Re-export and metabolism of xylem-delivered ABA in attached maize leaves under different transpirational fluxes and xylem ABA concentrations

By feeding radioactive³H-ABA into attached maize leaves, there-export and metabolism of xylem-delivered ABA and their relationshipswith xylem ABA transpirational fluxes and concentrations wereinvestigated. ABA entering leaves in the transpirational streamwas re-exported out of leaves slowly. Within 24 h the proportionof fed radioactivity that was re-exported was less than 45%.When different concentrations of ³H-ABA (100 nM versus 500 nM)was fed, no difference between the two concentrations was foundin their rates of re-export of the fed radioactivity duringthe first 5 h. After 5 h, very little fed radioactivity wasre-exported in leaves that were fed with 100 nM ³H-ABA, whileleaves that were fed with 500 nM ³H-ABA continued to re-exportsuch that the final proportion remaining in leaves after 24h was less as a result, suggesting a concentration-stimulatedre-export. When ³H-ABA was fed at two different transpirationrates which were induced by different air humidity, a 4-folddifference in transpirational fluxes did not produce any differencein terms of re-exportation of fed radioactivity. The rate ofcatabol-ism of xylem-fed ³H-ABA in the attached leaves was muchfaster than that of re-export. On average fed ³H-ABA had a half-lifeof 2.2 h and only 8% remained unmodified after 24 h of incubation,suggesting that re-exported radioactivity might not be the intactform of ABA at all. Using the parameters obtained from the feeding experiment, wecalculated that in a real soil-drying situation the possiblemaximum amount of xylem-delivered ABA that could accumulatein leaves during a day. It was found that the proportion ofdaily accumulated ABA was only 5% of the leaf ABA in well-wateredplants. In soil-dried plants the maximum amount of daily accumulationby xylem ABA could reach 20% of the leaf ABA at the beginningof soil drying, but it soon declined to about 5% again. Thedeclined contribution was mainly due to a reduced transpirationand an increased total leaf ABA as a result of aggravated leafwater deficit. A tight relationship between leaf conductanceand the accumulation of xylem-delivered ABA was not found. Key words: Abscisic acid, ABA, ABA export, ABA metabolism, xylem-delivered ABA, maize     

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.     

JA、ABA在海棠幼苗和枝条中的传输分布以及与水分的关系

采用示踪技术探索了JA和ABA在海棠幼苗和技条中的传输分布以及与水分的关系,发现从的行径主要由韧皮部自上而下运输,ABA主要通过木质部由下向上运输。两者均对土壤水分变化敏感,水分亏缺会加速JA向下运输,促进ABA向上运输,推测JA下运除影响根系生长外,可能与根系对根际环境变化的感应以及根源ABA的合成有关。     

Xylem Sap pH Increase: A Drought Signal Received at the Apoplastic Face of the Guard Cell That Involves the Suppression of Saturable Abscisic Acid Uptake by the Epidermal Symplast

Drought increased the pH of Commelina communis xylem sap from 6.1 to 6.7. Conductances of transpiring leaves were 50% lower in pH 7.0 than in pH 6.0 buffers, but bulk leaf abscisic acid (ABA) concentration and shoot water status were unaffected by pH. Stomatal apertures of isolated abaxial epidermis incubated on simple buffers increased with external pH, so in vivo this must be overridden by alternative pH effects. Reductions in leaf transpiration rate at pH 7.0 were dependent on the presence of 10-8 mol dm-3 ABA in the xylem stream. We inferred that at pH 7.0 leaf apoplastic ABA concentrations increased: pH did not affect distributions of ABA among leaf tissues, but isolated epidermis and mesophyll tissue took up more 3H-ABA from pH 6.0 than from pH 7.0 buffers. The apoplastic ABA increase at pH 7.0 may result from reduced symplastic sequestration. A portion of 3H-ABA uptake by the epidermis was saturable at pH 6.0 but not at pH 7.0. An ABA uptake carrier may contribute to ABA sequestration by the leaf symplast of well-watered plants, and its inactivity at pH 7.0 may favor apoplastic ABA accumulation in draughted plants. Effects of external pH on stomatal apertures in the isolated epidermis indicate that published data supporting a role for internal guard cell ABA receptors should be reassessed.     

