Leaf hydraulic safety margin and safety–efficiency trade-off across angiosperm woody species

Leaf hydraulic conductance and the vulnerability to water deficits have profound effects on plant distribution and mortality. In this study, we compiled a leaf hydraulic trait dataset with 311 species-at-site combinations from biomes worldwide. These traits included maximum leaf hydraulic conductance (K_leaf), water potential at 50% loss of K_leaf (P50_leaf), and minimum leaf water potential (Ψ_min). Leaf hydraulic safety margin (HSM_leaf) was calculated as the difference between Ψ_min and P50_leaf. Our results indicated that 70% of the studied species had a narrow HSM_leaf (less than 1 MPa), which was consistent with the global pattern of stem hydraulic safety margin. There was a positive relationship between HSM_leaf and aridity index (the ratio of mean annual precipitation to potential evapotranspiration), as species from humid sites tended to have larger HSM_leaf. We found a significant relationship between K_leaf and P50_leaf across global angiosperm woody species and within each of the different plant groups. This global analysis of leaf hydraulic traits improves our understanding of plant hydraulic response to environmental change.     

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.     

Plasticity of vulnerability to leaf hydraulic dysfunction during acclimation to drought in grapevines: an osmotic‐mediated process

Previous studies have reported correlation of leaf hydraulic vulnerability with pressure–volume parameters related to cell turgor. This link has been explained on the basis of the effects of turgor on connectivity among cells and tissue structural integrity, which affect leaf water transport. In this study, we tested the hypothesis that osmotic adjustment to water stress would shift the leaf vulnerability curve toward more negative water potential (Ψ_leaf) by increasing turgor at low Ψ_leaf. We measured leaf hydraulic conductance (K_leaf), K_leaf vulnerability [50 and 80% loss of K_leaf (P₅₀ and P₈₀); |Ψ_leaf| at 50 and 80% loss of K_leaf, respectively), bulk leaf water relations, leaf gas exchange and sap flow in two Vitis vinifera cultivars (Tempranillo and Grenache), under two water treatments. We found that P₅₀, P₈₀ and maximum K_leaf decreased seasonally by more than 20% in both cultivars and watering treatments. However, K_leaf at ?2 MPa increased threefold, while osmotic potential at full turgor and turgor loss point decreased. Our results indicate that leaf resistance to hydraulic dysfunction is seasonally plastic, and this plasticity may be mediated by osmotic adjustment.     

Leaf hydraulic vulnerability to drought is linked to site water availability across a broad range of species and climates

Background and Aims

Vulnerability of the leaf hydraulic pathway to water-stress-induced dysfunction is a key component of drought tolerance in plants and may be important in defining species'' climatic range. However, the generality of the association between leaf hydraulic vulnerability and climate across species and sites remains to be tested.

Methods

Leaf hydraulic vulnerability to drought (P50_leaf, the water potential inducing 50 % loss in hydraulic function) was measured in a diverse group of 92 woody, mostly evergreen angiosperms from sites across a wide range of habitats. These new data together with some previously published were tested against key climate indices related to water availability. Differences in within-site variability in P50_leaf between sites were also examined.

Key Results

Values of hydraulic vulnerability to drought in leaves decreased strongly (i.e. became more negative) with decreasing annual rainfall and increasing aridity across sites. The standard deviation in P50_leaf values recorded within each site was positively correlated with increasing aridity. P50_leaf was also a good indicator of the climatic envelope across each species'' distributional range as well as their dry-end distributional limits within Australia, although this relationship was not consistently detectable within sites.

Conclusions

The findings indicate that species sorting processes have influenced distributional patterns of P50_leaf across the rainfall spectrum, but alternative strategies for dealing with water deficit exist within sites. The strong link to aridity suggests leaf hydraulic vulnerability may influence plant distributions under future climates.     

Differential coordination of stomatal conductance,mesophyll conductance,and leaf hydraulic conductance in response to changing light across species

Stomatal conductance (g_s) and mesophyll conductance (g_m) represent major constraints to photosynthetic rate (A), and these traits are expected to coordinate with leaf hydraulic conductance (K_leaf) across species, under both steady‐state and dynamic conditions. However, empirical information about their coordination is scarce. In this study, K_leaf, gas exchange, stomatal kinetics, and leaf anatomy in 10 species including ferns, gymnosperms, and angiosperms were investigated to elucidate the correlation of H₂O and CO₂ diffusion inside leaves under varying light conditions. Gas exchange, K_leaf, and anatomical traits varied widely across species. Under light‐saturated conditions, the A, g_s, g_m, and K_leaf were strongly correlated across species. However, the response patterns of A, g_s, g_m, and K_leaf to varying light intensities were highly species dependent. Moreover, stomatal opening upon light exposure of dark‐adapted leaves in the studied ferns and gymnosperms was generally faster than in the angiosperms; however, stomatal closing in light‐adapted leaves after darkening was faster in angiosperms. The present results show that there is a large variability in the coordination of leaf hydraulic and gas exchange parameters across terrestrial plant species, as well as in their responses to changing light.     

