Cell Pressure Probe Research Articles

The effect of exogenous abscisic acid on stomatal development, stomatal mechanics, and leaf gas exchange in Tradescantia virginiana.

Gas exchange parameters and stomatal physical properties were measured in Tradescantia virginiana plants grown under well-watered conditions and treated daily with either distilled water (control) or 3.0 mM abscisic acid (ABA). Photosynthetic capacity (CO(2) assimilation rate for any given leaf intercellular CO(2) concentration [c(i)]) and relative stomatal sensitivity to leaf-to-air vapor-pressure difference were unaffected by the ABA treatment. However, at an ambient CO(2) concentration (c(a)) of 350 micromol mol(-1), ABA-treated plants operated with significantly lower c(i). ABA-treated plants had significantly smaller stomata and higher stomatal density in their lower epidermis. Stomatal aperture versus guard cell pressure (P(g)) characteristics measured with a cell pressure probe showed that although the form of the relationship was similar in control and ABA-treated plants, stomata of ABA-treated plants exhibited more complete closure at P(g) = 0 MPa and less than half the aperture of stomata in control plants at any given P(g). Scaling from stomatal aperture versus P(g) to stomatal conductance versus P(g) showed that plants grown under ABA treatment would have had significantly lower maximum stomatal conductance and would have operated with lower stomatal conductance for any given guard cell turgor. This is consistent with the observation of lower c(i)/c(a) in ABA-treated plants with a c(a) of 350 micromol mol(-1). It is proposed that the ABA-induced changes in stomatal mechanics and stomatal conductance versus P(g) characteristics constitute an improvement in water-use efficiency that may be invoked under prolonged drought conditions.

Transient variations of water transfer induced by HgCl2 in excised roots of young maize plants: new data on the inhibition process

Effects of low concentrations of HgCl2 on water transport in excised maize (Zea mays L.) roots have been monitored using a reliable system that permits continuous measurement of xylem flux variations. The sap flow of exuding roots treated with 11 M HgCl2 decreased by 80–90% in 10 min at 25˚C. Reversal of this inhibition was obtained using a sulfhydryl [β-mercaptoethanol (β-ME)], or non-sulfhydryl [ethylenediaminetetraacetate disodium salt (EDTA) and ferricyanide (FeCy)] reagents. The time course of reversal was not the same in the three cases. β-ME reversed quickly but not sustainably, whereas EDTA or FeCy reversed slowly and sustainably. Using a cell pressure probe, turgor was measured in the epidermis, and the first layer of cells in the cortex. Turgor was considerably decreased in root epidermal cells after HgCl2 treatment, suggesting that a normal root water-transfer required an optimal turgor in these cells. Recovery of cell turgor, was obtained satisfactorily with FeCy. In parallel with flux measurements, histochemical analyses revealed a localization of Hg only in peripheral root cells, suggesting that Hg targets are localized in the first root cell layers. Involvement of water channels and/or ion transport in the regulation of root water transport is discussed in the light of our data.

Radial hydraulic conductivity along developing onion roots.

Although most studies have shown that water uptake varies along the length of a developing root, there is no consistent correlation of this pattern with root anatomy. In the present study, water movement into three zones of onion roots was measured by a series of mini-potometers. Uptake was least in the youngest zone (mean hydraulic conductivity, Lpr = 1.5 x 10(-7) +/- 0.34 x 10(-7) m MPa-1 s-1; +/- SE, n = 10 roots) in which the endodermis had developed only Casparian bands and the exodermis was immature. Uptake was significantly greater in the middle zone (Lpr = 2.4 x 10(-7) +/- 0.43 x 10(-7) m MPa-1 s-1; +/- SE, n = 10 roots) which had a mature exodermis with both Casparian bands and suberin lamellae, and continued at this level in the oldest zone in which the endodermis had also developed suberin lamellae (Lpr = 2.8 x 10(-7) +/- 0.30 x 10(-7) m MPa-1 s-1; +/- SE, n = 10 roots). Measurements of the hydraulic conductivities of individual cells (Lp) in the outer cortex using a cell pressure probe indicated that this parameter was uniform in all three zones tested (Lp = 1.3 x 10(-6) +/- 0.01 x 10(-6) m MPa-1 s-1; +/- SE, n = 60 cells). Lp of the youngest zone was lowered by mercuric chloride treatment, indicating the involvement of mercury-sensitive water channels (aquaporins). Water flow in the older two root zones measured by mini-potometers was also inhibited by mercuric chloride, despite the demonstrated impermeability of their exodermal layers to this substance. Thus, water channels in the epidermis and/or exodermis of the older regions were especially significant for water flow. The results of this and previous studies are discussed in terms of two models. The first, which describes maize root with an immature exodermis, is the 'uniform resistance model' where hydraulic resistances are evenly distributed across the root cylinder. The second, which describes the onion root with a mature exodermis, is the 'non-uniform resistance model' where resistances can be variable and are concentrated in a certain layer(s) on the radial path.

