Dynamics of spatial heterogeneity of stomatal closure in Tradescantia virginiana altered by growth at high relative air humidity

The spatial heterogeneity of stomatal closure in response to rapid desiccation of excised well-watered Tradescantia virginiana leaves grown at moderate (55%) or high (90%) relative air humidity (RH) was studied using a chlorophyll fluorescence imaging system under non-photorespiratory conditions. Following rapid desiccation, excised leaves grown at high RH had both a greater heterogeneity and a higher average value of PSII efficiency (Phi(PSII)) compared with leaves grown at moderate RH. Larger decreases in relative water content resulted in smaller decreases in water potential and Phi(PSII) of high RH-grown leaves compared with moderate RH-grown leaves. Moreover, the Phi(PSII) of excised high RH-grown leaves decreased less with decreasing water potential, implying that the stomata of high RH-grown leaves are less sensitive to decreases in leaf water potential compared with moderate RH-grown leaves. After desiccation, some non-closing stomata were distributed around the main vein in high RH-grown leaves. Direct measurements of stomatal aperture showed 77% stomatal closure in the margins after 2 h desiccation compared with 40% closure of stomata in the main-vein areas in high RH-grown leaves. Faster closure of stomata in leaf margins compared with main-vein areas of leaves grown at high RH was related to substantially lower relative water content in these areas of the leaves.     

The role of abscisic acid in disturbed stomatal response characteristics of Tradescantia virginiana during growth at high relative air humidity

In this study, the role of abscisic acid (ABA) in altered stomatal responses of Tradescantia virginiana leaves grown at high relative air humidity (RH) was investigated. A lower ABA concentration was found in leaves grown at high RH compared with leaves grown at moderate RH. As a result of a daily application of 20 microM ABA to leaves for 3 weeks during growth at high RH, the stomata of ABA-treated leaves grown at high RH showed the same behaviour as did the stomata of leaves grown at moderate RH. For example, they closed rapidly when exposed to desiccation. Providing a high RH around a single leaf of a plant during growth at moderate RH changed the stomatal responses of this leaf. The stomata in this leaf grown at high RH did not close completely in response to desiccation in contrast to the stomata of the other leaves from the same plant. The ABA concentration on a fresh weight basis, though not on a dry weight basis, of this leaf was significantly lower than that of the others. Moreover, less closure of stomata was found in the older leaves of plants grown at high RH in response to desiccation compared with younger leaves. This was correlated with a lower ABA concentration in these leaves on a fresh weight basis, though not on a dry weight basis. Stomata of leaves grown at moderate RH closed in response to short-term application of ABA or sodium nitroprusside (SNP), while for leaves grown at high RH there was a clear difference in stomatal responses between the leaf margins and main-vein areas. The stomatal aperture in response to short-term application of ABA or SNP at the leaf margins of leaves grown at high RH remained significantly wider than in the main-vein areas. It was concluded that: (i) a long-term low ABA concentration in well-watered plants during growth at high RH could be a reason for less or no stomatal closure under conditions of drought stress; and (ii) the long-term ABA concentration on a fresh weight basis rather than on a dry weight basis is likely to be responsible for structural or physiological changes in stomata during leaf growth.     

Stomatal response characteristics of Tradescantia virginiana grown at high relative air humidity

Plants produced at high relative air humidity (RH) show poor control of water loss after transferring to low RH, a phenomenon which is thought to be due to their stomatal behaviour. The stomatal anatomy and responses of moderate (55%) and high (90%) RH grown Tradescantia virginiana plants to treatments that normally induce stomatal closure, i.e. desiccation, abscisic acid (ABA) application and exposure to darkness were studied using attached or detached young, fully expanded leaves. Compared with plants grown at moderate RH the transpiration rate, stomatal conductance and aperture of high RH grown plants measured at the same condition (40% RH) were, respectively, 112, 139 and 132% in light and 141, 188 and 370% in darkness. Besides the differences in stomatal size (guard cell length was 56.7 and 73.3 µm for moderate and high RH grown plants, respectively), there was a clear difference in stomatal behaviour. The stomata responded to desiccation, ABA and darkness in both moderate and high RH grown plants, but the high variability of stomatal closure in high RH grown plants was striking. Some stomata developed at high RH closed in response to darkness or to a decrease in relative water content to the same extent as did stomata from moderate RH grown plants, whereas others closed only partly or did not close at all. Evidently, some as yet unidentified physiological or anatomical changes during development disrupt the normal functioning of some stomata in leaves grown at high RH. The failure of some stomata to close fully in response to ABA suggests that ABA deficiency was not responsible for the lack of stomatal closure in response to desiccation.     

