). Results show that A _ves of kiwifruit twigs was ~2.5-fold of that in olive twigs, and the hydraulically weighted mean diameter was up to threefold that of the olive ones. One-year-old olive twigs had lower hydraulic conductivity (k) than the kiwifruit, while values of leaf-specific conductivity (i.e. k normalised per unit leaf area) were higher than the kiwifruit (i.e. ~49 and 29 × 10^?6 kg m^?1 s^?1 MPa^?1, respectively). In the field experiment, the flux of sap (heat balance method) and differences in water potential through the soil–plant system (ΔP) were used for both species to calculate the whole-plant conductance that was normalised per unit leaf area (leaf-specific whole-plant conductance, K _plant,LA). Values of K _plant,LA are attributable to the combined effect of the ΔP and anatomical features of conduits. Olive species showed a larger ΔP (2.4 MPa at midday) than the kiwifruit (0.5 MPa) which contributed to lower K _plant,LA in Olea than the Actinidia plants. This information, combined with vessel density data, contributes to explain differences amidst olive and kiwifruit species, in terms of susceptibility to some drought-related hydraulic impairments induced by the Mediterranean environment.     

Water Transport in the Liana Bauhinia fassoglensis (Fabaceae)

To determine the efficiency of xylem conductance in the liana (woody vine) Bauhinia fassoglensis Kotschy ex Schweinf., we measured hydraulic conductance per unit stem length (measured K_h), leaf-specific conductivity (LSC = K_h/distal leaf area), transpiration rate (E), xylem water potential (ε), vessel number, and vessel diameter. The measured K_h was 49% (se = 7%) of the predicted K_h from Poiseuille's law. The mean LSC for unbranched stem segments was 1.10 × 10⁻⁸ square meters per megapascal per second (se = 0.07). LSCs were much lower (about 0.2) at branch junctions. At midday, with E at 7 × 10⁻⁸ meters per second, the measured drop in ε was about 0.08 megapascal per meter along the stems and branches and about 0.27 megapascal in going from stem to leaf. In addition, there was a drop of about 0.20 megapascal at branch junctions as predicted by E/LSC. In diurnal measurements leaf ε never dropped below about −1.2 megapascal. For long (e.g. 16 meters) stems, the predicted mid-day drop in ε through the xylem transport system might be great enough to have substantial physiological impact.     

Dynamic changes in petiole specific conductivity in red maple (Acer rubrum L.), tulip tree (Liriodendron tulipifera L.) and northern fox grape (Vitis labrusca L.)

Diurnal variation in petiole specific hydraulic conductivity and simultaneous measurements of leaf water potential were recorded in red maple, tulip tree and fox grape. Petiole specific conductivity was determined from in situ measurements of water flow into the distal (leaf‐bearing) end of an attached petiole as a function of applied hydrostatic pressure and petiole dimensions. The hydraulic properties of the petiole dominated the measurements, indicating that this technique can be used for rapid estimates of petiole hydraulic conductivity. There was a significant decrease in petiole specific conductivity associated with increasingly more negative leaf water potentials in maple and tulip tree, but not in grape. Petiole specific conductivity increased during the afternoon while the plant was actively transpiring and the xylem sap was under tension. The recovery of petiole conductivity during the afternoon suggests that hydraulic conductivity reflects a dynamic balance between a loss of hydraulic conductivity with increasing water stress, and its restoration as tension within the xylem decreases. Three experimental manipulations were applied to red maple and tulip tree to examine the sensitivity of diurnal changes in petiole conductivity to various physiological perturbations. Both phloem girdling and application of HgCl₂ to the transpiration stream resulted in a marked decrease in the degree to which petiole specific conductivity recovered as xylem tension relaxed during the afternoon. Delivery of a surfactant to the xylem, however, did not significantly alter the relation between leaf water potential and petiole hydraulic conductivity.     

Sodium transport in basil

In order to diversify the production of plants with pharmacological interest, it is important to understand the mechanisms involved in their tolerance to environmental constraints, such as salinity. Basil (Ocimum basilicum), known for its therapeutic uses, has been claimed to be salt tolerant, but physiological aspects of this behavior remain unknown. Since salt tolerance is known to be associated with several characteristics concerning Na⁺ transport to leaves, we studied this function in hydroponically grown basil. We analyzed the response of 30-day-old seedlings to 25–50 mM NaCl applied for 15 days. Growth was poorly affected, indicating that these concentrations corresponded to the tolerated salinity range. Leaves accumulated Na⁺ at relatively high concentration, without dehydrating. Potassium concentration in leaf tissues was maintained close to control level, indicating that K⁺ was 15- to 25-fold preferred over Na⁺ for ion transport and deposition. Collection of xylem sap on detopped plants revealed that this preference was only 10-fold for ion introduction into root xylem sap. Short-term (24 h) changes in Na⁺ distribution between organs after stem (steam) girdling suggested that Na⁺ downward recirculation by phloem occurred. Although modest, this transport might have augmented K⁺ selectivity of ion deposition in leaves.     

