Involvement of root ABA and hydraulic conductivity in the control of water relations in wheat plants exposed to increased evaporative demand

We studied the possible involvement of ABA in the control of water relations under conditions of increased evaporative demand. Warming the air by 3°C increased stomatal conductance and raised transpiration rates of hydroponically grown Triticum durum plants while bringing about a temporary loss of relative water content (RWC) and immediate cessation of leaf extension. However, both RWC and extension growth recovered within 30 min although transpiration remained high. The restoration of leaf hydration and growth were enabled by increased root hydraulic conductivity after increasing the air temperature. The use of mercuric chloride (an inhibitor of water channels) to interfere with the rise on root hydraulic conductivity hindered the restoration of extension growth. Air warming increased ABA content in roots and decreased it in shoots. We propose this redistribution of ABA in favour of the roots which increased the root hydraulic conductivity sufficiently to permit rapid recovery of shoot hydration and leaf elongation rates without the involvement of stomatal closure. This proposal is based on known ability of ABA to increase hydraulic conductivity confirmed in these experiments by measuring the effect of exogenous ABA on osmotically driven flow of xylem sap from the roots. Accumulation of root ABA was mainly the outcome of increased export from the shoots. When phloem transport in air-warmed plants was inhibited by cooling the shoot base this prevented ABA enrichment of the roots and favoured an accumulation of ABA in the shoot. As a consequence, stomata closed.     

Effect of increased irradiance on the hormone content,water relations,and leaf elongation in wheat seedlings

In wheat (Triticum durum Desf., cv. Bezenchukskaya 139) seedlings, an increase in irradiance from 20 to 400 μmol/(m² s) PAR enhanced transpiration and increased stomatal conductance by three times on the background of reduced relative water content (RWC). After this treatment, leaves quickly ceased to grow and became even shrunk later. In 40 or 50 min, leaf growth was resumed. At this period, we observed an increase in hydraulic conductivity and RWC and also in leaf extensibility. As soon as 10 min after treatment, some changes in hormone content were noted. In the zones of leaf growth and its mature part, zeatin and zeatin riboside were accumulated, whereas ABA accumulation was observed in the zone of leaf growth and in the roots. The results obtained indicate that leaf expansion at increased irradiance was related to changes in cell-wall extensibility and hydraulic conductivity. The first effect could be due to cytokinin accumulation, whereas the second one, to ABA accumulation.     

Unusual stomatal behaviour on partial root excision in wheat seedlings

The excision of four out of five primary roots of wheat (Triticum durum Desf.) seedlings often leads to an enhanced rate of transpiration. Surprisingly this enhancement could be maintained for several hours after root excision and was particularly likely to occur at low irradiances or high atmospheric humidity. This long‐term enhancement could not be explained in terms of conventional hydropassive stomatal effects. Elevated rates of transpiration were associated with and possibly caused by increased cytokinin concentrations in shoots of plants with partially excised roots. The single root remaining after excision was able to maintain an adequate water uptake for the continued enhanced transpiration, after only a short transient reduction in leaf water content. The enhanced capacity for water uptake by the remaining root was confirmed by measuring the water flow from detached roots at negative hydrostatic pressure. Even without additional suction, flow from the reduced root system increased about 1.5 h after the start of treatment, suggesting an increase in membrane permeability for water. Although abscisic acid (ABA) concentrations in the roots increased after the root excision treatment, there was no evidence for any enhanced concentration in the xylem sap. The possible role that this accumulation of ABA in roots may have in the apparent increase in hydraulic conductivity after root excision is discussed.     

Effect of partial root excision on transpiration, root hydraulic conductance and leaf growth in wheat seedlings.

Removal of four out of five roots did not lower transpiration and stomatal conductivity of wheat (Triticum durum Desf.) seedlings. Water content of mature expanded leaf lamina remained constant at control levels. The results suggest that the only remaining root was capable to supply the shoot with water. This was evidenced by an increase in hydraulic conductivity of the root system following partial root excision measured at low subatmospheric pressures induced by vacuum. In the absence of a hydrostatic gradient, water flow from reduced root system was initially not higher than from an intact system, but increased subsequently. ABA content was increased in roots 1 h after partial root excision, which might contribute to the increase in hydraulic conductivity.     