Leaf hydraulic vulnerability influences species’ bioclimatic limits in a diverse group of woody angiosperms

The ability of plants to maintain water flow through leaves under water stress-induced tension (assessed as the leaf hydraulic vulnerability; P50 _leaf) is intimately linked with survival. We examined the significance of P50 _leaf as an adaptive trait in influencing the dry-end distributional limits of cool temperate woody angiosperm species. We also examined differences in within-site variability in P50 _leaf between two high-rainfall montane rainforest sites in Tasmania and Peru, respectively. A significant relationship between P50 _leaf and the 5th percentile of mean annual rainfall across each species distribution was found in Tasmania, suggesting that P50 _leaf influences species climatic limits. Furthermore, a strong correlation between P50 _leaf and the minimum rainfall availability was found using five phylogenetically independent species pairs in wet and dry evergreen tree species, suggesting that rainfall is an important selective agent in the evolution of leaf hydraulic vulnerability. Greater within-site variability in P50 _leaf was found among dominant montane rainforest species in Tasmania than in Peru and this result is discussed within the context of differences in spatial and temporal environmental heterogeneity and parochial historical ecology.     

Leaf physiology does not predict leaf habit; examples from tropical dry forest

Leaf structure and physiology are thought to be closely linked to leaf longevity and leaf habit. Here we compare the seasonal variation in leaf hydraulic conductance (k_leaf) and water potential of two evergreen tree species with contrasting leaf life spans, and two species with similar leaf longevity but contrasting leaf habit, one being deciduous and the other evergreen. One of the evergreen species, Simarouba glauca, produced relatively short-lived leaves that maintained high hydraulic conductance year round by periodic flushing. The other evergreen species, Quercus oleoides, produced longer-lived leaves with lower k_leaf and as a result minimum leaf water potential was much lower than in S. glauca (–2.8 MPa vs –1.6 MPa). Associated with exposure to lower water potentials, Q. oleoides leaves were harder, had a higher modulus of elasticity, and were less vulnerable to cavitation than S. glauca leaves. Both species operate at water potentials capable of inducing 20 (S. glauca) to 50% (Q. oleoides) loss of k_leaf during the dry season although no evidence of cumulative losses in k_leaf were observed in either species suggesting regular repair of embolisms. Leaf longevity in the deciduous species Rhedera trinervis is similar to that of S. glauca, although maximum k_leaf was lower. Furthermore, a decline in leaf water potential at the onset of the dry season led to cumulative losses in k_leaf in R. trinervis that culminated in leaf shedding.     

Diurnal depression of leaf hydraulic conductance in a tropical tree species

Diurnal patterns of hydraulic conductance of the leaf lamina (K_leaf) were monitored in a field‐grown tropical tree species in an attempt to ascertain whether the dynamics of stomatal conductance (g_s) and CO₂ uptake (A_leaf) were associated with short‐term changes in K_leaf. On days of high evaporative demand mid‐day depression of K_leaf to between 40 and 50% of pre‐dawn values was followed by a rapid recovery after 1500 h. Leaf water potential during the recovery stage was less than ?1 MPa implying a refilling mechanism, or that loss of K_leaf was not linked to cavitation. Laboratory measurement of the response of K_leaf to Ψ_leaf confirmed that leaves in the field were operating at water potentials within the depressed region of the leaf ‘vulnerability curve’. Diurnal courses of K_leaf and Ψ_leaf predicted from measured transpiration, xylem water potential and the K_leaf vulnerability function, yielded good agreement with observed trends in both leaf parameters. Close correlation between depression of K_leaf, g_s and A_leaf suggests that xylem dysfunction in the leaf may lead to mid‐day depression of gas exchange in this species.     

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.     

