Osmotic adjustment in leaves of sorghum in response to water deficits

The relationships among the total water potential, osmotic potential, turgor potential, and relative water content were determined for leaves of sorghum (Sorghum bicolor [L.] Moench cvs. `RS 610' and `Shallu') with three different histories of water stress. Plants were adequately watered (control), or the soil was allowed to dry slowly until the predawn leaf water potential reached either −0.4 megapascal (MPa) (treatment A) or −1.6 MPa (treatment B). Severe soil and plant water deficits developed sooner after cessation of watering in `Shallu' than in `RS 610', but no significant differences in osmotic adjustment or tissue water relations were observed between the two cultivars. In both cultivars, the stress treatments altered the relationship between leaf water potential and relative water content, resulting in the previously stressed plants maintaining higher tissue water contents than control plants at the same leaf water potential. The osmotic potential at full turgor in the control sorghum was −0.7 MPa: stress pretreatment significantly lowered the osmotic potential to −1.1 and −1.6 MPa in stress treatments A and B, respectively. As a result of this osmotic adjustment, leaf turgor potentials at a given value of leaf water potential exceeded those of the control plants by 0.15 to 0.30 MPa in treatment A and by 0.5 to 0.65 MPa in treatment B. However, zero turgor potential occurred at approximately the same value of relative water content (94%) irrespective of previous stress history. From the relationship between turgor potential and relative water content there was an approximate doubling of the volumetric elastic modulus, i.e. a halving of tissue elasticity, as a result of stress preconditioning. The influence of stress preconditioning on the moisture release curve is discussed.     

Osmotic adjustment in indoor grown cassava in response to water stress

The water content-water potential relation in stressed and unstressed cassava ( Man-ihot species) was examined to ascertain (i) the magnitude of osmotic adjustment in response to water stress and (ii) the mechanisms of such adjustments.
Water stress resulted in a displacement of the water content-potential relation such that at any leaf water potential the water content was higher in the stressed plants. The osmotic potentials of turgid leaves (100% relative water content) were -0.97 and -1.00 MPa in the unstressed cultivars CMC 9 and MCOL 113 respectively. In the stressed plants, the values were-1.13 MPa (CMC 9) and-1.14 MPa (MCOL 113). The 0.14 to 0.16 MPa osmotic potential difference between the stressed and unstressed plants suggests that a stress-induced osmotic adjustment occurred in both cultivars. The biiSk volumetric elastic moduli at turgor pressures above 0.10 MPa were 9.84 MPa (CMC 9) and 13.58 MPa (MCOL 113) in the unstressed plants. Tbe higher values found in the stressed plants, 14.56 MPa in CMC 9 and 16.91 MPa in MCOL 113, suggest a stress-induced decrease in cell wall elasticity. Hence, the observed shift in the wafer content-potential relations in the cassava involved both an osmotic adjustment and a decrease in cell wall elasticity. Increasing the number of stress cycles per plant did not cause a further displacement of the water content-potential curves.     

Osmotic relations of the drought-tolerant shrub Artemisia tridentata in response to water stress

Turgor maintenance, solute content and recovery from water stress were examined in the drought-tolerant shrub Artemisia tridentata. Predawn water potentials of shrubs receiving supplemental water remained above ?2 MPa throughout summer, while predawn water potentials of untreated shrubs decreased to ?5 MPa. Osmotic potentials decreased in conjunction with water potentials maintaining turgor pressures above 0 MPa. The decreases in osmotic potentials were not the result of osmotic adjustment (i.e. solute accumulation). Leaf solute contents decreased during drought, but leaf water volumes decreased more than 75% from spring to summer, thereby passively concentrating solutes within the leaves. The maintenance of positive turgor pressures despite decreases in leaf water volumes is consistent with other studies of species with elastic cell walls. Inorganic ion, organic acid, and carbohydrate contents of leaves declined during drought. The only solutes accumulating in leaves of A. tridentata with water stress were proline and a cyclitol, both considered compatible solutes. Total and osmotic potentials recovered rapidly following rewatering of shrubs; solute contents did not change except for a decrease in proline. Maintaining turgor through the passive concentration of solutes may be advantageous compared to synthesis of new solutes for osmotic adjustment in arid environments.     

