Evaluation of water stress control with polyethylene glycols by analysis of guttation

The water relations of pepper plants (Capsicum frutescens L.) under conditions conducive to guttation were studied to evaluate the control of plant water stress with polyethylene glycols. The addition of polyethylene glycol 6000 to the nutrient solution resulted in water relations similar to those expected in soil at the same water potentials. Specifically, xylem pressure potential in the root and leaf became more negative during a 24-hour treatment period, while osmotic potential of the root xylem sap remained constant. The decrease in pressure potential was closely correlated with the decrease in osmotic potential of the nutrient solution. In contrast, the addition of polyethylene glycol 400 to the nutrient medium resulted in a reduction of osmotic potential in the root xylem sap; this osmotic adjustment in the xylem was large enough to establish an osmotic gradient for entry of water and cause guttation at a nutrient solution osmotic potential of −4.8 bars. Pressure potential in the root and leaf xylem became negative only at nutrient solution osmotic potentials lower than −4.8 bars. About half of the xylem osmotic adjustment in the presence of polyethylene glycol 400 was caused by increased accumulation of K⁺, Na⁺, Ca²⁺, and Mg²⁺ in the root xylem. These studies indicate that larger polyethylene glycol molecules such as polyethylene glycol 6000 are more useful for simulating soil water stress than smaller molecules such as polyethylene glycol 400.     

Water stress induced by PEG decreases the maximum growth pressure of the roots of pea seedlings

Roots of plants growing in dry soil often experience large mechanical impedance because the decreased soil water content is associated with increased in soil strength. The combined effect of mechanical impedance and water stress hinders the establishment of seedlings in many soils, but little is known about the interaction between these two stresses. A method has been designed that, for the first time, measured the maximum axial force exerted by a root growing under controlled water stress. Using this technique the axial force exerted by a pea radicle was measured using a shear beam, while the seedling was suspended in an aerate solution of polyethylene glycol 20 000 at osmotic potentials between 0 and -0.45 MPa. The maximum growth force was then divided by the cross-sectional area of the root to give the maximum axial growth pressure. The value of maximum axial growth pressure decreased linearly from 0.66 and 0.35 MPa as the osmotic potentials of the solution of PEG decreased from 0 to -0.45 MPa. In dry soil, therefore, the maximum strength of soil that a root can penetrate is decreased because of the decrease in maximum growth pressure. The elongation rates of unimpeded roots were similar whether the roots were subject to either a matric potential in soil or to an osmotic potential in a solution of PEG.Key words: Pisum sativum L, pea, mechanical impedance, axial growth pressure, water stress, PEG 20 000.      

Effect of Water Stress on Turgor Differences and C-Assimilate Movement in Phloem of Ecballium elaterium

The effects of water stress on pressure differences and ¹⁴C-assimilate translocation in sieve tubes of squirting cucumber Ecballium elaterium A. Rich were studied. Water stress was induced by transfer of plants from culture solution to a polyethylene glycol 6,000 solution having an osmotic potential of −18.2 atm. Sieve tube turgor, turgor differences between source and sink, and translocation rate were decreased. After 260 minutes of translocation, only 19% of the total fixed ¹⁴CO₂ had moved out of the leaf, compared to the control value of 62% after the same period of time. The results suggest that water stress slows translocation by lowering sieve tube turgor differences, which are essential for the pressure flow mechanism of conduction.     

Leaf age effects on solute accumulation in water-stressed grapevines

The effects of leaf age on water relations, organic solute, and total ion accumulation were studied in mature and immature leaves of two-year-old grapevines (Vitis vinifera L., cv. Savatiano) grown under water stress conditions. Osmotic potential at full turgor decreased significantly in leaves of stressed plants, irrespective of leaf age, indicating the occurrence of an active osmotic adjustment. The apoplastic water fraction (A) increased during leaf ontogeny in both control and stressed plants. However, the values of A were lower in stressed plants. Starch concentration decreased significantly in both mature and immature leaves during the drought cycle, while the relative proportion of monosaccharides and sucrose was markedly different in immature leaves compared to mature. The accumulation of total inorganic ions, induced by drought, was also age dependent, increasing significantly with leaf age, while there were no significant differences in total amino acids content. Inorganic ions and carbohydrates seem to be the major component of osmotic adjustment in mature and immature grapevine leaves, respectively.     

