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.     

Abscisic Acid is not the only stomatal inhibitor in the transpiration stream of wheat plants

Xylem sap was collected from the transpiration stream of wheat (Triticum aestivum L.) plants and assayed for the presence of an inhibitor of transpiration using leaves detached from well-watered plants. Transpiration of detached leaves was reduced by nearly 60% by sap collected from plants in drying soil, and to a lesser extent (about 25%) by sap from plants in well-watered soil. As the soil dried the abscisic acid (ABA) concentration in the sap increased by about 50 times to 5 × 10⁻⁸ molar. However, the ABA in the sap did not cause its inhibitory activity. Synthetic ABA of one hundred times this concentration was needed to reduce transpiration rates of detached leaves to the same extent. Furthermore, inhibitory activity of the sap was retained after its passage through an immunoaffinity column to remove ABA. Xylem sap was also collected by applying pressure to the roots of plants whose leaf water status was kept high as the soil dried. Sap collected from these plants reduced transpiration to a lesser extent than sap from nonpressurised plants. This suggests that the inhibitory activity was triggered partly by leaf water deficit and partly by root water deficit.     

Transpiration inhibition by stored xylem sap from well-watered maize plants

There is increasing evidence that a chemical signal exists in xylem sap of plants subjected to water deficits which influences physiological responses in plant shoots. An important method of studying this signal is the transpiration response of excised leaves exposed to xylem sap collected from plants. However, Munns et al [Plant, Cell & Environment 16, 867–877] cautioned that transpiration inhibition is observed when xylem sap collected from wheat and barley is stored before determining physiological activity. The objective of the study reported here was to determine if transpiration inhibition develops in maize sap collected from well-watered plants when the sap is stored under various conditions. It was found that storage of maize sap collected from well-watered plants for only 1 d at -20°C resulted in the development of substantial transpiration inhibition in bioassay leaves. Storage of sap at 4°C resulted in the development of the effect after 2 weeks, while storage at ?86°C showed only small transpiration inhibition after 3 weeks. The major source of the transpiration inhibition was the development of a substance in the stored sap that resulted in physical blockage of the transpiration stream in bioassay leaves. However, a small signal component may also have developed in the stored sap. Because of the possibility of ionic activity under freezing conditions at ?20°C, calcium was studied for its potential involvement in the transpiration inhibition. However, the calcium concentrations found to inhibit transpiration were nearly an order of magnitude larger than the calcium concentrations observed in xylem sap.     

Do increases in xylem sap pH and/or ABA concentration mediate stomatal closure following nitrate deprivation?

Stomatal conductance (g(s)) of pepper (Capsicum annuum L.) plants decreased during the second photoperiod (day 2) after withholding nitrate (N). Stomatal closure of N-deprived plants was not associated with a decreased shoot water potential (Psi(shoot)); conversely Psi(shoot) was lower in N-supplied plants. N deprivation transiently (days 2 and 3) alkalized (0.2-0.3 pH units) xylem sap exuded from de-topped root systems under root pressure, and xylem sap expressed from excised shoots by pressurization. The ABA concentration of expressed sap increased 3-4-fold when measured on days 2 and 4. On day 2, leaves detached from N-deprived and N-supplied plants showed decreased transpiration rates when fed an alkaline (pH 7) artificial xylem (AX) solution, independent of the ABA concentration (10-100 nM) supplied. Thus changes in xylem sap composition following N deprivation can potentially close stomata. However, the lower transpiration rate of detached N-deprived leaves relative to N-supplied leaves shows that factors residing within N-deprived leaves also mediate stomatal closure, and that these factors assume greater importance as the duration of N deprivation increases.     

What information is conveyed by an ABA signal from maize roots in drying field soil?

During two seasons, ABA concentrations were monitored in roots, leaves and xylem sap of field-grown maize. The water status of soil and plant was also measured. Plants were grown on plots with compacted or non-compacted soil, which were irrigated or remained unwatered. ABA concentration in the xylem sap before dawn and in the roots increases 25-fold and five-fold, respectively, as the soil dried, with a close correlation with the soil water status, but with no clear effect of the soil structure. In contrast to the results of several laboratory experiments, no appreciable increase in xylem [ABA] and reduction in stomatal conductance were observed with dehydration of the part of the root system located in soil upper layers. These responses only occurred when the water reserve of the whole soil profile was close to depletion and the transpiration declined. Xylem [ABA] measured during the day was appreciably higher in the compacted treatment than in non-compacted treatment, unlike that measured before dawn. Since a mechanical message is unlikely to undergo such day-night alterations, we suggest that this was due to a faster decrease in root water potential and water flux in the compacted treatment, linked to the root spatial arrangement. These results raise the possibility that ABA concentration in the xylem sap could be controlled by two coexisting mechanisms: (1) the rate of ABA synthesis in the roots linked to the soil or root water status, as shown in laboratory experiments; (2) the dilution of ABA in the water flow from roots, which could be an overriding mechanism in field conditions. This second mechanism would allow the plant to sense the water flux through the root system.     