Chemical composition of apoplastic transport barriers in relation to radial hydraulic conductivity of corn roots (Zea mays L.).

The hydraulic conductivity of roots (Lp(r)) of 6- to 8-d-old maize seedlings has been related to the chemical composition of apoplastic transport barriers in the endodermis and hypodermis (exodermis), and to the hydraulic conductivity of root cortical cells. Roots were cultivated in two different ways. When grown in aeroponic culture, they developed an exodermis (Casparian band in the hypodermal layer), which was missing in roots from hydroponics. The development of Casparian bands and suberin lamellae was observed by staining with berberin-aniline-blue and Sudan-III. The compositions of suberin and lignin were analyzed quantitatively and qualitatively after depolymerization (BF(3)/methanol-transesterification, thioacidolysis) using gas chromatography/mass spectrometry. Root Lp(r) was measured using the root pressure probe, and the hydraulic conductivity of cortical cells (Lp) using the cell pressure probe. Roots from the two cultivation methods differed significantly in (i) the Lp(r) evaluated from hydrostatic relaxations (factor of 1.5), and (ii) the amounts of lignin and aliphatic suberin in the hypodermal layer of the apical root zone. Aliphatic suberin is thought to be the major reason for the hydrophobic properties of apoplastic barriers and for their relatively low permeability to water. No differences were found in the amounts of suberin in the hypodermal layers of basal root zones and in the endodermal layer. In order to verify that changes in root Lp(r) were not caused by changes in hydraulic conductivity at the membrane level, cell Lp was measured as well. No differences were found in the Lp values of cells from roots cultivated by the two different methods. It was concluded that changes in the hydraulic conductivity of the apoplastic rather than of the cell-to-cell path were causing the observed changes in root Lp(r).

The plant cell pressure probe

The pressure probe is a micro manometer for the simultaneous direct recording and manipulation of plant cell hydrostatic pressure. It is used to map in space and time the turgor pressures of individual cells within tissues and organs of intact plants. This is used to study the hydraulic architecture of tissues, tissue movement and the responses of tissues to water stress. The approach can be augmented by simultaneous measurement of individual cell osmotic pressure. This permits the hydraulic driving forces across selectively permeable membranes and walls to be assessed fully. By manipulating manually the pressure, cell wall elasticity and its properties can also be mapped. Under some conditions this can be extended to plastic behaviour.

Direct measurement of xylem pressure in leaves of intact maize plants. A test of the cohesion-tension theory taking hydraulic architecture into consideration

The water relations of maize (Zea mays L. cv Helix) were documented in terms of hydraulic architecture and xylem pressure. A high-pressure flowmeter was used to characterize the hydraulic resistances of the root, stalk, and leaves. Xylem pressure measurements were made with a Scholander-Hammel pressure bomb and with a cell pressure probe. Evaporation rates were measured by gas exchange and by gravimetric measurements. Xylem pressure was altered by changing the light intensity, by controlling irrigation, or by gas pressure applied to the soil mass (using a root pressure bomb). Xylem pressure measured by the cell pressure probe and by the pressure bomb agreed over the entire measured range of 0 to -0.7 MPa. Experiments were consistent with the cohesion-tension theory. Xylem pressure changed rapidly and reversibly with changes in light intensity and root-bomb pressure. Increasing the root-bomb pressure increased the evaporation rate slightly when xylem pressure was negative and increased water flow rate through the shoots dramatically when xylem pressure was positive and guttation was observed. The hydraulic architecture model could predict all observed changes in water flow rate and xylem. We measured the cavitation threshold for oil- and water-filled pressure probes and provide some suggestions for improvement.

