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.     

Correlation between the Maintenance of Photosynthesis and in Situ Protoplast Volume at Low Water Potentials in Droughted Wheat

Studies were undertaken to examine the relationship between water deficit effects on photosynthesis and the extent of protoplast volume reduction which occurs in leaves at low water potential (Ψ_w). This relationship was monitored in two cultivars (`Condor' and `Capelle Desprez') of cultivated wheat (Triticum aestivum) that differed in sensitivity to drought, and in a wild relative of cultivated wheat (Triticum kotschyi) that has been previously found to be `drought resistant.' When subjected to periods of water stress, Condor and T. kotschyi plants underwent osmotic adjustment; Capelle plants did not. Photosynthetic capacity was maintained to different extents in the three genotypes as leaf Ψ_w declined during stress; Capelle plants were most severely affected. Calculations of internal leaf [CO₂] and stomatal conductance from gas exchange measurements indicated that differences in photosynthetic inhibition at low Ψ_w among the genotypes were primarily due to nonstomatal effects. The extent of protoplast volume reduction that occurred in leaves at low Ψ_w was also found to be different in the three genotypes; maintenance of protoplast volume and photosynthetic capacity in stressed plants of the genotypes appeared to be correlated. When the extent of water stress-induced inhibition of photosynthesis was plotted as a function of declining protoplast volume, this relationship appeared identical for the three genotypes. It was concluded that there is a correlative association between protoplast volume and photosynthetic capacity in leaves of wheat plants subjected to periods of water stress.     

Photophosphorylation in Attached Leaves of Helianthus annuus at Low Water Potentials

The in situ response of photophosphorylation and coupling factor activity to low leaf water potential (ψ_L) was investigated using kinetic spectroscopy to measure the flash-induced electrochromic absorption change in attached sunflower (Helianthus annuus L. cv IS894) leaves. The electrochromic change is caused by the formation of an electric potential across the thylakoid membrane associated with proton uptake. Since depolarization of the thylakoid membrane following flash excitation is normally dominated by proton efflux through the coupling factor during ATP formation, this measurement can provide direct information about the catalytic activity of the coupling factor. Under low ψ_L conditions in which a clear nonstomatal limitation of net photosynthesis could be demonstrated, we found a strong inhibition of coupling factor activity in dark-adapted leaves which was probably caused by an increase in the energetic threshold for the activation of the enzyme at low ψ_L. While this result supported earlier in vitro findings, we further discovered that the light-dependent reduction of coupling factor reversed any observable effect of low ψ_L on the energetics of activation or on photophosphorylation competence. Furthermore, coupling factor was reduced, even in severely droughted sunflower, almost immediately upon illumination. Based on these measurements, we conclude that the nonstomatal limitation of photosynthesis observed by us and others in droughted plants cannot be explained by impaired coupling factor activity.     

Limitations of Photosynthesis in Pinus taeda L. (Loblolly Pine) at Low Soil Temperatures

The relative importance of stomatal and nonstomatal limitations to net photosynthesis (A) and possible signals responsible for stomatal limitations were investigated in unhardened Pinus taeda seedlings at low soil temperatures. After 2 days at soil temperatures between 13 and 7°C, A was reduced by 20 to 50%, respectively. The reduction in A at these moderate root-chilling conditions appeared to be the result of stomatal limitations, based on the decrease in intercellular CO₂ concentrations (c_i). This conclusion was supported by A versus c_i analysis and measurements of O₂ evolution at saturating CO₂, which suggested increases in stomatal but not biochemical limitations at these soil temperatures. Nonuniform stomatal apertures, which were demonstrated with abscisic acid, were not apparent 2 days after root chilling, and results of our A versus c_i analysis appear valid. Bulk shoot water potential (ψ) declined as soil temperature dropped below 16°C. When half the root system of seedlings was chilled, shoot ψ and gas-exchange rates did not decline. Thus, nonhydraulic root-shoot signals were not implicated in stomatal limitations. The initial decrease in leaf conductance to water vapor after root chilling appeared to precede any detectable decrease in bulk fascicle ψ, but may be in response to a decrease in turgor of epidermal cells. These reductions in leaf conductance to water vapor, which occurred within 30 minutes of root chilling, could be delayed and temporarily reversed by reducing the leaf-to-air vapor-pressure deficit, suggesting that hydraulic signals may be involved in initiating stomatal closure. By independently manipulating the leaf-to-air vapor-pressure deficit of individual fascicles, we could induce uptake of water vapor through stomata, suggesting that nonsaturated conditions occur in the intercellular airspaces. There was an anomaly in our results on seedlings maintained for 2 days at soil temperatures below 7°C. Lower A appeared primarily the result of nonstomatal limitations, based on large increases in calculated c_i and A versus c_i analysis. In contrast, measurements of O₂ evolution at saturating CO₂ concentrations implied nonstomatal limitations per se did not increase at these temperatures. One explanation for this paradox is that calculations of c_i are unreliable at very low gas-exchange rates because of inadequate measurement resolution, and limitations of A are predominantly stomatal. An alternative interpretation is that increases in c_i are real and the results from O₂-evolution measurements are in error. The high CO₂ concentration used in O₂-evolution measurements (15%) may have overcome nonstomatal limitations by enzymes that were down-regulated by a feedback mechanism. In this scenario, carbohydrate feedback limitations may be responsible for nonstomatal reductions in A after 2 days at soil temperatures below 7°C.     

