Changes in Stomatal Conductance in Intact Ageing Wheat Leaves in Response to Abscisic Acid

The characteristics of ABA-induced changes in the fluxes ofCO₂ and water vapour from whole leaves of spring wheat (Triticumaestivum cv. Wembley) were examined. Aqueous solutions of ABAwere supplied via the transpiration stream to intact leavesof different ages mounted within a gas exchange cuvette. ABA caused a reduction in stomatal conductance (g) that wasproportional to the concentration in the solution fed to theleaf. For the maintenance of a reduction in g there was a requirementfor a continual supply of ABA. At concentrations greater than10^–2 mol m^–3 ABA reduced g by at least 50% of thecontrol value, while 1.0 mol m^–3 closed stomata within2 h. Concentrations as low as 10^–3 mol m^–3 produceda 20% reduction in g. As leaves aged they became less responsiveto applied ABA. The possibility that the stomatal response may change aftera leaf has previously experienced a pulse of ABA was exploredby repeating the exposure of a leaf to 10^–2 mol m^–3ABA. The first pulse of ABA produced a greater reduction ing than a subsequent exposure the following day. This declinein response of g to ABA on repeated exposure was maintainedwith leaves of different ages. The characteristics of the stomatal response to ABA are discussedin the context of what is known about the location of receptorsfor the hormone. It seems likely that a failure to respond toABA that has previously accumulated in the guard cells shouldbe viewed by means of maximizing the sensitivity to the currentsupply of ABA. It is suggested that the smaller response ofthe stomata of older leaves to ABA makes them more susceptibleto water stress, so that they can act as sensors for decliningwater potentials to give early protection to younger, metabolicallyactive leaves. Key words: Abscisic acid, leaf age, stomatal conductance, Triticum aestivum     

Abscisic Acid and Apical Dominance in Phaseolus coccineus L. The Role of Tissue Age

Abscisic acid (ABA) applied to the decapitated second internodeof runner bean plants enhanced outgrowth of lateral buds onlywhen internode stumps were no longer elongating. Applied toelongating internodes of slightly younger plants, ABA causesinhibition of bud outgrowth. Together with 10^–4 M indol-3-ylacetic acid (IAA), ABA stimulated internode elongation and interactedadditively in the inhibition of bud outgrowth. A mixture of10^–5 M ABA and 10^–6 M gibberellic acid (GA₃ ) causedsimilar effects on internode growth as IAA + ABA, but was mutuallyantagonistic in effect on growth of the lateral buds. Abscisic acid, apical dominance, gibberellic acid, indol-3yl acetic acid, Phaseolus coccineus, bean     

The Metabolism of Abscisic Acid

The light-catalysed isomerization of (+)-abscisic acid (ABA)to its trans isomer during isolation from leaves was monitoredby the addition of (±)-[2-¹⁴C]ABA to the extraction medium.(+)Trans-abscisic acid (t-ABA) was found to occur naturallyin rose (Rosa arvensis) leaves at 20µg/kg fresh weight;(+)-ABA was present at 594µg/kg. (±)-[2-¹⁴D]Trans-abscisicacid was not isomerized enzymically to ABA in tomato shoots. (±)-Abscisic acid was converted by tomato shoots to awater-soluble neutral product, ‘Metabolite B’, whichwas identified as abscisyl-ß-D-glucopyranoside. When(±)-[2-¹⁴C]trans-abscisic acid in an equimolar mixturewith (±)-[2-¹⁴C}ABA was fed to tomato shoots it was convertedto its glucose ester 10 times faster than was ABA. Trans-abscisyl-ß-D-glucopyrano8ide only was formedfrom (±)-[2-¹⁴C]t-ABA in experiments lasting up to 30h. Glucosyl abscisate was formed slowly from ABA and the freeacid fraction contained an excess of the unnatural (–).ABAas did the ABA released from abscisyl-ß-D-glucopyranosideby alkaline hydrolysis. The (+).ABA appeared to be the solesource of the acidic ‘Metabolite C" previously noted. The concentrations of endogenous (+)-, (+)-[2-¹⁴C]-, and (–)-[2-¹⁴C]ABAremaining as free acid, and also in the hydrolysate of abscisyl-ß-D-glucopyranoside,were measured by the ORD, UV absorption, and scintillation spectrometryof highly purified extracts of ABA from tomato shoots whichhad been supplied with racemic [2-^l4C]ABA.     