Concentrations of abscisic acid and other solutes in xylem sap from root systems of tomato and castor-oil plants are distorted by wounding and variable sap flow rates

We investigated if concentrations of abscisic acid (ABA) andother solutes measured in the first few droplets of xylem sapfrom detopped root systems, are good estimates of those in thetranspiration stream as it enters the shoot-base of whole plants.Xylem sap from root systems of pot-grown tomato plants (Lycopersiconesculentum Mill., cv. Ailsa Craig), at the seven-leaf stage,was obtained by placing root systems in chambers pressurizedto 0.3 MPa with air. The first sample was taken from the cut-surfaceof the hypo-cotyl stump within 30 s of removing the shoot. ABA,sucrose and other osmolytes were more concentrated in the initial100–200 mm³ of xylem sap than in subsequent samples. Thissuggested the sap was contaminated and not unchanged transpirationfluid. The effect was reproduced on the same plant, severaltimes, by recutting the hypocotyl prior to reassembling thesap collecting set-up and repressurizing. Similar results werefound with castor-oil plants (Ricinus communis L., cv. Gibsonii).However, neither release of ABA from the cut surface of thehypocotyl stump, nor the effects of pressure to the roots causedthe contamination. Instead, small radial pressures exerted bya rubber sleeve attached to the hypocotyl stump, for collectingthe sap, were responsible. The effect was reproduced by lightlysqueezing the hypocotyl by hand. The possibility was examined that reliable estimates of ABAconcentrations in transpiration stream fluid may be obtainedfrom sap samples taken immediately after rejecting the initial,contaminated 200 mm³. However, ABA concentrations in these latersamples were also unsatisfactory since they changed with rateof sap flow. The problem may be overcome by analysing sap inducedto flow through detached root systems at rates close to thoseof whole-plant transpiration. Key words: Tomato, Lycopersicon esculentum Mill., Castor-oil plant, Ricinus communis L., roots, root to shoot communication, xylem sap, abscisic acid, sucrose, transpiration stream     

Anti-transpirant activity in xylem sap from flooded tomato (Lycopersicon esculentum Mill.) plants is not due to pH-mediated redistributions of root- or shoot-sourced ABA

In flooded soils, the rapid effects of decreasing oxygen availability on root metabolic activity are likely to generate many potential chemical signals that may impact on stomatal apertures. Detached leaf transpiration tests showed that filtered xylem sap, collected at realistic flow rates from plants flooded for 2 h and 4 h, contained one or more factors that reduced stomatal apertures. The closure could not be attributed to increased root output of the glucose ester of abscisic acid (ABA-GE), since concentrations and deliveries of ABA conjugates were unaffected by soil flooding. Although xylem sap collected from the shoot base of detopped flooded plants became more alkaline within 2 h of flooding, this rapid pH change of 0.5 units did not alter partitioning of root-sourced ABA sufficiently to prompt a transient increase in xylem ABA delivery. More shoot-sourced ABA was detected in the xylem when excised petiole sections were perfused with pH 7 buffer, compared with pH 6 buffer. Sap collected from the fifth oldest leaf of "intact" well-drained plants and plants flooded for 3 h was more alkaline, by approximately 0.4 pH units, than sap collected from the shoot base. Accordingly, xylem [ABA] was increased 2-fold in sap collected from the fifth oldest petiole compared with the shoot base of flooded plants. However, water loss from transpiring, detached leaves was not reduced when the pH of the feeding solution containing 3-h-flooded [ABA] was increased from 6.7 to 7.1 Thus, the extent of the pH-mediated, shoot-sourced ABA redistribution was not sufficient to raise xylem [ABA] to physiologically active levels. Using a detached epidermis bioassay, significant non-ABA anti-transpirant activity was also detected in xylem sap collected at intervals during the first 24 h of soil flooding.     