Osmotic Adjustment in Leaves of VA Mycorrhizal and Nonmycorrhizal Rose Plants in Response to Drought Stress

Osmotic adjustment in Rosa hybrida L. cv Samantha was characterized by the pressure-volume approach in drought-acclimated and unacclimated plants brought to the same level of drought strain, as assayed by stomatal closure. Plants were colonized by either of the vesicular-arbuscular mycorrhizal fungi Glomus deserticola Trappe, Bloss and Menge or G. intraradices Schenck and Smith, or were nonmycorrhizal. Both the acclimation and the mycorrhizal treatments decreased the osmotic potential (Ψ_π) of leaves at full turgor and at the turgor loss point, with a corresponding increase in pressure potential at full turgor. Mycorrhizae enabled plants to maintain leaf turgor and conductance at greater tissue water deficits, and lower leaf and soil water potentials, when compared with nonmycorrhizal plants. As indicated by the Ψ_π at the turgor loss point, the active Ψ_π depression which attended mycorrhizal colonization alone was 0.4 to 0.6 megapascals, and mycorrhizal colonization and acclimation in concert 0.6 to 0.9 megapascals, relative to unacclimated controls without mycorrhizae. Colonization levels and sporulation were higher in plants subjected to acclimation. In unacclimated hosts, leaf water potential, water saturation deficit, and soil water potential at a particular level of drought strain were affected most by G. intraradices. G. deserticola had the greater effect after drought preconditioning.     

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.     

Risk-taking plants: Anisohydric behavior as a stress-resistance trait

Water scarcity is a critical limitation for agricultural systems. Two different water management strategies have evolved in plants: an isohydric strategy and an anisohydric strategy. Isohydric plants maintain a constant midday leaf water potential (Ψ_leaf) when water is abundant, as well as under drought conditions, by reducing stomatal conductance as necessary to limit transpiration. Anisohydric plants have more variable Ψ_leaf and keep their stomata open and photosynthetic rates high for longer periods, even in the presence of decreasing leaf water potential. This risk-taking behavior of anisohydric plants might be beneficial when water is abundant, as well as under moderately stressful conditions. However, under conditions of intense drought, this behavior might endanger the plant. We will discuss the advantages and disadvantages of these two water-usage strategies and their effects on the plant’s ability to tolerate abiotic and biotic stress. The involvement of plant tonoplast AQPs in this process will also be discussed.     