Effects of Abscisic Acid on the Growth Pattern of the Shoot Apical Meristem and on Flowering in Chenopodium rubrum L.

Abscisic acid (ABA) at 1 x 10^–4 M or 3 x 10^–4 Mwas applied to the apical buds of Chenopodium rubrum plantsexposed to different photoperiodic treatments and showing differentpatterns of floral differentiation. Stimulation of growth inwidth of the apical meristem of the shoot and/or inhibitionof growth in length was obtained under all photoperiodic treatments.This change of growth pattern was followed by different effectson flowering. In non-induced plants grown under continuous light ABA stimulatedpericlinal divisions in the peripheral zone and the initiationof leaves as well as the growth in width of bud primordia. Inplants induced by two short days reduced growth of the meristemcoincided with ABA application. Longitudinal growth of the meristemwas inhibited in this case and only a temporary stimulationof inflorescence formation took place. In plants induced ata very early stage, ABA exerted a strong inhibitory effect onflowering. A permanent and reproducible stimulatory effect onflowering was obtained in plants induced by three sub-criticalphotoperiodic cycles if ABA was applied to apices released fromapical dominance. In this case formation of lateral organs andinternodes was promoted by ABA and was followed by stimulatedinflorescence formation. Gibberellic acid (GA2) at 1x 10^–4M or 3 x 10^–4 M brought about a similar effect on floweringas ABA, although the primary growth effect was different, i.e.GA₂ stimulated longitudinal growth. The effects of ABA and GA₂ on floral differentiation have beencompared with earlier results obtained from auxin and kinetinapplications. These growth hormones are believed to regulateflowering by changing cellular growth within the shoot apex.Depending on the actual state of the meristem identical growthresponses may result in different patterns of organogenesisand even in opposite effects on flowering. Shoot apex, flowering, photoperiodic induction, abscisic acid, gibberellic acid, Chenopodium rubrum L.     

The Effect of Abscisic Acid on Amino Nitrogen and Protein Content of Potato Plants in Relation to the Inhibition of Nitrate Reductase Activity

Treatment of potato plants grown in nutrient solution with 3.8µM ABA resulted in reduced soluble protein in roots andin leaves at 24 h, but not in stems. This treatment reducedin vivo nitrate reductase activity in all organs for about 48h with the most pronounced reduction occurring in the roots.Excised root and leaf segments from plants treated with ABAfor 24, 48 and 72 h absorbed significantly more ¹⁴C leucine,compared to the control but the percent incorporation into proteinwas not altered in roots. In response to ABA total free amino nitrogen in leaves was lowerat 5 and 72 h and in stems at 72 h. Amino nitrogen content ofroots was enhanced by ABA at 5, 24 and 72 h due to generallyhigher levels of aspartate, serine, glutamate, proline and ammonia.There was no consistent relationship between ABA suppressionof nitrate reductase activity and ammonia or specific aminoacid (except proline) levels in leaves and stems. The increasedfree amino nitrogen levels in response to the hormone may bethe result of impaired NO₃– reduction rather than thecause. The results of protein synthesis studies and solubleprotein content suggest that ABA inhibition of nitrate reductaseis not due to general inhibition of protein synthesis and mayinvolve specific inhibition of nitrate reductase protein synthesis. ¹ Contribution No. 684, Department of Plant Science, Universityof Manitoba.     

