Most stomatal closure in woody species under moderate drought can be explained by stomatal responses to leaf turgor.

Reduced stomatal conductance (gs ) during soil drought in angiosperms may result from effects of leaf turgor on stomata and/or factors that do not directly depend on leaf turgor, including root-derived abscisic acid (ABA) signals. To quantify the roles of leaf turgor-mediated and leaf turgor-independent mechanisms in gs decline during drought, we measured drought responses of gs and water relations in three woody species (almond, grapevine and olive) under a range of conditions designed to generate independent variation in leaf and root turgor, including diurnal variation in evaporative demand and changes in plant hydraulic conductance and leaf osmotic pressure. We then applied these data to a process-based gs model and used a novel method to partition observed declines in gs during drought into contributions from each parameter in the model. Soil drought reduced gs by 63-84% across species, and the model reproduced these changes well (r(2) = 0.91, P < 0.0001, n = 44) despite having only a single fitted parameter. Our analysis concluded that responses mediated by leaf turgor could explain over 87% of the observed decline in gs across species, adding to a growing body of evidence that challenges the root ABA-centric model of stomatal responses to drought.

Leaf-to-branch scaling of C-gain in field-grown almond trees under different soil moisture regimes.

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 CO2 assimilation was quantified by a global scaling factor (fg), defined as the product of two specific scaling factors: (i) a structural scaling factor (fs), determined under well-watered conditions, mainly involving leaf mutual shading; and (ii) a water stress scaling factor (fws,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 fs ∼0.33) and close to the projected-to-total leaf area ratio of almond branches (ε = 0.31), while the contribution of water stress mechanisms was moderate (mean fws,b ∼0.85), thus supplying an fg 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.

Effects of water stress on irradiance acclimation of leaf traits in almond trees.

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

Disentangling the contributions of ontogeny and water stress to photosynthetic limitations in almond trees.

Very few studies have attempted to disentangle the respective role of ontogeny and water stress on leaf photosynthetic attributes. The relative significance of both effects on photosynthetic attributes has been investigated in leaves of field-grown almond trees [Prunus dulcis (Mill.) D. A. Webb] during four growth cycles. Leaf ontogeny resulted in enhanced leaf dry weight per unit area (W(a)), greater leaf dry-to-fresh weight ratio and lower N content per unit of leaf dry weight (N(w)). Concomitantly, area-based maximum carboxylation rate (V(cmax)), maximum electron transport rate (J(max)), mesophyll conductance to CO2 diffusion (gm)' and light-saturated net photosynthesis (A(max)) declined in both well-watered and water-stressed almond leaves. Although g(m) and stomatal conductance (g(s)) seemed to be co-ordinated, a much stronger coordination in response to ontogeny and prolonged water stress was observed between g(m) and the leaf photosynthetic capacity. Under unrestricted water supply, the leaf age-related decline of A(max) was equally driven by diffusional and biochemical limitations. Under restricted soil water availability, A(max) was mainly limited by g(s) and, to a lesser extent, by photosynthetic capacity and g(m). When both ontogeny and water stress effects were combined, diffusional limitations was the main determinant of photosynthesis limitation, while stomatal and biochemical limitations contributed similarly.

Partial rootzone drying improves almond tree leaf-level water use efficiency and afternoon water status compared with regulated deficit irrigation.

To determine whether partial rootzone drying (PRD) optimised leaf gas exchange and soil-plant water relations in almond (Prunus dulcis (Mill.) D.A. Webb) compared with regulated deficit irrigation (RDI), a 2 year trial was conducted on field-grown trees in a semiarid climate. Five irrigation treatments were established: full irrigation (FI) where the trees were irrigated at 100% of the standard crop evapotranspiration (ETc); three PRD treatments (PRD70, PRD50 and PRD30) that applied 70, 50 and 30% ETc, respectively; and a commercially practiced RDI treatment that applied 50% ETc during the kernel-filling stage and 100% ETc during the remainder of the growth season. Measurements of volumetric soil moisture content in the soil profile (0-100cm), predawn leaf water potential (Ψpd), midday stem water potential (Ψms), midday leaf gas exchange and trunk diameter fluctuations (TDF) were made during two growing seasons. The diurnal patterns of leaf gas exchange and stem water potential (Ψs) were appraised during the kernel-filling stage in all irrigation regimes. When tree water relations were assessed at solar noon, PRD did not show differences in either leaf gas exchange or tree water status compared with RDI. At similar average soil moisture status (adjudged by similar Ψpd), PRD50 trees had higher water status than RDI trees in the afternoon, as confirmed by Ψs and TDF. Although irrigation placement showed no effects on diurnal stomatal regulation, diurnal leaf net photosynthesis (Al) was substantially less limited in PRD50 than in RDI trees, indicating that PRD improved leaf-level water use efficiency.

Root water potential integrates discrete soil physical properties to influence ABA signalling during partial rootzone drying.