Avoiding high relative air humidity during critical stages of leaf ontogeny is decisive for stomatal functioning

Plants of several species, if grown at high relative air humidity (RH ≥85%), develop stomata that fail to close fully in case of low leaf water potential. We studied the effect of a reciprocal change in RH, at different stages of leaf expansion of Rosa hybrida grown at moderate (60%) or high (95%) RH, on the stomatal closing ability. This was assessed by measuring the leaf transpiration rate in response to desiccation once the leaves had fully expanded. For leaves that started expanding at high RH but completed their expansion after transfer to moderate RH, the earlier this switch took place the better the stomatal functioning. Leaves initially expanding at moderate RH and transferred to high RH exhibited poor stomatal functioning, even when this transfer occurred very late during leaf expansion. Applying a daily abscisic acid (ABA) solution to the leaves of plants grown at continuous high RH was effective in inducing stomatal closure at low water potential, if done before full leaf expansion (FLE). After FLE, stomatal functioning was no longer affected either by the RH or ABA level. The results indicate that the degree of stomatal adaptation depends on both the timing and duration of exposure to high RH. It is concluded that stomatal functionality is strongly dependent on the humidity at which the leaf completed its expansion. The data also show that the effect of ambient RH and the alleviating role of ABA are restricted to the period of leaf expansion.     

Foliar abscisic acid content underlies genotypic variation in stomatal responsiveness after growth at high relative air humidity

Background and Aims

Stomata formed at high relative air humidity (RH) respond less to abscisic acid (ABA), an effect that varies widely between cultivars. This study tested the hypotheses that this genotypic variation in stomatal responsiveness originates from differential impairment in intermediates of the ABA signalling pathway during closure and differences in leaf ABA concentration during growth.

Methods

Stomatal anatomical features and stomatal responsiveness to desiccation, feeding with ABA, three transduction elements of its signalling pathway (H₂O₂, NO, Ca²⁺) and elicitors of these elements were determined in four rose cultivars grown at moderate (60 %) and high (90 %) RH. Leaf ABA concentration was assessed throughout the photoperiod and following mild desiccation (10 % leaf weight loss).

Key Results

Stomatal responsiveness to desiccation and ABA feeding was little affected by high RH in two cultivars, whereas it was considerably attenuated in two other cultivars (thus termed sensitive). Leaf ABA concentration was lower in plants grown at high RH, an effect that was more pronounced in the sensitive cultivars. Mild desiccation triggered an increase in leaf ABA concentration and equalized differences between leaves grown at moderate and high RH. High RH impaired stomatal responses to all transduction elements, but cultivar differences were not observed.

Conclusions

High RH resulted in decreased leaf ABA concentration during growth as a result of lack of water deficit, since desiccation induced ABA accumulation. Sensitive cultivars underwent a larger decrease in leaf ABA concentration rather than having a higher ABA concentration threshold for inducing stomatal functioning. However, cultivar differences in stomatal closure following ABA feeding were not apparent in response to H₂O₂ and downstream elements, indicating that signalling events prior to H₂O₂ generation are involved in the observed genotypic variation.     

Smaller stomata require less severe leaf drying to close: A case study in Rosa hydrida

Stomata formed at high relative air humidity (RH) close less as leaf dries; an effect that varies depending on the genotype. We here quantified the contribution of each stomatal response characteristic to the higher water loss of high RH-grown plants, and assessed the relationship between response characteristics and intraspecific variation in stomatal size. Stomatal size (length multiplied by width), density and responsiveness to desiccation, as well as pore dimensions were analyzed in ten rose cultivars grown at moderate (60%) or high (85%) RH. Leaf morphological components and transpiration at growth conditions were also assessed. High growth RH resulted in thinner (11%) leaves with larger area. A strong positive genetic correlation of daytime and nighttime transpiration at either RH was observed. Stomatal size determined pore area (r = 0.7) and varied by a factor of two, as a result of proportional changes in length and width. Size and density of stomata were not related. Following desiccation, high RH resulted in a significantly lower (6–19%) decline of transpiration in three cultivars, whereas the relative water content (RWC) of high RH-expanded leaflets was lower (29–297%) in seven cultivars. The lower RWC of these leaflets was caused by (a) higher (33–72%) stable transpiration and/or (b) lower (12–143%) RWC at which this stable transpiration occurred, depending on the cultivar. Stomatal size was significantly correlated with both characteristics (r = 0.5 and -0.7, respectively). These results indicate that stomatal size explains much of the intraspecific variation in the regulation of transpiration upon water deprivation on rose.     

