Steady-state Stomatal Conductance Research Articles

NaCl affects photosynthetic and stomatal dynamics by osmotic effects and reduces photosynthetic capacity by ionic effects in tomato.

NaCl stress affects stomatal behavior and photosynthesis by a combination of osmotic and ionic components, but it is unknown how these components affect stomatal and photosynthetic dynamics. Tomato (Solanum lycopersicum) plants were grown in a reference nutrient solution [control; electrical conductivity (EC)=2.3 dS m-1], a solution containing additional macronutrients (osmotic effect; EC=12.6 dS m-1), or a solution with additional 100mM NaCl (osmotic and ionic effects; EC=12.8 dS m-1). Steady-state and dynamic photosynthesis, and leaf biochemistry, were characterized throughout leaf development. The osmotic effect decreased steady-state stomatal conductance while speeding up stomatal responses to light intensity shifts. After 19 d of treatment, photosynthetic induction was reduced by the osmotic effect, which was attributable to lower initial stomatal conductance due to faster stomatal closing under low light. Ionic effects of NaCl were barely observed in dynamic stomatal and photosynthetic behavior, but led to a reduction in leaf photosynthetic capacity, CO2 carboxylation rate, and stomatal conductance in old leaves after 26 d of treatment. With increasing leaf age, rates of light-triggered stomatal movement and photosynthetic induction decreased across treatments. We conclude that NaCl impacts dynamic stomatal and photosynthetic kinetics by osmotic effects and reduces photosynthetic capacity by ionic effects.

Combined action of guard cell plasma membrane rapid- and slow-type anion channels in stomatal regulation

Initiation of stomatal closure by various stimuli requires activation of guard cell plasma membrane anion channels, which are defined as rapid (R)- and slow (S)-type. The single-gene loss-of-function mutants of these proteins are well characterized. However, the impact of suppressing both the S- and R-type channels has not been studied. Here, by generating and studying double and triple Arabidopsis thaliana mutants of SLOW ANION CHANNEL1 (SLAC1), SLAC1 HOMOLOG3 (SLAH3), and ALUMINUM-ACTIVATED MALATE TRANSPORTER 12/QUICK-ACTIVATING ANION CHANNEL 1 (QUAC1), we show that impairment of R- and S-type channels gradually increased whole-plant steady-state stomatal conductance. Ozone-induced cell death also increased gradually in higher-order mutants with the highest levels observed in the quac1 slac1 slah3 triple mutant. Strikingly, while single mutants retained stomatal responsiveness to abscisic acid, darkness, reduced air humidity, and elevated CO2, the double mutant lacking SLAC1 and QUAC1 was nearly insensitive to these stimuli, indicating the need for coordinated activation of both R- and S-type anion channels in stomatal closure.

Mild reductions in guard cell sucrose synthase 2 expression leads to slower stomatal opening and decreased whole plant transpiration in Nicotiana tabacum L

The understanding of the dynamics of stomatal movements has increased substantially through genetic manipulation of plant metabolism either at the whole plant level or specifically in guard cells. However, the regulation of stomatal speediness remains not completely elucidated. Here we shown that reduced expression of guard cell sucrose synthase 2 (NtSUS2) of Nicotiana tabacum L. altered the topology and the connectivity of the guard cell metabolic network and the accumulation of metabolites positively correlated with stomatal speediness during dark-to-light transition. This leads to a slower light-induced stomatal opening, lower steady-state stomatal conductance and a strong reduction (up to 44 %) in daily whole plant transpiration in the transgenics, when compared to wild type plants. Furthermore, the transgenic lines transpired more or have lower reduction in whole plant transpiration under short water deficit periods, indicating a higher effective use of water under this condition. Our results collectively suggest that the regulation of stomatal movement and speediness involve a complex modulation of the guard cell metabolic network, in which NtSUS2 has an important role. The results are discussed on the role of guard cell metabolism for the regulation of both stomatal speediness and whole plant transpiration.

Dynamic thermal imaging confirms local but not fast systemic ABA responses.

