Determination of standardized transpiration used as an indirect method for considering stomatal aperture under field conditions

An indirect method for determination of stomata apertures is described. It is based on comparison of two simultaneous curves : the one obtained under natural conditions and the second obtained under standard conditions with minimum air humidity which can be considered as a criterion of stomata apertures.     

Stomatal Diffusion Resistance of Snap Beans. II. Effect of Light

The effect of light on the stomatal resistance of abaxial and adaxial leaf surfaces of snap beans (Phaseolus vulgaris L.) was studied in the growth chamber and in the field. The adaxial stomata required more light to open than the abaxial stomata; the abaxial stomatal apertures were still about 50% open at 1% full sunlight and light-induced closure was never observed under daytime field conditions. A given value of abaxial stomatal resistance was obtained at a given illumination of the abaxial guard cells whether illumination was adaxial or abaxial.     

In situ observation of stomatal movements and gas exchange of Aegopodium podagraria L. in the understorey

Observations of stomata in situ while simultaneously measuring CO(2) gas exchange and transpiration were made in field experiments with Aegopodium podagraria in a highly variable light climate in the understorey of trees. The low background photosynthetic photon flux density (PPFD) caused a slight opening of the stomata and no visible response to sporadic lightflecks. However, if lightflecks were frequent and brighter, slow opening movements were observed. Small apertures were sufficient to allow maximal photosynthetic rates. Therefore, the small apertures observed in low light usually only caused minor stomatal limitations of lightfleck photosynthesis. The response of stomata to step-wise changes in PPFD under different levels of leaf to air vapour pressure difference (Delta(W)) was observed under controlled conditions. High Delta(W) influenced the stomatal response only slightly by reducing stomatal aperture in low light and causing a slight reduction in the initial capacity to utilize high PPFD levels. Under continuous high PPFD, however, stomata opened to the same degree irrespective of Delta(W). Under high Delta(W), opening and closing responses to PPFD-changes were faster, which enabled a rapid removal of the small stomatal limitations of photosynthesis initially present in high Delta(W) after longer periods in low light. It is concluded that A. podagraria maintains a superoptimal aperture in low light which leads to a low instantaneous water use efficiency, but allows an efficient utilization of randomly occurring lightflecks.     

北京山区落叶阔叶林优势种叶片特点及其生理生态特性

荆条、山杏、辽东栎、北京丁香、大叶白蜡、核桃楸等６种植物在不同光照条件下的气孔导度,不同生境下叶片形态参数和叶片解剖特征进行了研究。扫描电子显微镜图片显示这片植物的气孔全着生在远轴面,气孔密度大小是：辽东栎〉山杏〉北京丁香〉核桃楸〉大叶白蜡,方差分析结果显示这５种植物叶片上气孔密度存在极显著差异,且叶片外表微观上差异比较明显。生境不仅影响叶片的气孔密度,而且也导气孔导度的差异,全光照条件下,山杏、     

Statistical Distribution of Stomatal Apertures of Vicia faba and Hordeum vulgare and the Spannungsphase of Stomatal Opening

Plants of Vicia faba and Hordeum vulgare were grown in growthboxes with 7 mW cm^–2 PAR, 14 h day/10 h night, at 22/20°C. Stomata of attached leaves were measured under controlledconditions by means of an optical microscope and the distributionfunctions of the widths of pores were established. For Viciafaba they appeared to be symmetrical bell-shaped functions.In the process of stomatal opening or closure the shape of thedistribution remained constant, its maximum sliding left andright along the aperture axis. This result has been interpretedto mean that increments or decrements of apertures were equalfor all stomata independent of their individual apertures. Theconclusion has been drawn that the ‘driving force’is evenly distributed, equal for all stomata, and varies withinwider limits than is possible for stomatal apertures. Stomatalopening is limited by the closed state from below and by ananatomically possible maximum aperture from above.     

Some counteractive effects of antitranspirants

Some previously unexpected effects of antitranspirants on stomata and plant growth are described, and their implications are discussed. Antitranspirant films increase the resistance to diffusion of water vapor from stomata. This increases leaf water potential and results in wider stomatal apertures immediately under the film, as well as on those parts of a partially treated leaf which are not covered by the film. Antitranspirants are expected to curtail growth via reduced photosynthesis. However, by increasing plant water potential they can enhance the growth of fruits and shoots.     