Influence of Water Stress on the Vacuole/Extravacuole Distribution of Proline in Protoplasts of Nicotiana rustica

Tobacco (Nicotiana rustica) plants were stressed by addition of polyethylene glycol solution (−20 bar) to the growth medium. The proline contents and concentrations in total protoplasts, vacuoles, and extravacuolar fractions of these plants have been determined and compared with protoplasts and cell fractions of well-watered plants. As compared to the control, the stress treatment of intact plants results in a 7-fold increase of the proline content in the extravacuolar fraction while the vacuolar content was enriched only 2.6-fold. In protoplasts of control plants, a proline concentration ratio (extravacuole to vacuole) of 1 was measured. In protoplasts of stressed plants, this ratio was nearly 3. Thus, water stress seems to have an effect on a tonoplast-localized transport system for proline.     

Water Content-Potential Relationship in Soya Bean: Changes in Component Potentials for Mature and Immature Leaves under Field Conditions

Changes in turgor and osmotic potentials of soya bean leaves(Glycine max.) with changes in water content were measured throughouta season using the pressure-volume technique. Two distinct reponsesto water loss were found. When water was expressed from leavesin the pressure chamber their osmotic behavior was describedby a concentration effect based on the osmotic volume. The osmoticfraction of the total water content averaged 0·72 and0·84 for mature and immature leaves, respectively. Thechanges in turgor pressure in the chamber were described bya volumetric modulus of elasticity which increased linearlywith turgor pressure. The changes in total potential at highturgor pressures were almost exclusively due to changes in turgordue to the high modulus (high tissue rigidity) in that range.Responses were different, however, for leaves drying in thefield. For these, the osmotic changes were always large anddominated by solute adjustment. Diurnal changes in osmotic potentialwere as much as 5 bars (500 kPa), or around 50 per cent, andwere about the same magnitude as the changes in turgor pressurefor both mature and immature leaves. The elastic modulus atthe time of sampling showed the normal turgor dependence forimmature leaves but for mature leaves the initial modulus wasapparently constant at about 180 bars. The different behaviourin the pressure bomb and the field is interpreted in terms ofa rate dependence for turgor and osmotic response to water loss.     

Effect of water deficit and sulphur dioxide on total soluble proteins,nitrate reductase activity and free proline content in sunflower leaves

Sunflower (Helianthus annus L. cv. PSH-7) plants were subjected to different osmotic potentials, using polyethylene glycol-6000 (PEG-6000), after, prior to and during SO₂ fumigation. Total soluble proteins and nitrate reductase activity (NRA) decreased, and free proline content increased with the increasing water stress. These biochemical parameters were more adversely affected in fumigated plants than in non-fumigated ones, when mild water stress was provided prior to and during fumigation. When severe water stress was given prior to and during fumigation, total soluble proteins, NRA and free proline content were nearly the same in fumigated and non-fumigated water-stressed plants; it is because the stomatal closure was observed in water-stressed plants. The leaf water potential decreased with the increasing water stress; however, it was not significantly affected due to SO₂ fumigation.     