Measurement of Leaf Hydraulic Conductance and Stomatal Conductance and Their Responses to Irradiance and Dehydration Using the Evaporative Flux Method (EFM)

Water is a key resource, and the plant water transport system sets limits on maximum growth and drought tolerance. When plants open their stomata to achieve a high stomatal conductance (g_s) to capture CO₂ for photosynthesis, water is lost by transpiration^1,2. Water evaporating from the airspaces is replaced from cell walls, in turn drawing water from the xylem of leaf veins, in turn drawing from xylem in the stems and roots. As water is pulled through the system, it experiences hydraulic resistance, creating tension throughout the system and a low leaf water potential (Ψ_leaf). The leaf itself is a critical bottleneck in the whole plant system, accounting for on average 30% of the plant hydraulic resistance³. Leaf hydraulic conductance (K_leaf = 1/ leaf hydraulic resistance) is the ratio of the water flow rate to the water potential gradient across the leaf, and summarizes the behavior of a complex system: water moves through the petiole and through several orders of veins, exits into the bundle sheath and passes through or around mesophyll cells before evaporating into the airspace and being transpired from the stomata. K_leaf is of strong interest as an important physiological trait to compare species, quantifying the effectiveness of the leaf structure and physiology for water transport, and a key variable to investigate for its relationship to variation in structure (e.g., in leaf venation architecture) and its impacts on photosynthetic gas exchange. Further, K_leaf responds strongly to the internal and external leaf environment³. K_leaf can increase dramatically with irradiance apparently due to changes in the expression and activation of aquaporins, the proteins involved in water transport through membranes⁴, and K_leaf declines strongly during drought, due to cavitation and/or collapse of xylem conduits, and/or loss of permeability in the extra-xylem tissues due to mesophyll and bundle sheath cell shrinkage or aquaporin deactivation^5-10. Because K_leaf can constrain g_s and photosynthetic rate across species in well watered conditions and during drought, and thus limit whole-plant performance they may possibly determine species distributions especially as droughts increase in frequency and severity^11-14.We present a simple method for simultaneous determination of K_leaf and g_s on excised leaves. A transpiring leaf is connected by its petiole to tubing running to a water source on a balance. The loss of water from the balance is recorded to calculate the flow rate through the leaf. When steady state transpiration (E, mmol • m^-2 • s^-1) is reached, g_s is determined by dividing by vapor pressure deficit, and K_leaf by dividing by the water potential driving force determined using a pressure chamber (K_leaf= E /- Δψ_leaf, MPa)¹⁵.This method can be used to assess K_leaf responses to different irradiances and the vulnerability of K_leaf to dehydration^14,16,17.     

Hydraulic Failure Defines the Recovery and Point of Death in Water-Stressed Conifers

This study combines existing hydraulic principles with recently developed methods for probing leaf hydraulic function to determine whether xylem physiology can explain the dynamic response of gas exchange both during drought and in the recovery phase after rewatering. Four conifer species from wet and dry forests were exposed to a range of water stresses by withholding water and then rewatering to observe the recovery process. During both phases midday transpiration and leaf water potential (Ψ_leaf) were monitored. Stomatal responses to Ψ_leaf were established for each species and these relationships used to evaluate whether the recovery of gas exchange after drought was limited by postembolism hydraulic repair in leaves. Furthermore, the timing of gas-exchange recovery was used to determine the maximum survivable water stress for each species and this index compared with data for both leaf and stem vulnerability to water-stress-induced dysfunction measured for each species. Recovery of gas exchange after water stress took between 1 and >100 d and during this period all species showed strong 1:1 conformity to a combined hydraulic-stomatal limitation model (r² = 0.70 across all plants). Gas-exchange recovery time showed two distinct phases, a rapid overnight recovery in plants stressed to <50% loss of leaf hydraulic conductance (K_leaf) and a highly Ψ_leaf-dependent phase in plants stressed to >50% loss of K_leaf. Maximum recoverable water stress (Ψ_min) corresponded to a 95% loss of K_leaf. Thus, we conclude that xylem hydraulics represents a direct limit to the drought tolerance of these conifer species.     