Leaf gas exchange and water potential responses to drought in nine poplar (Populus spp.) clones with contrasting drought tolerance

The dynamic responses of stomatal conductance (g _s) net photosynthesis (A) and leaf water potential (Ψ_leaf) to a progressive drought were examined in nine poplar clones (Populus spp.) with contrasting drought tolerance from the Canadian Prairies, a region prone to frequent droughts. Plants were grown in a greenhouse and either well-watered or drought preconditioned (5–6 cycles of drought) for 8 weeks. At the end of the last cycle, plants were watered to saturation then progressively dried-down (−1.25 MPa Ψ_soil) during which A, g _s and Ψ_leaf were measured. Drought tolerant Okanese reached the lowest combined Ψ_leaf while sensitive clones (Assiniboine and Imperial) had the highest (−1.6 vs. −1.1 MPa). Steady state g _s (measured under well watered conditions) was lower in tolerant (Okanese and Tristis SBC#1) than in sensitive clones. Preconditioning reduced steady state g _s in all clones, lowered the threshold Ψ_leaf for stomatal closure and the minimum Ψ_leaf in most clones but did not affect the steady state A. Tolerant and some moderately tolerant clones maintained higher A at lower Ψ_leaf than the other clones. Stomatal closure was gradual in tolerant clones and in moderately tolerant Northwest but rapid in the other clones. Stomata in the sensitive clones closed at the highest Ψ_leaf, Okanese closed at the lowest. The substantial range in gas exchange and Ψ_leaf responses observed here represented both drought tolerance and taxonomic (Aegiros or Tacamahaca sections) traits which could play a role in the survival and productivity in environments with limited water or during periods of drought.     

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.     

Stomatal and nonstomatal limitations of photosynthesis in 19 temperate tree species on contrasting sites during wet and dry years

A unique approach was used to evaluate stomatal and nonstomatal constraints to photosynthesis in 19 naturally occurring, deciduous tree species on xeric, mesic and wetmesic sites in central Pennsylvania, USA, during relatively wet (1990) and dry (1991) growing seasons. All species exhibited significantly decreased stomatal conductance to CO₂ (g_c) in 1991 compared to 1990. The mesic species had drought related decreases in photosynthesis (A) attributed primarily to increased absolute stomatal limitation to A (L_g), whereas in the wet-mesic species, the absolute mesophyll limitation (Lm) was at least as important as L_g in limiting A during drought. The xeric species maintained relatively high A during drought despite decreased g_c. In the xeric and mesic species, L_m decreased and L_g increased during drought due to stomatal closure. From xeric to mesic to wet-mesic, the relative stomatal limitation (I_g) generally decreased faster, and relative mesophyll limitations to A increased faster, with increasing g_c suggesting greater photosynthetic capacity (i.e. greater potential maximum A) with increasing drought tolerance rank of species. Few species exhibited a significant drought-related decrease in photosynthetic capacity. The results of this landscape-based study indicate that the interaction of stomatal and nonstomatal limitations of A vary in a manner consistent with species' drought tolerance and site conditions, and that nonstomatal constraints to A in field plants during a moderate, season-long drought were generally not as severe as reported in controlled studies.     

Drought stress revealed physiological,biochemical and gene-expressional variations in ‘Yoshihime’ peach (Prunus Persica L) cultivar

It is indispensable to comprehend the mechanism that regulates plant responses to drought conditions to intensify the water use efficiency of stone fruits. The physiological, biochemical and molecular responses of drought-treated peach leaves were investigated. Results revealed that drought-treated plants manifested a significant attenuation in water potential as compared to control plants. Furthermore, sorbitol and proline contents were accumulated contrary to glucose, fructose, and sucrose that were dwindled significantly throughout the drought period. Similarly, the activities of antioxidant enzymes and expression pattern of related genes were hoisted to counter the lipid peroxidation in drought-treated plants. Moreover, reduced stomatal conductance has repressed the photosynthesis process and linked genes during drought stress. The expression level of regulatory genes (dehydration-responsive element-bindings and WRKYs) exhibited up-regulation in the drought-treated group. Overall, this study asserts that ‘Yoshihime’ peach cultivar possesses unique physiological, biochemical, and molecular responses under different spells of drought stress.     