Abscisic acid in apoplastic sap can account for the restriction in leaf conductance of white lupins during moderate soil drying and after rewatering

We studied the effects of drought on leaf conductance (g) and on the concentration of abscisic acid (ABA) in the apoplastic sap of Lupinus albus L. leaves. Withholding watering for 5d resulted in complete stomatal closure and in severe leaf water deficit. Leaf water potential fully recovered immediately after rewatering, but the aftereffect of drought on stomata persisted for 2d. ABA and sucrose were quantified in pressurized leaf xylem extrudates. We assumed that the xylem sucrose concentration is negligible and hence that the presence of sucrose in leaf extrudates indicated that they were contaminated by phloem. To eliminate this interference, the concentration of ABA in leaf apoplast was estimated by extrapolation to zero sucrose concentration, using the regression between ABA and sucrose concentrations. The estimated apoplastic ABA concentration increased by 100-fold with soil drying and did not return to pre-stress values immediately following rewatering. g was closely related to the concentration of ABA in leaf apoplast. Furthermore, the feeding of exogenous ABA to leaves detached from well-watered plants brought about the same degree of depression in g as resulted from the drought-induced increase in ABA concentration. We therefore conclude that the observed changes in the concentration of ABA in leaf apoplast were quantitatively adequate to explain drought-induced stomatal closure and the delay in stomatal reopening following rewatering.     

Manipulation of the apoplastic pH of intact plants mimics stomatal and growth responses to water availability and microclimatic variation

The apoplastic pH of intact Forsythiaxintermedia (cv. Lynwood) and tomato (Solanum lycopersicum) plants has been manipulated using buffered foliar sprays, and thereby stomatal conductance (g(s)), leaf growth rate, and plant water loss have been controlled. The more alkaline the pH of the foliar spray, the lower the g(s) and/or leaf growth rate subsequently measured. The most alkaline pH that was applied corresponds to that measured in sap extracted from shoots of tomato and Forsythia plants experiencing, respectively, soil drying or a relatively high photon flux density (PFD), vapour pressure deficit (VPD), and temperature in the leaf microclimate. The negative correlation between PFD/VPD/temperature and g(s) determined in well-watered Forsythia plants exposed to a naturally varying summer microclimate was eliminated by spraying the plants with relatively alkaline but not acidic buffers, providing evidence for a novel pH-based signalling mechanism linking the aerial microclimate with stomatal aperture. Increasing the pH of the foliar spray only reduced g(s) in plants of the abscisic acid (ABA)-deficient flacca mutant of tomato when ABA was simultaneously sprayed onto leaves or injected into stems. In well-watered Forsythia plants exposed to a naturally varying summer microclimate (variable PFD, VPD, and temperature), xylem pH and leaf ABA concentration fluctuated but were positively correlated. Manipulation of foliar apoplastic pH also affected the response of g(s) and leaf growth to ABA injected into stems of intact Forsythia plants. The techniques used here to control physiology and water use in intact growing plants could easily be applied in a horticultural context.     

Root and bacterial secretions regulate the interaction between plants and PGPR leading to distinct plant growth promotion effects

Background and aims

Long distance signals in xylem from roots to leaves are important in plant response to drought stress. Abscisic acid (ABA) plays a key role in drought signaling in plants but apoplastic pH may modulate its effect by distributing ABA into various compartments in leaves. We aimed to reveal the dynamics of changes in sap pH and its relationships with the transport of inorganic and organic ions in eight herbaceous plant species under continuously declining soil water content. We tested several hypotheses related to the mechanism of pH changes in xylem.