Abscisic acid (ABA) signals regulating stomatal aperture and water loss are usually studied in detached leaves or isolated epidermal peels and at infrequent timepoints. Measuring stomatal ABA responses in attached leaves across a time course enables the study of stomatal behaviour in the physiological context of the plant. Infrared thermal imaging is often used to characterize steady-state stomatal conductance via comparisons of leaf surface temperature but is rarely used to capture stomatal responses over time or across different leaf surfaces. We used dynamic thermal imaging as a robust, but sensitive, tool to observe stomatal ABA responses in a whole plant context. We detected stomatal responses to low levels of ABA in both monocots and dicots and identified differences between the responses of different leaves. Using whole plant thermal imaging, stomata did not always behave as described previously for detached samples: in Arabidopsis, we found no evidence for fast systemic ABA-induced stomatal closure, and in barley, we observed no requirement for exogenous nitrate during ABA-induced stomatal closure. Thus, we recommend dynamic thermal imaging as a useful approach to complement detached sample assays for the study of local and systemic stomatal responses and molecular mechanisms underlying stomatal responses to ABA in the whole plant context.

Steady-state stomatal responses of C3 and C4 species to blue light fraction: Interactions with CO2 concentration.

Blue light induced stomatal opening has been studied by applying a short pulse (~5 to 60 s) of blue light to a background of saturating photosynthetic red photons, but little is known about steady-state stomatal responses. Here we report stomatal responses to blue light at high and low CO2 concentrations. Steady-state stomatal conductance (gs ) of C3 plants increased asymptotically with increasing blue light to a maximum at 20% blue (120 μmol m-2 s-1 ). This response was consistent from 200 to 800 μmol mol-1 atmospheric CO2 (Ca ). In contrast, blue light induced only a transient stomatal opening (~5 min) in C4 species above a Ca of 400 μmol mol-1 . Steady-state gs of C4 plants generally decreased with increasing blue intensity. The net photosynthetic rate of all species decreased above 20% blue because blue photons have lower quantum yield (moles carbon fixed per mole photons absorbed) than red photons. Our findings indicate that photosynthesis, rather than a blue light signal, plays a dominant role in stomatal regulation in C4 species. Additionally, we found that blue light affected only stomata on the illuminated side of the leaf. Contrary to widely held belief, the blue light-induced stomatal opening minimally enhanced photosynthesis and consistently decreased water use efficiency.

Ozone Amplifies Water Loss from Mature Trees in the Short Term But Decreases It in the Long Term

We measured whole-tree transpiration of mature Fagus sylvatica and Picea abies trees exposed to ambient and twice-ambient O3 regimes (1xO3 and 2xO3 free-air fumigation). After eight years, mean daily total transpiration did not vary with the O3 regime over the 31 days of our study, even though individual daily values increased with increasing daily O3 peaks in both species. Although the environmental parameters were similar at 1xO3 and 2xO3, the main factors affecting daily transpiration were vapour pressure deficit in 2xO3 spruce and O3 peaks in beech. For a mechanistic explanation, we measured O3-induced sluggish stomatal responses to variable light (sunflecks) by means of leaf-level gas exchange measurements only in the species where O3 was a significant factor for transpiration, i.e., beech. Stomata were always slower in closing than in opening. The 2xO3 stomata were slower in opening and mostly in closing than 1xO3 stomata, so that O3 uptake and water loss were amplified before a steady state was reached. Such delay in the stomatal reaction suggests caution when assessing stomatal conductance under O3 pollution, because recording gas exchange at the time photosynthesis reached an equilibrium resulted in a significant overestimation of stomatal conductance when stomata were closing (ab. 90% at 1xO3 and 250% at 2xO3). Sun and shade leaves showed similar sluggish responses, thus suggesting that sluggishness may occur within the entire crown. The fact that total transpiration was similar at 1xO3 and 2xO3, however, suggests that the higher water loss due to stomatal sluggishness was offset by lower steady-state stomatal conductance at 2xO3. In conclusion, O3 exposure amplified short-term water loss from mature beech trees by slowing stomatal dynamics, while decreased long-term water loss because of lower steady-state stomatal conductance. Over the short term of this experiment, the two responses offset each other and no effect on total transpiration was observed.

Nitrogen Can Alleviate the Inhibition of Photosynthesis Caused by High Temperature Stress under Both Steady-State and Flecked Irradiance.