New evidence for a role of vessel-associated cells and phloem in the rapid xylem refilling of cavitated stems of Laurus nobilis L.

Xylem recovery from embolism was studied in Laurus nobilis L. stems that were induced to cavitate by combining negative xylem pressure potentials (P_X = ?1.1 MPa) with positive air pressures (P_C) applied using a pressure collar. Xylem refilling was measured by recording the percentage loss of hydraulic conductance (PLC) with respect to the maximum 2 min, 20 min and 15 h after pressure release. Sodium orthovanadate (an inhibitor of many ATP‐ases) strongly inhibited xylem refilling while fusicoccin (a stimulator of the plasma membrane H⁺‐ATPase) promoted complete embolism reversal. So, the refilling process was interpreted to result from energy‐dependent mechanisms. Stem girdling induced progressively larger inhibition to refilling the nearer to the embolized stem segment phloem was removed. The starch content of wood parenchyma was estimated as percentages of ray and vasicentric cells with high starch content with respect to the total, before and after stem embolism was induced. A closely linear positive relationship was found to exist between recovery from PLC and starch hydrolysis. This, was especially evident in vasicentric cells. A mechanism for xylem refilling based upon starch to sugar conversion and transport into embolized conduits, assisted by phloem pressure‐driven radial mass flow is proposed.     

Hydraulic adjustment of maple saplings to canopy gap formation

The leaf-specific hydraulic conductivity (K _L) of plant stems can control leaf water supply. This property is influenced by variation in leaf/sapwood area ratio (A _L/A _S) and the specific hydraulic conductivity of xylem tissue (K _S). In environments with high atmospheric vapor pressure deficit (VPD), K _L may increase to support higher transpiration rates. We predicted that saplings of Acerrubrum and A.pensylvanicum grown in forest canopy gaps, under high light and VPD, would have higher K _L and lower A _L/A _S than similar sized saplings in the understory. Leaf-specific hydraulic conductivity and K _S increased with sapling size for both species. In A. rubrum, K _S did not differ between the two environments but lower A _L/A _S (P=0.05, ANCOVA) led to higher K _L for gap-grown saplings (P < 0.05, ANCOVA). In A. pensylvanicum, neither K _S, A _L/A _S, nor K_L differed between environments. In a second experiment, we examined the impact of sapling size on the water relations and carbon assimilation of A.pensylvanicum. Maximum stomatal conductance for A.pensylvanicum increased with K _L (r ²=0.75, P < 0.05). A hypothetical large A. pensylvanicum sapling (2 m tall) had 2.4 times higher K _L and 22 times greater daily carbon assimilation than a small (1 m tall) sapling. Size-related hydraulic limitations in A.pensylvanicum caused a 68% reduction in daily carbon assimilation in small saplings. Mid-day water potential increased with A.pensylvanicum sapling size (r ²=0.69, P < 0.05). Calculations indicated that small A.pensylvanicum saplings (low K _L) could not transpire at the rate of large saplings (high K _L) without reaching theoretical thresholds for xylem embolism induction. The coordination between K _L and stomatal conductance in saplings may prevent xylem water potential from reaching levels that cause embolism but also limits transpiration. The K _S of the xylem did not vary across environments, suggesting that altering biomass allocation is the primary mechanism of increasing K _L. However, the ability to alter aboveground biomass allocation in response to canopy gaps is species-specific. As a result of the increase in K _L and K _S with sapling size for both species, hydraulic limitation of water flux may impose a greater restriction on daily carbon assimilation for small saplings in the gap environment. Received: 18 February 1997 / Accepted: 23 June 1997     