A comparison of the water relations characteristics of Helianthus annuus and Helianthus petiolaris when subjected to water deficits

The effect of water deficits on the water relations and stomatal responses of Helianthus annuus and Helianthus petiolaris were compared in plants growing in the glasshouse under controlled conditions. Unirrigated plants of both genotypes were subjected to two different stress rates in which predawn leaf water potentials declined steadily at either 0.15 MPa day^?1 or 0.50 MPa day^?1. In both genotypes water stress induced a gradual and similar decrease in leaf conductance from 1.6 to 0.3 cm s^?1 as water potential decreased from-0.5 to-2.0 MPa. The relationship between leaf conductance and leaf water potential was not affected by the rate of stress development. Development of predawn leaf water potentials of-1.3 MPa had no significant effect on the relative water content at zero turgor, the apoplastic water content or the volumetric elastic modulus of whole leaves in either species, but decreased the osmotic potential at full turgor and zero turgor by 0.22 MPa and decreased the turgid weight: dry weight ratio from 10.6 to 8.4 in H. annuus, but not in H. petiolaris. In H. annuus leaves expanded during stress development, changes in the osmotic potential at full turgor induced by water deficits did not disappear on rewatering.     

Tissue water relations and leaf growth of tropical corn cultivars under water deficits

Abstract This study reports on the effect of water deficit on the tissue water relations and leaf growth of six corn cultivars, growing in glasshouse conditions, in order to understand growth responses to drought of tropical corn. A mild water-stress treatment was imposed slowly; plants reached a minimum pre-dawn leaf water potential of about –1.5 MPa by day 12 after watering was withheld. Analysis of the water relation characteristics of growing leaves using the pressure–volume technique demonstrated that under water deficits all the cultivars changed their moisture-release curves compared with irrigated plants. Osmotic potential at full turgor was lowered in water-stressed plants of all the genotypes and the degree of such change was between 0.34 MPa and 0.58 MPa. Thus, turgor pressure was lost at a lower water potential in water-stressed plants than in irrigated plants of all the varieties. Volumetric elastic moduli were also increased under water deficits and the increase ranged between 10% and 141% among the cultivars. In all the genotypes, the stress imposed led to a reduction of leaf area and dry matter accumulation. Leaf expansion was very sensitive to low turgor pressure and it ceased when turgor reached 0.2 MPa. Thus, varieties able to maintain a higher degree of turgor pressure (i.e. by osmotic adjustment) under water deficits may be able to prolong leaf growth.     

Changes in free amino acids and osmotic adjustment in leaves of water-stressed bean

Bean ( Phaseolus vulgaris L. cv. Topcrop) plants were raised in a growth chamber in small pots or flower boxes and kept at full water regime until the full development of primary leaves (14–16 days). Both potted and flower box-grown plants were subjected to a gradually increased water stress of about 60–70 kPa day⁻¹ leaf water potential (stressed plants) or full water regime (control). The water potential, osmotic potential and turgor pressure in freshly detached primary leaves and osmotic potential at zero turgor were calculated using pressure/volume curves. Changes in free amino acids and amides were also measured in parallel trials. Water relation parameters documented that in the stressed leaves there was moderate osmotic adjustment, which was more evident in the potted plants. If considered 0% ionised, the accumulation of free amino acids and amides (μmol g⁻¹ H₂O) accounts for a van't Hoff's value of about 10.2 kPa in the small pot-grown and 5.5 kPa in the box-grown plants. The values are twice as high if considered 100% ionised. Proline accumulation accounted for about 6.4% of the pool enlargement in the potted plants and 22.3% in the flower box plants.     