Disruption of a gene encoding C4-dicarboxylate transporter-like protein increases ozone sensitivity through deregulation of the stomatal response in Arabidopsis thaliana

To understand better the plant response to ozone, we isolated and characterized an ozone-sensitive (ozs1) mutant strain from a set of T-DNA-tagged Arabidopsis thaliana ecotype Columbia. The mutant plants show enhanced sensitivity to ozone, desiccation and sulfur dioxide, but have normal sensitivity to hydrogen peroxide, low temperature and high light levels. The T-DNA was inserted at a single locus which is linked to ozone sensitivity. Identification of the genomic sequences flanking the T-DNA insertion revealed disruption of a gene encoding a transporter-like protein of the tellurite resistance/C(4)-dicarboxylate transporter family. Plants with either of two different T-DNA insertions in this gene were also sensitive to ozone, and these plants failed to complement ozs1. Transpiration levels, stomatal conductance levels and the size of stomatal apertures were greater in ozs1 mutant plants than in the wild type. The stomatal apertures of ozs1 mutant plants responded to light fluctuations but were always larger than those of the wild-type plants under the same conditions. The stomata of the mutant and wild-type plants responded similarly to stimuli such as light, abscisic acid, high concentrations of carbon dioxide and ozone. These results suggest that OZS1 helps to close stomata, being not involved in the responses to these signals.     

Stomatal oscillations at small apertures: indications for a fundamental insufficiency of stomatal feedback-control inherent in the stomatal turgor mechanism.

Continuous measurements of stomatal aperture simultaneously with gas exchange during periods of stomatal oscillations are reported for the first time. Measurements were performed in the field on attached leaves of undisturbed Sambucus nigra L. plants which were subjected to step-wise increases of PPFD. Oscillations only occurred when stomatal apertures were small under high water vapour mole fraction difference between leaf and atmosphere (DeltaW). They consisted of periodically repeated opening movements transiently leading to very small apertures. Measurements of the area of the stomatal complex in parallel to the determination of aperture were used to record volume changes of guard cells even if stomata were closed. Stomatal opening upon a light stimulus required an antecedent guard cell swelling before a slit occurred. After opening of the slit the guard cells again began to shrink which, with some delay, led to complete closure. Opening and closing were rhythmically repeated. The time-lag until initial opening was different for each individual stoma. This led to counteracting movements of closely adjacent stomata. The tendency to oscillate at small apertures is interpreted as being a failure of smoothly damped feedback regulation at the point of stomatal opening: Volume changes are ineffective for transpiration if stomata are still closed; however, at the point of initial opening transpiration rate rises steeply. This discontinuity together with the rather long time constants inherent in the stomatal turgor mechanism makes oscillatory overshooting responses likely if at high DeltaW the 'nominal value' of gas exchange demands a small aperture.     

Light quality and osmoregulation in vicia guard cells : evidence for involvement of three metabolic pathways

Osmoregulation in opening stomata of epidermal peels from Vicia faba L. leaves was investigated under a variety of experimental conditions. The K⁺ content of stomatal guard cells and the starch content of guard cell chloroplasts were examined with cobaltinitrite and iodine-potassium iodide stains, respectively; stomatal apertures were measured microscopically. Red light (50 micromoles per square meter per second) irradiation caused a net increase of 3.1 micrometers in aperture and a decrease of −0.4 megapascals in guard cell osmotic potential over a 5 hour incubation, but histochemical observations showed no increase in guard cell K⁺ content or starch degradation in guard cell chloroplasts. At 10 micromoles per square meter per second, blue light caused a net 6.8 micrometer increase in aperture over 5 hours and there was a substantial decrease in starch content of chloroplasts but no increase in guard cell K⁺ content. At 25 micromoles per square meter per second of blue light, apertures increased faster (net gain of 5.7 micrometers after 1 hour) and starch content decreased. About 80% of guard cells had a higher K⁺ content after 1 hour of incubation but that fraction decreased to 10% after 5 hours. In the absence of KCl in the incubation medium, stomata opened slowly in response to 25 micomoles per square meter per second of blue light, without any K⁺ gain or starch loss. In dual beam experiments, stomata irradiated with 50 micomoles per square meter per second of red light for 3 hours opened without detectable starch loss or K⁺ gain; addition of 25 micomoles per square meter per second of blue light caused a further net gain of 4.4 micometers in aperture accompanied by substantial K⁺ uptake and starch loss. Comparison of K⁺ content in guard cells of opened stomata in epidermal peels with those induced to open in leaf discs showed a substantially higher K⁺ content in the intact tissue than in isolated peels. These results are not consistent with K⁺ (and its counterions) as the universal osmoticum in guard cells of open stomata under all conditions; rather, the data point to sugars arising from photosynthesis and from starch degradation as additional osmotica. Biochemical confirmation of these findings would indicate that osmoregulation during stomatal opening is the result of three key metabolic processes: ion transport, photosynthesis, and sugar metabolism.     

