The effect of pH on stomatal sensitivity to abscisic acid

Abstract. The sensitivity of stomata of Commelina communis L. to abscisic acid (ABA) was evaluated by analysing the initial rates of response to the compound at different hormone concentrations. This was carried out at pH 6.8 and pH 5.5. The data were modelled and statistically analyzed by means of a computer program employing non-linear regression techniques and step-down analysis of variance. The response kinetics as quantified in terms of three sensitivity parameters were found to differ significantly between the two pH values. This finding is discussed in relation to previous research on purified ABA-binding proteins.     

Phytochrome effects on K+ fluxes in guard cells of Commelina communis

The effect of phytochrome on K⁺ transport in guard cells of Commelina communis L. was studied following stomatal movement and using the K⁺−channel blockers tetraethylammonium (TEA), Cs⁺ and quinidine. TEA and quinidine prevented stomatal opening and closure in red light, but not when it was supplemented with far-red. This indicates that channels that can be blocked by TEA and quinidine are regulated by phytochrome. Evidence for a phytochrome effect on K⁺ leakage through other membranal compartments was also found. These phytochrome effects are modified by temperature. Elevated temperature decreases the involvement of channels and increases K⁺ transport through other membrane compartments, while low temperature causes channel opening and diminishes K⁺ leakage. The interaction between phytochrome effects and those of temperature is discussed.     

The effect of blue light on energy levels in epidermal strips

Red light applied together with blue enhanced stomatal opening in epidermal strips of Commelina communis L. more than red light alone. In red light, stomatal opening was enhanced by exogenously applied ATP and was inhibited by 3-(3,4-dichlorophe-nyl)-l,l-dimethylurea (DCMU), while in the presence of blue light external ATP was almost without effect, and DCMU stimulated stomatal opening. Blue light increased the ATP levels in the epidermal strips. DCMU diminished the amount of ATP in both red light and red + blue light treatments, but did not abolish the stimulatory effect of blue light. Blue light also stimulated the respiration rate of the epidermal strips. Rotenone, which inhibited stomatal opening and respiration rate, abolished the effect of blue light in both processes. These results imply that blue light increases the ATP levels by stimulation of oxidative phosphorylation.     

Effect of boron on stomatal opening in epidermal strips of Commelina communis

Boron enhances stomalal opening in epidermal strips of Commelina communis L. under conditions conductive to opening, i.e . light and KCI. This effect is obtained when boron is given together or before KCI application. Since boron increases ⁸⁶Rb net uptake in both illuminated epidermal strips and illuminated isolated guard cells, it is suggested that boron acts either by reducing potassium leakage or by increasing its uptake, thus leading to increased osmotic potential and, as a result, to stomatal opening.     

Possible involvement of calmodulin in the regulation of ATPase activity in guard cells

Calcium ions play an important role in the regulation of stomatal movement and the mechanism underlying this action is yet to be determined. It is suggested that guard cell plasma membrane ATPase is a target for calcium action and that this effect is mediated by calmodulin. In this study, the effects of calcium and two calmodulin antagonists on ATPase activity in a crude homogenate of Commelina communis L. guard cell protoplasts were examined. The homogenate contained Mg²⁺-dependent, K⁺-simulated ATPase activity, which was inhibited by CaCl² while stimulated by the calmodulin antagonists, compound 48/80 and chlorpromazine. The calmodulin antagonists partially reversed the inhibitory effect of calcium ions. The results support the possibility of calmodulin involvement in the regulation of guard cell ATPase activity by calcium ions.     

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; )     

The site of action of abscisic acid at the guard cell plasmalemma of Valerianella locusta

Abstract. Abscisic acid (ABA) is taken up by guard cells of isolated epidermata of Valerianella locusta only at low external pH values. At pH 8.0, when nearly all ABA molecules are present as the union of ABA (ABA⁻), no uptake can be observed. ABA-dependent movement of stomata was tested at external pH values between 5.0 and 8.0. Independent of the external pH, ABA induced stomatal closure at all tested ABA concentrations. It is concluded that ABA need not be taken up into the cytosol of the guard cells in order to induce slomatal closure. The primary site of ABA action at the guard cell plasmalemma must be located either at the outer surface of the plasmalemma or at least be easily accessible from outside. ABA− is as effective as undissociated ABA (ABAH).     