Leaf age and salinity influence water relations of pepper leaves

Plant growth is reduced under saline conditions even when turgor in mature leaves is maintained by osmotic adjustment. The objective of this study was to determine if young leaves from salt-affected plants were also osmotically adjusted. Pepper plants (Capsicum annuum L. cv. California Wonder) were grown in several levels of solution osmotic potential and various components of the plants' water relations were measured to determine if young, rapidly growing leaves could accumulate solutes rapidly enough to maintain turgor for normal cell enlargement. Psychrometric measurements indicated that osmotic adjustment is similar for both young and mature leaves although osmotic potential is slightly lower for young leaves. Total water potential is also lower for young leaves, particularly at dawn for the saline treatments. The result is reduced turgor under saline conditions at dawn for young but not mature leaves. This reduced turgor at dawn, and presumably low night value, is possibly a cause of reduced growth under saline conditions. No differences in leaf turgor occur at midday. Porometer measurements indicated that young leaves at a given salinity level have a higher stomatal conductance than mature leaves, regardless of the time of day. The result of stomatal closure is a linear reduction of transpiration.     

Phloem water relations and translocation

Satisfactory measurements of phloem water potential of trees can be obtained with the Richards and Ogata psychrometer and the vapor equilibration techniques, although corrections for loss of dry weight and for heating by respiration are required for the vapor equilibrium values. The psychrometer technique is the more satisfactory of the 2 because it requires less time for equilibration, less tissue, and less handling of tissue. Phloem water potential of a yellow-poplar tree followed a diurnal pattern quite similar to that of leaves, except that the values were higher (less negative) and changed less than in the leaves.

The psychrometer technique permits a different approach to the study of translocation in trees. Measurements of water potential of phloem discs followed by freezing of samples and determination of osmotic potential allows estimation of turgor pressure in various parts of trees as the difference between osmotic potential and total water potential. This technique was used in evaluating gradients in water potential, osmotic potential, and turgor pressure in red maple trees. The expected gradients in osmotic potential were observed in the phloem, osmotic potential of the cell sap increasing (sap becoming more dilute) down the trunk. However, values of water potential were such that a gradient in turgor pressure apparently did not exist at a time when rate of translocation was expected to be high. These results do not support the mass flow theory of translocation favored by many workers.

     

Effects of water stress on the water relations of Phaseolus vulgaris and the drought resistant Phaseolus acutifolius

The tepary bean ( Phaseolus acutifolius Gray var. latifolius ), a drought resistant species, was compared under water stress conditions with the more drought susceptible P. vulgaris L. cvs Pinto and White Half Runner (WHR). In order to better understand the basis for the superior drought resistance of tepary, this study was designed to determine the relationships among leaf water potential, osmotic potential, turgor potential, and relative water content (RWC).
Plants were prestressed by withholding irrigation water. These stress pretreatments changed the relation between leaf water potential and relative water content of both species so that prestressed plants had lower water potentials than controls at the same leaf RWC. Tepary had lower water potentials at given RWC levels than Pinto or WHR; this can account for part of the superior resistance of tepary. In all genotypes, prestressed plants maintained osmotic potentials approximately 0.2 MPa lower than controls. Tepary reached osmotic potentials that were significantly lower (0.15 to 0.25 MPa) than Pinto or WHR. Both control and prestressed tepary plants had 0.05 to 0.25 MPa more turgor than Pinto or WHR at RWC values between 65 and 80%. Both prestressed and control tepary plants had greater elasticity (a lower elastic modulus) than Pinto or WHR. This greater turgor of tepary at low RWC values could be caused by several factors including greater tissue elasticity, active accumulation of solutes, or greater solute concentration.
Tepary had significantly lower osmotic potentials than the P. vulgaris cultivars, but there was little difference in osmotic potential between Pinto and WHR. Knowledge of differences in osmotic and turgor potentials among and within species could be useful in breeding for drought resistance in Phaseolus.     

