Stomatal responses to rapidly imposed water stress and light/dark transition in norflurazon-treated wheat leaves

Hoglund, H. O. and Klockare, R. 1987. Stomatal responses to rapidly imposed water stress and light/dark transition in norflurazon-treated wheat leaves.
Stomatal responses to rapidly imposed water stress and to light/dark transition were studied in leaves of wheat ( Triticum aestivum L. cv. Starke II) treated with nor-flurazon (NF) which is known to inhibit abscisic acid (ABA) accumulation. The stomatal response was studied in an open air flow system. It was shown that these plants have the ability to respond to externally added ABA. When the water potential in the nutrient solution was rapidly reduced, stomata in green plants responded with a transient opening followed by a strongly decreased aperture. NF-treated plants responded with a similar rapid opening of stomata, but the following closure was strongly reduced. Transfer from light to darkness induced a rapid closure of stomata in green plants but the closing response was strongly delayed in NF-treated plants. These results indicate that NF affects one or more regulators involved in the closure of stomata under rapidly imposed water stress and in the light/dark transition. The possibility that this regulator is ABA is discussed.     

亚低温与干旱胁迫对番茄植株水分传输和形态解剖结构的影响

为探讨亚低温和干旱对植株水分传输的影响机制,以番茄幼苗为试材,利用人工气候室设置常温(昼25 ℃/夜18 ℃)和亚低温(昼15 ℃/夜8 ℃)环境,采用盆栽进行正常灌水(75%～85%田间持水量)和干旱处理(55%～65%田间持水量),分析了温度和土壤水分对番茄植株水分传输、气孔和木质部导管形态解剖结构的影响。结果表明: 与常温正常灌水处理相比,干旱处理使番茄叶水势、蒸腾速率、气孔导度、水力导度、茎流速率、气孔长度和叶、茎、根导管直径显著减小,而使叶、茎、根导管细胞壁厚度和抗栓塞能力增强;亚低温处理下番茄叶水势、蒸腾速率、气孔导度、水力导度和叶、茎、根导管直径显著降低,但气孔变大,叶、根导管细胞壁厚度和叶、茎、根抗栓塞能力显著升高。亚低温条件下土壤水分状况对番茄叶水势、蒸腾速率、气孔导度、水力导度、气孔形态、叶、根导管结构均无显著影响。总之,干旱处理下番茄通过协同调控叶、茎、根结构使植株水分关系重新达到稳态;亚低温处理下番茄植株水分关系的调控主要通过改变叶和根导管结构实现,且受土壤水分状况的影响较小。     

Different blocking effects of HgCl2 and NaCl on aquaporins of pepper plants

In this study we have compared the short-term effects of both NaCl and HgCl₂ on aquaporins of Capsicum annuum L. plants, in order to determine whether or not they are similar. Stomatal conductance, turgor, root hydraulic conductance and water status were measured after 0.5, 2, 4 and 6 h of NaCl (60 mmol/L) or HgCl₂ (50 μmol/L) treatment. When 60 mmol/L NaCl was added to the nutrient solution, a large decrease in stomatal conductance was observed after 2 h. However, when HgCl₂ (50 μmol/L) was added, the decrease occurred after 4 h. The number of open stomata closed was always lower in plants treated with HgCl₂ than in plants treated with NaCl. The water content of the Hg²⁺-treated plants was decreased, compared with controls and NaCl-treated. The root hydraulic conductance decreased after HgCl₂ and NaCl treatment plants. Turgor of leaf epidermal cells was greatly reduced in plants treated with HgCl₂, but remained constant in the NaCl treatment, compared with control plants. The fact that the stomatal conductance was reduced more rapidly after NaCl addition, followed by the stomatal closure, and that both water content and turgor did not differ from the control suggests that in NaCl-treated plants there must be a signal moving from root to shoot. Therefore, the control of plant homeostasis through a combined regulation of root and stomatal exchanges may be dependent on aquaporin regulation.     