Factors Affecting the Biosynthesis of Abscisic Acid

Incorporation of labelled mevalonate into abscisic acid (ABA)has been demonstrated in the cotyledons of mature avocado seeds,embryos and endosperms of developing wheat seeds, and avocadostems. The increase in ABA concentration on wilting parallelsthe increased incorporation of [2^–14C)mevalonate intoABA in avocado leaves and stems, suggesting that the increasein ABA content occurs by synthesis rather than by release froma stored precursor. Incorporation of [2^–14C]mevalonateby avocado mesocarp segments is unaffected by an 18 per centwater loss. The ABA content of roots was hardly affected bya 30 per cent water loss, indicating that the wilt-activatedmechanism is not fully operative in these tissues. Submerged Ceratophyllum plants and submerged parts of Callitricheshoots show a twofold increase in ABA content on wilting whereasthe aerial rosettes of the latter plant show a sixfold increase.This suggests that the occurrence of the wilt-induced mechanismis affected by previous growth conditions as well as by themorphology of the tissue.     

Abscisic Acid Content and Effects During Dehydration of Detached Leaves of Desiccation Tolerant Plants

Borya nitida is an angiospcrm whose detached leaves developcomplete tolerance to dehydration when they are equilibratedto air of 96% r.h. This treatment causes leaves to yellow aschlorophyll is destroyed, and abscisic acid contents increaseseveral-fold. Exogenous ABA (at 0.038–0.38 mol m^–3)promoted desiccation tolerance (a) in leaves undergoing toleranceinduction at 96% r.h., (b) only slightly during rapid dryingat rates which are normally injurious, and (c) considerablyin turgid tissue treated with ABA 48 h before rapid drying. ABA content also increased with intense water stress in Myrothamnusflabellifolia, a desiccation tolerant angiosperm which, unlikeBorya, retains most of its chlorophyll when dehydrated. Preliminaryincubation in ABA of detached leaves of this ‘resurrectionplant’ also promoted survival during rapid drying. Theability of ABA to substitute for the normal induction periodsuggests that this hormone participates in the development ofdesiccation tolerance. Key words: Abscisic acid, ABA, Drought tolerance, Resurrection plant     

Correlative Inhibition of Lateral Bud Growth in Pisum sativum L. and Phaseolus vulgaris L.: Studies of the Role of Abscisic Acid

The possibility has been investigated that abscisic acid (ABA)might act as a correlative inhibitor of lateral bud growth inPisum sativum and Phaseolus vulgaris. Application of ABA insmall quantities (2µg) to axillary buds on decapitatedplants of P. sativum caused appreciable inhibition of theirgrowth, and induced a compensatory growth of the bud on an adjacentnode. Application of this same quantity of ABA to axillary budson decapitated plants of Phaseolus vulgaris was without effect,but a high concentration in lanolin (1 mg g^–1) did substantiallyreduce bud outgrowth. Endogenous ABA-like substances in Phaseolusvulgaris, detected by bioassay and electron capture g.l.c.,were present in similar concentrations in shoot tips, lateralbuds on intact plants and lateral buds on plants decapitated24 h earlier. The effects of applied ABA suggested that it might be involvedin the mechanism of correlative inhibition in Pisum sativum,but it was not possible to test this hypothesis by determiningendogenous ABA levels in axillary buds because of their smallsize. The evidence presented here suggests that ABA is not acorrelative inhibitor in Phaseolus vulgaris even though at highconcentration it can inhibit the growth of axillary buds.     

Metabolism of xylem-delivered ABA in relation to ABA flux and concentration in leaves of maize and Commelina communis

When detached maize leaves were fed with an ABA solution viathe xylem, the relationship between the relative stomatal inhibitionand ABA concentrations was similar under different humidityconditions, but the relationship between such inhibition andABA flux was different according to changes of humidity. Tounderstand whether such stomatal behaviour was related to theway through which xylem-delivered ABA was metabolized, detachedleaves of maize and Commelina were fed with tritium-labelled(³H)-ABA at concentrations similar to that found in xylem ofdroughted plants and it was found that xylem-delivered ABA wasmetabolized rapidly in both species. The half-life of ABA metabolism,calculated from the time-related ABA disappearance curve, was42 and 64 min for maize and Commelina, respectively. The veryshort half-life suggests that there is a large capacity in leavesto metabolize xylem-delivered ABA and that metabolism is a majorfactor in the control of ABA accumulation in leaves. When ABAwas fed at different fluxes, either through changing the feedingconcentrations or through manipulating the rates of leaf transpiration(i.e. the volume flux), ABA was metabolized at rates that wereproportional to the amount that was delivered. The absoluterate of ABA metabolism was, therefore, linearly related to theamount of ABA that had arrived. It was found that xylem-deliveredABA reached the epidermis of Commelina, and was metabolizedat the same pattern as that in mesophyll tissues, i.e. at asimilar half-life and at rates constantly related to the amountthat was delivered. The role of the rapid ABA metabolism wasdiscussed in the context of stomatal control by either concentrationor flux of xylem-carried ABA. Key words: Abscisic acid, ABA metabolism, xylem-delivered ABA, maize, Commelina     