Characterization of the effects of metabolic inhibitors, ATPase inhibitors and a potassium-channel blocker on stomatal opening and closing in isolated epidermis of Commelina communis L.

Abstract. The specific effects of hypoxia and various inhibitors on stomatal opening in the light and closing in the dark were characterized in isolated epidermis from Commelina communis L. Reducing the guard cell metabolism with hypoxia and the uncoupler carbonyl cyanide-m-chloro-phenyl-hydrazone, CCCP, respectively, inhibited both stomatal opening and closing. Stomatal closing was very efficiently blocked by CCCP and this effect could be readily reversed by washing out the inhibitor. The authors were unable to inhibit stomatal opening with ATPase-inhibitors, without also affecting closing. Orthovanadate, up to 2 mol m⁻³, affected neither opening nor closing. Dicyclohexylcarbodiimide, DCCD, and diethylstilbestrol, DES, inhibited opening as well as closing to about 50%. The K⁺ -channel blocker tetraethylammonium chloride, TEA-Cl, inhibited both stomatal opening and closing, as did phenyl acetic acid, PAA, a compound considered to interfere with blue light induced stomatal opening. The results are discussed in the view that the uncontrolled K⁺ leakage from the guard cells is low, that K⁺ efflux during stomatal closing, as well as K⁺ influx during opening, occurs through specific K⁺-channels and that ATP and/or a membrane potential seems to be needed to keep these channels open.     

Effect of leaf water status and xylem pH on metabolism of xylem-transported abscisic acid

Metabolism and distribution of xylem-fed ABA were investigated in leaves of maize (Zea mays) and Commelina communis when water stress and xylem pH manipulation were applied. ³H-ABA was fed to excised leaves via the transpiration stream. Water stress was applied through either a previous soil-drying before leaves were excised, or a quick dehydration after leaves were fed with ABA. Xylem-delivered ABA was metabolised rapidly in the leaves (half-life 0.7 h and 1.02 h for maize and Commelina respectively), but a previous soil-drying or a post-feeding dehydration significantly extended the half-life of fed ABA in both species. In the first few hours after ABA was fed into the detached leaves, percentages of applied ABA remaining unmodified were always higher in leaves which received water stress treatments than in control leaves. However the percentage decreased to below the control levels several hours later in leaves which received a previous soil-drying treatment prior to excision, but had then been rehydrated by the xylem-feeding process itself. One possible explanation for this could be a changed pattern of compartmentalisation for xylem-carried ABA. A post-feeding dehydration treatment also changed the distribution of xylem-fed ABA within the leaves: more ABA was found in the epidermis of Commelina leaves which had been dehydrated rapidly after ABA had been fed, compared to the controls. The levels of xylem-delivered ABA remaining unmodified increased as the pH of the feeding solution increased from 5 to 8. The results support the hypothesis that water stress and a putative stress-induced xylem pH change may modify stomatal sensitivity to ABA by changing the actual ABA content of the leaf epidermis.     

Variation in stomatal characteristics over the lower surface of Commelina communis leaves

Abstract. The silicone rubber impression technique was used to measure stomatal apertures in 9 mm² sampling areas covering the entire lower surface of leaves of Commelina communis L. The data were analysed using a computer program which produced 'iso-aperture' contours illustrating local differences in mean stomatal aperture. Little consistency was seen in the iso-aperture patterns among sampling times, although the stomata were always relatively closed at the leaf tip and base. When stomata in the middle of the lamina were open, those near to the leaf margin tended to be relatively closed. In places, gradients of mean stomatal aperture were as high as 1 μm mm⁻¹. Measurements along a transect across the lower epidermis revealed no correlation of stomatal aperture with the presence of major veins in the mesophyll tissue. Variation in guard-cell size and stomatal frequency on the lower leaf surface was also analysed. The guard cells were smallest and the stomata more frequent near to the leaf margins. The significance of the results is discussed in relation to measurements of leaf conductance and models of stomatal function.     