Nitrogen is one of the most important elements for plants and is closely related to photosynthesis. High temperature stress significantly inhibits photosynthesis under both steady-state and flecked irradiance. However, it is not known whether nitrogen can affect the decrease in photosynthesis caused by high temperature, especially under flecked irradiance. In the present study, a pot experiment was conducted under two nitrogen (N) supplies with rice plants, and the steady-state and dynamic photosynthesis rates were measured under 28 and 40°C. High temperature significantly increased leaf hydraulic conductance (Kleaf) under high N supply (HN) but not under low N supply (LN). The increased Kleaf maintained a constant leaf water potential (Ψleaf) and steady-state stomatal conductance (gs,sat) under HN, while the Ψleaf and gs,sat significantly decreased under high temperature in LN conditions. This resulted in a more severe decrease in steady-state photosynthesis (Asat) under high temperature in the LN conditions. After shifting from low to high light, high temperature significantly delayed the recovery of photosynthesis, which resulted in more carbon loss under flecked irradiance. These effects were obtained under HN to a lesser extent than under LN supply. Therefore, it is concluded that nitrogen can alleviate the inhibition of photosynthesis caused by high temperature stress under both steady-state and flecked irradiance.

Stomatal dynamics are limited by leaf hydraulics in ferns and conifers: results from simultaneous measurements of liquid and vapour fluxes in leaves.

Stomatal responsiveness to vapour pressure deficit (VPD) results in continuous regulation of daytime gas-exchange directly influencing leaf water status and carbon gain. Current models can reasonably predict steady-state stomatal conductance (gs ) to changes in VPD but the gs dynamics between steady-states are poorly known. Here, we used a diverse sample of conifers and ferns to show that leaf hydraulic architecture, in particular leaf capacitance, has a major role in determining the gs response time to perturbations in VPD. By using simultaneous measurements of liquid and vapour fluxes into and out of leaves, the in situ fluctuations in leaf water balance were calculated and appeared to be closely tracked by changes in gs thus supporting a passive model of stomatal control. Indeed, good agreement was found between observed and predicted gs when using a hydropassive model based on hydraulic traits. We contend that a simple passive hydraulic control of stomata in response to changes in leaf water status provides for efficient stomatal responses to VPD in ferns and conifers, leading to closure rates as fast or faster than those seen in most angiosperms.

Determinants of stomatal sluggishness in ozone-exposed deciduous tree species

Our knowledge of ozone effects on dynamic stomatal response is still limited, especially in Asian tree species. We thus examined ozone effects on steady-state leaf gas exchange and stomatal dynamics in three common tree species of China (Ailanthus altissima, Fraxinus chinensis and Platanus orientalis). Seedlings were grown and were exposed to three levels of ozone in open-top chambers (42, 69, 100nmolmol−1 daylight average, from 09:00 to 18:00). At steady-state, ozone exposure induced an uncoupling of photosynthesis and stomatal conductance, as the former decreased while the latter did not. Dynamic stomatal response was investigated by cutting the leaf petiole after a steady-state stomatal conductance was reached. Ozone exposure increased stomatal sluggishness, i.e., slowed stomatal response after leaf cutting, in the following order of sensitivity, F. chinensis>A. altissima>P. orientalis. A restriction of stomatal ozone flux reduced the ozone-induced sluggishness in P. orientalis. The ozone-induced impairment of stomatal control was better explained by stomatal ozone flux per net photosynthesis rather than by stomatal ozone flux only. This suggests that ozone injury to stomatal control depends both on the amount of ozone entering a leaf and on the capacity for biochemical detoxification or repair. Leaf mass per area and the density of stomata did not affect stomatal sluggishness.

PYR/RCAR Receptors Contribute to Ozone-, Reduced Air Humidity-, Darkness-, and CO2-Induced Stomatal Regulation