Hydraulic conductance and water potential gradients in squash leaves showing mycorrhiza-induced increases in stomatal conductance

Stomatal conductance (g _s) and transpiration rates vary widely across plant species. Leaf hydraulic conductance (k _leaf) tends to change with g _s, to maintain hydraulic homeostasis and prevent wide and potentially harmful fluctuations in transpiration-induced water potential gradients across the leaf (ΔΨ _leaf). Because arbuscular mycorrhizal (AM) symbiosis often increases g _s in the plant host, we tested whether the symbiosis affects leaf hydraulic homeostasis. Specifically, we tested whether k _leaf changes with g _s to maintain ΔΨ _leaf or whether ΔΨ _leaf differs when g _s differs in AM and non-AM plants. Colonization of squash plants with Glomus intraradices resulted in increased g _s relative to non-AM controls, by an average of 27% under amply watered, unstressed conditions. Stomatal conductance was similar in AM and non-AM plants with exposure to NaCl stress. Across all AM and NaCl treatments, k _leaf did change in synchrony with g _s (positive correlation of g _s and k _leaf), corroborating leaf tendency toward hydraulic homeostasis under varying rates of transpirational water loss. However, k _leaf did not increase in AM plants to compensate for the higher g _s of unstressed AM plants relative to non-AM plants. Consequently, ΔΨ _leaf did tend to be higher in AM leaves. A trend toward slightly higher ΔΨ _leaf has been observed recently in more highly evolved plant taxa having higher productivity. Higher ΔΨ _leaf in leaves of mycorrhizal plants would therefore be consistent with the higher rates of gas exchange that often accompany mycorrhizal symbiosis and that are presumed to be necessary to supply the carbon needs of the fungal symbiont.     

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.     

Stomatal responses to changes in vapor pressure deficit reflect tissue‐specific differences in hydraulic conductance

The vapor pressure deficit (D) of the atmosphere can negatively affect plant growth as plants reduce stomatal conductance to water vapor (g_wv) in response to increasing D, limiting the ability of plants to assimilate carbon. The sensitivity of g_wv to changes in D varies among species and has been correlated with the hydraulic conductance of leaves (K_leaf), but the hydraulic conductance of other tissues has also been implicated in plant responses to changing D. Among the 19 grass species, we found that K_leaf was correlated with the hydraulic conductance of large longitudinal veins (K_lv, r² = 0.81), but was not related to K_root (r² = 0.01). Stomatal sensitivity to D was correlated with K_leaf relative to total leaf area (r² = 0.50), and did not differ between C₃ and C₄ species. Transpiration (E) increased in response to D, but 8 of the 19 plants showed a decline in E at high D, indicative of an ‘apparent feedforward’ response. For these individuals, E began to decline at lower values of D in plants with low K_root (r² = 0.72). These results show the significance of both leaf and root hydraulic conductance as drivers of plant responses to evaporative demand.     

Soybean leaf hydraulic conductance does not acclimate to growth at elevated [CO2] or temperature in growth chambers or in the field

Background and Aims

Leaf hydraulic properties are strongly linked with transpiration and photosynthesis in many species. However, it is not known if gas exchange and hydraulics will have co-ordinated responses to climate change. The objective of this study was to investigate the responses of leaf hydraulic conductance (K_leaf) in Glycine max (soybean) to growth at elevated [CO₂] and increased temperature compared with the responses of leaf gas exchange and leaf water status.

Methods

Two controlled-environment growth chamber experiments were conducted with soybean to measure K_leaf, stomatal conductance (g_s) and photosynthesis (A) during growth at elevated [CO₂] and temperature relative to ambient levels. These results were validated with field experiments on soybean grown under free-air elevated [CO₂] (FACE) and canopy warming.

Key results

In chamber studies, K_leaf did not acclimate to growth at elevated [CO₂], even though stomatal conductance decreased and photosynthesis increased. Growth at elevated temperature also did not affect K_leaf, although g_s and A showed significant but inconsistent decreases. The lack of response of K_leaf to growth at increased [CO₂] and temperature in chamber-grown plants was confirmed with field-grown soybean at a FACE facility.

Conclusions

Leaf hydraulic and leaf gas exchange responses to these two climate change factors were not strongly linked in soybean, although g_s responded to [CO₂] and increased temperature as previously reported. This differential behaviour could lead to an imbalance between hydraulic supply and transpiration demand under extreme environmental conditions likely to become more common as global climate continues to change.     

Hydraulic architecture of leaf blades: where is the main resistance?

The hydraulic architecture of Laurus nobilis L. and Juglans regia L. leaves was studied using three different approaches: (1) hydraulic measurements of both intact leaves and of leaves subjected to treatments aimed at removing the extra‐vascular resistance; (2) direct measurements of the vascular pressure with a pressure probe; and (3) modelling the hydraulic architecture of leaf venation system on the basis of measurements of vein densities and conductivities. The hydraulic resistance of leaves (R_leaf) either cut, boiled or frozen–thawed was reduced by about 60 and 85% with respect to control leaves for laurel and walnut, respectively. Direct pressure drop measurements suggested that 88% of the resistance resided outside the vascular system in walnut. Model simulations were in agreement with these results provided vein hydraulic conductance was 0.12–0.28 that of the conductance predicted by Poiseuille's law. The results suggest that R_leaf is dominated by substantial extra‐vascular resistances and therefore contrast with the conclusions of recent studies dealing with the hydraulic architecture of the leaf. The present study confirms the ‘classical’ view of the hydraulic architecture of leaves as composed by a low‐resistance component (the venation) and a high‐resistance component (the mesophyll).     