Acclimation of photosynthesis to low leaf water potentials

Photosynthesis is reduced at low leaf water potentials (Ψ_l) but repeated water deficits can decrease this reduction, resulting in photosynthetic acclimation. The contribution of the stomata and the chloroplasts to this acclimation is unknown. We evaluated stomatal and chloroplast contributions when soil-grown sunflower (Helianthus annuus L.) plants were subjected to water deficit pretreatments for 2 weeks. The relationship between photosynthesis and Ψ_l, determined from gas-exchange and isopiestic thermocouple psychometry, was shifted 3 to 4 bars towards lower Ψ_l, in pretreated plants. Leaf diffusive resistance was similarly affected. Chloroplast activity, demonstrated in situ with measurements of quantum yield and the capacity to fix CO₂ at all partial pressures of CO₂, and in vitro by photosystem II activity of isolated organelles, was inhibited at low Ψ_l but less in pretreated plants than in control plants. The magnitude of this inhibition indicated that decreases in chloroplast activity contributed more than closure of stomata both to losses in photosynthesis and to the acclimation of photosynthesis to low Ψ_l.     

Effects of Water-Deficit Stress on Photosynthesis, Its Components and Component Limitations, and on Water Use Efficiency in Wheat (Triticum aestivum L.)

It is of theoretical as well as practical interest to identify the components of the photosynthetic machinery that govern variability in photosynthesis rate (A) and water-use efficiency (WUE), and to define the extent by which the component processes limit A and WUE during developing water-deficit stress. For that purpose, leaf exchange of CO₂ and H₂O was determined in two growth-chamber-grown wheat cultivars (Triticum aestivum L. cv TAM W-101 and cv Sturdy), and the capacity of A was determined and broken down into carboxylation efficiency (c.e.), light- and CO₂-saturated A, and stomatal conductance (g_s) components. The limitations on A measured at ambient CO₂ concentration (A₃₅₀) were estimated. No cultivar difference was observed when A₃₅₀ was plotted versus leaf water potential (Ψ_w). Light- and CO₂-saturated A, c.e., and g_s decreased with decreasing leaf Ψ_w, but of the corresponding photosynthesis limitations only those caused by insufficient c.e. and g_s increased. Thus, reduced stomatal aperture and Calvin cycle activity, but not electron transport/photophosphorylation, appeared to be major reasons for drought stress-induced inhibition of A₃₅₀. WUE measured as A₃₅₀/g_s first increased with stomatal closure down to a g_s of about 0.25 mol H₂O m⁻² s⁻¹ (Ψ_w = −1.6 MPa). However, it was predicted that A₃₅₀/g_s would decrease with more severe stress due to inhibition of c.e.     

Leaf water potential, stomatal resistance, and photosynthetic response to water stress in peach seedlings

Individual groups of peach (Prunus persica [L.] Batsch) seedlings stressed to −17, −26 and −36 bars recovered to control levels within 1, 3, and 4 days, respectively. Stomatal resistance was significantly correlated with both leaf water potential and net photosynthesis. In seedlings stressed to −52 bars, leaf water potential and stomatal resistance recovered sooner than net photosynthesis, despite recovery of 0₂ evolution at a rate similar to leaf water potential. Therefore, some nonstomatal factor other than reduction in photochemical activity must be responsible for the lag in recovery of CO₂ assimilation following irrigation.     

Ecology of SO2 resistance: II. Photosynthetic changes of shrubs in relation to SO2 absorption and stomatal behavior

Summary In an effort to predict SO₂ sensitivity of plants from their morphological and physiological features, the effects of SO₂ on photosynthesis were partitioned between stomatal and nonstomatal components for a drought deciduous shrub, Diplacus aurantiacus, and an evergreen shrub, Heteromeles arbutifolia. As predicted, the drought deciduous shrub had the higher gas conductance, and hence SO₂ absorptance. However, nonstomatal components also play a role in determining SO₂ sensitivity. Apparently a plant with a high intrinsic photosynthetic capacity will be more sensitive to SO₂ than one with a lower capacity.     

Chlorophyll a Fluorescence and Photosynthetic and Growth Responses of Pinus radiata to Phosphorus Deficiency, Drought Stress, and High CO(2)