Rapid stomatal closure induced by changes in the environment, such as elevation of CO2, reduction of air humidity, darkness, and pulses of the air pollutant ozone (O3), involves the SLOW ANION CHANNEL1 (SLAC1). SLAC1 is activated by OPEN STOMATA1 (OST1) and Ca(2+)-dependent protein kinases. OST1 activation is controlled through abscisic acid (ABA)-induced inhibition of type 2 protein phosphatases (PP2C) by PYRABACTIN RESISTANCE/REGULATORY COMPONENTS OF ABA RECEPTOR (PYR/RCAR) receptor proteins. To address the role of signaling through PYR/RCARs for whole-plant steady-state stomatal conductance and stomatal closure induced by environmental factors, we used a set of Arabidopsis (Arabidopsis thaliana) mutants defective in ABA metabolism/signaling. The stomatal conductance values varied severalfold among the studied mutants, indicating that basal ABA signaling through PYR/RCAR receptors plays a fundamental role in controlling whole-plant water loss through stomata. PYR/RCAR-dependent inhibition of PP2Cs was clearly required for rapid stomatal regulation in response to darkness, reduced air humidity, and O3. Furthermore, PYR/RCAR proteins seem to function in a dose-dependent manner, and there is a functional diversity among them. Although a rapid stomatal response to elevated CO2 was evident in all but slac1 and ost1 mutants, the bicarbonate-induced activation of S-type anion channels was reduced in the dominant active PP2C mutants abi1-1 and abi2-1. Further experiments with a wider range of CO2 concentrations and analyses of stomatal response kinetics suggested that the ABA signalosome partially affects the CO2-induced stomatal response. Thus, we show that PYR/RCAR receptors play an important role for the whole-plant stomatal adjustments and responses to low humidity, darkness, and O3 and are involved in responses to elevated CO2.

Whole-tree water use efficiency is decreased by ambient ozone and not affected by O3-induced stomatal sluggishness.

Steady-state and dynamic gas exchange responses to ozone visible injury were investigated in an ozone-sensitive poplar clone under field conditions. The results were translated into whole tree water loss and carbon assimilation by comparing trees exposed to ambient ozone and trees treated with the ozone-protectant ethylenediurea (EDU). Steady-state stomatal conductance and photosynthesis linearly decreased with increasing ozone visible injury. Dynamic responses simulated by severing of a leaf revealed that stomatal sluggishness increased until a threshold of 5% injury and was then fairly constant. Sluggishness resulted from longer time to respond to the closing signal and slower rate of closing. Changes in photosynthesis were driven by the dynamics of stomata. Whole-tree carbon assimilation and water loss were lower in trees exposed to ambient O3 than in trees protected by EDU, both under steady-state and dynamic conditions. Although stomatal sluggishness is expected to increase water loss, lower stomatal conductance and premature leaf shedding of injured leaves aggravated O3 effects on whole tree carbon gain, while compensating for water loss. On average, WUE of trees exposed to ambient ozone was 2–4% lower than that of EDU-protected control trees in September and 6–8% lower in October.

Ozone-induced stomatal sluggishness develops progressively in Siebold's beech (Fagus crenata)

We investigated the effects of ozone and leaf senescence on steady-state stomatal conductance and stomatal response to light variation. Measurements were carried out in a free-air ozone exposure experiment on a representative deciduous broadleaved tree species in Japan (Fagus crenata). Both steady-state and dynamic stomatal response to light variation varied intrinsically with season due to leaf senescence. Ozone induced the decrease in steady-state leaf gas exchange and the sluggish stomatal closure progressively. These findings suggest that ozone reduces the ability of plants to adapt to a fluctuating light environment under natural conditions, and therefore impairs plant growth and ability to control water loss.

Interspecific variability of plant stomatal response to step changes of [CO2

The kinetics of a stomatal response to sudden increases or decreases of CO2 concentrations ([CO2]) was studied in 13 plant species growing in the field. Plants were well supplied with water. In each plant, gas exchange measurements were made on a fully developed leaf that was first left to achieve steady-state stomatal conductance (gs) at 400μmol(CO2)mol−1) and then exposed to a step change of [CO2] (to 700μmolmol−1 in one experiment; and to 700 and back to 400μmolmol−1 in a second experiment). Porometric data were captured in intervals of 3s until a new steady state was reached.A comparison of t1/2, the half-time needed to achieve new gs, indicates similar responses of stomata in grasses when compared to herbs. The stomata of C4 plants responded in approximately 5min, the highest closure rate was detected in Echinochloa crus-galli and Digitaria sanguinalis. Opening rates were similar to closing rates and the response as a whole was rather symmetric. In C3 plants, the full response of stomata was much slower. Analysis revealed differences in absolute rates of gs change between C3 and C4 plants. These differences can be related to the specificities of the type of photosynthetic metabolism. C4 photosynthesis enables plants to reduce gs, which can hasten further changes of diffusivity in response to the environmental signals. A possible coupling of C4 metabolism to the regulation of guard cells also has to be taken into account when explaining the observed results.