Effects of water stress and rewatering on leaf water relations of lemon plants

Potted two-year-old lemon plants (Citrus limon (L.) Burm. fil.) cv. Fino, growing under field conditions were subjected to drought by withholding irrigation for 13 d. After that, plants were re-irrigated and the recovery was studied for 5 d. Control plants were daily irrigated maintaining the soil matric potential at about -30 kPa. Young leaves of control plants presented higher leaf conductance (g1) and lower midday leaf water potential (Ψmd) than mature ones. Young leaves also showed higher leaf water potential at the turgor loss point (Ψtlp) than mature leaves. In both leaf types g1 decreased with increased vapour pressure deficit of the atmosphere. From day 1 of the withholding water, predawn and midday leaf water potentials (Ψpd and Ψmd) decreased, reaching in both cases minimum values of -5.5 MPa, with no significant differences between mature and young leaves. Water stress induced stomatal closure, leaf rolling and partial defoliation. No osmotic adjustment was found in response to water stress in either leaf type, but both were able to enhance the cell wall elasticity (elastic adjustment). After rewatering, leaf water potential recovered quickly (within 2 d) but g1 did not. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effect of Soil Water Stress on Osmotic Adjustment and Elongation Growth of Wheat Leaves

The results of the experiment showed that leaf elongation rate in two wheat cultivars decreased under soil water stress. Rewatering after water stress, growth restoration.of “Changle No.5” was faster than that of “Lumai No.5”. The osmotic adjustment ability of leaves in these two wheat cultivars increased to 0.41MPa for “Changle No.5” and 0.33MPa for “Lumai No.5” as water potential decreased. At the same leaf elongation rate water potential and osmotic potential of “Changle No5” decreased more than that of “Lumai No.5” Leaf elongation rate fell to zero as water potential and osmotic potential were –1.50MPa and –1.70MPa for “Changle No.5” and –1.20MPa and –1.30MPa for “Lumai No.5” The threshold turgor pressure of elongation growth in leaf cell was different being 0.22MPa for “Changle No.5’ and 0.15MPa for “Lumai No.5”. The difference in the gross extensible coefficient of growing leaf was very small.     

Genotypic variation in tissue water relations of leaves and roots of black walnut (Juglans nigra) seedlings

Genetic variation in the drought response of leaf and root tissue water relations of seedlings of eight sources of black walnut ( Juglans nigra L.) was investigated using the pressure-volume technique. Tissue water relations were characterized at three stages of a drying cycle during which well-watered plants were allowed to desiccate and then were reirrigated.
Sources varied both in the capacity for and degree of leaf and root osmotic adjustment, and in the mechanism by which it was achieved. A decrease in osmotic potential at the turgor loss point (ψ_πp) of 0.4 MPa was attributable to increased leaf tissue elasticity in seedlings of four sources, while seedlings of an Ontario source exhibited a 0.7–0.8 MPa decline in ψ_πp as a result of both increased solute content and increased leaf tissue elasticity. Seedlings of a New York source showed no detectable osmotic adjustment.
In roots, decreased ψ_πp (osmotic potential at full hydration) and ψ_πp were observed under drought. Sources that exhibited significant leaf osmotic adjustment also generally showed a similar response in roots. Tissue elasticity and ψ_πp of roots were higher than those of shoots, whereas ψ_πp of the two organs was similar for most sources. Because of greater elasticity, roots exhibited a more gradual decline in turgor and total water potential than did leaves as tissue relative water content decreased.     

Does turgor limit growth in tall trees?

The gravitational component of water potential contributes a standing 0.01 MPa m^?1 to the xylem tension gradient in plants. In tall trees, this contribution can significantly reduce the water potential near the tree tops. The turgor of cells in buds and leaves is expected to decrease in direct proportion with leaf water potential along a height gradient unless osmotic adjustment occurs. The pressure–volume technique was used to characterize height‐dependent variation in leaf tissue water relations and shoot growth characteristics in young and old Douglas‐fir trees to determine the extent to which growth limitation with increasing height may be linked to the influence of the gravitational water potential gradient on leaf turgor. Values of leaf water potential (Ψ_l), bulk osmotic potential at full and zero turgor, and other key tissue water relations characteristics were estimated on foliage obtained at 13.5 m near the tops of young (approximately 25‐year‐old) trees and at 34.7, 44.2 and 55.6 m in the crowns of old‐growth (approximately 450‐year‐old) trees during portions of three consecutive growing seasons. The sampling periods coincided with bud swelling, expansion and maturation of new foliage. Vertical gradients of Ψ_l and pressure–volume analyses indicated that turgor decreased with increasing height, particularly during the late spring when vegetative buds began to swell. Vertical trends in branch elongation, leaf dimensions and leaf mass per area were consistent with increasing turgor limitation on shoot growth with increasing height. During the late spring (May), no osmotic adjustment to compensate for the gravitational gradient of Ψ_l was observed. By July, osmotic adjustment had occurred, but it was not sufficient to fully compensate for the vertical gradient of Ψ_l. In tall trees, the gravitational component of Ψ_l is superimposed on phenologically driven changes in leaf water relations characteristics, imposing potential constraints on turgor that may be indistinguishable from those associated with soil water deficits.     