Effects of preconditioning on electrolyte leakasge and lipid composition in black spruce (Picea mariand) stressed with polyethylene giycol

Black spruce ( Picea mariana Mill. B. S. P.) rooted cuttings were grown in solution culture and preconditioned by osmotically stressing plants with polyethylene glycol. After relief from preconditioning stress, water relations, membrane leakiness, and the composition of lipids and fatty acids were compared in preconditioned and control, unconditioned plants. Both groups of plants were subsequently subjected to a severe osmotic stress with polyethylene glycol and examined again. Osmotic stress decreased shoot water potentials and increased the leakage of electrolytes from shoots of stressed, compared with unstressed, plants. However, both unstressed and stressed preconditioned plants leaked less electrolytes compared with unconditioned plants. Changes in sterol, phospholipid and glycolipid composition were observed in preconditioned unstressed and stressed plants. Sterol and phospholipid levels de clined as a result of stress, and preconditioning resulted in a decline in sterol: phospholipid ratios in plants.     

Physiological Changes in Cultured Sorghum Cells in Response to Induced Water Stress : II. Soluble Carbohydrates and Organic Acids

Eight cultivars Sorghum bicolor (L.) Moench were grown as callus cultures under induced, prolonged water stress (8 weeks), with polyethylene glycol in the medium. Concentrations of soluble carbohydrates and organic acids in callus were measured at the end of the growth period to determine differences in response to prolonged water stress. Sucrose, glucose, fructose, and malate were the predominant solutes detected in all callus at all water potentials. All cultivars had high levels of solutes in the absence of water stress and low levels in the presence of prolonged water stress. However, at low water potentials, low levels of solutes were observed in drought-tolerant cultivar callus and high solute levels were observed in drought-susceptible cultivar callus. Estimated sucrose concentrations were significantly higher in water-stressed, susceptible cultivar callus. Large solute concentrations in susceptible cultivar callus were attributed to osmotic adjustment and/or reduced growth during water stress.     

Water relations of cotton plants under nitrogen deficiency

Nitrogen deficiency in cotton plants (Gossypium hirsutum L.) increased the threshold water potentials for both stomatal closure and leaf senescence (defined as loss of chlorophyll and protein) during drought. These studies attempted to answer two questions: (1) What is the basis for the N/water interaction on senescence? (2) Is there a direct relationship between stomatal closure and senescence? Young and old leaves from N-deficient and N-sufficient plants maintained their relative sensitivities to water stress when excised leaf discs were floated on solutions of polyethylene glycol in dim light. In this leaf disc system, both leaf aging and N deficiency increased the threshold water potential for senescence. Leaf aging and N deficiency also decreased the concentration of exogenous abscisic acid necessary to initiate senescence in discs. A role for cytokinins in controlling senescence could not be clearly shown. In young leaves of both N-deficient and N-sufficient plants, stomata closed at water potentials much higher than those causing senescence. During leaf aging, the water potentials causing senescence increased more than those causing stomatal closure. The two processes thus occurred at about the same potentials in the oldest leaves. These data argue against a general cause-and-effect relationship between stomatal closure and senescence. Rather, each process apparently responded independently to absicsic acid accumulated during drought.     

Leaf Elongation in Relation to Leaf Water Potential in Soybean

Leaf water potential, turgor pressure, and leaf elongation ratewere measured in soybeans growing in controlled environmentchambers, greenhouses, and outdoors. Plants in chambers hadthe highest water potentials and turgor pressures, and plantsoutdoors the lowest. In all three environments there was a linearrelationship between elongation rate and turgor pressure. Leavesof plants in drier environments required less turgor for elongation,and showed a greater increase in elongation rate per unit increasein turgor. Elongation rates over a 72 h period were equal inthe three environments. Leaves reached the largest final sizein the greenhouse (intermediate in water potential). Epidermalcells were larger in chamber- and greenhouse-grown leaves thanin leaves of plants grown outdoors. The number of epidermalcells per leaf was greater in the greenhouse and outdoors thanin the chamber. Leaf elongation characteristics of greenhouseplants were duplicated by mildly stressing chamber plants, andleaf elongation characteristics of field plants were duplicatedby more severely stressing chamber plants. Leaves of mildlystressed chamber plants also reached a larger final size thanleaves of more severely stressed chamber plants, or leaves ofcontrol plants in the chamber. Water stress in the chamber increasedthe number of epidermal cells per leaf. More severe water stressin the chamber reduced epidermal cell size. Based on the waterstress experiments it is concluded that the differences in plantwater status in the chamber, greenhouse, and field caused differencesin elongation characteristics, and were responsible for thedifferences in leaf size.     