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     

Hydraulic and Chemical Responses of Citrus Seedlings to Drought and Osmotic Stress

In this work we investigated the function of abscisic acid (ABA) as a long-distance chemical signal communicating water shortage from the root to the shoot in citrus plants. Experiments indicated that stomatal conductance, transpiration rates, and leaf water potential decline progressively with drought. ABA content in roots, leaves, and xylem sap was also increased by the drought stress treatment three- to sevenfold. The addition of norflurazon, an inhibitor of ABA biosynthesis, significantly decreased the intensity of the responses and reduced ABA content in roots and xylem fluid, but not in leaves. Polyethylene glycol (PEG)-induced osmotic stress caused similar effects and, in general, was counteracted only by norflurazon at the lowest concentration (10%). Partial defoliation was able to diminish only leaf ABA content (22.5%) at the highest PEG concentration (30%), probably through a reduction of the active sites of biosynthesis. At least under moderate drought (3–6 days without irrigation), mechanisms other than leaf ABA concentration were required to explain stomatal closure in response to limited soil water supply. Measurements of xylem sap pH revealed a progressive alkalinization through the drought condition (6.4 vs. 7.1), that was not counteracted with the addition of norflurazon. Moreover, in vitro treatment of detached leaves with buffers iso-osmotically adjusted at pH 7.1 significantly decreased stomatal conductance (more than 30%) as much as 70% when supplemented with ABA. Taken together, our results suggest that increased pH generated in drought-stressed roots is transmitted by the xylem sap to the leaves, triggering reductions in shoot water loss. The parallel rise in ABA concentration may act synergistically with pH alkalinization in xylem sap, with an initial response generated from the roots and further promotion by the stressed leaves.     

Control of Leaf Expansion Rate of Droughted Maize Plants under Fluctuating Evaporative Demand (A Superposition of Hydraulic and Chemical Messages?)

We have analyzed the possibility that chemical signaling does not entirely account for the effect of water deficit on the maize (Zea mays L.) leaf elongation rate (LER) under high evaporative demand. We followed time courses of LER (0.2-h interval) and spatial distribution of elongation rate in leaves of either water-deficient or abscisic acid (ABA)-fed plants subjected to varying transpiration rates in the field, in the greenhouse, and in the growth chamber. At low transpiration rates the effect of the soil water status on LER was related to the concentration of ABA in the xylem sap and could be mimicked by feeding artificial ABA. Transpiring plants experienced a further reduction in LER, directly linked to the transpiration rate or leaf water status. Leaf zones located at more than 20 mm from the ligule stopped expanding during the day and renewed expansion during the night. Neither ABA concentration in the xylem sap, which did not appreciably vary during the day, nor ABA flux into shoots could account for the effect of evaporative demand. In particular, maximum LER was observed simultaneously with a minimum ABA flux in the droughted plants, but with a maximum ABA flux in ABA-fed plants. All data were interpreted as the superposition of two additive effects: the first involved ABA signaling and was observed during the night and in ABA-fed plants, and the second involved the transpiration rate and was observed even in well-watered plants. We suggest that a hydraulic signal is the most likely candidate for this second effect.     

Delivery rates of abscisic acid in xylem sap of Ricinus communis L. plants subjected to part-drying of the soil

Abscisic acid (ABA) moving from roots to shoots in the transpirationstream is a potential hormonal message integrating perceptionof a root stress with adaptive changes in the shoot. A twinroot system was used to study ways of estimating effects ofdroughting the upper roots of Ricinus communis L. on abscisicacid (ABA) transport to the shoot in the transpiration stream.Droughted plants transpired more slowly than controls. Droughtingalso increased concentrations of ABA up to I I-fold in sap inducedto flow from the roots of freshly decapitated plants at ratesof whole plant transpiration. However, because of dilution effectsarising from the different sap flows in control and droughtedplants, these changes in ABA concentration in the xylem sapdid not accurately reflect amounts of ABA transported. To overcomethis problem, delivery rates were calculated by multiplyingconcentration with sap flow rate to generate ABA delivery interms of µmol s–1 per plant. Droughting for 24 hor more increased ABA delivery from roots to shoots by 5-fold.Since droughting can alter the relative sizes of the roots andshoots and also the root:shoot ratio these delivery rates wererefined in several ways to reflect both the amount of root generatingthe ABA message and the size of the recipient shoot system. Key words: Abscisic acid, Ricinus communis L., soil drying, xylem sap     

Transpiration, CO2 assimilation, WUE, and stomatal aperture in leaves of Viscum album (L.): Effect of abscisic acid (ABA) in the xylem sap of its host (Populus x euamericana).