Effect of abscisic acid on stomatal opening in isolated epidermal strips of abi mutants of Arabidopsis thaliana

Abscisic acid-insensitive mutants of Arabidopsis thaliana L. var. Landsberg erecta were selected for their decreased sensitivity to ABA during germination. Two of these mutants, abi-1 and abi-2 , display a wilty phenotype as adult plants, indicating disturbed water relations. Experiments were undertaken to find out if this results from insensitivity of mutant stomates to ABA.
Growth conditions and methods to isolate epidermal strips were optimized to study stomatal movement. Wild type stomates required external ionic conditions comparable to those found for other species such as Commelina communis . The largest light-induced opening of A. thaliana stomates was found at an external KCl concentration of 50 m M . Stomatal apertures were increased by lowering external Ca²⁺ to 0.05 m M . The apertures of stomates incubated with 10 μ M ABA were not altered by changes in Ca²⁺ from 0.05 to 1.0 m M .
Stomates of all abi mutants showed a light-stimulated stomatal opening. The opening of wild type and abi-3 stomates was inhibited by ABA, while stomates of abi-1 and abi-2 did not respond to ABA. The insensitivity of abi-1 and abi-2 stomates to ABA may thus explain the observed disturbed water relations.     

The effect of lanthanide chloride on abscisic acid and electron-transport activity of some crops

The abscisic acid (ABA) content of the root tips of four crops grown in lanthanide chloride solution and their root lengths had been determined. At lanthanide concentrations of 5 and 10 ppm, these crops all grew well and the ABA decreased. At higher lanthanide concentrations (100–500 ppm), the ABA is increased again. At these concentrations of lanthanum chloride, the photosystems I and II (PSI and PSII) and whole electron chain transport activities were inhibited. PSII was more sensitive than PSI, and it is concluded that La³⁺ acts on the diphenylcarbazide (DPC) action place of PSII oxidizing site.     

Abscisic add levels of individual leaf cells

Abscisic acid (ABA) concentrations of guard cells, subsidiary cells and epidermis cells of Comrnelina communis L. and Tradescantia virginiana L. were determined in cell sap samples extracted by means of micro glass capillaries. Concentrations up to 6.7 m M were indicated by commercial immunoassay test kits. A gradient of ABA concentration was found between guard cells (2,49 ± 1.81 m M , n = 25), subsidiary cells (1.25 ± 1.46 m M , n = 21) and epidermis cells (0.86 ± 0.76 m M , n = 20; mean values ± SD).     

盾负泥虫的食性研究

鸣跖草Ｃｏｍｍｅｌｉｎａ ｃｏｍｍｕｎｉｓ Ｌ．是顽固性杂草。质负泥虫Ｌｅｍａ ｓｃｕｔｅｌｌａｒｉｓ １年发生１代,以成虫滞育。用与鸭跖草同科及近缘科、属的植物对其食性进行测定,结果发现其食性单一,只取食鸭跖草,质负泥虫的发生期与鸭跖草的苗期同步,幼虫的喜食与鸭跖草的多分枝特性相吻合,取食营养位与粘液细胞在鸭跖草植株上的分布及细胞的含糖量关系密切。     

The non-osmotic volume of Commelina guard cells

Abstract. The non-osmotic volume (NOV) of Connnelina communis L. guard cells was estimated by observing the volumes of guard cell protoplasts incubated in mannitol solutions of different solute potential, and applying the Boyle-van't Hoff relation to the results. NOV values of between 517 and 1782 μm³ were obtained for different batches of protoplasts. There was a negative correlation between NOV and apparent protoplast solute contents, and the NOV and solute content were observed to alter when pretreatments affecting stomatal aperture were given. H is hypothesized that changes in guard cell chloroplast starch levels could account for variation in NOV and solute content.
For closed stomata, it is calculated that the NOV could reduce the proportion of the total guard cell volume which is osmotically active by over 40%. Serious inaccuracy may thus result if the NOV is not taken into account in the estimation of guard cell solute potential or solute concentration from measurements of solute levels per cell. The error is maximal at low stomatal apertures.     

Bacterial effectors manipulate plant abscisic acid signaling for creation of an aqueous apoplast

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Droopy: a wilty mutant of potato deficient in abscisic acid

Abstract. Droopy mutant of potato ( Solanum tubero-sum L., group Pliureja ) wilts because of excessive stomatal opening (Waggoner & Simmonds, 1966). Progeny of the cross between potato clones C.P.C. 4461 and C.P.C. 4463 showed characteristics similar to those of the original droopy potato. These plants wilted at high vapour pressure deficit and their stomatal conductances in the light and the dark were higher than those of normal plants. Conductances were reduced by applied abscisic acid (ABA), but stomata remained partially open even when guard cells were plasmolysed. Leaves of droopy plants accumulated very little ABA when water-stressed.