Acclimation of CO(2) Assimilation in Cotton Leaves to Water Stress and Salinity

Cotton (Gossypium hirsutum L. cv Acala SJ2) plants were exposed to three levels of osmotic or matric potentials. The first was obtained by salt and the latter by withholding irrigation water. Plants were acclimated to the two stress types by reducing the rate of stress development by a factor of 4 to 7. CO₂ assimilation was then determined on acclimated and nonacclimated plants. The decrease of CO₂ assimilation in salinity-exposed plants was significantly less in acclimated as compared with nonacclimated plants. Such a difference was not found under water stress at ambient CO₂ partial pressure. The slopes of net CO₂ assimilation versus intercellular CO₂ partial pressure, for the initial linear portion of this relationship, were increased in plants acclimated to salinity of −0.3 and −0.6 megapascal but not in nonacclimated plants. In plants acclimated to water stress, this change in slopes was not significant. Leaf osmotic potential was reduced much more in acclimated than in nonacclimated plants, resulting in turgor maintenance even at −0.9 megapascal. In nonacclimated plants, turgor pressure reached zero at approximately −0.5 megapascal. The accumulation of Cl⁻ and Na⁺ in the salinity-acclimated plants fully accounted for the decrease in leaf osmotic potential. The rise in concentration of organic solutes comprised only 5% of the total increase in solutes in salinity-acclimated and 10 to 20% in water-stress-acclimated plants. This acclimation was interpreted in light of the higher protein content per unit leaf area and the enhanced ribulose bisphosphate carboxylase activity. At saturating CO₂ partial pressure, the declined inhibition in CO₂ assimilation of stress-acclimated plants was found for both salinity and water stress.     

Polyribosome metabolism, growth and water status in the growing tissues of osmotically stressed plant seedlings

Relationships between growth of osmotically stressed intact seedlings and polyribosome levels and water status of growing tissues were examined. Sudden exposure of barley (Hordeum vulgare L. cv. Arivat) roots to a solution of ?0.8 MPa polyethylene glycol caused leaf growth to stop almost immedately, but growth resumed at a much lower rate after 0.5–1 h. In the growing region of leaves, the polyribosome: total ribosome ratio of free (non-membrane-bound) ribosomes was significantly reduced after 15 min stress, but a decrease in the large polyribosome:total polyribosome ratio occurred only after 1–2 h. Membrane-bound and free polyribosome levels both decreased to 70% of unstressed control values after 4 h stress. Recovery of total polyribosomes occurred within 1 h after relief of 4 h stress, but required 3 h after relief of 24 h stress. Stress detectably reduced the water potential and osmotic potential of growing tissue within 0.5–1.0 h, and osmotic adjustment continued for up to 10 h. Recovery of water status was incomplete after 1 h relief of a 4 h stress. In contrast, expanded blade tissues of stressed plants underwent minor changes in water status and slow decreases in polyribosomes levels. These results confirm that growing tissues of barley leaves are selectively responsive to stress, and suggest that changes in growth, water status and polyribosome levels may be initiated by the same signal. Measurements of seedling growth, polyribosome levels and water status of growing tissues of barley and wheat (Triticum aestivum L. cv. Zaragoza) leaves, etiolated pea (Pisum sativum L. cv. Alaska) epicotyl and etiolated squash (Cucurbita pepo L. cv. Elite) hypocotyl stressed with polyethylene glycol solutions of ?0.3 to ?0.8 MPa for 12 h or more showed that polyribosome levels were highly correlated with seedling growth rate as well as with tissue water and osmotic potentials, while turgor remained unchanged. These results suggest that long-term growth of osmotically stressed plants may be limited by a reduced capacity for protein synthesis in growing tissues and is not dictated by turgor loss.     