Ethylene and 1 -Aminocyclopropane-1-Carboxylic Acid Production in flacca, a Wilty Mutant of Tomato, Subjected to Water Deficiency and Pre-treatment with Abscisic Acid

Neill, S. J., McGaw, B. A. and Horgan, R. 1986. Ethylene and1-aminocyclopropane-l-carboxylic acid production in flacca,a wilty mutant of tomato, subjected to water deficiency andpretreatment with abscisic acid —J. exp. Bot. 37: 535–541. Plants of Lycoperstcon esculentum Mill. cv. Ailsa Craig wildtype and flacca (flc) were sprayed daily with H²O or 2?10^–2mol m^–3 abscisic acid (ABA). ABA treatment effected apartial phenotypic reversion of flc shoots; leaf areas wereincreased and transpiration rates decreased. Leaf expansionof wild type shoots was inhibited by ABA. Indoleacetic acid (IAA), ABA and l-aminocyclopropane-l-carboxylicacid (ACC) concentrations were determined by combined gas chromatography-massspectrometry using deuterium-labelled internal standards ABAtreatment for 30 d resulted in greatly elevated internal ABAlevels, increasing from 1?0 to 4?3 and from 0?45 to 4?9 nmolg^–1 fr. wt. in wild type and flc leaves respectively.Endogenous IAA and ACC concentrations were much lower than thoseof ABA. IAA content ranged from 0?05 to 0?1 nmol g^–1 andACC content from 0?07 to 0?24 nmol g^–1 Ethylene emanationrates were similar for wild type and flc shoots. Wilting of detached leaves induced a substantial increase inethylene and ACC accumulation in all plants, regardless of treatmentor type. Ethylene and ACC levels were no greater in flc leavescompared to the wild type. ABA pretreatment did not preventthe wilting-induced increase in ACC and ethylene synthesis. Key words: ABA, ACC, ethylene, wilting, wilty mutants     

Changes in endogenous hormones in apple during bud burst induced by defoliation

Under the tropical conditions of East Java, terminal buds of apple burst at any time of the year in response to removal of the subtending leaves. Following two such defoliations, two weeks apart on separate trees, there was a decrease in abscisic acid (ABA), a three-fold increase in gibberellin-like substances (GAs) and only a slight increase in cytokinin-like substances (CKs) in the apex tissue of closed buds. These changes preceded bud opening and the associated increases in fresh and dry weight, and may be causally related to bud burst. In open buds (i.e. young expanding leaves) the concentration of CKs was greater, and the concentrations of ABA and GAs less, than the concentrations in closed buds. As the leaves expanded, ABA increased and GAs and CKs decreased in concentration. The decrease in concentration of GAs and CKs, however, was due to the rise in dry weight of the expanding tissue; the amounts of all three hormones (per apex) increased. During bud burst there was a concurrent decrease in the CKs of subtending stems, suggesting a transfer into the expanding bud tissues. Removal of the old leaves by defoliation may remove the source of ABA and allow the amount of GAs in the apex to rise, bud burst following. Stem CKs may be utilized in the expansion of the new leaves in the bursting buds.     