Drought tolerance in faster- and slower-growing black spruce (Picea mariana) progenies: II. Osmotic adjustment and changes of soluble carbohydrates and amino acids under osmotic stress

The possibility was considered that osmotic adjustment, the ability to accumulate solutes in response to water stress, may contribute to growth rate differences among closely-related genotypes of trees. Progeny variation in osmotic adjustment and turgor regulation was investigated by comparing changes in osmotic and pressure potentials, soluble carbohydrates, and amino acids in osmotically stressed seedlings in 4 full-sib progenies of black spruce [ Picea mariana (Mill.) B. S. P.] that differed in growth rate under drought. Osmotic stress was induced by a stepwise increase in the concentration of polyethylene glycol (PEG)-3350 from 10 (w/v) to 18 and 25%, which provided osmotic potentials in solution culture of -0.4, -1.0 and -2.0 MPa each for 3 days. All 4 progenies maintained a positive cell turgor even at 25% PEG, due to a significant decline in osmotic potential. Although total amino acids, principally proline, increased, ca 60% of the decrease in osmotic potential was attributable to soluble carbohydrates and glucose was the major osmoregulating solute. There was little progeny variation in any of measured parameters in unstressed seedlings. Compared to two slower-growing progenies, the two progenies capable of more vigorous growth under drought in the field accumulated more soluble carbohydrates (mainly glucose and fructose), developed lower osmotic potential and maintained higher turgor pressure when osmotically-stressed in solution culture. The ability to adjust osmotically and maintain turgor under drought stress could thus be a useful criterion for the early selection of faster-growing, drought-tolerant genotypes.     

The role of solute accumulation, osmotic adjustment and changes in cell wall elasticity in drought tolerance in Ziziphus mauritiana (Lamk.)

Ber (Ziziphus mauritiana Lamk.) is a major fruit tree crop of the north-west Indian arid zone. In a study of the physiological basis of drought tolerance in this species, two glasshouse experiments were conducted in which trees were droughted during single stress-cycles. In the first experiment, during a 13 d drying cycle, pre-dawn leaf water (leaf) and osmotic () potentials in droughted trees declined from -0.5 and -1.4 MPa to -1.7 and -2.2 MPa, respectively, for a decrease in relative water content () of 14%. During drought stress, changes in sugar metabolism were associated with significant increases in concentrations of hexose sugars (3.8-fold), cyclitol (scyllo-inositol; 1.5-fold), and proline (35-fold; expressed per unit dry weight), suggesting that altered solute partitioning may be an important factor in drought tolerance of Ziziphus. On rewatering pre-dawn leaf and recovered fully, but remained depressed by 0.4 MPa relative to control values, indicating that solute concentration per unit water content had changed during the drought cycle.Evidence for osmotic adjustment was provided from a second study during which a gradual drought was imposed. Pressure-volume analysis revealed a 0.7 MPa reduction in osmotic potential at full turgor, with leaf at turgor loss depressed by 1 MPa in drought-stressed leaves. Coupled with osmotic adjustment, during gradual drought, was a 65% increase in bulk tissue elastic modulus (wall rigidity) which resulted in turgor loss at the same in both stressed and unstressed leaves. The possible ecological significance of maintenance of turgor potential and cell volume at low water potentials for drought tolerance in Ziziphus is discussed.Keywords: Ziziphus mauritiana, drought, solute accumulation, osmotic adjustment, proline.      