Carbon Balance of Sorghum Plants during Osmotic Adjustment to Water Stress

The daily (24-hour) carbon balances of whole sorghum plants (Sorghum bicolor L. Moench cv BTX616) were continuously measured throughout 15 days of water stress, followed by rewatering and 4 more days of measurements. The plants were grown under controlled environment conditions typical of warm, humid, sunny days. During the first 12 days, osmotic potentials decreased in parallel with decreased water potentials to maintain pressure potentials near 0.5 kilojoules per kilogram (5 bars). Immediately before rewatering on day 15, the water potential was −3.0 kilojoules per kilogram. Osmotic adjustment at this point was 1.0 kilojoules per kilogram, as measured by the decrease in the water potential at zero turgor from its initial value of −1.4 kilojoules per kilogram.

Gross input of carbon was less but the fraction retained was greater because a smaller fraction was lost through respiration in stressed plants than in unstressed plants. This was attributed to a lower rate of biomass synthesis, and conversely a higher rate of storage of photosynthate, due to inhibition of leaf expansion. The reduction in the cost associated with biomass synthesis more than balanced any metabolic cost of osmotic adjustment. The net daily gain of carbon was always positive in the stressed plants.

There was a large burst of respiration on rewatering, due to renewed synthesis of biomass from stored photosynthate. Over the next 3 days, osmotic adjustment was lost and the daily carbon balance returned to that typical of nonstressed plants. Thus, osmotic adjustment allowed the stressed plants to accumulate biomass carbon throughout the cycle, with little additional metabolic cost. Carbon stored during stress was immediately available for biomass synthesis on rewatering.

     

Effects of Different Levels of Water Stress on Leaf Water Potential, Stomatal Resistance, Protein and Chlorophyll Content and Certain Anti-oxidative Enzymes in Tomato Plants

A greenhouse experlment was performed In order to Investigate the effects of dlfferent levels of water stress on leaf water potentlal （ψw）, stomatal resistance （rs）, protein content and chlorophyll （Chl） content of tomato plants （Lycoperslcon esculentum Mill. cv. Nlkita）. Water stress was Induced by addlng polyethylene glycol （PEG 6 000） to the nutrlent solution to reduce the osmotlc potential （ψs）. We Investlgated the behavlor of antl-oxldant enzymes, such as catalase （CAT） and superoxide dlsmutase （SOD）, durlng the development of water stress. Moderate and severe water stress （i.e. ψs= -0.51 and -1.22 MPa, respectlvely） caused a decrease In ψw for all treated （water-stressed） plants compared with control plants, wlth the reductlon belng more pronounced for severely stressed plants. In addltion, rs was slgnlflcantly affected by the Induced water stress and a decrease in leaf soluble protelns and Chl content was observed. Whereas CAT actlvlty remained constant, SOD actlvlty was increased in water-stressed plants compared wlth unstressed plants. These results Indicate the possible role of SOD as an anti-oxidant protector system for plants under water stress condltlons. Moreover, It suggests the possibllity of using this enzyme as an addltional screening crlterlon for detecting water stress in plants.     

Water Stress Reduces Ozone Injury via a Stomatal Mechanism

Various studies have shown that water-stressed plants are more tolerant of ozone exposures than are unstressed plants. Two probable explanations for this tolerance are (a) stomatal closure which reduces ozone uptake and (b) biochemical or anatomical changes within the leaves. Phaseolus vulgaris cv Pinto bean plants were established and transferred to membrane systems which controlled the osmotic potential around the roots at −35 or −80 kilopascals for 5 days prior to ozone treatment (0 or 1.0 microliters per liter for 2 hours). Both water-stressed and unstressed plants were sprayed with various concentrations of abscisic acid to close the stomata or with fusicoccin to induce stomata opening. The abaxial stomatal resistances of primary and trifoliate leaves were measured just prior to ozone exposure. Plant response to ozone was determined by stress ethylene production and chlorophyll loss. Both water stress and abscisic acid induced stomatal closure and reduced ozone injury. In water-stressed plants, fusicoccin induced stomatal opening and those plants were as sensitive to ozone as were the non-water-stressed plants. These data suggest that water stress protects plants from ozone injury mainly through its influence on stomatal aperture rather than through biochemical or anatomical changes.     