Bundle sheath lignification mediates the linkage of leaf hydraulics and venation

The lignification of the leaf vein bundle sheath (BS) has been observed in many species and would reduce conductance from xylem to mesophyll. We hypothesized that lignification of the BS in lower‐order veins would provide benefits for water delivery through the vein hierarchy but that the lignification of higher‐order veins would limit transport capacity from xylem to mesophyll and leaf hydraulic conductance (K_leaf). We further hypothesized that BS lignification would mediate the relationship of K_leaf to vein length per area. We analysed the dependence of K_leaf, and its light response, on the lignification of the BS across vein orders for 11 angiosperm tree species. Eight of 11 species had lignin deposits in the BS of the midrib, and two species additionally only in their secondary veins, and for six species up to their minor veins. Species with lignification of minor veins had a lower hydraulic conductance of xylem and outside‐xylem pathways and lower K_leaf. K_leaf could be strongly predicted by vein length per area and highest lignified vein order (R² = .69). The light‐response of K_leaf was statistically independent of BS lignification. The lignification of the BS is an important determinant of species variation in leaf and thus whole plant water transport.     

Photosynthetic pathway alters xylem structure and hydraulic function in herbaceous plants

Plants using the C₄ photosynthetic pathway have greater water use efficiency (WUE) than C₃ plants of similar ecological function. Consequently, for equivalent rates of photosynthesis in identical climates, C₄ plants do not need to acquire and transport as much water as C₃ species. Because the structure of xylem tissue reflects hydraulic demand by the leaf canopy, a reduction in water transport requirements due to C₄ photosynthesis should affect the evolution of xylem characteristics in C₄ plants. In a comparison of stem hydraulic conductivity and vascular anatomy between eight C₃ and eight C₄ herbaceous species, C₄ plants had lower hydraulic conductivity per unit leaf area (K_L) than C₃ species of similar life form. When averages from all the species were pooled together, the mean K_L for the C₄ species was 1.60 × 10^?4 kg m^?1 s^?1 MPa^?1, which was only one‐third of the mean K_L of 4.65 × 10^?4 kg m^?1 s^?1 MPa^?1 determined for the C₃ species. The differences in K_L between C₃ and C₄ species corresponded to the two‐ to three‐fold differences in WUE observed between C₃ and C₄ plants. In the C₄ species from arid regions, the difference in K_L was associated with a lower hydraulic conductivity per xylem area, smaller and shorter vessels, and less vulnerable xylem to cavitation, indicating the C₄ species had evolved safer xylem than the C₃ species. In the plants from resource‐rich areas, such as the C₄ weed Amaranthus retroflexus, hydraulic conductivity per xylem area and xylem anatomy were similar to that of the C₃ species, but the C₄ plants had greater leaf area per xylem area. The results indicate the WUE advantage of C₄ photosynthesis allows for greater flexibility in hydraulic design and potential fitness. In resource‐rich environments in which competition is high, an existing hydraulic design can support greater leaf area, allowing for higher carbon gain, growth and competitive potential. In arid regions, C₄ plants evolved safer xylem, which can increase survival and performance during drought events.     

Size-dependent variability of leaf and shoot hydraulic conductance in silver birch

Variation in leaf and shoot hydraulic conductance was examined on detached shoots of silver birch (Betula pendula Roth), cut from the lower third (shade leaves) and upper third of the crown (sun leaves) of large trees growing in a natural temperate forest stand. Hydraulic conductances of whole shoots (K _S), leaf blades (K _lb), petioles (K _P) and branches (i.e. leafless stem; K _B) were determined by water perfusion using a high-pressure flow meter in quasi-steady state mode. The shoots were exposed to irradiance of photosynthetic photon flux density of 200–250 μmol m⁻² s⁻¹, using different light sources. K _lb depended significantly (P < 0.001) on light quality, canopy position and leaf blade area (A _L). K _lb increased from crown base to tree top, in parallel with vertical patterns of A _L. However, the analysis of data on shade and sun leaves separately revealed an opposite trend: the bigger the A _L the higher K _lb. Leaf anatomical study of birch saplings revealed that this trend is attributable to enhanced vascular development with increasing leaf area. Hydraulic traits (K _S, K _B, K _lb) of sun shoots were well co-ordinated and more strongly correlated with characteristics of shoot size than those of shade shoots, reflecting their greater evaporative load and need for stricter adjustment of hydraulic capacity with shoot size. K _S increased with increasing xylem cross-sectional area to leaf area ratio (Huber value; P < 0.01), suggesting a preferential investment in water-conducting tissue (sapwood) relative to transpiring tissue (leaves), and most likely contributing to the functional stability of the hydraulic system, essential for fast-growing pioneer species.