Stomatal malfunctioning under low VPD conditions: induced by alterations in stomatal morphology and leaf anatomy or in the ABA signaling?

Exposing plants to low VPD reduces leaf capacity to maintain adequate water status thereafter. To find the impact of VPD on functioning of stomata, stomatal morphology and leaf anatomy, fava bean plants were grown at low (L, 0.23 kPa) or moderate (M, 1.17 kPa) VPDs and some plants that developed their leaves at moderate VPD were then transferred for 4 days to low VPD (M→L). Part of the M→L‐plants were sprayed with ABA (abscisic acid) during exposure to L. L‐plants showed bigger stomata, larger pore area, thinner leaves and less spongy cells compared with M‐plants. Stomatal morphology (except aperture) and leaf anatomy of the M→L‐plants were almost similar to the M‐plants, while their transpiration rate and stomatal conductance were identical to that of L‐plants. The stomatal response to ABA was lost in L‐plants, but also after 1‐day exposure of M‐plants to low VPD. The level of foliar ABA sharply decreased within 1‐day exposure to L, while the level of ABA‐GE (ABA‐glucose ester) was not affected. Spraying ABA during the exposure to L prevented loss of stomatal closing response thereafter. The effect of low VPD was largely depending on exposure time: the stomatal responsiveness to ABA was lost after 1‐day exposure to low VPD, while the responsiveness to desiccation was gradually lost during 4‐day exposure to low VPD. Leaf anatomical and stomatal morphological alterations due to low VPD were not the main cause of loss of stomatal closure response to closing stimuli.     

Stomata of Micropropagated DelphiniumPlants Respond to ABA, CO2, Light and Water Potential, but Fail to Close Fully

Morphological and physiological characteristics of micropropagatedplants of Delphinium cv. Princess Caroline were studied. Leavesproduced in vitro showed poor control of water loss which appearsto result from restricted responses by stomata and not frompoor cuticular development. Stomata of leaves produced in vitrowere larger and more frequent than those produced during acclimatization.Despite the fact that stomata from isolated epidermis of leavesproduced in vitro reduced their apertures when exposed to turgor-reducingtreatments, they did not close fully. This, together with highstomatal frequencies might explain the poor control of waterloss shown by intact leaves produced in culture when exposedto dry air. While leaves from acclimatized plants showed almostcomplete closure with ABA, low water potentials, darkness andCO₂, stomata from leaves produced in vitro reduced their apertureswhen exposed to those factors, but only to a limit. Therefore,stomata from leaves cultured in vitro seem to be partially functional,but some physiological or anatomical alteration prevents themfrom closing fully. Stomata from leaves produced in vitro wereparticularly insensitive to ABA which appears to be partly associatedwith the high cytokinin concentration in the culture medium.In the long-term, this stomatal insensitivity to ABA might contributeto plant losses when micropropagated plantlets are transferredto soil. Key words: Micropropagation, stomatal physiology, dehydration, PEG, ABA, BAP, darkness, CO₂, Delphinium     

Stomatal Responses to Water Deficits and Abscisic Acid in Leaves of Sunflower Plants (Helianthus annuus L.) Grown under Different Conditions

The decrease in diffusive conductance of a leaf exposed to waterstress or to exogenous abscisic acid (ABA) was smaller in leavesof sunflower plants (Helianthus annuus L. cv. NK285) that hadbeen grown in a phytotron in humid air than in leaves of sunflowersgrown outdoors. Stomata of the phytotron-grown plants were slowerto close after detachment of a leaf than those of the outdoorplants. When stomata closed rapidly, as they did in detachedleaves and after treatment with ABA, the extent of closure wasvaried over the leaf's surface, in particular in the case ofphytotron-grown plants, and the extent of the heterogeneitywas greater in the phytotrongrown plants than in the outdoorplants. When stomata closed gradually, for example, under conditionsof limited moisture in the soil, closure occurred uniformlyover leaves of plants of both types. The smaller decrease indiffusive conductance of leaves from phytotron-grown plantsafter treatment with ABA resulted from the presence of patcheson the surface in which stomata remained open. The smaller decreaseof diffusive conductance in the phytotron-grown plants underconditions of limited moisture in the soil resulted from theuniformly lower responsiveness of stomata on a leaf to the decreasein water potential. When estimates are made of the intercellularconcentration of CO₂ (Ci) from gas-exchange measurements, heterogeneityin stomatal closure should be monitored when stomata close rapidly,in particular in plants grown in humid air, because heterogeneousstomatal closure can lead to overestimates of Ci. (Received April 18, 1994; Accepted May 25, 1995)     