Effects of the Ionic Composition and Water Potential of Aqueous Solution on the Activity and Survival of Orrina phyllobia

The activity and survival of Orrina phyllobia fourth-stage juveniles (J4) were examined in aqueous solutions representing 96 combinations of eight predominant soil solution ions at total concentrations of 100, 200, and 1,000 meq/liter. Various water potentials were imposed by the addition of mannitol or polyethylene glycol to ionic solutions. Nematode longevity increased as water potential was decreased. Longevity was approximately doubled at a water potential of -23 × 10⁵ Pa and more than tripled at -60 × 10⁵ Pa. No combination oflons at 200 meq/liter was lethal after a 6-day exposure. Several ion combinations significantly increased longevity at -10 and -23 × 10⁵ Pa. Single cation Na⁺ solutions consistently inhibited activity and more than doubled nematode longevity.     

The biophysics of leaf growth in salt-stressed barley. A study at the cell level

Biophysical parameters potentially involved in growth regulation were studied at the single-cell level in the third leaf of barley (Hordeum vulgare) after exposure to various degrees of NaCl stress for 3 to 5 d. Gradients of elongation growth were measured, and turgor pressure, osmolality, and water potentials (psi) were determined (pressure probe and picoliter osmometry) in epidermal cells of the elongation zone and the mature blade. Cells in the elongation zone adjusted to decreasing external psi through increases in cell osmolality that were accomplished by increased solute loads and reduced water contents. Cell turgor changed only slightly. In contrast, decreases in turgor also contributed significantly to psi adjustment in the mature blade. Solute deposition rates in the elongation zone increased at moderate stress levels as compared with control conditions, but decreased again at more severe NaCl exposure. Growth-associated psi gradients between expanding epidermal cells and the xylem were significant under control and moderate stress conditions (75 mM NaCl) but seemed negligible at severe stress (120 mM NaCl). We conclude that leaf cell elongation in NaCl-treated barley is probably limited by the rate at which solutes can be taken up to generate turgor, particularly at high NaCl levels.     

Semipermeable membrane system for subjecting plants to water stress

A system was evaluated for growing plants at reproducible levels of water stress. Beans (Phaseolus vulgaris L.) were grown in vermiculite, transferred to a semipermeable membrane system that encased the root-vermiculite mass, and then placed into nutrient solutions to which various amounts of polyethylene glycol (PEG) 20M were added to control solution water potential. The membrane (Spectrapor 1) had a minimum molecular weight cutoff that excluded the PEG 20M. The plants equilibrated with the nutrient solution within 1 to 4 days, and exhibited normal diurnal water relations. Use of the semipermeable membrane system to induce water stress reduces many of the problems associated with hydroponic media.     

Characterization of water stress and low temperature effects on flower induction in citrus

Experiments were conducted with containerized `Tahiti' lime (Citrus latifolia Tan.) trees in order to define conditions needed to induce flowering. Cyclical or continuous water stress for 4 to 5 weeks induced flowering. Moderate (−2.25 megapascals, midday) or severe (−3.5 megapascals, midday) water stress as measured by leaf xylem pressure potential, for as little as 2 weeks induced flowering, but the response was more significant in severely stressed trees. Low temperature (18°C day/10°C night) induced a time dependent flowering response much like that of moderate water stress. Significantly negative leaf xylem pressure potentials as compared to controls were found only under water stress treatment, suggesting that a common stress-linked event, separate from low plant water potential is involved in floral induction. Leafless, immature cuttings from mature, field-grown trees were induced to flower by water stress treatment, suggesting that leaves are not essential for a flower inductive response.