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。     

Changes in the Water Balance and Photosynthesis of Onion, Bean and Cotton Plants under Saline Conditions

The osmotic concentration (osmotic potential) of onion leaf sap did not adjust to chloride salinity, and consequently water potential, turgor, stomatal aperture and transpiration were reduced. Although osmotic concentration of bean and cotton leaf sap did adjust to a saline root medium and turgor was no less in the salinized plants than in the controls, stomata of the salinized plants remained only partly open and transpiration was reduced. Net photosynthesis of onion plants was reduced by salinity (this effect being much enhanced in a hot dry atmosphere) but it could be rapidly raised to the level of the controls by inducing elevated leaf turgor. Stomatal closure was initially responsible for most of the ～30 % reduction in photosynthesis of salinized beans. This was due to interference with CO₂ diffusion and could be overcome by raising the CO₂ concentration in the air. At a later stage of growth, salinity affected the light reaction of bean photosynthesis, and elevation of the air CO₂ had little effect. Closure of stomata of salinized cotton plants had only a relatively small effect on net photosynthesis. Light intensity and CO₂ concentration experiments showed that salinity was reducing the photosynthesis of cotton leaves mainly by affecting the light reaction of photosynthesis. It is concluded that chloride salinity does affect the water balance and rate of photosynthesis of plants and that the nature and degree of the effects will depend upon climatic conditions and may be very different between plant species and in the same species at different periods of growth.     

Effects of flooding and drought on stomatal activity, transpiration, photosynthesis, water potential and water channel activity in strawberry stolons and leaves

Transpiration, xylem water potential and water channel activity were studied in developing stolons and leaves of strawberry (Fragaria × ananassa Duch.) subjected to drought or flooding, together with morphological studies of their stomata and other surface structures. Stolons had 0.12 stomata mm^–2 and a transpiration rate of 0.6 mmol H₂O m^–2 s^–1, while the leaves had 300 stomata mm^–2 and a transpiration rate of 5.6 mmol H₂O m^–2 s^–1. Midday water potentials of stolons were always less negative than in leaves enabling nutrient ion and water transport via or to the strawberry stolons. Drought stress, but not flooding, decreased stolon and leaf water potential from –0.7 to –1 MPa and from –1 to –2 MPa, respectively, with a concomitant reduction in stomatal conductance from 75 to 30 mmol H₂O m^–2 s^–1. However, leaf water potentials remained unchanged after flooding. Similarly, membrane vesicles derived from stolons of flooded strawberry plants showed no change in water channel activity. In these stolons, turgor may be preserved by maintaining root pressure, an electrochemical and ion gradient and xylem differentiation, assuming water channels remain open. By contrast, water channel activity was reduced in stolons of drought stressed strawberry plants. In every case, the effect of flooding on water relations of strawberry stolons and leaves was less pronounced than that of drought which cannot be explained by increased ABA. Stomatal closure under drought could be attributed to increased delivery of ABA from roots to the leaves. However, stomata closed more rapidly in leaves of flooded strawberry despite ABA delivery from the roots in the xylem to the leaves being strongly depressed. This stomatal closure under flooding may be due to release of stress ethylene. In the relative absence of stomata from the stolons, cellular (apoplastic) water transport in strawberry stolons was primarily driven by water channel activity with a gradient from the tip of the stolon to the base, concomitant with xylem differentiation and decreased water transport potential from the stolon tip to its base. Reduced water potential in the stolons under drought are discussed with respect to reduced putative water channel activity.     

The Effect of 2-Chloroethyltrimethyl Ammonium Chloride (CCC) on Stomatal Diffusive Resistance in Tomato

CCC (2-chloroethyltrimethyl ammonium chloride) at a concentration of 6.3 mM was applied to tomato plants (cv. Grosse Lisse) grown in a controlled environment. There was an increase in adaxial leaf diffusive resistance but not in abaxial resistance, the effect being apparent before any growth retardation was measurable. The partial closure of adaxial stomata in response to CCC reduced transpiration from that leaf surface. In plants deprived of water, leaf water potential was higher when CCC was applied and both adaxial and abaxial stomatal closure was delayed. The data do not suggest that CCC influenced the relationship between leaf water potential and conductance for either abaxial or adaxial stomata.     