Ineffectiveness of Abscisic Acid in Stomatal Closure of Yellow Lupin, Lupinus luteus var. Weiko III

Stomata of yellow lupin leaves are remarkably insensitive toabscisic acid (ABA). Stomatal resistance was monitored usingboth a viscous now porometer and a diffusion porometer. Resultswere confirmed with scanning electron microscopy. When exogenousABA solutions were supplied via petioles, 10^–6 M solutionshad no effect on stomatal resistance. Upper (adaxial) stomatawere not affected by 10^–5 M ABA but lower stomata showed3-fold more resistance after 2 h. Stomata of both surfaces closedafter 30 min in 10^–4 M ABA. Isolated epidermal peels of lupin leaves were floated on ABAsolutions yet upper surface peels showed no stomatal closingin 10^–4 M ABA, while lower surface stomata closed to abarely significant extent. Stomata of intact leaves were not very sensitive to darkness,showing at most a doubling in resistance after 6 h darkness.Complete stomatal closure, however, was readily produced bywilting leaves. Hence, lupin stomata are physically capableof closing. Endogenous ABA levels of water-stressed leaves increased approximately10-fold, which corresponds to concentrations below 10 µMABA. It is concluded that ABA is unlikely to play a role incontrolling short-term stomatal response of lupins.     

Stomatal Functioning of In Vitro and Greenhouse Apple Leaves in Darkness, Mannitol, ABA, and CO2

Leaves from in vitro and greenhouse cultured plants of Malusdomestica (Borkh.) cv. Mark were subjected to 4 h of darkness;4 h of 1 M mannitol induced water stress; 1 h of 10^–4M to 10^–7 M cis-trans abscisic acid (ABA) treatment; 1h of 0.12% atmospheric CO₂. Stomatal closure was determinedby microscopic examination of leaf imprints. In all treatments,less than 5% of the stomata from leaves of in vitro culturedplants were closed. The diameter of open stomata on leaves fromin vitro culture remained at 8 µm. In contrast, an averageof 96% of the stomata on leaves of greenhouse grown plants wereclosed after 4 h in darkness; 56% after 4 h of mannitol inducedwater stress; 90% after 1 h of 10^–4 M ABA treatment; 61%after 1 h in an atmosphere of 0.12% CO₂. Stomata of in vitroapple leaves did not seem to have a closure mechanism, but acquiredone during acclimatization to the greenhouse environment. Thelack of stomatal closure in in vitro plants was the main causeof rapid water loss during transfer to low relative humidity.     

A comprehensive analysis of the physiological and anatomical components involved in higher water loss rates after leaf development at high humidity

To better understand the poor regulation of water loss after leaf development at high relative air humidity (RH), the relative importance of the physiological and anatomical components was analyzed focusing on cultivars with a contrasting sensitivity to elevated RH. The stomatal responsiveness to three closing stimuli (desiccation, abscisic acid feeding, light/dark transition), as well as several stomatal features (density, index, size and pore dimensions) and the cuticular transpiration rate (CTR) were determined in four rose cultivars, grown under moderate (60%) and high (95%) RH. Moreover, the effects of changes in stomatal density and pore dimensions on the stomatal conductance (g_s) were quantified using a modified version of the Brown and Escombe equation. Higher water loss, as a result of plant growth at high RH, was primarily caused by an increase in residual g_s, and to a lesser extent due to higher CTR. It was estimated that in leaflets subjected to desiccation the enhanced g_s in high RH- as compared to moderate RH-grown plants was mostly due to poor stomatal functionality and to a lesser extent the combined result of higher stomatal density and longer pore length. It is concluded that the reduced degree and, specially, the reduced rate of stomatal closure are the primary causes of the large genotypic variation in the control of water loss in high RH-grown plants. Furthermore, it was found that although changes in stomatal length have no influence on stomatal functionality, changed anatomical features per se represent a significant and direct contribution to the increased water loss.     