Hormonal Control of the Shoot-to-Root Ratio Is Not Related to Water Deficiency in Wheat Plants under Mineral Deficiency

We measured the content of hormones, the rate of growth, and some parameters of water regime (water content, transpiration, and stomatal and hydraulic conductivities) one and two days after wheat plant transfer from 10 to 1% Hoagland-Arnon nutrient medium. It was shown that, a day after dilution of nutrient solution, the content of various cytokinin forms decreased in the xylem sap, shoots, and roots. This decrease was most pronounced in the case of zeatin in the xylem sap and zeatin riboside in the mature zone of the first leaf. ABA was found to accumulate in shoots. A day after dilution of nutrient solution, we observed root elongation evidently induced by mineral nutrient deficiency, and this accelerated root growth was maintained later. Two days after dilution of nutrient solution, we observed the slowing of shoot weight accumulation, whereas root weight remained unchanged. Plant growth response could be related to ABA accumulation in shoots and cytokinin depletion in the whole plant. A reduced hydraulic conductivity and water content in the growing leaf zone was detected only two days after dilution of nutrient solution. Thus, changes in the growth rates and hormone contents could not result from disturbances in water regime induced by mineral nutrient deficiency.     

Effect of Changes of Temperature Around Roots in Relation to Water Uptake by Roots and Leaf Transpiration

Effects of changes in temperature around roots on water uptake by roots and leaf transpiration were studied in Leucaena leucocephala (Lam.) de Wit., a subtropical woody plant species, and in Zea mays L. When the temperature around roots was rapidly lowered from 25 ℃ to 15 ℃, the water uptake by the roots and leaf transpiration were stimulated significantly within a short period ( 14 min). However, this effect did not occur when the cooling time was prolonged neither did if occur when the temperature around the roots was resumed from 15 ℃ to 25 ℃. Both the hydraulic conductivity of roots and leaf transpiration were increased substantially at first (within 20 min)and then decreased steadily to a level lower than those of the control in which the roots were continuous exposed to a low temperature ( 15 ℃ ). Low temperature also promoted the biosynthesis of ABA in roots and enhanced the xylem ABA concentration, but such stimulation did not occur untill about 30 min after cooling treatment, leaf transpiration was reduced markedly, but the hydraulic conductivity of roots increased when the root system was treated with exogenous ABA. It was suggested that some mechanisms other than ABA may be involved in the short-time cryostimulation of water uptake by roots and leaf transpiration.     

Effect of partial root excision on shoot water relations

Removing 4 out of 5 serminal roots from 7-day-old wheat seedlings arrested leaf elongation for 1.5 h. This effect can be explained by an initial decrease in foliar water content resulting from the smaller root surface area available for water uptake. Subsequently, leaf hydration increased with time and came to equal that of intact plants within 2 h. The rehydration was seemingly effected by an increasing conductivity of the one remaining root axis, since transpiration of the partially de-rooted plants did not fall below that of controls. With time, leaf elongation resumed, but at a slower rate than in intact plants. This slower growth may be attributed to a decrease in leaf extensibility since this was found to be reduced when measured by a counterweight technique involving linear displacement transducers. Loss of extensibility was associated with decreased IAA concentration in the leaf elongation zone.     

土壤干旱条件下氮素营养对玉米内源激素含量影响

在田间持水量分别保持于35％、55％和75％±5％的土壤水分条件下,利用盆栽实验研究了土壤干旱和氮素营养对玉米内源激素和气孔导度的影响．结果表明,土壤干旱下氮素营养明显降低了玉米根系木质部汁液ABA浓度,而正常供水下施氮处理间则无显著差异(施氮处理仍较低),同时测定的叶片ABA浓度则呈相反的变化趋势,表现为干旱下施氮处理要高于不施氮处理;施氮处理木质部汁液中ZRs浓度应低于相应的不施氮处理,在调控气孔行为方面并未表现拮抗ABA作用;3种土壤水分条件下,施氮玉米叶片的气孔导度均高于不施氮处理,与木质部汁液ABA浓度呈负相关,说明施氮处理较低的根源ABA浓度是导致其气孔导度较大的主要原因．     

The Role of Hormones in Fast Growth Responses of Wheat Plants to Osmotic and Cold Shocks