Anatomical adaptations of Astragalus gombiformis Pomel. under drought stress

The present study was designed to study the effect of drought on root, stem and leaf anatomy of Astragalus gombiformis Pomel. Several root, stem and leaf anatomical parameters (cross section diameter, cortex, root cortical cells, pith, leaf lamina and mesophyll thickness) were reduced under moderate to severe water deficit (20–30 days of withheld irrigation). The stele/cross section root ratio increased under moderate water deficit. The root’s and stems vascular systems showed reduced xylem vessel diameter and increased wall thickness under water deficit. In addition, the root xylem vessel density was increased in these drought conditions while it was unchanged in the stems. The stomata density was increased under prolonged drought conditions whereas the stomata size was untouched. The leaf vascular system showed reduced xylem and phloem tissue thickness in the main vein under moderate to severe water deficit. However, in the lamina the vascular tissue and the distance between vascular bundle were unaffected. Our findings suggest a complex network of anatomical adaptations such as a reduced vessel size with increased wall thickness, lesser cortical and mesophyll parenchyma formation and increased stomata density. These proprieties are required for the maintenance of water potential and energy storage under water stress which can improve the resistance of A. gombiformis to survive in arid areas.     

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。     

Root to Shoot Communication in Maize Plants of the Effects of Soil Drying

Seedlings of Zea mays L. (John Innes hybrid) were grown withroots divided between two containers such that part of the rootsystem could reduce the water potential of the soil in its immediatevicinity while the rest of the root system was well suppliedwith water. When compared to plants rooted in two pots of moistsoil, drying of part of the root system resulted in partialclosure of stomata, even though leaf water potential, turgorand abscisic acid (ABA) content remained unaffected. When leafpieces were removed from the two groups of plants and incubatedunder conditions favourable for stomatal opening, stomata ofthe ‘half-watered’ plants still showed restrictedapertures. Incubation in kinetin (10 mmol m^–3) or zeatin(100 mmol m^–3) reversed the closure of stomata stimulatedby soil drying. These results suggest that a continuous supplyof cytokinin from roots may be necessary to sustain maximalstomatal opening and an interruption of this supply due to soildrying may act as an indicator of inhibited root activity, resultingin restricted stomatal opening and thereby restricted wateruse. Key words: Zea mays L., Soil drying, Stomata, Roots     

Responses of stomata of barley and maize to phenylmercuric acetate

A reduction of stomatal aperture in light was found in leaves of maize after they had been treated with 10“3-5 m phenylmercuric acetate (PMA). Complete closure of the stomata in darkness was prevented, whilst there was total closure in the controls. Higher PMA concentrations had bigger effects. The relative water content (RWC) of barley tissues was slightly reduced 12 hours after treatment with PMA. The transpiration rate observed on PMA-treated barley plants was lower in light and higher in darkness than in untreated plants. Water saturation deficit (WSD) was higher by about 5%, and water holding capacity (WHC) lower (25%) than in untreated plants. The results suggest that the concentration of PMA normally applied as an antitranspirant is unfavourable for healthy growth of maize and barley.     

Early stomatal closure in waterlogged pea plants is mediated by abscisic acid in the absence of foliar water deficits

Abstract Soil waterlogging decreased leaf conductance (interpreted as stomatal closure) of vegetative pea plants (Pisuin sativum L. cv. ‘Sprite’) approximately 24 h after the start of flooding, i.e. from the beginning of the second 16 h-long photo-period. Both adaxial and abaxial surfaces of leaves of various ages and the stipules were affected. Stomatal closure was sustained for at least 3 d with no decrease in foliar hydration measured as water content per unit area, leaf water potential or leaf water saturation deficit. Instead, leaves became increasingly hydrated in association with slower transpiration. These changes in the waterlogged plants over 3 d were accompanied by up to 10-fold increases in the concentration of endogenous abscisic acid (ABA). Waterlogging also increased foliar hydration and ABA concentrations in the dark. Leaves detached from non-waterlogged plants and maintained in vials of water for up to 3 d behaved in a similar way to leaves on flooded plants, i.e. stomata closed in the absence of a water deficit but in association with increased ABA content. Applying ABA through the transpiration stream to freshly detached leaflets partially closed stomata within 15 min. The extractable concentrations of ABA associated with this closure were similar to those found in flooded plants. When an ABA-deficient ‘wilty’ mutant of pea was waterlogged, the extent of stomatal closure was less pronounced than that in ordinary non-mutant plants, and the associated increase in foliar ABA was correspondingly smaller. Similarly, waterlogging closed stomata of tomato plants within 24 h, but no such closure was seen in ‘flacca’, a corresponding ABA-deficient mutant. The results provide an example of stomatal closure brought about by stress in the root environment in the absence of water deficiency. The correlative factor operating between the roots and shoots appeared to be an inhibition of ABA transport out of the shoots of flooded plants, causing the hormone to accumulate in the leaves.     