Needles from phosphorus deficient seedlings of Pinus radiata D. Don grown for 8 weeks at either 330 or 660 microliters CO₂ per liter displayed chlorophyll a fluorescence induction kinetics characteristic of structural changes within the thylakoid chloroplast membrane, i.e. constant yield fluorescence (F_O) was increased and induced fluorescence ([F_P-F_I]/F_O) was reduced. The effect was greatest in the undroughted plants grown at 660 μl CO₂ L⁻¹. By week 22 at 330 μl CO₂ L⁻¹ acclimation to P deficiency had occurred as shown by the similarity in the fluorescence characteristics and maximum rates of photosynthesis of the needles from the two P treatments. However, acclimation did not occur in the plants grown at 660 μl CO₂ L⁻¹. The light saturated rate of photosynthesis of needles with adequate P was higher at 660 μl CO₂ L⁻¹ than at 330 μl CO₂ L⁻¹, whereas photosynthesis of P deficient plants showed no increase when grown at the higher CO₂ concentration. The average growth increase due to CO₂ enrichment was 14% in P deficient plants and 32% when P was adequate. In drought stressed plants grown at 330 μl CO₂ L⁻¹, there was a reduction in the maximal rate of quenching of fluorescence (R_Q) after the major peak. Constant yield fluorescence was unaffected but induced fluorescence was lower. These results indicate that electron flow subsequent to photosystem II was affected by drought stress. At 660 μl CO₂ L⁻¹ this response was eliminated showing that CO₂ enrichment improved the ability of the seedlings to acclimate to drought stress. The average growth increase with CO₂ enrichment was 37% in drought stressed plants and 19% in unstressed plants.     

Stomatal and nonstomatal limitations of photosynthesis in relation to the drought and shade tolerance of tree species in open and understory environments

 Light saturated photosynthesis (A) in field saplings of shade tolerant, intermediate, and intolerant tree species was analyzed for stomatal and nonstomatal limitations to test differences between species and sun and shade phenotypes during drought. Throughout the study, photosynthesis was highest and mesophyll limitations of A (L_m) lowest in the intolerant species in both open and understory habitats. The shade tolerant species exhibited the only drought-related decreased A and increased L_m in the open, and the greatest drought-related decreased A and increased L_m in the understory. Few species exhibited significant habitat or drought-related differences in stomatal conductance to CO₂ (g_c), but even slight decreases in g_c during drought were associated with large increases in stomatal limitations to A (L_g). Combined changes in L_m and L_g resulted in increased relative stomatal limitation to A (l _g) in several species during drought. Nevertheless, the overall lack of stomatal closure allowed for nonstomatal limitations to play a major role in reduced A during drought. Higher leaf N was associated with shallower slope of the l _g versus g_c relationship, an indication of greater A capacity. Photosynthetic capacity tended to be greater in the intolerant species than the tolerant species, and it tended to decrease during drought primarily in the shade tolerant species in the understory. Findings in the literature suggest that carbon reduction reactions may be more susceptible to drought than photosynthetic light reactions. If so, reduced carbon reduction capacity of shade tolerant species or shade phenotypes may predispose them to drought conditions, which suggests a mechanism behind the well-recognized tradeoff between drought tolerance and shade tolerance of temperate tree species. Received: 20 October 1995 / Accepted: 20 February 1996     

Osmotic adjustment, symplast volume, and nonstomatally mediated water stress inhibition of photosynthesis in wheat

At low water potential (ψ_w), dehydration reduces the symplast volume of leaf tissue. The effect of this reduction on photosynthetic capacity was investigated. The influence of osmotic adjustment on this relationship was also examined. To examine these relationships, comparative studies were undertaken on two wheat cultivars, one that osmotically adjusts in response to water deficits (`Condor'), and one that lacks this capacity (`Capelle Desprez'). During a 9-day stress cycle, when water was withheld from plants grown in a growth chamber, the relative water content of leaves declined by 30% in both cultivars. Leaf osmotic potential (ψ_s) declined to a greater degree in Condor plants. Measuring ψ_s at full turgor indicated that osmotic adjustment occurred in stressed Condor, but not in Capelle plants. Two methods were used to examine the degree of symplast (i.e. protoplast) volume reduction in tissue rapidly equilibrated to increasingly low ψ_w. Both techniques gave similar results. With well-watered plants, symplast volume reduction from the maximum (found at high ψ_w for each cultivar) was the same for Condor and Capelle. After a stress cycle, volume was maintained to a greater degree at low ψ_w in Condor leaf tissue than in Capelle. Nonstomatally controlled photosynthesis was inhibited to the same degree at low ψ_w in leaf tissue prepared from well-watered Condor and Capelle plants. However, photosynthetic capacity was maintained to a greater degree at low ψ_w in tissue prepared from stressed Condor plants than in tissue from stressed Capelle plants. Net CO₂ uptake in attached leaves was monitored using an infrared gas analyzer. These studies indicated that in water stressed plants, photosynthesis was 106.5% higher in Condor than Capelle at ambient [CO₂] and 21.8% higher at elevated external [CO₂]. The results presented in this report were interpreted as consistent with the hypothesis that there is a causal association between protoplast (and presumably chloroplast) volume reduction at low ψ_w and low ψ_w inhibition of photosynthesis. Also, the data indicate that osmotic adjustment allows for maintenance of relatively greater volume at low ψ_w, thus reducing low ψ_w inhibition of chloroplast photosynthetic potential.     