The effects of nutrient-solution cooling and PEG addition to the nutrient solution on the phytohormone content, the rate of leaf growth, leaf extensibility under the influence of external mechanical action, osmotic potential, and transpiration were studied in seven-day-old wheat plants. Leaf growth rapidly ceased, and the transpiration rate was reduced in both treatments. Growth cessation induced by PEG was transient, and growth resumption was preceded by an increase in the leaf extensibility. The functional role of auxin accumulation in plant shoots in the control of extensibility as well as the relationship between the ABA accumulation and a decrease in the cytokinin content, on the one hand, and reduced transpiration, on the other hand, under stress conditions are discussed.     

Accumulation rate of ABA in detached maize roots correlates with root water potential regardless of age and branching order

How much ABA can be supplied by the roots is a key issue for modelling the ABA-mediated influence of drought on shoot physiology. We quantified accumulation rates of ABA ( S _ABA) in maize roots that were detached from well-watered plants and dehydrated to various extents by air-drying. S _ABA was estimated from changes in ABA content in root segments incubated at constant relative water content (RWC). Categories of root segments, differing in age and branching order, were compared (root branches, and nodal roots subdivided into root tips, subapical unbranched sections, and mature sections). All categories of roots accumulated ABA, including turgid and mature tissues containing no apex. S _ABA measured in turgid roots changed with root age and among root categories. This variability was largely accounted for by differences in water content among different categories of turgid roots. The response of S _ABA to changes in root water potential ( Ψ _root) induced by dehydration was common to root tips, nodal roots and branches of several ages, while this was not the case if root dehydration was expressed in terms of RWC. Differences among root categories in the response of S _ABA to RWC were due to different RWC values among categories at a given Ψ _root, and not to differences in the response of S _ABA to Ψ _root.     

Cytokinin producing bacteria enhance plant growth in drying soil

Cytokinins can promote stomatal opening, stimulate shoot growth and decrease root growth. When soil is drying, natural cytokinin concentrations decrease in association with stomatal closure and a redirection of growth away from the shoots to the roots. We asked if decreased cytokinin concentrations mediate these adaptive responses by lessening water loss and promoting root growth thereby favouring exploration for soil water. Our approach was to follow the consequences for 12-d-old lettuce seedlings of inoculating the growing medium with cytokinin-producing bacteria under conditions of water sufficiency and deficit. Inoculation increased shoot cytokinins as assessed by immunoassay and mass spectrometry. Inoculation also promoted the accumulation of shoot mass and shortened roots while having a smaller effect on root mass. Inoculation did not raise stomatal conductance. The possible promoting effect of these cytokinins on stomatal conductance was seemingly hampered by increases in shoot ABA that inoculation also induced. Inoculation lowered root/shoot ratios by stimulating shoot growth. The effect was greater in non-droughted plants but remained sufficiently strong for shoot mass of inoculated droughted plants to exceed that of well-watered non-inoculated plants. We conclude that compensating for the loss of natural cytokinins in droughted plants interferes with the suppression of shoot growth and the enhancement of root elongation normally seen in droughted plants.     

The Effect of Rapid Temperature Increase on the Growth Rate of Wheat Leaves

The growth rate of the first leaf of eight-day-old wheat plants was measured using a DLT-2 highly sensitive linear displacement transducer. Leaf extensibility was evaluated from the growth rate under the increase in the pulling force by 2 g. An increase in the air temperature resulted in the doubling of the transpiration rate and immediate slowing of the leaf growth followed by the leaf shrinkage. However, growth was later resumed almost completely. Heat treatment did not induce any changes in the leaf extensibility, indicating that cell-wall mechanical properties were not changed. Growth retardation was supposed to result from a decrease in the water content in the leaf tissues because the balance between water influx from roots and its loss through transpiration was shifted toward the water loss. An initial drop in the relative water content (RWC) indicates such a misbalance. Subsequent growth resumption coincided with a decreased water deficiency. Since the rate of transpiration was not reduced, RWC and growth rate restoring evidently occurred due to the activated water uptake by roots, which can be explained by the increased hydraulic permeability detected in our experiments.     