Soil water deficits decrease the internal conductance to CO2 transfer but atmospheric water deficits do not

The internal conductance to CO2 supply from substomatal cavities to sites of carboxylation poses a large limitation to photosynthesis. It is known that internal conductance is decreased by soil water deficits, but it is not known if it is affected by atmospheric water deficits (i.e. leaf to air vapour pressure deficit, VPD). The aim of this paper was to examine the responses of internal conductance to atmospheric and soil water deficits in seedlings of the evergreen perennial Eucalyptus regnans F. Muell and the herbaceous plants Solanum lycopersicum (formerly Lycopersicon esculentum) Mill. and Phaseolus vulgaris L. Internal conductance was estimated with the variable J method from concurrent measurements of gas exchange and fluorescence. In all three species steady-state stomatal conductance decreased by approximately 30% as VPD increased from 1 kPa to 2 kPa. In no species was internal conductance affected by VPD despite large effects on stomatal conductance. In contrast, soil water deficits decreased stomatal conductance and internal conductance of all three species. Decreases in stomatal and internal conductance under water deficit were proportional, but this proportionality differed among species, and thus the relationship between stomatal and internal conductance differed among species. These findings indicate that soil water deficits affect internal conductance while atmospheric water deficits do not. The reasons for this distinction are unknown but are consistent with soil and atmospheric water deficits having differing effects on leaf physiology and/or root-shoot communication.

Higher rates of leaf gas exchange are associated with higher leaf hydrodynamic pressure gradients

Steady-state leaf gas-exchange parameters and leaf hydraulic conductance were measured on 10 vascular plant species, grown under high light and well-watered conditions, in order to test for evidence of a departure from hydraulic homeostasis within leaves as hydraulic conductance varied across species. The plants ranged from herbaceous crop plants to mature forest trees. Across species, under standardized environmental conditions (saturating light, well watered), mean steady-state stomatal conductance to water vapour (g(w)) was highly correlated with mean rate of CO2 assimilation (A) and mean leaf hydraulic conductance normalized to leaf area (k(leaf)). The relationship between A and g(w) was well described by a power function, while that between A and k(leaf) was highly linear. Non-linearity in the relationship between g(w) and k(leaf) contributed to an increase in the hydrodynamic (transpiration-induced) water potential drawdown across the leaf (delta psi(leaf)) as k(leaf) increased across species, although across the 10 species the total increase in delta psi(leaf) was slightly more than twofold for an almost 30-fold increase in g(w). Higher rates of leaf gas exchange were therefore associated with higher k(leaf) and higher leaf hydrodynamic pressure gradients. A mechanistic model incorporating the stomatal hydromechanical feedback loop is used to predict the relationship between delta psi(leaf) and k(leaf), and to explore the coordination of stomatal and leaf hydraulic properties in supporting higher rates of leaf gas exchange.

Stomatal conductance does not correlate with photosynthetic capacity in transgenic tobacco with reduced amounts of Rubisco

High-resolution imaging of chlorophyll a fluorescence from intact tobacco leaves was used to compare the quantum yield of PSII electron transport in the chloroplasts of guard cells with that in the underlying mesophyll cells. Transgenic tobacco plants with reduced amounts of Rubisco (anti-Rubisco plants) were compared with wild-type tobacco plants. The quantum yield of PSII in both guard cells and underlying mesophyll cells was less in anti-Rubisco plants than in wild-type plants, but closely matched between the two cell types regardless of genotype. CO2 assimilation rates of anti-Rubisco plants were 4.4 micromol m(-2) s(-1) compared with 17.3 micromol m(-2) s(-1) for the wild type, when measured at a photon irradiance of 1000 micromol m(-2) s(-1) and ambient CO2 of 380 micromol mol(-1). Despite the large difference in photosynthetic capacity between the anti-Rubisco and wild-type plants, there was no discernible difference in the rate of stomatal opening, steady-state stomatal conductance or response of stomatal conductance to ambient CO2 concentration. These data demonstrate clearly that the commonly observed correlation between photosynthetic capacity and stomatal conductance can be disrupted in the long term by manipulation of photosynthetic capacity via antisense RNA technology. It was concluded that stomatal conductance is not directly determined by the photosynthetic capacity of guard cells or the leaf mesophyll.