Age-related Changes in Stomatal Response to Cytokinins and Abscisic Acid

Kinetin and zeatin(100 mmol m^–3)reversald the ABA-mediated(100mmol ^m-2)closure of stomata of young maize leaves but did notaffect stomatal apertures of these leaves when applied alone.As leaves aged, kinetin or zeatin alone promoted increased stomatalapertures, while abscisic acid (ABA) applied alone had a reducedeffect on stomata. Even with older leaves, cytokinins reversadthe effect of ABA on stomata. Maize, stomata, abscisic acid, kineusc, zeatin, Zea mays     

The Effects of Temperature and ABA on Stomata of Zea mays L.

Epidermal fragments were removed from maize leaves by tearingparallel to the veins. These were incubated at a number of differenttemperatures in several concentrations of ABA. The sensitivityof stomata to temperature was dependent upon the technique usedto incubate epidermis. Generally, the widest apertures wererecorded at around 25°C. In all experiments, stomata incubatedat low (10°C) temperatures on 5.6 x 10^–4 M ABA showedwider apertures than those incubated on distilled water. ThisABA-stimulated stomatal opening was accompanied by an increasein the intensity of potassium staining in the guard cells. At25 °C, epidermis incubated on several concentrations ofABA showed some stomatal closure, decreased potassium stainingin the guard cells and increased potassium staining in the subsidiarycells.     

Are diurnal patterns of stomatal movement the result of alternating metabolism of endogenous guard cell ABA and accumulation of ABA delivered to the apoplast around guard cells by transpiration?

Abscisic acid (ABA) prevents opening of closed stomata and causes open stomata to close. A dual-source model is proposed linking ABA to diurnal stomatal movements. Darkness would favour guard cell biosynthesis of endogenous ABA and disfavour ABA catabolism. At first light, xanthophyll cycling, isomerization of ABA precursors, and activation of a cytochrome P450 mono-oxygenase (CytP450) would deplete endogenous guard cell ABA. The NADPH-requiring CytP450 would be activated by elevated O2 and reduced CO2 concentrations resulting from mesophyll photosynthesis. An increased O2-to-CO2 ratio would limit the Calvin cycle in guard cells, diverting NADPH produced by photosynthetic electron transport to the cytosol where, along with elevated O2, it would activate CytP450. Depletion of endogenous ABA would liberate guard cells to extrude protons and accumulate the ions and water needed to increase guard cell turgor and open stomata. By midday, stomata would be regulated by steady-state concentrations of ABA delivered to the apoplast around guard cells by transpiration. In temperate conditions, ABA would reach concentrations high enough to trigger ion efflux from guard cells, but too low to defeat the accumulation of sugars used to maintain opening. In dry conditions, ABA would reach effective concentrations by midday, high enough to trigger ion efflux and inhibit sugar uptake, reducing apertures for the rest of the day. At sunset, conditions would again favour biosynthesis and disfavour catabolism of endogenous guard cell ABA. The model can be used to reconcile proposed cellular mechanisms for guard cell signal transduction with patterns of stomatal movements in leaves.     