Diurnal variations in hydraulic conductivity and root pressure can be correlated with the expression of putative aquaporins in the roots of lotus japonicus

The hydraulic conductivity of excised roots (Lp(r)) of the legume Lotus japonicus (Regel) K. Larsen grown in mist (aeroponic) and sand cultures, was found to vary over a 5-fold range during a day/night cycle. This behaviour was seen when Lp(r) was measured in roots exuding, either under root pressure (osmotic driving force), or under an applied hydrostatic pressure of 0.4 MPa which produced a rate of water flow similar to that in a transpiring plant. A similar daily pattern of variation was seen in plants grown in natural daylight or in controlled-environment rooms, in plants transpiring at ambient rates or at greatly reduced rates, and in plants grown in either aeroponic or sand culture. When detached root systems were connected to a root pressure probe, a marked diurnal variation was seen in the root pressure generated. After excision, this circadian rhythm continued for some days. The hydraulic conductivity of the plasma membrane of individual root cells was measured during the diurnal cycle using a cell pressure probe. Measurements were made on the first four cell layers of the cortex, but no evidence of any diurnal fluctuation could be found. It was concluded that the conductance of membranes of endodermal and stelar cells may be responsible for the observed diurnal rhythm in root Lp(r). When mRNAs from roots were probed with cDNA from the Arabidopsis aquaporin AthPIP1a gene, an abundant transcript was found to vary in abundance diurnally under high-stringency conditions. The pattern of fluctuations resembled closely the diurnal pattern of variation in root Lp(r). The plasma membranes of root cells were found to contain an abundant hydrophobic protein with a molecular weight of about 31 kDa which cross-reacted strongly to an antibody raised against the evolutionarily conserved N-terminal amino acid sequence of AthPIP1a.

A study of stomatal mechanics using the cell pressure probe

The relationship between stomatal aperture (a) and guard cell pressure (Pg) was measured directly in four different species (Vicia faba, Tradescantia virginiana, Ginkgo biloba and Nephrolepis exaltata) using a special cell pressure probe technique. The effect of epidermal turgor (Pep) on this relationship was also measured in T. virginiana. The relationship was sigmoidal for V. faba and T. virginiana, but entirely convex for G. biloba and N. exaltata. Epidermal turgor was found to have a pronounced closing effect on stomata of T. virginiana. Maximum aperture with full epidermal turgor (0·92 MPa) was about half that with zero epidermal turgor. Also, with full epidermal turgor stomata of T. virginiana did not begin to open until Pg was more than 1·25 MPa. These characteristics were used to develop an expression for a as a function of Pg and Pep. Results for the different species are compared and discussed in terms of possible advantages and limitations of water economy.

Why do leaves and leaf cells of N-limited barley elongate at reduced rates?