Methods

We used a pressure chamber to collect xylem sap and to measure of leaf/stem water potential at various stages of soil drying. We measured pH and concentrations of the most abundant inorganic (NO₃ ^?, SO₄ ^2?, PO₄ ^3? and Cl^?) and organic (malate and citrate) anions in xylem sap.

Results

Species differed considerably in the dynamics of pH changes in xylem in drying soil. Changes in xylem sap pH during drying did not relate to the nitrogen assimilation strategy but may be affected by sap flow rate. Simultaneous changes in the concentrations of inorganic and organic anions were highly species-specific.

Conclusions

High variability among species in the observed relationships in response to drought indicates that comparisons among different studies and the generalization of results should be made with caution.

     

Stomatal Reactions of Two Different Maize Lines to Osmotically Induced Drought Stress

Two maize lines differing in drought resistance were grown at different drought stress induced by polyethylene glycol (PEG 10 000) solutions with osmotic potentials of –0.20, –0.40 and –0.80 MPa in the semipermeable membrane system. During the five days soil water content decreased (from 0.43 to 0.29, 0.25 and 0.23 g cm^–3 for three PEG solutions, respectively) as well as leaf water potentials (_w; from – 0.54 to –0.76, –1.06 and –1.46 MPa). These values were not significantly different between the investigated lines, indicating that a controlled and consistent soil moisture stress was achieved. Soil drying induced an increase in the ABA content of leaves and xylem of both lines and the effects on stomatal conductance were greater in drought susceptible line (B-432) compared to drought resistant line (ZPBL-1304). To test possible difference in stomatal sensitivity to xylem ABA between lines and to assess any ABA vs. _w interaction, roots were fed with 10, 50 and 100 mmol m^–3 ABA solutions in another set of experiments. These results showed that manipulation of xylem ABA affected stomata of both lines similarly. Comparison of stomatal sensitivity to drought-induced and applied ABA demonstrated that drought treatment affected stomata of investigated lines by differentially increasing their sensitivity to xylem ABA, thus confirming an interaction between chemical signalling and hydraulic signalling.     

Effects of Xylem pH on Transpiration from Wild-Type and flacca Tomato Leaves : A Vital Role for Abscisic Acid in Preventing Excessive Water Loss Even from Well-Watered Plants