Abaxial and adaxial stomata from Pima cotton (Gossypium barbadense L.) differ in their pigment content and sensitivity to light quality

Sensitivity to light quality and pigment composition were analysed and compared in abaxial and adaxial stomata of Gossypium barbadense L. (Pima cotton). In most plants, abaxial (lower) stomatal conductances are higher than adaxial (upper) ones, and stomatal opening is more sensitive to blue light than to red. In greenhouse-grown Pima cotton, abaxial stomatal conductances were two to three times higher than adaxial ones. In contrast, adaxial stomatal conductances were 1·5 to two times higher than abaxial ones in leaves from growth chamber-grown plants. To establish whether light quality was a factor in the regulation of the relationship between abaxial and adaxial stomatal conductances, growth-chamber-grown plants were exposed to solar radiation outdoors and to increased red light in the growth chamber. In both cases, the ratios of adaxial to abaxial stomatal conductance reverted to those typical of greenhouse plants. We investigated the hypothesis that adaxial stomata are more sensitive to blue light and abaxial stomata are more sensitive to red light. Measurements of stomatal apertures in mechanically isolated epidermal peels from growth chamber and greenhouse plants showed that adaxial stomata opened more under blue light than under red light, while abaxial stomata had the opposite response. Using HPLC, we quantified the chlorophylls and carotenoids extracted from isolated adaxial and abaxial guard cells. All pigments analysed were more abundant in the adaxial than in the abaxial guard cells. Antheraxanthin and β-carotene contents were 2·3 times higher in adaxial than in abaxial guard cells, comparing with ad/ab ratios of 1·5–1·9 for the other pigments. We conclude that adaxial and abaxial stomata from Pima cotton have a differential sensitivity to light quality and their distinct responses are correlated with different pigment content.     

Central Roles for Potassium and Sucrose in Guard-Cell Osmoregulation

Osmoregulation in guard cells of intact, attached Vicia faba leaves grown under growth chamber and greenhouse conditions was studied over a daily light cycle of stomatal movements. Under both growth conditions guard cells had two distinct osmoregulatory phases. In the first (morning) phase, opening was correlated with K+ uptake and, to a lesser extent, sucrose accumulation. In the second (afternoon) phase, in which apertures were maximal, K+ content declined and sucrose became the dominant osmoticum. Reopening of the stomata after a CO2-induced closure was accompanied by accumulation of either K+ or sucrose, depending on the time of day, indicating that a single environmental signal may use multiple osmoregulatory pathways. Malate accumulation, correlated with K+ uptake, was detected under growth chamber but not greenhouse conditions, whereas Cl- was the main K+ counterion in the greenhouse. These results indicate that guard-cell osmoregulation in the intact leaf depends on at least two different osmoregulatory pathways, K+ transport and sucrose metabolism. Furthermore, the relative importance of the K+ counterions malate and Cl- appears to be environment-dependent.     

外源ABA、NO和H2O2对葱莲花瓣及叶片表皮气孔开闭的影响

以葱莲(Zephyranthes candida)为材料,研究不同浓度外源脱落酸、硝普钠(sodium nitroprusside,SNP)及过氧化氢对花瓣和叶片表皮气孔开闭的影响,以期为三者在切花保鲜中的应用提供新的依据。实验结果表明,10~1000 μmol/L脱落酸和硝普钠均能不同程度地引起花瓣和叶片表皮气孔关闭,且花瓣气孔较叶片气孔有更高的敏感性。过氧化氢对叶片表皮气孔开闭的影响大于对花瓣气孔的影响,花瓣表皮的气孔孔径仅在1000 μmol/L处理时变化显著。这说明在外源信号物质延缓切花衰老的过程中,花瓣表皮气孔的运动也可能起到了一定的作用。适当外源信号物质处理能诱导花瓣表皮气孔关闭,从而使花瓣的蒸腾作用减小,维持植物体内水势,延缓切花衰老。     

Cadmium effects on leaf transpiration of sugar beet (Beta vulgaris)

Seedlings of sugar beet ( Beta vulgaris L. cv Monohill) were cultivated for 4 weeks in nutrient solution containing different concentrations of CdCl₂ (0 to 10 μ M ). The effects of Cd on appearance and function of stomata and leaf cuticle were investigated by water loss measurements and microscopy. The leaf transpiration rate increased with increasing Cd concentrations while the sum total of stomatal aperture area per unit leaf area decreased. Already at low Cd levels. an increase of defective and undeveloped stomata was found in Cd treated plants. These stomata are closed or have small apertures and probably lack a functional closing mechanism. The number of intact stomata per unit leaf area was lower in leaves of Cd treated plants than in controls, and Cd induced closure of intact stomata. The total number of stomata per leaf area slightly increases with increasing Cd concentration. as does the percentage of small stomata. Furthermore. specific leaf area increased, while the density of leaf structure was decreased by Cd. From this observation we conclude that the increase in transpiration rate caused by Cd is primarily due to effects on the permeability of the leaf cuticle to water.     