The objective of the present study was to assess whether, in barley, nitrogen supply limits the rate of leaf elongation through a reduction in (relative) cell elongation rate and whether this is attributable to a reduced turgor, a reduced availability of osmolytes or, by implication, changed wall properties. Plants were grown on full-strength Hoagland solution (“Hoagland”-plants), or on N-deficient Hoagland solution while receiving N at a relative addition rate of 16 or 8% N · plant-N−1 · d−1 (“16%-” and “8%-plants”). Hoagland-plants were demand-limited, whereas 16%- and 8%-plants were supply-limited in N. Third leaves were analysed for leaf elongation rate and final epidermal cell length, and, within the basal growing region, for the spatial distribution of relative segmental elongation rates (RSER, pin-pricking method), epidermal cell turgor (cell-pressure probe), osmotic pressure (OP, picolitre osmometry) and water potential (Ψ). During the development of the third leaf, plants grew at relative growth rates (relative increase in fresh weight ) of 18.2, 15.6 and 8.1% · d−1 (Hoagland-, 16%- and 8%-plants, respectively). Final leaf length and leaf elongation rate were highest in Hoagland plants (ca. 34.1 cm and 2.33–2.60 mm · h−1, respectively), intermediate in 16%- plants (31.0 cm and 1.89–1.96 mm · h−1) and lowest in 8%-plants (29.4 cm and 1.41–1.58 mm · h−1). These differences were accompanied by only small differences in final cell length, but large differences in cell-flux rates (146, 187 and 201 cells · cell-file−1 · d−1 in 8%-, 16%- and Hoagland-plants, respectively). The length of the growth zone (32–38 mm) was not much affected by N-levels (and nutrient technique). A decrease in RSER in the growth zone distal to 10 mm produced the significant effect of N-levels on leaf elongation rate. In all treatments, cell turgor was almost constant throughout the growing region, as were cell OP and Ψ in 16%- and 8%-plants. In Hoagland-plants, however, cell OP increased by ca. 0.1 MPa within the zone of highest elongation rates and, as a consequence, cell Ψ decreased simultaneously by 0.1 MPa. Cell Ψ increased considerably where elongation ceased. Within the zone where differences in RSERs were highest between treatments (10–34 mm from base) average turgor was lowest, OP highest and Ψ most negative in Hoagland- compared to 8%- and 16%-plants (P < 0.001), but not significantly different between 8%- and 16%-plants.

The function of water channels in Chara : The temperature dependence of water and solute flows provides evidence for composite membrane transport and for a slippage of small organic solutes across water channels

Using the cell pressure probe, the effects of temperature on hydraulic conductivity (Lp; osmotic water permeability), solute permeability (permeability coefficient, Ps), and reflection coefficients (σs) were measured on internodes of Chara corallina, Klein ex Willd., em R.D.W.. For the first time, complete sets of transport coefficients were obtained in the range between 10 and 35 °C which provided evidence about pathways of water and solutes as they move across the plasma membrane (water channel and bilayer arrays). Test solutes used to check for the selectivity of water channels were monohydric alcohols of different molecular size and shape (ethanol, n-propanol, iso-propanol, and tert-butanol) and heavy water (HDO). Within the limits of accuracy, Q10 values for Lp and for the diffusive water permeability (Pd) were identical (Q10 for Lp = 1.29 ± 0.17 (± SD; n = 15 cells) and Q10 for Pd = 1.25 ± 0.16 (n = 5 cells)). The Q10 values were equivalent to activation energies of Ea = 16.8 ± 6.4 and 16.6 ± 10.0 kJ · mol−1, respectively, which is similar to that of self-diffusion or of viscous flow of water. The Q10 values and activation energies for Ps of the alcohols were significantly larger (ethanol: Q10 = 1.68 ± 0.16, Ea = 37.1 ± 5.9 kJ · mol−1; n-propanol: Q10 = 1.75 ± 0.40, Ea = 43.1 ± 15.3 kJ · mol−1; iso-propanol: Q10 = 2.12 ± 0.42, Ea = 52.2 ± 14.6 kJ · mol−1; tert-butanol: Q10 = 2.13 ± 0.56, Ea = 51.6 ± 17.1 kJ · mol−1; ±SD; n = 5 to 6 cells). Effects of temperature on reflection coefficients were most pronounced. With increasing temperature, σs values of the alcohols decreased and those of HDO increased. The data indicate that water and solutes use different pathways when crossing the membrane. Ordinary and isotopic water use water channels and the other test solutes use the bilayer array (composite transport model of membrane). Changes in σs values with temperature were found to be a sensitive measure for the open/closed state of water channels. The decrease of σs with temperature was theoretically predicted from the temperature dependence of Ps and Lp. Differences between predicted and measured values of σs allowed estimation of the bypass flow (slippage) of solutes through water channels which did not completely exclude test solutes. The permeability of channels depended on the structure and size of test solutes. It is concluded that water channels are much less selective than is usually thought. Since water channels represent single-file or no-pass pores, solutes drag along considerable amounts of water as they diffuse across channels. This results in low overall values of σs. The σs of HDO was extremely low. Its response to temperature was opposite to that for the σs of the alcohols. This suggested a stronger effect of temperature on the hydraulic (osmotic) than on the diffusive water flow across individual water channels, i.e. a differential sensitivity of different mechanisms to temperature.