The pH of xylem sap from tomato (Lycopersicon esculentum) plants increased from pH 5.0 to 8.0 as the soil dried. Detached wild-type but not flacca leaves exhibited reduced transpiration rates when the artificial xylem sap (AS) pH was increased. When a well-watered concentration of abscisic acid (0.03 μm) was provided in the AS, the wild-type transpirational response to pH was restored to flacca leaves. Transpiration from flacca but not from wild-type leaves actually increased in some cases when the pH of the AS was increased from 6.75 to 7.75, demonstrating an absolute requirement for abscisic acid in preventing stomatal opening and excessive water loss from plants growing in many different environments.Jones (1980) and Cowan (1982) were the first to suggest that plants can “measure” soil water status independently of shoot water status via the transfer of chemical information from roots to shoots. Dehydrating roots in drying soil synthesize ABA more rapidly than fully turgid tissue, and resultant increases in the ABA concentration of xylem sap flowing toward the still-turgid shoot constitutes a chemical signal to the leaves (for review, see Davies and Zhang, 1991): the xylem vessels give up their contents to the leaf apoplast, thereby increasing the ABA concentration in this compartment. ABA receptors on the external surface of stomatal guard cells respond to the apoplastic ABA concentration (Hartung, 1983; Anderson et al., 1994; but see Schwartz et al., 1994). When bound, the receptors transduce a reduction in guard cell turgor, which leads to stomatal closure (Assmann, 1993). This maintains shoot water potential despite the reduction in soil water availability.Another chemical change related to soil drying in the absence of a reduction in shoot water status is an increase in the pH of the xylem sap flowing from the roots (Schurr et al., 1992). The pH of the xylem and/or apoplastic sap of plants can also change dramatically in response to soil flooding, diurnal or annual rhythms, and mineral nutrient supply (Table ​(TableI)I) in the absence of concomitant changes in either root or shoot water status. We already know that, like the increase in xylem ABA concentration described above, an increase in xylem pH can also act as a signal to leaves to close their stomata (Wilkinson and Davies, 1997). Since the conditions that affect xylem/apoplastic pH can also affect transpiration (light intensity [Cowan et al., 1982]; soil drying [Davies and Zhang, 1991]; nitrate supply [Clarkson and Touraine, 1994]; soil flooding [Else, 1996]), the possibility exists that the pH change that they induce could be the means by which they alter stomatal aperture. Table IpH changes that occur in plant xylem or apoplastic sap under various conditions It was originally suggested that an increase in xylem sap pH could putatively enhance stomatal closure by changing the distribution of the ABA that is present in all nonstressed plants at a low “background” concentration, without requiring de novo ABA synthesis (Schurr et al., 1992; Slovik and Hartung, 1992a, 1992b). This hypotheses is built on the well-known fact that weak acids such as ABA accumulate in more alkaline compartments (Kaiser and Hartung, 1981). More recently, Wilkinson and Davies (1997) and Thompson et al. (1997) directly demonstrated that increases in xylem sap pH reduced rates of water loss from Commelina communis and tomato (Lycopersicon esculentum) leaves detached from well-watered plants. This was found to be mediated by the relatively low endogenous concentration of ABA (about 0.01 mmol m⁻³) contained in the xylem vessels and apoplast of these leaves, a concentration of ABA that did not itself affect transpiration at a well-watered sap pH of 6.0. The mechanism by which the combination of high sap pH and such a low concentration of ABA was able to increase the apoplastic ABA concentration sufficiently to close stomata was also elucidated: the mesophyll and epidermis cells of these leaves had a greatly reduced ability to sequester ABA away from the apoplast when the pH of the latter was increased by the incoming xylem sap (Wilkinson and Davies, 1997).In contrast to the indirect ABA-mediated effect of pH on stomata, it was also demonstrated that increasing the pH of the external solution (from 5.0 to 7.0) bathing isolated abaxial epidermis tissue peeled from well-watered C. communis leaves actually increased stomatal aperture (Wilkinson and Davies, 1997). Mechanisms for this direct effect of pH on guard cells have been speculated on by Thompson et al. (1997). If this process were to occur in vivo, environments that increase xylem sap pH could potentially induce excessive water loss from the plants experiencing them, over and above rates of transpiration occurring in unstressed plants. The latter may contain stomata with apertures smaller than the maximum that is possible, even under favorable local conditions. It was assumed that high-pH-induced apoplastic ABA accumulation in C. communis in vivo was sufficient to override the direct stomatal opening effect seen in the isolated tissue (Wilkinson and Davies, 1997). To test these possibilities, effects of pH on transpiration rates from leaves of the flacca mutant of tomato were investigated. flacca does not synthesize ABA as efficiently as wild-type tomato (Parry et al., 1988; Taylor et al., 1988). It contains a very low endogenous ABA concentration (Tal and Nevo, 1973), although it retains the ability to respond to an application of this hormone (Imber and Tal, 1970). The results demonstrate not only that ABA mediates high xylem sap pH-induced stomatal closure but also that it is necessary to prevent high xylem sap pH-induced stomatal opening and dangerously excessive water loss.     