Characterisation of Non-Uniform Photosynthesis Induced by Abscisic Acid in Leaves Having Different Mesophyll Anatomies

The effects of abscisic acid (ABA) on photosynthesis in leavesof Helianthus annuus L. were compared with those in leaves ofVicia faba L. After the ABA treatment, the response of photosyntheticCO₂ assimilation rate, A, to calculated intercellular partialpressure of CO₂, P_i, (A(p_i) relationship) was markedly depressedin H. annuus. A less marked depression was also observed inV.faba. However, when the abaxial epidermes were removed fromthese leaves, neither the maximum rate nor the CO₂ responseof photosynthetic oxygen evolution was affected by the applicationof ABA. Starch-iodine tests revealed that photosynthesis was not uniformover the leaves of H. annuus treated with ABA. The starch contentwas diffferent in each bundle sheath extension compartment (thesmallest subdivision of mesophyll by veins with bundle sheathextensions, having an area of ca. 0.25 mm² and ca. 50 stomata).In some compartments, no starch was detected. The distributionof open stomata, examined using the silicone rubber impressiontechniques, was similar to the pattern of starch accumulation.In V.faba leaves, which lack bundle sheath extensions, distributionof starch was more homogeneous. These results indicate that the apparent non-stomatal inhibitionof photosynthesis by ABA deduced from the depression of A(pⁱ)relationship is an artifact which can be attributed to the non-uniformdistribution of transpiration and photosynthesis over the leaf.Intercellular gaseous environment in the ABA-treated leavesis discussed in relation to mesophyll anatomy. ¹ Present address: Department of Botany, Duke University, Durham,NC 27706, U.S.A. (Received September 30, 1987; Accepted January 13, 1988)     

Effect of abscisic acid on membrane-bound epidermal ATPase from tobacco leaves

Membrane-bound Mg⁺⁺-activated ATPase activity in epidermal stripsfrom tobacco leaves (Nicotiana tabacum L. Samsun NN) was stimulatedby abscisic acid (ABA) when the strips were floated on ABA solutionin light or in darkness. The optimum ABA concentrations in lightand in darkness were 10^–5 M and 10^–6 M, respectively.Carbonyl cyanide m-chlorophenylhydrazone (CCCP) and N, N'-dicyclohexylcarbodiimide(DCCD) completely blocked the basal level membrane-bound epidermalATPase activity. ABAinduced membrane-bound epidermal ATPaseactivity was completely inhibited by CCCP, but only partly byDCCD. H⁺-influx into epidermal strips on a solution in light was lowerthan that in darkness. ABA stimulated H⁺-influx into epidermalstrips in light and in darkness. CCCP suppressed basal levelH⁺-influx, whereas DCCD did not. CCCP also suppressed ABA-inducedH⁺-influx, whereas DCCD did not. Interaction between H⁺-influxand membranebound epidermal ATPase activity is discussed. (Received May 23, 1978; )     

Environmental and Chemical Control of Vegetative Dormant Bud Production in Hydrilla verticillata

Bud production was investigated using algal-free cultures ofthe aquatic weed, Hydrilla verticillata. Vegetative dormantbud production in hydrilla is stimulated by a shortened photoperiodand is not temperature-dependent between 15 and 30 C. The phytochromesystem appears to be involved in this process and ABA stimulatesbud production. Artificial illumination throughout the nightprevented dormant bud production. Ethylene, applied as ethephon,reduced bud production in greenhouse cultures by 80 per centwhen applied at a level of 0.1 mgl^–1 ethephon at 2-d intervals. Hydrilla verticillata, aquatic plants, dormant buds, ethephon, ethylene, abscisic acid     

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     

Epicormic Bud Development in Quercus robur L. Studies of Endogenous IAA, ABA, IAA Polar Transport and Water Potential in Cambial Tissues10