Phototropic Response of the Bean Pulvinus: Movement of Water and Ions

Abstract:Segmental analysis of the laminar pulvinus of Phaseolus vulgaris L. showed that its phototropic curvature is accompanied by efflux of inorganic ions and water from its contracting sector and a comparable influx into its expanding one. All the major ions, except Na+, contributed to this transport, suggesting that the response to light involves changes in the driving force, or conductivity of a wide range of solutes. During the curvature, K+ and CI− made the greatest and equivalent contributions to efflux, but only Cl− exhibited a matching influx into the expanding sector, while K+ influx was much less. Use of the cell pressure probe showed that, as the laminar angle of elevation changed between −40° to +40°, turgor pressure in the expanding motor cells increased by 0.48 MPa and decreased in the contracting cells by 0.32 MPa. Picoliter osmometry of single‐cell samples showed that during this movement vacuolar osmotic pressure remained constant. Thus, changes in turgor pressure resulted from changes in apoplastic, rather than the protoplastic osmotic pressure. Volumetric modulus of elasticity of pulvinar motor cells is very low, showing that their walls are very elastic. These properties increase the effectiveness of converting osmotic work into the large‐scale, reversible volume changes responsible for leaf movements.

Cell turgor, osmotic pressure and water potential in the upper epidermis of barley leaves in relation to cell location and in response to NaCl and air humidity

Previous single-cell studies on the upper epidermis of barley leaves have shown that cells differ systematically in their solute concentrations depending on their location relative to stomatal pores and veins and that during NaCI stress, gradients in osmotic pressure (π) develop (Fricke et al., 1995, 1996; Hinde, 1994). The objective of the present study was to address the question to which degree these intercellular differences in solute concentrations and π are associated with intercellular differences in turgor or water potential (ψ). Epidermal cells analysed were located at various positions within the ridge regions overlying large lateral or intermediate veins, in the trough regions between those veins or in between stomata (i.e. interstomatal cells). Turgor pressure of cells was measured using a cell pressure probe, and π of extracted cell sap was determined by picolitre osmometry. For both large and intermediate lateral veins, there were no systematic differences in turgor between cells located at the base, mid or top of ridges, regardless of whether plants were analysed at low or high PAR (≤ 10 or 300-400 μmol photons m -2 s -1 ). However, turgor within a ridge region was not necessarily uniform, but could vary by up to 0.14 MPa (1.4 bar) between adjacent cells. In 60 out of 63 plants, turgor of ridge cells was either slightly or significantly higher than turgor of trough (lowest turgor) or interstomatal cells (intermediate turgor). The significance and magnitude of turgor differences was higher in plants analysed under high PAR or local air flow than in plants analysed under low PAR. The largest (up to 0.41 MPa) and consistently significant differences in turgor were found in plants treated for 3-9 d prior to analysis with 100 mM NaCI. For both NaCI-treated and non-treated (control) plants, differences in turgor between cell types were mainly due to differences in π since differences in ψ were negligible (0.01-0.04 MPa). Epidermal cell ψ in NaCI-treated plants was about 0.38 MPa more negative than in control plants due to higher π. Turgor pressures were similar. Following a sudden change in rooting-medium ψ or air humidity, turgor of both ridge and trough cells responded within seconds and followed the same time-course of relaxation. The half time (T 1/2 ) of turgor relaxation was not limited by the cell's T 1/2 for water exchange.