High relative air humidity and continuous light reduce stomata functionality by affecting the ABA regulation in rose leaves

Plants developed under high (90%) relative air humidity (RH) have previously been shown to have large, malfunctioning stomata, which results in high water loss during desiccation and reduced dark induced closure. Stomatal movement is to a large extent regulated by abscisic acid (ABA). It has therefore been proposed that low ABA levels contribute to the development of malfunctioning stomata. In this study, we investigated the regulation of ABA content in rose leaves, through hormone analysis and β‐glucosidase quantification. Compared with high RH, rose plants developed in moderate RH (60%) and 20 h photoperiod contained higher levels of ABA and β‐glucosidase activity. Also, the amount of ABA increased during darkness simultaneously as the ABA‐glucose ester (GE) levels decreased. In contrast, plants developed under high RH with 20 h photoperiod showed no increase in ABA levels during darkness, and had low β‐glucosidase activity converting ABA‐GE to ABA. Continuous lighting (24 h) resulted in low levels of β‐glucosidase activity irrespective of RH, indicating that a dark period is essential to activate β‐glucosidase. Our results provide new insight into the regulation of ABA under different humidities and photoperiods, and clearly show that β‐glucosidase is a key enzyme regulating the ABA pool in rose plants.     

Drought-induced Closure of Phaseolus vulgaris L. Stomata Precedes Leaf Water Deficit and any Increase in Xylem ABA Concentration

Young seedlings of two cultivars of Phaseolus vulgaris L. (cv.Cacahuate-72 and Michoacan-12A3) were subjected to a soil dryingtreatment. Stomata started to close before any leaf water deficitcould be detected. Soil drying promoted a small degree of ABAaccumulation in roots, but, at the time stomatal closure wasinitiated, at least in one cultivar, xylem ABA and bulk leafABA concentration were not enhanced. The mechanism of the stomatalresponse is discussed in terms of an as-yet unidentified regulatorof stomatal behaviour, redistribution of existing ABA in plants,and a high sensitivity of stomata to small changes in ABA concentration.     

Rapid low temperature-induced stomatal closure occurs in cold-tolerant Commelina communis leaves but not in cold-sensitive tobacco leaves, via a mechanism that involves apoplastic calcium but not abscisic acid

Commelina communis stomata closed within 1 h of transferring intact plants from 27 degrees C to 7 degrees C, whereas tobacco (Nicotiana rustica) stomata did not until the leaves wilted. Abscisic acid (ABA) did not mediate cold-induced C. communis stomatal closure: At low temperatures, bulk leaf ABA did not increase; ABA did not preferentially accumulate in the epidermis; its flux into detached leaves was lower; its release from isolated epidermis was not greater; and stomata in epidermal strips were less sensitive to exogenous ABA. Stomata of both species in epidermal strips on large volumes of cold KCl failed to close unless calcium was supplied. Therefore, the following cannot be triggers for cold-induced stomatal closure in C. communis: direct effects of temperature on guard or epidermal cells, long-distance signals, and effects of temperature on photosynthesis. Low temperature increased stomatal sensitivity to external CaCl(2) by 50% in C. communis but only by 20% in tobacco. C. communis stomata were 300- to 1,000-fold more sensitive to calcium at low temperature than tobacco stomata, but tobacco epidermis only released 13.6-fold more calcium into bathing solutions than C. communis. Stomata in C. communis epidermis incubated on ever-decreasing volumes of cold calcium-free KCl closed on the lowest volume (0.2 cm(3)) because the epidermal apoplast contained enough calcium to mediate closure if this was not over diluted. We propose that the basis of cold-induced stomatal closure exhibited by intact C. communis leaves is increased apoplastic calcium uptake by guard cells. Such responses do not occur in chill-sensitive tobacco leaves.     

The Effects of Cytokinins and ABA on Stomatal Behaviour of Maize and Commelina

Epidermal strips and leaf fragments of Commelina and leaf fragmentsof maize were incubated on solutions containing naturally-occurringor synthetic cytokinins and/or ABA. The effects of these treatmentson stomatal behaviour were assessed. Cytokinins alone did notpromote stomatal opening in either species but concentrationsof both zeatin and kinetin from 10^–3 to 10^–1 molm^–3 caused some reversal of ABA-stimulated closure ofmaize stomata. The reversal of the ABA effect increased withincreasing cytokinin concentration. Cytokinins had no effecton ABA-stimulated closure of Commelina stomata. When appliedalone, at high concentration (10^–1 mol m^–3), toCommelina epidermis or leaf pieces both zeatin and kinetin restrictedstomatal opening. Key words: ABA, Cytokinins, Stomata, Maize, Commelina     

Some Effects of Abscisic Acid and Water Stress on Stomata of Vicia faba L.