Changes in the concentration of ABA in xylem sap as a function of changing soil water status can account for changes in leaf conductance and growth

Abstract. Maize seedlings ( Zea mays L. John Innes F1 hybrid) were grown in a greenhouse in l-m-long tubes of soil. When the plants were well established, water was withheld from half of the tubes. Control plants were watered every day during the 20-d experimental period. The soil drying treatment resulted in a substantial restriction of stomatal conductance and a limitation in shoot growth, even though there was no detectable difference in the water relations of watered and unwatered plants. From day 7 of the soil drying treatment, xylem ABA concentrations (measured using the sap exuded from detopped plants) were substantially increased in unwatered plants compared to values recorded with sap from plants watered every day. Measurements of water potential through the profile of unwatered soil suggest that xylem ABA concentrations reflects the extent of soil drying. Leaf ABA content was a much less sensitive indicator of the effect of soil drying and during the whole of experimental period there was no significant difference between ABA concentration in leaves of well watered and unwatered plants. In a second set of experiments, ABA was fed to part of the roots of potted maize plants to manipulate xylem ABA concentration. These manipulations suggested that the increases in ABA concentration in xylem sap, which resulted from soil drying, were adequate to explain the observed variation in stomatal conductance and might also explain the restriction in leaf growth rate. These results are discussed in the light of recent work which suggests that stomatal responses to soil drying are partly attributable to an as-yet unidentified inhibitor of stomatal opening.     

Compartmental distribution and redistribution of abscisic acid in intact leaves

Using a computer model written for whole leaves (Slovik et al. 1992, Planta 187, 14–25) we present in this paper calculations of abscisic acid (ABA) redistribution among different leaf tissues and their compartments in relation to stomatal regulation under drought stress. The model calculations are based on experimental data and biophysical laws. They yield the following results and postulates: (i) Under stress, compartmental pH-shifts come about as a consequence of the inhibition of the pH component of proton-motive forces at the plasmalemma. There is a decrease of net proton fluxes by about 8.6 nmol · s^–1 · m^–2. (ii) Using stress-induced pH-shifts we demonstrate how stress intensities can be quantified on a molecular basis. (iii) As the weak acid ABA is the only phytohormone which behaves in vivo and in vitro ideally according to the Henderson-Hasselbalch equation, pH-shifts induce a complicated redistribution amongst compartments in the model leaf. (iv) The final accumulation of ABA in guard-cell walls is intensive: up to 16.1-fold compared with only up to 3.4-fold in the guard-cell cytosol. We propose that the binding site of the guard-cell ABA receptor faces the apoplasm. (v) A twoto three-fold ABA accumulation in guard-cell walls is sufficient to induce closure of stomata. (vi) The minimum time lag until stomata start to close is 1–5 min; it depends on the stress intensity and on the guard-cell sensitivity to ABA: the more moderate the stress is, the later stomata start to close or they do not close at all. (vii) In the short term, there is almost no influence of the velocity of pH-shifts on the velocity of the ABA redistribution, (viii) Six hours after the termination of stress there is still an ABA concentration 1.4-fold the initial level in the guard-cell cytosol (delay of ABA relaxation, aftereffect), (ix) The observed induction of net ABA synthesis after onset of stress may be explained by a decrease in cytosolic ABA degradation. About 1 h after onset of stress the model leaf would start to synthesise ABA (and its conjugates) automatically, (x) This ABA net synthesis serves to inform roots via an increased ABA concentration in the phloem sap. The stress-induced ABA redistribution is per se not sufficient to feed the ploem sap with ABA. (xi) The primary target membrane of stress is the plasmalemma, not thylakoids. (xii) The effective stress sensor, which induces the proposed signal chain finally leading to stomatal closure, is located in epidermal cells. Mesophyll cells are not capable of creating a significant ABA signal to guard cells if the epidermal plasmalemma conductance to undissociated molecular species of ABA (HABA) is indeed higher than the plasmalemma conductance of the mesophyll (plasmodesmata open), (xiii) All model conclusions which can be compared with independent experimental data quantitatively fit to them. We conclude that the basic experimental data of the model are consistent. A stress-induced ABA redistribution in the leaf lamina elicits stomatal closure.Abbreviations ABA abscisic acid - CON vacuolar ABA conjugates We are grateful to Prof. U. Heber (Lehrstuhl Botanik I, University of Würzburg, FRG) for stimulating discussions. This work has been performed within the research program of the Sonderforschungsbereich 251 (TP 3 and 4) of the University of Würzburg. It has been also supported by the Fonds der Chemischen Industrie.     