Abscisic acid accumulation in the roots of nutrient-limited plants: its impact on the differential growth of roots and shoots

We describe the involvement of abscisic acid (ABA) in the control of differential growth of roots and shoots of nutrient limited durum wheat plants. A ten-fold dilution of the optimal concentration of nutrient solution inhibited shoot growth, while root growth remained unchanged, resulting in a decreased shoot/root ratio. Addition of fluridone (inhibitor of ABA synthesis) prevented growth allocation in favour of the roots. This suggests the involvement of ABA in the redirecting of growth in favour of roots under limited nutrient supply. The ABA content was greater in shoots and growing apical root parts of starved plants than in nutrient sufficient plants. Accumulation of ABA in shoots of nutrient deficient plants was linked to a decrease in leaf turgor. Increased flow of ABA in the phloem apparently contributed to the accumulation of ABA in the apical part of the roots. Thus, partitioning of growth between roots and shoots of wheat plants limited in mineral nutrients appears to be modulated by accumulation of ABA in roots. This ABA may originate in the shoots, where its synthesis is stimulated by the loss of leaf turgor.     

Effects of Water Stress and Abscisic Acid on Transpiration Regulation in Wheat

The transpiration response to recurrent light periods was studied'n water-stressed wheat seedlings. Seedlings were stressed by three methods: addition of mannitol to the root medium, root cooling and drving of the roots in air. All three methods induced almost equal effects on transpiration regulation during alternating dark and light intervals. Exogenous abscisic acid supplied to the shoots of excised plants had qualitatively the same effect as water stress. Water stress and ABA increased the time lapse between light-on and the onset of transpiration increase and lowered the amplitude of transpiration increase in light. Weak light introduced before strong light shortened the delay times.     

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.     

不同磷水平下小麦-蚕豆间作根系形态的变化及其与内源激素的关系

采用水培方法,研究了不同磷水平下小麦-蚕豆间作体系根系形态变化及其与内源激素的相关关系。结果表明: 与单作小麦相比,在低磷(1/2P)水平下,小麦-蚕豆间作能显著增加小麦的根长,显著减少小麦根系的平均直径,显著增加根系的表面积;在常规磷(P)水平下,间作能显著降低小麦根系的平均直径,有增加小麦根长和根表面积的趋势;与单作蚕豆相比,间作能明显促进蚕豆根系的增长,同时增加蚕豆根表面积。在1/2P水平下,间作能显著提高小麦和蚕豆根系中的生长素(IAA)、脱落酸(ABA)、水杨酸(SA)和茉莉酸(JA)含量;在P水平下,间作能显著提高小麦根系中的IAA、ABA和JA含量,单、间作小麦根系中的SA含量没有显著差异,间作显著增加了蚕豆根系中ABA和SA含量,单、间作蚕豆根系中的IAA和JA含量无显著差异。单作条件下,小麦和蚕豆根系中的内源激素(IAA、ABA、SA和JA)含量与其根系形态(根长、根平均直径和根表面积)无显著相关性;间作条件下,小麦和蚕豆根系中的IAA含量与根长和根表面积之间存在明显的正相关关系。由此可见,小麦-蚕豆间作能够诱导小麦和蚕豆根系IAA的增加。这种变化可能是驱动间作系统根系形态变化的重要因子。     

Early stomatal closure in waterlogged pea plants is mediated by abscisic acid in the absence of foliar water deficits

Abstract Soil waterlogging decreased leaf conductance (interpreted as stomatal closure) of vegetative pea plants (Pisuin sativum L. cv. ‘Sprite’) approximately 24 h after the start of flooding, i.e. from the beginning of the second 16 h-long photo-period. Both adaxial and abaxial surfaces of leaves of various ages and the stipules were affected. Stomatal closure was sustained for at least 3 d with no decrease in foliar hydration measured as water content per unit area, leaf water potential or leaf water saturation deficit. Instead, leaves became increasingly hydrated in association with slower transpiration. These changes in the waterlogged plants over 3 d were accompanied by up to 10-fold increases in the concentration of endogenous abscisic acid (ABA). Waterlogging also increased foliar hydration and ABA concentrations in the dark. Leaves detached from non-waterlogged plants and maintained in vials of water for up to 3 d behaved in a similar way to leaves on flooded plants, i.e. stomata closed in the absence of a water deficit but in association with increased ABA content. Applying ABA through the transpiration stream to freshly detached leaflets partially closed stomata within 15 min. The extractable concentrations of ABA associated with this closure were similar to those found in flooded plants. When an ABA-deficient ‘wilty’ mutant of pea was waterlogged, the extent of stomatal closure was less pronounced than that in ordinary non-mutant plants, and the associated increase in foliar ABA was correspondingly smaller. Similarly, waterlogging closed stomata of tomato plants within 24 h, but no such closure was seen in ‘flacca’, a corresponding ABA-deficient mutant. The results provide an example of stomatal closure brought about by stress in the root environment in the absence of water deficiency. The correlative factor operating between the roots and shoots appeared to be an inhibition of ABA transport out of the shoots of flooded plants, causing the hormone to accumulate in the leaves.     