Water channels inChara corallina

Water relations parameters of Chara corallina internodes were measured using the single cell pressure probe. The effect of mercurials, which are recognized as non-specific water channel inhibitors, was examined. HgCI2 concentrations greater than 5 mmol m 3 were found to inhibit hydraulic conductivity (Lp) close to 90%, whereas pCMPS was found to have no effect on Lp. The activation energy of water flow was increased significantly from 21.0 kJ mol~1 to 45.6 kJ mol 1, following the application of HgCI 2. These results are in accordance with evidence for Hg2+-sensitive water channels in the plasma membrane of charophytes (Henzler and Steudle, 1995; Tazawa et a/., 1996). The metabolic effects must, however, be considered in view of the rapid inhibition of respiration and the depolarization of the membrane potential with HgCI2 concentrations lower than those found to affect Lp. It was possible to measure simultaneously water relations and membrane PD, in order to examine the contribution of potassium channels to Lp. Cells were induced into a K+ permeable state. The K+ channels, assumed to be open, were subsequently blocked by various blockers. No significant difference in Lp was found for any of these treatments. Finally, the permeability of C. corallina membranes to ethanol was examined. HgCI2 was found to cause a decrease in reflection coefficient, coinciding with a decrease in Lp, but there was no change in the ethanol permeability coefficient. This has been interpreted in terms of both the frictional model and composite model of non-electrolyte membrane transport.

A New Turgor/Membrane Potential Probe Simultaneously Measures Turgor and Electrical Membrane Potential

Abstract: A new combined turgor/membrane potential probe (T‐EP probe) monitored cell turgor and membrane potential simultaneously in single giant cells. The new probe consisted of a silicone oil‐filled micropipette (oil‐microelectrode), which conducted electric current. Measurements of turgor and hydraulic conductivity were performed as with the conventional cell pressure probe besides the membrane potential. In internodal cells of Chara corallina, steady state turgor (0.5‐0.7 MPa) and resting potentials (‐200 to −220 mV) in APW, and hydraulic conductivity (0.07 to 0.21 × 10∼5 m s−1 MPa−1) were measured with the new probe, and cells exhibited healthy cytoplasmic streaming for at least 24 h during measurements. When internodal cells of Chara corallina were treated with 30, 20, 10, and 5 mM KCI, turgor responded immediately to all concentrations, and the osmotic changes in the medium were measured. Action potentials, which brought the membrane potential to a steady depolarization that measured the concentration difference of K+ in the medium, were induced in a concentration — dependent delay and occurred only 30, 20, and 10 mM of KCl. When the solution was changed back to APW, the repolarization of membrane potential consisted of a quick and a following slow phase. During the quick phase, which took place immediately and lasted 1 to 3 min, the plasma membrane remained activated. The membrane was gradually deactivated in the slow phase, and entirely deactivated when the membrane potential recovered to the resting potential in APW. Although the activated plasma membrane was permeable to K+, no major ion channels were activated on the tonoplast, and therefore, internodal cells of Chara corallina did not regulate turgor when osmotic potential changed in the surrounding medium.

Control of leaf expansion in sunflower (Helianthus annuusL.) by nitrogen nutrition

Seedlings of Helianthus annuus L. were grown at an initially high relative nitrate supply rate (0.27 mol N mol N–1 d–1). The supply was subsequently reduced to a low rate (0.04 mol N mol N–1 d–1). The response of leaf area development to this abrupt decrease in nitrate availability was characterized by following the expansion of the primary and secondary leaf pairs. The timing of the drop in nitrate supply was when cell division in the epidermis of the primary leaf pair was largely complete. Reducing the availability of nitrate had a strong effect on leaf area expansion. The final leaf size of the primary leaf pair was affected indicating an effect of nitrate availability on cell expansion. By the end of the experiment the secondary leaf pair was only one-third the area of that on control seedlings. The role of epidermal cell turgor pressure in this growth response was assessed by direct measurements with a miniature cell pressure probe. No reduction in cell turgor pressure following the decrease in nitrate availability was detected. It is concluded that a reduction in turgor pressure was not responsible for the reduction in leaf area expansion and it is suggested that reduced cell expansion was due to changes in cell wall properties. Concentrations of leaf and root abscisic acid increased following the reduction in nitrate availability.