Foliar application of nitrate or ammonium as sole nitrogen supply in Ricinus communis. II. The flows of cations, chloride and abscisic acid

Following a precultivation with pedospheric nitrogen nutrition, Ricinus plants were supplied with nitrogen solely by spraying nitrate or ammonium solution onto the leaves during the experimental period. The chemical composition of tissues, xylem and phloem exudates was determined and on the basis of the previously determined nitrogen flows (Peuke et al., New Phytologist (1998), 138 , 657–687) the flows of potassium, sodium, magnesium, calcium, chloride and ABA were modelled. These data, which permit quantification of net-uptake, transport in xylem and phloem, and utilization in shoot and root, were compared with results obtained in plants with pedospherically-supplied nitrate or ammonium and data in the literature. Although the overall effects on the chemical composition of supplying ammonium to the leaves were not as pronounced as in pedospherically supplied plants, there were some typical responses of plants fed with ammonium (ammonium syndrome). In particular, in ammonium-sprayed plants uptake and transport of magnesium decreased and chloride uptake was increased compared with nitrate-sprayed plants. Furthermore, acropetal ABA transport in the xylem in ammonium-sprayed Ricinus was threefold higher than in nitrate-sprayed plants. Additionally, concentrations of anions were more or less increased in tissues, particularly in the roots, and transport fluids. The overall signal from ammonium-sprayed leaves without a direct effect of ammonium ions on uptake and transport systems in the root is discussed.     

Sequential response of whole plant water relations to prolonged soil drying and the involvement of xylem sap ABA in the regulation of stomatal behaviour of sunflower plants

Sunflower plants ( Helianihus animus cv. Tall Single Yellow} were grown in the greenhouse in drain pipes (100 mm inside diameter and 1 m long) rilled with John Innes No. 2 compost. When the fifth leaf had emerged, half of the plants were left unwatered for 6 days, rewatered for 2 days and then not watered for another 12 days. Measurements of water relations and abaxial stomatal conductance were made at each leaf position at regular intervals during the experimental period. Estimates were also made of soil water potentials along the soil profile and of ABA concentrations in xylem sap and leaves.
Soil drying led to some reduction in stomatal conductance alter only 3 days but leaf turgors were not reduced until day 13 (6 days after rewatering). When the water relations of leaves did change, older leases became substantially dehydrated while high turgors were recorded in younger leaves. Leaf ABA content measured on the third youngest leaf hardly changed over the first 13 days of the experiment, despite substantial soil drying, while xylem ABA concentrations changed very significantly and dynamically as soil water status varied, even when there was no effect of soil drying on leaf water relations. We argue that the highest ABA concentrations in the xylem, found as a result of substantial soil drying, arise from synthesis in both the roots and the older leaves, and act to delay the development of water deficit in younger leases.
In other experiments ABA solutions were watered on to the root systems of sunflower plants to increase ABA concentrations in xylem sap. The stomatal response to applied ABA was quantitatively very similar to that to ABA generated as a result of soil drying. There was a log-linear relationship between the reduction of leaf conductance and the increase of ABA concentration m xylem sap.     