Localization of mechanisms involved in hydropassive and hydroactive stomatal responses of Sambucus nigra to dry air

The response of stomata to a reduction of air humidity is composed of a hydropassive opening followed by active closure. Whereas the mechanisms behind the hydropassive opening are largely understood, the location and physiological basis of the sensing mechanisms leading to active closure are not yet known. This study attempts to evaluate the importance of a single pore's transpiration on its own response and that of adjacent pores. Selected stomata on attached intact leaves of Sambucus nigra were sealed with mineral oil and the response to a reduction of humidity was continuously observed in situ. Blocking a pore's transpiration had no appreciable effect on hydropassive opening and subsequent stomatal closure. If the adjacent stomata were additionally sealed, the closing response was reduced, but not the hydropassive opening. On the other hand, sealing the entire leaf surface, except a small area including the observed stomata, also reduced stomatal closure. These results indicate that strictly local processes triggered by a pore's own transpiration are not required to induce stomatal closure. To describe the effect of one pore's transpiration on the hydropassive and hydroactive responses of neighboring stomata, a simple spatial model was constructed. It suggests that 90% of the closing effect covers an area of approximately 0.5 mm2, whereas the effect on hydropassive opening affects an area of approximately 1 mm2. This divergence may suggest mechanisms other than or in addition to those involving changes of local leaf water potential.     

Water Stress Reduces Ozone Injury via a Stomatal Mechanism

Various studies have shown that water-stressed plants are more tolerant of ozone exposures than are unstressed plants. Two probable explanations for this tolerance are (a) stomatal closure which reduces ozone uptake and (b) biochemical or anatomical changes within the leaves. Phaseolus vulgaris cv Pinto bean plants were established and transferred to membrane systems which controlled the osmotic potential around the roots at −35 or −80 kilopascals for 5 days prior to ozone treatment (0 or 1.0 microliters per liter for 2 hours). Both water-stressed and unstressed plants were sprayed with various concentrations of abscisic acid to close the stomata or with fusicoccin to induce stomata opening. The abaxial stomatal resistances of primary and trifoliate leaves were measured just prior to ozone exposure. Plant response to ozone was determined by stress ethylene production and chlorophyll loss. Both water stress and abscisic acid induced stomatal closure and reduced ozone injury. In water-stressed plants, fusicoccin induced stomatal opening and those plants were as sensitive to ozone as were the non-water-stressed plants. These data suggest that water stress protects plants from ozone injury mainly through its influence on stomatal aperture rather than through biochemical or anatomical changes.     

Soybean adaptation to water stress at selected stages of growth

Soybean (Glycine max [L.] Merr. cv Braxton) plants were grown in sandy soil with only natural rainfall (N) or with supplemental irrigation (I). Water-stressed plants grew more extensive root systems, whereas irrigated plants developed larger shoots and smaller root systems. Maximum stomatal apertures were observed at the beginning of each photoperiod. Partial stomatal closure occurred each afternoon, but stomata of I plants remained open longer than those of N plants. Significant reductions in net carbon fixation rate generally accompanied decreases in stomatal aperture, which coincided with periods of high temperature, low relative humidity, maximum solar radiation, and water stress. Leaf water potential decreased from morning to afternoon, with a greater decrease observed for N plants. Midafternoon stomatal closure did not occur in N plants with very large root systems following a heavy rain which saturated the soil profile. With smaller root systems and greater evaporative demand from larger shoots, the I plants continued to show midafternoon stress following the heavy rain. The large root systems of the N plants absorbed sufficient water to meet shoot evaporative demand for several days following the rain. Root soil system resistance apparently contributed to the afternoon water stress in the I plants.     