Leaves of the mistletoe Viscum album (L.) show a high rate of transpiration, even when the host is under severe drought stress. The hypothesis that a strong control of ABA influx from the xylem sap of the host into the mistletoe prevents stomatal closure in mistletoe leaves was tested under the following conditions: sections of poplar twigs carrying a mistletoe were perfused with artificial xylem sap that contained different ABA concentrations and both transpiration and ABA levels were analysed in mistletoe leaves. Despite variation by a factor of 10(4), the ABA content of the host xylem did not affect ABA levels, leaf transpiration, CO(2) assimilation, WUE, or the degree of stomatal aperture in mistletoe leaves. These observations support the hypothesis of a strong control of ABA influx from the host of the xylem into the mistletoe, although degradation of ABA before it enters the mistletoe leaves cannot be excluded. This mechanism may ensure a water and nutritional status favourable for the mistletoe, even if the water status of the host is impaired. Despite the lack of short-term sensitivity of ABA levels in mistletoe leaves to even strong changes of ABA levels in the xylem sap of the host, ABA levels in mistletoe leaves were relatively high compared to ABA levels in the leaves of several tree species including poplar. Since significant transpiration of the mistletoe leaves was observed despite high ABA levels, a diminished sensitivity of the stomata of mistletoe leaves to ABA has to be concluded. The stomatal density of adaxial Viscum leaves of 89+/-23 stomata per mm is lower than those reported in a study performed at the end of the 19th century.     

Xylem ABA controls the stomatal conductance of field-grown maize subjected to soil compaction or soil drying

Stomatal conductance of individual leaves was measured in a maize field, together with leaf water potential, leaf turgor, xylem ABA concentration and leaf ABA concentration in the same leaves. Stomatal conductance showed a tight relationship with xylem ABA, but not with the current leaf water status or with the concentration of ABA in the bulk leaf. The relationship between stomatal conductance and xylem [ABA] was common for variations in xylem [ABA] linked to the decline with time of the soil water reserve, to simultaneous differences between plants grown on compacted, non-compacted and irrigated soil, and to plant-to-plant variability. Therefore, this relationship is unlikely to be fortuitous or due to synchronous variations. These results suggest that increased concentration of ABA in the xylem sap in response to stress can control the gas exchange of plants under field conditions.     

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     

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.     

The Flux and Distribution of Xylem Sap Calcium to Adaxial and Abaxial Epidermal Tissue 8

Inherent differences in the responses of stomata on abaxialand adaxial epidermal surfaces of leaves of Commelina communishave previously been suggested to be due to differences in theconcentrations of apoplastic Ca. Adaxial stomata have also beenreported to be more sensitive than abaxial stomata to appliedabscisic acid (ABA). The aims of these experiments were to determinethe validity of these conclusions and to see if xylem sap Cahas a role in determining the response of stomata to ABA. It can be shown from measurements of relative stomatal resistance(determined with a viscous flow porometer) and stomatal conductancethat stomata were more open in plants grown on 8-0 mol m^–3Ca, than with those grown on 2-0 mol m^–3 Ca. When attachedleaves were fed with ABA via the transpiration stream neitherthe extent nor the rate with which conductance declined wasdependent on Ca nutrition. The extent of Ca accumulation within both epidermes was relatedto the concentration of Ca in the rhizosphere and in the xylemsap. It did not, however, appear to reflect the apparent differencesin the flux of the transpiration stream between the two epidermes.Plants growing at the lower Ca concentration accumulated proportionallymore epidermal Ca relative to Ca in xylem sap. The evidencepresented suggests that Ca movement from the xylem to the epidermiscannot be simply described by a mass flow model, and that thedistribution of Ca is not an adequate explanation of the differencesin the behaviour of adaxial and abaxial stomata. The potentialrole for changes in xylem sap Ca to act as a regulator of stomatalbehaviour are discussed. Key words: Abscisic acid, calcium, Commelina communis L., stomatal conductance     

ABA xylem concentrations determine maximum daily leaf conductance of field-grown Vitis vinifera L. plants

Differences in maximum leaf conductance in grapevine plants growing in soils with contrasting water availabilities during mid-summer in Portugal could be accounted for by differences in the concentration of ABA in xylem sap. This conclusion is reinforced by the observation that the relationship between leaf conductance and endogenous ABA concentration can be mimicked by the application of exogenous ABA to leaves detached from irrigated plants. During the day, leaf conductance decreased after a morning peak, even when the leaves remained in a constant environment at a moderate temperature and leaf-to-air vapour pressure difference. This decline in leaf conductance was not a consequence of an increase in the xylem ABA concentration or the rate of delivery of this compound by the transpiratory stream. The afternoon depression in leaf conductance was associated with an apparent limitation in stomatal opening potential, which persisted even when detached leaves were fed with water and rehydrated. The reason for this inhibition has still to be identified.     