Patterns of auxin and abscisic acid movement in the tips of gravistimulated primary roots of maize

Because both abscisic acid (ABA) and auxin (IAA) have been suggested as possible chemical mediators of differential growth during root gravitropism, we compared with redistribution of label from applied ³H-IAA and ³H-ABA during maize root gravitropism and examined the relative basipetal movement of ³H-IAA and ³H-ABA applied to the caps of vertical roots. Lateral movement of ³H-ABA across the tips of vertical roots was non-polar and about 2-fold greater than lateral movement of ³H-IAA (also non-polar). The greater movement of ABA was not due to enhanced uptake since the uptake of ³H-IAA was greater than that of ³H-ABA. Basipetal movement of label from ³H-IAA or ³H-ABA applied to the root cap was determined by measuring radioactivity in successive 1 mm sections behind the tip 90 minutes after application. ABA remained largely in the first mm (point of application) whereas IAA was concentrated in the region 2–4 mm from the tip with substantial levels found 7–8 mm from the tip. Pretreatment with inhibitors of polar auxin transport decreased both gravicurvature and the basipetal movement of IAA. When roots were placed horizontally, the movement of ³H-IAA from top to bottom across the cap was enhanced relative to movement from bottom to top whereas the pattern of movement of label from ³H-ABA was unaffected. These results are consistent with the hypothesis that IAA plays a role in root gravitropism but contrary to the idea that gravi-induced asymmetric distribution of ABA contributes to the response.     

Chemical signals and their regulations on the plant growth and water use efficiency of cotton seedlings under partial root-zone drying and different nitrogen applications

Partial root-zone drying during irrigation (PRD) has been shown effective in enhancing plant water use efficiency (WUE), however, the roles of chemical signals from root and shoot that are involved and the possible interactions affected by nitrogen nutrition are not clear. Pot-grown cotton (Gossypium spp.) seedlings were treated with three levels of N fertilization and PRD. The concentrations of nitrate (NO₃⁻), abscisic acid (ABA) and the pH value of leaf and root xylem saps, biomass and WUE were measured. Results showed that PRD plants produced larger biomass and higher WUE than non-PRD plants, with significant changes in leaf xylem ABA, leaf and root xylem NO₃⁻ concentrations and pH values, under heterogeneous soil moisture conditions. Simultaneously, high-N treated plants displayed larger changes in leaf xylem ABA and higher root xylem NO₃⁻ concentrations, than in the medium- or low-N treated plants. However, the WUE of plants in the low-N treatment was higher than that of those in the high- and medium-N treatments. PRD and nitrogen levels respectively induced signaling responses of ABA/NO₃⁻ and pH in leaf or root xylem to affect WUE and biomass under different watering levels, although significant interactions of PRD and nitrogen levels were found when these signal molecules responded to soil drying. We conclude that these signaling chemicals are regulated by interaction of PRD and nitrogen status to regulate stomatal behavior, either directly or indirectly, and thus increase PRD plant WUE under less irrigation.     

Signalling mechanisms involved in the response of two varieties of Humulus lupulus L. to soil drying: II. changes in the concentration of abscisic acid catabolites and stress-induced phytohormones

Abscisic acid (ABA) is one of the most common stress signals that appear in plant organs in response to soil drying. Equilibrium between ABA biosynthesis and catabolism regulates ABA accumulation in plants under water stress. The aim of our work was to explore the dynamics of changes in ABA metabolites as well as other stress-induced phytohormones such as jasmonic acid, indole-3-acetic acid, and their respective metabolites in hop [Humulus lupulus (L.)] plants during drying and to identify among them potential signals involved in drought signalling. We showed that the concentrations of all ABA metabolites (except the concentration of ABA glucosyl ester in leaves) increased in the same manner in leaves and xylem sap approximately at the same level of soil water content when the relative water content of leaves decreased. The predominant metabolites in leaves and xylem sap were phaseic acid and dihydroxyphaseic acid. ABA glucosyl ester was not a source of the increased concentration of ABA in leaves and xylem sap because of its considerably lower concentration compared to ABA. The concentration of jasmonates decreased in leaves of hop plants. Changes in auxin concentration suggest that this hormone is involved in the response of hop plants to soil drying.     