Vicia faba seedlings grown under a plastic tent in the laboratorywere either watered well throughout their growth period or weresubjected to a water stress treatment for several days priorto an experimental treatment. The effects of a further waterstress treatment or an application of an aqueous solution ofabscisic acid (ABA) on the stomata of these plants were determined.Stomata of previously water-stressed plants proved to be moresensitive than stomata of well watered plants to ABA appliedthrough the petiole via the transpiration stream and sprayedonto leaf surfaces. Stomata of previously water-stressed plantsclosed more rapidly and to a greater degree than stomata ofwell watered plants. The hormone had only a small effect whenapplied directly to epidermal fragments removed from both groupsof plants. Stomata of plants which had received a water stresspretreatment were less sensitive to a subsequent period of waterstress than were stomata of previously well watered plants.It is proposed that stomatal adaptation to water stress maybe related to changes in the hormonal balance of the plant.     

Diurnal change in the relationship between stomatal conductance and abscisic acid in the xylem sap of field-grown peach trees

To evaluate whether abscisic acid (ABA) in the xylem sap playsan important role in controlling stomatal aperture of field-grownPrunus persica trees under drought conditions, stomatal conductance(g) and xylem ABA concentrations were monitored both in irrigatedand non-irrigated trees, on two consecutive summer days (threetimes a day). Stomata1 conductance of non-irrigated trees hada morning maximum and declined afterwards. The changes in gduring the day, rather than resulting from variations in theconcentrations of ABA in the xylem sap or the delivery rateof this compound to the leaves, were associated with changesin the relationship between g and xylem ABA. The stomata ofwater-stressed trees opened during the first hours of the day,despite the occurrence of a high concentration of ABA in thexylem sap. However, stomatal responsiveness to ABA in the xylemwas enhanced throughout the day. As a result, a tight inverserelationship between g and the logarithm of xylem ABA concentrationwas found both at midday and in the afternoon. A similar relationshipbetween g and ABA was found when exogenous ABA was fed to leavesdetached from well-watered trees. These results indicate thatABA derived from the xylem may account for the differences ing observed between field-grown peach trees growing with differentsoil water availabilities. Several possible explanations forthe apparent low stomatal sensitivity to xylem ABA in the morning,are discussed, such as high leaf water potential, low temperatureand high cytokinin activity. Key words: Prunus persica L., stomata, xylem ABA, water deficits, root-to-shoot communication     

The CO(2) response of Vicia guard cells acclimates to growth environment

Stomata of growth chamber-grown Vicia faba leaves have an enhanced CO(2) response, measured as change in stomatal aperture, compared to stomata of greenhouse-grown leaves. Reciprocal transfer experiments showed that the stomatal response to CO(2) acclimated to the growing environment. Stomata of growth chamber-grown leaves transferred to a greenhouse lost their high CO(2) sensitivity within 2-3 d while stomata of greenhouse-grown leaves transferred to a growth chamber acquired a high CO(2) sensitivity within 5-7 d. Experiments measuring the CO(2) responses of stomata in detached epidermis showed that growth chamber and greenhouse-grown stomata have the same contrasting CO(2) sensitivity observed in the intact leaf, indicating that the responses reflect intrinsic guard cell properties. The acclimation properties of the CO(2) response of guard cells have implications for the understanding of stomatal function under the predicted increases in atmospheric CO(2).     

Potassium Loss from Stomatal Guard Cells at Low Water Potentials

The potassium content of guard cells and the resistance to viscousflow of air through the leaf were determined in sunflower (Helianthusannuus) subjected to low leaf water potentials under illuminatedconditions. In intact plants desiccated slowly by withholdingwater from the soil, large losses in guard cell K occurred asleaf water potentials decreased. Leaf viscous resistance increased,indicating stomatal closure. Similar results were obtained whendetached leaf segments were desiccated rapidly. Upon rehydrationof leaves, no stomatal opening was observed initially, despiteleaf water potentials at predesiccated levels. After severalhours, however, re-entry of K occurred and stomata became fullyopen. Turgid leaf segments floated on an ABA solution showedlosses of guard cell K and closure of stomata as rapidly andcompletely as those brought about by desiccation. It is concludedthat stomatal closure at low water potentials under illuminatedconditions is not controlled solely by water loss from the tissuebut involves the loss of osmoticum from the guard cells as well.This in turn decreases the turgor difference between the guardcells and the surrounding cells, and closing occurs.