外源ABA、NO和H2O2对葱莲花瓣及叶片表皮气孔开闭的影响

以葱莲（Zephyranthes candida）为材料,研究不同浓度外源脱落酸、硝普钠（sodium nitroprusside,SNP）及过氧化氢对花瓣和叶片表皮气孔开闭的影响,以期为三者在切花保鲜中的应用提供新的依据。实验结果表明,10～1000μmol/L脱落酸和硝普钠均能不同程度地引起花瓣和叶片表皮气孔关闭,且花瓣气孔较叶片气孔有更高的敏感性。过氧化氢对叶片表皮气孔开闭的影响大于对花瓣气孔的影响,花瓣表皮的气孔孔径仅在1000μmol/L处理时变化显著。这说明在外源信号物质延缓切花衰老的过程中,花瓣表皮气孔的运动也可能起到了一定的作用。适当外源信号物质处理能诱导花瓣表皮气孔关闭,从而使花瓣的蒸腾作用减小,维持植物体内水势,延缓切花衰老。     

Correlations Between the Unbound Water Content of Guard Cells and Stomatal Aperture in Tradescantia virginiana L.

When the apertures of stomata in an epidermal strip changedthere was found to be a simultaneous change in the unbound watercontent of the tissue. This occurred even when only the guardcells were living. It was concluded that the change in unboundwater occurred in the guard cells, and possibly the guard cellwalls.     

Responses of stomata to environmental factors-experiments with isolated epidermal strips of Polypodium vulgare

Summary Stomatal apertures of isolated and suitably conditioned epidermal strips of Polypodium vulgare are described as the stomata respond to the influences of temperature, air humidity, and water potential at the epidermal inner walls. Water stress as a result of reduced water potential in the substomatal airspace leads to narrower stomatal pores when water potential falls below -8 bar. Water potentials above this threshold value show minor influence. Stomatal responses to such water stress strongly interact with the responses to humidity changes in ambient air and to temperature. The linear dependence of stomatal apertures on the vapor saturation deficit of the air (closing) is shifted to lower values (more closed) by lower leaf bulk water potentials.Stomatal behavior depending on the temperature factor seems to be reversed by higher water stress. Without water stress, rising temperatures between 20 and 28° C are accompanied by further opening of the pores, whereas an increase of temperature within this range leads to narrowing of the stomata under the influence of lower water potentials within the substomatal airspace. It can be demonstrated that stomatal aperture values of Polypodium vulgare depending on temperature always describe optimum curves. With no water stress, closing does not occur before rather high temperatures are reached and above a broad range of maximal opening. Water stress, on the other hand, results in more pronounced narrowing of stomatal pores and shifts the onset to considerably lower temperatures.     

Light-Induced Stomatal Opening Is Affected by the Guard Cell Protein Kinase APK1b

Guard cells allow land plants to survive under restricted or fluctuating water availability. They control the exchange of gases between the external environment and the interior of the plant by regulating the aperture of stomatal pores in response to environmental stimuli such as light intensity, and are important regulators of plant productivity. Their turgor driven movements are under the control of a signalling network that is not yet fully characterised. A reporter gene fusion confirmed that the Arabidopsis APK1b protein kinase gene is predominantly expressed in guard cells. Infrared gas analysis and stomatal aperture measurements indicated that plants lacking APK1b are impaired in their ability to open their stomata on exposure to light, but retain the ability to adjust their stomatal apertures in response to darkness, abscisic acid or lack of carbon dioxide. Stomatal opening was not specifically impaired in response to either red or blue light as both of these stimuli caused some increase in stomatal conductance. Consistent with the reduction in maximum stomatal conductance, the relative water content of plants lacking APK1b was significantly increased under both well-watered and drought conditions. We conclude that APK1b is required for full stomatal opening in the light but is not required for stomatal closure.