Seasonal measurements of IAA,³ made using GC-MS, ⁴ indicatedthat in Q. robur the spring initiation of cambial activity andonset of visible bud outgrowth in the canopy is preceded byan increase in cambial region IAA. The effects of notch-girdlescut into the bole indicated that IAA in the cambial region laterwas present in separate physiological pools, with only the polar-transportedfraction affecting epicormic bud outgrowth. The stage in thespring when the epicormic buds grew out coincided with an increaseboth in cambial region IAA and in the capacity of cambial explantsfor IAA polar transport. Thus the stimulus needed by the epicormicbuds to overcome inhibition by polar-transported IAA appearedto be self-generated. The observed effects of exogenous hormoneson epicormic bud outgrowth from stem explants indicated thatthis stimulus might be cytokinin. The seasonal changes detectedin cambial region ABA³ were consistent with a role for stress-inducedABA in the induction of epicormic bud dormancy after canopydevelopment during the summer. No consistent effects of standthinning on cambial region IAA, ABA, water potentials or watercontents were detected, although polar transport of exogenousIAA by cambial region explants removed in the spring was reducedby thinning. Key words: Epicormic buds, cambium, hormones     

The uptake and flow of C, N and ions between roots and shoots in Ricinus communis L. III. Long-distance transport of abscisic acid depending on nitrogen nutrition and salt stress

Seedlings of Ricinus communis L. were cultivated in quartz sandand supplied with media which contained either different concentrationsof nitrate or ammonium nitrogen and were treated with a lowsalt stress. The concentration of ABA was determined in tissuesand in xylem and phloem saps. Between 41 and 51 day after sowing,abscisic acid (ABA) flows between roots and shoots were modelled.Long-distance transport of ABA was not stimulated under conditionsof nitrate deficiency (0.2 mol m^–3). However, when ammoniumwas given as the only N source (1.0 mol m^–3), ABA transportin both xylem and phloem was increased significantly. Mild saltstress (40 mol m^–3 NaCl) increased ABA transport in nitrate-fedplants, but not in ammonium-fed plants. The leaf conductancewas lowered by salt treatment with both nitrogen sources, butit was always lower in ammonium-fed compared to nitrate-fedplants. A negative correlation of leaf conductance to ABA levelsin leaves or flow in xylem was found only in comparison of ammonium-fedto nitrate-fed plants. Key words: Abscisic acid, ammonium, Ricinus communis, phloem, xylem, transport, nitrate, nitrogen nutrition     

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.     

The Water Relations of Young Olive Trees in a Mediterranean Winter: Measurements of Evaporation from Leaves and Water Conduction in Wood

We report measurements of evaporation rate, leaf resistanceto evaporation and water conduction in the stems of young olivetrees (Olea europea L.) growing in Messina, Italy, during thewinter and early spring. We have measured what Zimmermann calls‘leaf specific conductivity’ (LSC) of stem segmentsexcised from olive trees. The LSC is a measure of the specifichydraulic conductivity of stem segments normalized per unitarea of leaves supplied by the stem segment rather than perunit area of sapwood cross-sectional area. We find that theLSC's of primary stems were the largest followed in magnitudeby the LSC's of secondary stems and tertiary stems. Under winterand early spring conditions the maximum evaporative flux fromCoratina and Nocellara varieties of olive trees is about 2.6x 10^–5 kg 8^–1 m^–2. From this and the LSC measurementswe calculate that the pressure gradients needed to maintainthis rate of evaporation in the steady state is 65 kPa m^–1in primary stems, 170 kPa m^–1 in secondary stems and 560kPa m^–1 in tertiary stems. Olive, Olea europea L, evaporation, leaf specific conductivity, hydraulic conductivity, leaf resistance     

The Influence of Prior Water Stress and Abscisic Acid Foliar Spraying on Stomatal Responses to CO2, IAA, ABA, and Calcium in Leaves of Solanum melongena

The influence of a water stress or foliar ABA spraying pretreatmenton stomatal responses to water loss, exogenous ABA, IAA, Ca²⁺,and CO₂ were studied using excised leaves of Solanum melongena.Both pretreatments increased stomatal sensitivity of water loss,in the presence and absence of CO₂, but decreased stomatal sensitivityto exogenous ABA. CO₂ greatly reduced the effect of exogenouslyapplied ABA. IAA decreased leaf diffusion resistance for controland ABA sprayed leaves, but did not influence the LDR of previouslywater-stressed leaves. CA²⁺ did not influence LDR of any leavesof any treatments. Key words: Water stress, stomatal response, pretreatments