To investigate the influence of different growing substrates (two mineral, two organic) on root xylem ABA concentration ([ABA](root)) and the contribution of the drying root system to total sap flow during partial rootzone drying (PRD), sunflower (Helianthus annuus L.) shoots were grafted onto the root systems of two plants grown in separate pots. Sap flow through each hypocotyl was measured below the graft union when one pot ('wet') was watered and other ('dry') was not. Each substrate gave unique relationships between dry pot matric potential (Psi(soil)), volumetric water content ((v)) or penetrometer resistance (Q) and either the fraction of photoperiod sap flow from roots in drying soil or [ABA](root). However, decreased relative sap flow, and increased [ABA](root), from roots in drying soil varied with root water potential (Psi(root)) more similarly across a range of substrates. The gradient between Psi(soil) and Psi(root) was greater in substrates with high sand or peat proportions, which may have contributed to a more sensitive response of [ABA](root) to Psi(soil) in these substrates. Whole plant transpiration was most closely correlated with the mean Psi(soil) of both pots, and then with detached leaf xylem ABA concentration. Although Psi(root) best predicted decreased relative sap flow, and increased [ABA](root), from roots in drying soil across a range of substrates, the inaccessibility of this variable in field studies requires a better understanding of how measurable soil variables (Psi(soil), (v), Q) affect Psi(root).

Seasonal effects of deficit irrigation on leaf photosynthetic traits of fruiting and non-fruiting shoots in almond trees.

We investigated seasonal trends in, and relationships between, leaf structural properties, leaf nitrogen concentration, and maximum (A(m)) and potential (A(p)) leaf net CO(2) assimilation of 1-year-old fruiting (f) and current-year non-fruiting (nf) shoots in 5-year-old almond trees (Prunus dulcis (Mill.) D.A. Webb cv Marta). These trees had been subjected in the previous 4 years to either full irrigation (FI regime) or sustained deficit irrigation (DI) at 50% of standard crop evapotranspiration during the entire growing season (DI regime) in the semiarid climate of southeast Spain. Measurements were made during an entire growing season on sun-exposed leaves. Leaf dry mass per unit area (W(a)), area and dry-mass-based leaf N concentrations (N(a) and N(w), respectively), and area and dry-mass-based A(m) (A(ma) and A(mw), respectively) were lower in f-leaves than in nf-leaves. Changes in leaf structural attributes induced by DI were more pronounced in nf-leaves than in f-leaves, the latter being little affected. Over the entire growth season, A(m) and A(p) were correlated negatively with W(a) and positively with N(w) for both the leaf classes and the irrigation regimes. When calculated with respect to total leaf N concentration, maximum photosynthetic nitrogen-use efficiency (PNUE(m)) was significantly higher in f-leaves than in nf-leaves, with no significant differences between the leaf classes among the irrigation regimes. However, when PNUE(m) was calculated with respect to photosynthetic N, no significant effect of leaf class or irrigation regime was observed. Overall, our results showed that DI and FI trees exhibited similar seasonal patterns of leaf structural properties and maximum and potential leaf net CO(2) assimilation rates, but there were distinct N-allocation patterns between f- and nf-leaves. In the DI treatment, leaf structural adjustments appeared to operate to maintain a high N status in the leaves of fruit-bearing shoots, to the detriment of N resources allocated to vegetative shoots.

Accounting for sap flow from different parts of the root system improves the prediction of xylem ABA concentration in plants grown with heterogeneous soil moisture.

When soil moisture is heterogeneous, sap flow from, and ABA status of, different parts of the root system impact on leaf xylem ABA concentration ([X-ABA]leaf). The robustness of a model for predicting [X-ABA]leaf was assessed. 'Two root-one shoot' grafted sunflower (Helianthus annuus L.) plants received either deficit irrigation (DI, each root system received the same irrigation volumes) or partial rootzone drying (PRD, only one root system was watered and the other dried the soil). Irrespective of whether relative sap flow was assessed using sap flow sensors in vivo or by pressurization of de-topped roots, each root system contributed similarly to total sap flow during DI, while sap flow from roots in drying soil declined linearly with soil water potential (Psisoil) during PRD. Although Psisoil of the irrigated pot determined the threshold Psisoil at which sap flow from roots in drying soil decreased, the slope of this decrease was independent of the wet pot Psisoil. Irrespective of whether sap was collected from the wet or dry root system of PRD plants, or a DI plant, root xylem ABA concentration increased as Psisoil declined. The model, which weighted ABA contributions of each root system according to the sap flow from each, almost perfectly explained [X-ABA] immediately above the graft union. That the model overestimated measured [X-ABA]leaf may result from changes in [X-ABA] along the transport pathway or an artefact of collecting xylem sap from detached leaves. The implications of declining sap flow through partially dry roots during PRD for the control of stomatal behaviour and irrigation scheduling are discussed.

Abscisic acid signalling when soil moisture is heterogeneous: decreased photoperiod sap flow from drying roots limits abscisic acid export to the shoots.

To investigate the contribution of different parts of the root system to total sap flow and leaf xylem abscisic acid (ABA) concentration ([X-ABA](leaf)), individual sunflower (Helianthus annuus L.) shoots were grafted onto the root systems of two plants grown in separate pots and sap flow through each hypocotyl measured below the graft union. During deficit irrigation (DI), both pots received the same irrigation volumes, while during partial root zone drying (PRD) one pot ('wet') was watered and another ('dry') was not. During PRD, once soil water content (theta) decreased below a threshold, the fraction of sap flow from drying roots declined. As theta declined, root xylem ABA concentration increased in both irrigation treatments, and [X-ABA](leaf) increased in DI plants, but [X-ABA](leaf) of PRD plants actually decreased within a certain theta range. A simple model that weighted ABA contributions of wet and dry root systems to [X-ABA](leaf) according to the sap flow from each, better predicted [X-ABA](leaf) of PRD plants than either [X-ABA](dry), [X-ABA](wet) or their mean. Model simulations revealed that [X-ABA](leaf) during PRD exceeded that of DI with moderate soil drying, but continued soil drying (such that sap flow from roots in drying soil ceased) resulted in the opposite effect.