Measurement of a growth-induced water potential gradient in tall fescue leaves

Spatial distribution of cell turgor pressure, cell osmotic pressure and relative elemental growth rate were measured in growing tall fescue leaves ( Festuca arundinacea ). Cell turgor pressure (measured with a pressure probe) was c . 0.55 MPa in expanding cells but increased steeply (+0.3 MPa) in cells where elongation had stopped. However, cell osmotic pressure (measured with a picolitre osmometer) was almost constant at 0.85 MPa throughout the leaf. The water potential difference between the growth zone and the mature zone (0.3 MPa) was interpreted as a growth-induced water potential gradient. This and further implications for the mechanism of growth control are discussed.     

Water Potential-Water Content Relationships In Apple Leaves

Three methods for determining the relationship between xylempressure potential as measured in a pressure chamber (an estimateof leaf water potential) and leaf relative water content werecompared for apple leaves. A range of leaf water contents wasobtained either by sampling leaves in the field at differenttimes of day and on days with differing evaporative demand,or by allowing evaporation from excised leaves in the laboratory,or by expressing sap by overpressurization in a pressure chamber.The first two methods gave very similar results, but the lasttended to give rather lower water potentials at any given watercontent. A possible explanation for these results and theirimplications for the estimation of osmotic potentials usingpressure-volume curves are discussed. Some osmotic adjustmentwas observed in trees droughted for 3 months, with estimatedosmotic potentials, both at full turgor and zero turgor, beingnearly 0.3 MPa lower than in irrigated controls.     

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.     

Drought-induced variations of water relations parameters in Olea europaea

The effects of water stress on water potential components, tissue water content, mean elastic modulus and the osmoregulation capacity of olive (Olea europaea L. cv. Coratina) leaves was determined. Artificial rehydration of olive leaf tissues altered the P-V relationships so that a plateau phenomenon occurred. Points in the P-V curve in the region affected by the plateau, generally up to –0.5 MPa, were corrected for all the samples analyzed. In the corrected P-V relationship, an osmotic adjustment was found in drought-stressed leaf tissues. Osmotic potentials at full turgor (_{0 (sat)}) and osmotic potential at turgor-loss (_{0 (TVT)}) decreased from –2.06±0.01 MPa and –3.07±0.16 MPa in controls to –2.81±0.03 MPa and –3.85±0.12 MPa in most stressed plants. Osmotic adjustment values obtained from the P-V curves agreed with those obtained using an osmometer. An active osmotic adjustment of 1.42 MPa was also observed in 1–4 mm- diameter roots. Mannitol is the main carbohydrate involved in osmotic potential decrease in all treatments. The maximum elastic modulus increased from 11.6±0.95 MPa in the controls to 18.6±0.61 MPa in the most stressed plants.     

Cell water potential,osmotic potential,and turgor in the epidermis and mesophyll of transpiring leaves

Water potential, osmotic potential and turgor measurements obtained by using a cell pressure probe together with a nanoliter osmometer were compared with measurements obtained with an isopiestic psychrometer. Both types of measurements were conducted in the mature region of Tradescantia virginiana L. leaves under non-transpiring conditions in the dark, and gave similar values of all potentials. This finding indicates that the pressure probe and the osmometer provide accurate measurements of turgor, osmotic potentials and water potentials. Because the pressure probe does not require long equilibration times and can measure turgor of single cells in intact plants, the pressure probe together with the osmometer was used to determine in-situ cell water potentials, osmotic potentials and turgor of epidermal and mesophyll cells of transpiring leaves as functions of stomatal aperture and xylem water potential. When the xylem water potential was-0.1 MPa, the stomatal aperture was at its maximum, but turgor of both epidermal and mesophyll cells was relatively low. As the xylem water potential decreased, the stomatal aperture became gradually smaller, whereas turgor of both epidermal and mesophyll cells first increased and afterward decreased. Water potentials of the mesophyll cells were always lower than those of the epidermal cells. These findings indicate that evaporation of water is mainly occurring from mesophyll cells and that peristomatal transpiration could be less important than it has been proposed previously, although peristomatal transpiration may be directly related to regulation of turgor in the guard cells.