Root water flow and leaf stomatal conductance in aspen (Populus tremuloides) seedlings treated with abscisic acid

Exogenous abscisic acid (ABA) applied to the roots and excised shoots of aspen (Populus tremuloides Michx.) inhibited stomatal conductance. However, the effect of ABA on stomatal conductance was more pronounced in the excised shoots compared with the intact seedlings. Approximately 10% of the ABA concentration applied to the roots was found in the xylem exudates of root systems exposed to a hydrostatic pressure of 0.3 MPa. A similar concentration of ABA applied to the excised shoots produced a faster and greater reduction of stomatal conductance. ABA applied to the roots had no effect on root steady-state flow rate over the 5-h experimental period. Moreover, pre-incubating root systems of intact seedlings for 12 h with 5 x 10(-5) M ABA did not significantly reduce volume flow density. Similarly, ABA had no effect on root hydraulic conductivity and the activation energy of root water flow rates.     

Effect of root anaerobiosis on the water relations of several Pyrus species

Solution culture experiments were designed to investigate the plant water relations of 3 Pyrus species subjected to root anaerobiosis. Root anaerobiosis induced partial stomatal closure prior to alterations in leaf water potential (ΨLW) or root osmotic potential (ΨRπ). In contrast, stomatal closure was accompanied by a decline in root hydraulic conductivity (Lp). Anoxia markedly reduced ΨLW for Pyrus communis L. and eventually led to wilting and defoliation. Pyrus betulaefolia Bunge and Pyrus calleryana Decne, however, were less affected by root anaerobiosis. To delineate if the increased root resistance was in the radial or longitudinal direction, 10⁻⁴ M cistrans abscisic acid (ABA) was added to detopped root systems of P. communis in solution culture after steady-state rates of Lp were established. A consistent 25 to 30% promotion of Lp was observed 1.5 h after the addition of ABA for aerobically treated plants. ABA did not influence Lp when applied to roots previously deprived of O₂ for 4 days. Additional evidence against the limiting resistance being in the radial direction was obtained when water fluxes were compared through intact P. communis roots, roots with all feeder roots detached, and stems without root systems. Severing feeder roots from anaerobically treated plants did not increase water flux to rates observed for aerobically treated plants. Resistance progressed basipetally to eventually encompass the stem itself. These results can only be explained by occlusion of the xylem vessels.     

Exogenous zeatin accumulation in wheat root cells shows its role in regulation of cytokinin transport

Here we have shown that 24 hours after addition of zeatin to the nutrient solution the cytokinin content in xylem sap of wheat plants appears to be about 10 times lower that in the nutrient solution. Cytokinins accumulated mostly in roots and not in shoots of treated plants. These data demonstrate the existence of some barrier on cytokinin pathway from the nutrient solution to the plant shoot. With the help of Sudan III an increase in lignin and suberin deposition in the endodermis could be detected, being stronger with the increase in the distance from the root tip. The increase in deposition of suberin and lignin coincided with the decrease in cytokinin immunolabeling in root cells revealed with the help of monoclonal cytokinin antibodies and the second gold-labelled antibodies. Simultaneously exogenous cytokinins accumulated in root stele cells showing that the Casparian band was not only barrier on cytokinin pathway to plant shoot. It is concluded that high cytokinin immunolabe ling in the stele parenchyma cells around the stele vessels demonstrated accumulation of cytokinins by these cells, which could be important in regulation of cytokinin loading to the xylem vessels during there transport to the shoot. The role of cytokinin transporters is discussed.