南亚热带不同演替阶段森林优势树种叶片构建成本与机械抗性的协同关系

为探究不同演替阶段森林优势种叶片资源获取策略的差异以及叶片构建成本与机械抗性的关系,对我国南亚热带不同演替阶段森林14优势种的叶片构建成本、机械抗性、角质层厚度和比叶重等结构性状进行测定。结果表明,与演替早期相比,演替晚期优势种具有更高的单位面积叶片构建成本、叶片撕裂力以及穿透力,但其叶片最大光合速率较低;同时,单位面积叶片构建成本与机械抗性呈显著正相关关系,而叶片角质层厚度、比叶重等结构性状也与叶片构建成本、机械抗性均呈显著正相关。因此,从叶片能量投资策略上反映了南亚热带森林演替进程中叶片构建成本与机械抗性的协同关系。     

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.     

Circadian regulation of leaf hydraulic conductance in sunflower (Helianthus annuus L. cv Margot)

The circadian regulation of leaf hydraulic conductance (K_leaf) was investigated in Helianthus annuus L. (sunflower). K_leaf was measured with an high pressure flow meter during the light and dark period from plants growing at a photoperiod of 12 h. K_leaf was 4.0 e−4 kg s⁻¹ m⁻² MPa⁻¹ during the light period (LL) and 30–40% less during the dark period (DL). When photoperiod was inverted and leaves were measured for K_leaf at their subjective light or dark periods, K_leaf adjusted to the new conditions requiring 48 h for increasing from dark to light values and 4 d for the opposite transition. Plants put in continuous dark showed K_leaf oscillating from light to dark values in phase with their subjective photoperiod indicating that K_leaf changes were induced by the circadian clock. Several cuts through the minor veins reduced leaf hydraulic resistance (R_leaf) of both LL and DL to the same value (1.0 e + 3 MPa m² s kg⁻¹) that equalled the vascular resistance (R_v). The contribution of the non-vascular leaf resistance (R_nv) to R_leaf was of 71.9% in DL and of 58.4% in LL. The dominant R_nv was shown to be reversibly modulated by mercurials, suggesting that aquaporins play a role in diurnal changes of K_leaf.     

Effects of temperature on leaf hydraulic architecture of tobacco plants

Main Conclusion

Modifications in leaf anatomy of tobacco plants induced greater leaf water transport capacity, meeting greater transpirational demands and acclimating to warmer temperatures with a higher vapor pressure deficit. Temperature is one of the most important environmental factors affecting photosynthesis and growth of plants. However, it is not clear how it may alter leaf hydraulic architecture. We grew plants of tobacco (Nicotiana tabacum) ‘k326’ in separate glasshouse rooms set to different day/night temperature conditions: low (LT 24/18 °C), medium (MT 28/22 °C), or high (HT 32/26 °C). After 40 days of such treatment, their leaf anatomies, leaf hydraulics, photosynthetic rates, and instantaneous water-use efficiency (WUE_i) were measured. Compared with those under LT, plants exposed to HT or MT conditions had significantly higher values for minor vein density (MVD), stomatal density (SD), leaf area, leaf hydraulic conductance (K _leaf), and light-saturated photosynthetic rate (A _sat), but lower values for leaf water potential (ψ _l) and WUE_i. However, those parameters did not differ significantly between HT and MT conditions. Correlation analyses demonstrated that SD and K _leaf increased in parallel with MVD. Moreover, greater SD and K _leaf were partially associated with accelerated stomatal conductance. And then stomatal conductance was positively correlated with A _sat. Therefore, under well-watered, fertilized conditions, when relative humidity was optimal, changes in leaf anatomy seemed to facilitate the hydraulic acclimation to higher temperatures, meeting greater transpirational demands and contributing to the maintenance of great photosynthetic rates. Because transpiration rate increased more with temperature than photosynthetic rate, WUE_i reduced under warmer temperatures. Our results indicate that the modifications of leaf hydraulic architecture are important anatomical and physiological strategies for tobacco plants acclimating to warmer temperatures under a higher vapor pressure deficit.