Characterization of the photosynthetic induction response in a Populus species with stomata barely responding to light changes

The photosynthetic induction response is constrained by stomatal and biochemical limitations. However, leaves in some plants like Populus koreana x trichocarpa cv. Peace (a hybrid clone) may have little stomatal limitation because their stomata barely respond to changes in photon flux density (PFD). We examined the induction responses of leaves of well-watered and dehydrated P. koreana x trichocarpa plants grown in a high-light or a low-light regime. With an increase in PFD from 50 to 500 micromol m(-2) s(-1), steady-state stomatal conductance (g(s)) increased by only 0.25-8.2%, regardless of the initial g(s), but steady-state assimilation rate (A) increased by 550-1810%. Photosynthetic induction times required to reach 50% (IT50) and 90% (IT90) of A at high PFD were 60-90 s and 210-360 s, respectively. Examination of the dynamic relationships between A and g(s), and between A and intercellular CO2 concentration, indicated that the induction limitation was imposed completely by the biochemical components within 30-40 s after the PFD increase. Values of IT50 and IT90 were significantly higher in low-light leaves than in high-light leaves, whereas the induction state at 60 s and the induction efficiency at 60 and 120 s after the increase in PFD were lower in low-light leaves than in high-light leaves. Dehydration reduced leaf water potential (psi) significantly, resulting in a significantly decreased initial g(s). Leaf water potential had no significant effects on induction time in high-light leaves, but a low psi significantly reduced the induction time in low-light leaves. We conclude that the photosynthetic induction response was limited almost completely by biochemical components because the stomata barely responded to light changes. The biochemical limitation appeared to be higher in low-light leaves than in high-light leaves. Mild water stress may have reduced steady-state A and g(s), but it had little effect on the photosynthetic induction response in high-light leaves.

Consequences of an excess Al and a deficiency in Ca and Mg for stomatal functioning and net carbon assimilation of beech leaves

Stomatal function and photosynthesis were investigated in beech seedlings submitted to excess Al, or/and to a deficiency in Ca and Mg. Excess Al in the nutrient solution promoted a decrease of Ca and Mg leaf contents, while K was increased. Stomata l responses to darkness, ABA and ambient CO 2 remained normal. In contrast, steady-state stomatal conductance in light was signifi- cantly smaller and correlated to a lower accumulation of K in the guard cells. Similar stomatal responses were observed for Ca- Mg deficient plants. In response to combined Al stress and low Ca and Mg nutrition, stomata remained almost insensitive to the dif ferent stimuli. The constancy in K guard cell concentration revealed a disturbance in K fluxes. Lower CO 2 assimilation rates and chloro- phyll contents, on a leaf area basis, were recorded in response to all treatments. In conclusion, excess Al associated to low C a and Mg nutrition lead to a strong stomatal dysfonction and reduced photosynthesis of beech seedlings.

Sensitivity to far-red radiation in stomata of Phaseolus vulgaris L.: Rhythmic effects on conductance and photosynthesis

The influence of far-red (FR; 700-800 nm) radiation on steady-state stomatal conductance and net photosynthesis in P. vulgaris has been studied. Whereas FR radiation alone was relatively ineffective, addition of FR to a background of white light (WL; predominantly 400-700 nm) resulted in increased stomatal conductance. Stomata exhibited a marked diurnal sensitivity to FR. The action maximum for enhancing stomatal conductance was near 714 nm. A combination of FR and infra-red (IR; >800 nm) enhanced net photosynthesis when added to a background of WL. When IR alone was added to WL, there was a net decrease in photosynthesis, indicating that it is the FR waveband which is responsible for the observed photosynthetic effects. Naturally occurring levels of FR radiation (235 μmol·m(-2)·s(-1)) in vegetation-canopy shade enhanced net photosynthetic CO2 gain by 28% when added to a background of 55 μmol·m(-2)·s(-1) WL.