Internal CO(2) Measured Directly in Leaves : Abscisic Acid and Low Leaf Water Potential Cause Opposing Effects

Observations of nonuniform photosynthesis across leaves cast doubt on internal CO₂ partial pressures (p_i) calculated on the assumption of uniformity and can lead to incorrect conclusions about the stomatal control of photosynthesis. The problem can be avoided by measuring p_i directly because the assumptions of uniformity are not necessary. We therefore developed a method that allowed p_i to be measured continuously in situ for days at a time under growth conditions and used it to investigate intact leaves of sunflower (Helianthus annuus L.), soybean (Glycine max L. Merr.), and bush bean (Phaseolus vulgaris L.) subjected to high or low leaf water potentials (ψ_w) or high concentrations of abscisic acid (ABA). The leaves maintained a relatively constant differential (Δp) between ambient CO₂ and measured p_i throughout the light period when water was supplied. When water was withheld, ψ_w decreased and the stomata began to close, but measured p_i increased until the leaf reached a ψ_w of −1.76 (bush bean), −2.12 (sunflower) or −3.10 (soybean) megapascals, at which point Δp = 0. The increasing p_i indicated that stomata did not inhibit CO₂ uptake and a Δp of zero indicated that CO₂ uptake became zero despite the high availability of CO₂ inside the leaf. In contrast, when sunflower leaves at high ψ_w were treated with ABA, p_i did not increase and instead decreased rapidly and steadily for up to 8 hours even as ψ_w increased, as expected if ABA treatment primarily affected stomatal conductance. The accumulating CO₂ at low ψ_w and contrasting response to ABA indicates that photosynthetic biochemistry limited photosynthesis at low ψ_w but not at high ABA.     

Chemical and hydraulic signals regulate stomatal behavior and photosynthetic activity in maize during progressive drought

The objectives of this study were to investigate stomatal regulation in maize seedlings during progressive soil drying and to determine the impact of stomatal movement on photosynthetic activity. In well-watered and drought-stressed plants, leaf water potential (Ψ _leaf), relative water content (RWC), stomatal conductance (g _s), photosynthesis, chlorophyll fluorescence, leaf instantaneous water use efficiency (iWUE_leaf), and abscisic acid (ABA) and zeatin-riboside (ZR) accumulation were measured. Results showed that g _s decreased significantly with progressive drought and stomatal limitations were responsible for inhibiting photosynthesis in the initial stages of short-term drought. However, after 5 days of withholding water, non-stomatal limitations, such as damage to the PSII reaction center, became the main limiting factor. Stomatal behavior was correlated with changes in both hydraulic and chemical signals; however, changes in ABA and ZR occurred prior to any change in leaf water status. ABA in leaf and root tissue increased progressively during soil drying, and further analysis found that leaf ABA was negatively correlated with g _s (R ² = 0.907, p < 0.05). In contrast, leaf and root ZR decreased gradually. ZR in leaf tissue was positively correlated with g _s (R ² = 0.859, p < 0.05). These results indicate that ABA could induce stomatal closure, and ZR works antagonistically against ABA in stomatal behavior. In addition, the ABA/ZR ratio also had a strong correlation with g _s, suggesting that the combined chemical signal (the interaction between ABA and cytokinin) plays a role in coordinating stomatal behavior. In addition, Ψ _leaf and RWC decreased significantly after only 3 days of drought stress, also affecting stomatal behavior.