Effect of root anaerobiosis on the water relations of several Pyrus species

Solution culture experiments were designed to investigate the plant water relations of 3 Pyrus species subjected to root anaerobiosis. Root anaerobiosis induced partial stomatal closure prior to alterations in leaf water potential (ΨLW) or root osmotic potential (ΨRπ). In contrast, stomatal closure was accompanied by a decline in root hydraulic conductivity (Lp). Anoxia markedly reduced ΨLW for Pyrus communis L. and eventually led to wilting and defoliation. Pyrus betulaefolia Bunge and Pyrus calleryana Decne, however, were less affected by root anaerobiosis. To delineate if the increased root resistance was in the radial or longitudinal direction, 10⁻⁴ M cistrans abscisic acid (ABA) was added to detopped root systems of P. communis in solution culture after steady-state rates of Lp were established. A consistent 25 to 30% promotion of Lp was observed 1.5 h after the addition of ABA for aerobically treated plants. ABA did not influence Lp when applied to roots previously deprived of O₂ for 4 days. Additional evidence against the limiting resistance being in the radial direction was obtained when water fluxes were compared through intact P. communis roots, roots with all feeder roots detached, and stems without root systems. Severing feeder roots from anaerobically treated plants did not increase water flux to rates observed for aerobically treated plants. Resistance progressed basipetally to eventually encompass the stem itself. These results can only be explained by occlusion of the xylem vessels.     

Changes in water status during water stress at different stages of development in wheat

The dynamics of stomatal resistance and osmotic adjustment in response to plant water deficits and stage of physiological development was studied in the leaves of spring wheat ( Triticum aestivum L., GWO 1809). Plants were germinated and grown in pots in a growth chamber at the Duke University Phytotron to four physiological stages of development (4th leaf, 7th leaf, anthesis, and soft dough), during which time stomatal resistance, total water potential and osmotic potential were measured on the last fully developed leaf of water stressed and non-stressed plants. Pressure potential was obtained by difference. Stomatal closure of the abaxial and adaxial surfaces were independent of each other, each having a different critical total water potential. The total water potential required to close the stomata on the last fully developed leaf were different at different stages of physiological development, decreasing as the plants grew older. The development of osmoregulation in wheat allows the closure of stomata during the vegetative stage at a high total water potential, but insures that stomata remain open from anthesis through the ear filling period to a lower total water potential.     

The diurnal course of plant water potential,stomatal conductance and transpiration in a papyrus (Cyperus papyrus L.) canopy

Summary The diurnal course of water potential, stomatal conductance and transpiration was measured on mature umbels (the major evaporating surface) of papyrus (Cyperus papyrus L.) growing in a fringing swamp on Lake Naivasha, Kenya.Umbel water potential declined only slightly during the morning but fell rapidly after midday to a minimum value of-1.5 M Pa in early afternoon. The two main structures forming the umbels, the bracteoles and rays, showed similar patterns of change of stomatal conductance throughout the day. The values of conductance indicate major stomatal opening during the morning, partial midday closure and some recovery of opening during the afternoon.It appears that the increase in water vapour pressure deficit of the air is the major cause of the midday closure of the stomata and that plant water potential has little effect. The reason why transpiration is reduced at high vapour pressure deficits when water is freely available to the roots is not clear. However, it is speculated that the restricted water movement into the plant from the anaerobic root environment has the effect of reducing the uptake of toxic ferrous iron.The daily total of canopy transpiration is estimated to be 12.5 mm, twice the value previously reported for papyrus but similar to daily valus determined for other wetland communities.     

Zooming into sub-organellar localization of reactive oxygen species in guard cell chloroplasts during abscisic acid and methyl jasmonate treatments

Regulation of stomata movements is crucial for plants ability to cope with their changing environment. Guard cell’s (GC) water potential directs water flux inside/outside this cell, which eventually is causing the stoma to open or close, respectively. Some of the osmolytes which accumulates in the GC cytoplasm and are known to play a role in stomata opening are sugars, arising from chloroplast starch degradation. During stomata closure, the accumulated osmolytes are removed from the GC cytoplasm. Surprisingly little is known about prevention of starch degradation and forming additional sugars which may interfere with osmotic changes that are necessary for correct closure of stomata.   One of the early events leading to stomata closure is production of reactive oxygen species (ROS) in various sub-cellular sites and organelles of the stoma. Here we report that ROS production during abscisic acid (ABA) and methyl jasmonate (MJ) stimuli in Arabidopsis GC chloroplasts were more than tripled. Moreover, ROS were detected on the sub-organelle level in compartments that are typically occupied by starch grains. This observation leads us to suspect that ROS function in that particular location is necessary for stomata closure. We therefore hypothesize that these ROS are involved in redox control that lead to the inactivation of starch degradation that takes place in these compartments, thus contributing to the stoma closure in an additional way.