Calcium in Xylem Sap and the Regulation of its Delivery to the Shoot

Amounts of total and free calcium in root and shoot xylem sapwere quantified for a number of species grown in comparableenvironments and in a rooting medium not deficient in calcium.The potential for the shoot to sequester calcium was also examined,along with the ability for ABA to regulate calcium flux to theleaf. Xylem sap calcium showed considerable interspecific and diurnalvariation, even though the plants were grown with similar rhizosphericcalcium concentrations. The potential for the shoot to sequesterxylem sap calcium was also highly variable between species andimplied an ability, at least in some species, to regulate thecalcium reaching the shoot in the transpiration stream. Long distance transport of calcium in the xylem was not primarilyby mass flow, because neither calcium uptake nor distributionwere closely related to water uptake or transpiration. The diurnalchanges in xylem sap total ion concentration appeared to benegatively correlated with transpiration while, in contrast,the calcium ion concentration showed two peaks, one occurringin the dark and the other in the light period. The application of ABA to roots caused an increase in the rateof exudation from the xylem of detopped well-watered plants.These experiments suggest that changes in root water relationsdriven by ionic fluxes were the likely cause for enhanced sapexudation from ABA-treated roots. The steady-state concentrationof calcium in the xylem sap was unaffected by ABA when exudationrate increased and, consequently, the flux of calcium must alsohave increased. Key words: Abscisic acid, calcium, xylem sap, ionic fluxes     

Effect of partial rootzone drying on the concentration of zeatin-type cytokinins in tomato (Solanum lycopersicum L.) xylem sap and leaves

Decreased cytokinin (CK) export from roots in drying soil might provide a root-to-shoot signal impacting on shoot physiology. Although several studies show that soil drying decreases the CK concentration of xylem sap collected from the roots, it is not known whether this alters xylem CK concentration ([CK(xyl)]) in the leaves and bulk leaf CK concentration. Tomato (Solanum lycopersicum L.) plants were grown with roots split between two soil columns. During experiments, water was applied to both columns (well-watered; WW) or one (partial rootzone drying; PRD) column. Irrigation of WW plants aimed to replace transpirational losses every day, while PRD plants received half this amount. Xylem sap was collected by pressurizing detached leaves using a Scholander pressure chamber, and zeatin-type CKs were immunoassayed using specific antibodies raised against zeatin riboside after separating their different forms (free zeatin, its riboside, and nucleotide) by thin-layer chromatography. PRD decreased the whole plant transpiration rate by 22% and leaf water potential by 0.08 MPa, and increased xylem abscisic acid (ABA) concentration 2.5-fold. Although PRD caused no detectable change in [CK(xyl)], it decreased the CK concentration of fully expanded leaves by 46%. That [CK(xyl)] was maintained and not increased while transpiration decreased suggests that loading of CK into the xylem was also decreased as the soil dried. That leaf CK concentration did not decline proportionally with CK delivery suggests that other mechanisms such as CK metabolism influence leaf CK status of PRD plants. The causes and consequences of decreased shoot CK status are discussed.     

Does ABA in the Xylem Control the Rate of Leaf Growth in Soil-Dried Maize and Sunflower Plants?

Maize (Zea mays L.) and sunflower (Helianthus annuus L.) plantswere grown in large volumes of soil and leaf growth rate wasmonitored on a daily basis. Half the plants were given a soildrying treatment and when they showed a significant restrictionof growth rate (compared to both their daily growth rate beforedrying and the average growth rate of well-watered plants onthe same day), leaf water relations were measured and xylemsap was extracted using several techniques. There was a significant negative log-linear relationship betweenthe rate of leaf growth and the concentration of ABA in thexylem for both species. There was no clear relationship betweenleaf growth rate and leaf water potential or turgor for eitherspecies. Assessment of different methods for sampling xylemsap suggests that exudates collected from stem stumps or samplescollected by pressurizing the whole root system are suitablefor estimating ABA concentration in xylem, at least with largeplants of maize or sunflower, provided the first few hundredcubic millimetres of collected sap are used for the assay. Centrifugationof sections of stems resulted in dilution of ABA in the xylemsap with sap squeezed from parenchyma tissue. This is because,at least in plants subjected to mild soil drying, the concentrationof the ABA in the xylem is far higher than that in the cellsap of stem tissue. Results support the proposal that ABA plays a major role asa chemical signal involved in the root-to-shoot communicationof the effects of soil drying. The non-hydraulic restrictionof leaf growth by a chemical signal can be explained by theextra root-sourced ABA in the xylem and may be an importantcomponent of the modification of growth and development whichresults from prolonged soil drought. Key words: Soil drying, ABA, leaf growth, Zea mays L., Helianthus annuus L.