Changes of free and conjugated abscisic acid and phaseic acid in xylem sap of drought-stressed sunflower plants

Abscisic acid (ABA), conjugated abscisic acid, phaseic acid (PA), and conjugated phaseic acid were determined by enzyme-linked immunosorbent assay (ELISA) and gas chromatography (GC) in xylem sap of well-watered and drought-stressed sunflower plants. Conjugated ABA and conjugated PA were determined indirectly after chemical or enzymatic hydrolysis. Conjugated ABA was found to be the predominant ABA metabolite in xylem sap. In xylem sap from well-watered plants at least five, and in sap from drought-stressed plants at least six alkaline hydrolysable ABA conjugates were found. One of them corresponds chromatographically (HPLC) with abscisic acid glucose ester (ABAGE). Under drought conditions the concentrations of ABA, alkaline hydrolysable ABA conjugates, -glucosidase hydrolysable ABA conjugates, PA, and conjugated PA increased. After rewatering the drought-stressed plants, the ABA and the conjugated ABA content decreased. The possible function of the ABA conjugates in the xylem sap as a source of free ABA is discussed.     

Signalling mechanisms involved in the response of two varieties of Humulus lupulus L. to soil drying: I. changes in xylem sap pH and the concentrations of abscisic acid and anions

Background and aims

Soil drying leads to the generation of chemical signals in plants that regulate water use via control of the stomatal aperture. The aim of our work was to identify the presence and identity of potential chemical signals, their dynamics, and their relationship with transpiration rate during soil drying in hop (Humulus lupulus (L.)) plants.

Methods

We used pressure chamber technique for measurement of shoot water potential and collection of shoot xylem sap. We analyzed concentrations of abscisic acid (ABA), nitrate, phosphate, sulphate and malate in sap and also the rate of whole plant transpiration.

Results

Transpiration rate decreased prior to changes in shoot water potential. The concentration of ABA in xylem sap continuously increased from early to later stages of water stress, whereas in leaves it increased only at later stages. Shoot sap pH increased simultaneously with the decrease of transpiration rate. Xylem sap alkalization was in some cases accompanied by a decrease in nitrate concentration and an increase in malate concentration. Concentration of sulphate increased in xylem sap during drying and sulphate in combination with a higher ABA concentration enhanced stomatal closure.

Conclusions

Several early chemical signals appear in sap of hop plants during soil drying and their impact on transpiration may vary according to the stage of soil drying.     

Transport, synthesis and catabolism of abscisic acid (ABA) in intact plants of castor bean (Ricinus communis L.) under phosphate deficiency and moderate salinity

Flows of abscisic acid (ABA) were investigated in whole plantsof castor bean (Ricinus communis) grown in sand culture undereither phosphate deficiency or moderate salinity. Xylem transportof ABA in P-deficient plants was stimulated by a factor of 6whereas phloem transport was affected only very slightly. ABAdeposition into leaves of P-deficient plants was not appreciablydifferent from the controls because of strong net degradationin leaves. Since conjugation of ABA was strongly reduced inall organs of P-deficient plants ABA was presumably metabolizedmainly to phaseic acid and dihydrophaseic acid. The increasedimport of ABA occurred predominantly into fully differentiatedbut not senescent leaves and showed a good correlation withthe inhibition of leaf conductance under P deficiency. As with low-P-plants salt stress increased ABA synthesis inroots and associated transport in the xylem. However, salinitycaused a distinctly greater accumulation of ABA in the leaves,stem segments and the apex than in P-deficient plants. As opposedto P deficiency, ABA export in the phloem from the leaves wasstimulated by salinity. Modelling of ABA flows within an individualleaf over its life cycle showed that young growing leaves importedABA from both phloem and xylem, whereas the adult non-senescentleaves were a source of ABA and thus provided a potential shoot-to-rootstress signal as well as an acceptor for reciprocal signalsfrom root to shoot. In senescing leaves ABA flows and accumulationwere somewhat retarded and ABA was lost in net terms by exportfrom the leaf. Key words: Abscisic acid, phosphorus deficiency, salt stress, phloem and xylem transport