Turgor Changes in Morchella esculenta during Translocation and Sclerotial Formation

Amir, R., Steudle, E., Levanon, D., Hadar, Y., and Chet, I. 1995. Turgor changes in Morchella esculenta during translocation and sclerotial formation. Experimental Mycology 19, 129-136. Turgor pressure was measured during six stages of growth and pseudosclerotial formation in Morchella esculenta indirectly (by thermocouple psychrometer) and directly (by cell pressure probe). The fungus was grown on a split plate, enabling separation between mycelium growing on defined medium (water potential -0.5 MPa) and sclerotia which formed on glucose noble agar (water potential -2.1 MPa). Under these conditions, nutrients were translocated from the mycelium to the developing sclerotia. Direct turgor potential measurements showed that the gradient between the mycelium and the sclerotia increases during sclerotial development (reaching a maximum of 0.53 MPa), thereby suggesting that translocation is a turgor-driven mass flow. During sclerotial development, the turgor potential in the peripheral tips of the sclerotial hyphae must be high enough to bring about the growth of the numerous hyphae, which comprise the sclerotium, and simultaneously low enough in the primary hyphae, which carry the stream of nutrients, to attract translocation from the mycelium. Since sclerotial hyphae are too small for direct measurement by cell pressure probe, a psychrometer was used, revealing high turgor in the sclerotial tissue (1.2 MPa) during selerotial development. Direct measurement in the primary hyphae at this time gave a value of 0.7 MPa. Taken together, these measurements indicate the presence of a turgor gradient inside the sclerotial tissue, from the primary hyphae to the peripheral cells. The present study is the first to make use of a cell pressure probe to measure turgor gradients in a fungus during translocation followed by sclerotial morphogenesis.

Stimulation and inhibition of pine root growth by osmotic stress

summaryThe effect of osmotic stress on the regulation of extension of roots of pine seedlings (Pinus pinaster Ait.) was studied. Extension rate (r) was measured as increase in root length, and as local growth rate (LGR) along the growing region. Turgor pressure (P) was measured over the same profile using the cell pressure probe. Correlation of LGR and P allowed changes in cell wall mechanical properties to be followed.In unstressed plants, differences in LGR along the root apex were owing to variations in the mechanical properties of the cell walls since P remained constant at 0.5 MPa along the first 10 mm of the root tip. Contrary to expectation, a moderate water stress of 0.15 MPa stimulated root growth despite a small reduction in P to 0.4 MPa. The walls of the expanding cells became looser, allowing enhanced growth in spite of reduced turgor. Examination of LGR showed that these changes occurred uniformly over the whole elongation zone. The advantage to the plant of this is discussed. Higher stresses (0.45 and 0.66 MPa) decreased cell turgor to a greater degree (≤0.3 MPa) and decreased growth rate. At the highest stress (0.66 MPa), P was maximal in cells that were still elongating, resulting in a longitudinal gradient in turgor. In both treatments LGR remained unchanged at the apex of the growing zone. Wall loosening appeared to have occurred in this region. Pine root extension is therefore modulated by changes in cell wall properties, with cell wall loosening resulting in increased growth at moderate stress. It appears that growth under higher stresses is limited by inability to maintain turgor.

Rapid Response of the Yield Threshold and Turgor Regulation during Adjustment of Root Growth to Water Stress in Zea mays.

Responses of cortical cell turgor (P) following rapid changes in osmotic pressure ([pi]m) were measured throughout the elongation zone of maize (Zea mays L.) roots using a cell pressure probe and compared with simultaneously measured root elongation to evaluate: yield threshold (Y) (minimum P for growth), wall extensibility, growth-zone radial hydraulic conductivity (K), and turgor recovery rate. Small increases in [pi]m (0.1 MPa) temporarily decreased P and growth, which recovered fully in 5 to 10 min. Under stronger [pi]m (up to 0.6 MPa), elongation stopped for up to 30 min and then resumed at lower rates. Recoveries in P through solute accumulation and lowering of Y enabled growth under water stress. P recovery was as much as 0.3 MPa at [pi]m = 0.6 MPa, but recovery rate declined as water stress increased, suggesting turgor-sensitive solute transport into the growth zone. Under strong [pi]m, P did not recover in the basal part of the growth zone, in conjunction with a 30% shortening of the growth zone. Time courses showed Y beginning to decrease within several minutes after stress imposition, from about 0.65 MPa to a minimum of about 0.3 MPa in about 15 min. The data concerning Y were not confounded significantly by elastic shrinkage. K was high (1.3 x 10-10 m2 s-1 MPa-1), suggesting very small growth-induced water potential gradients.

Hydraulic propagation of pressure along immature and mature xylem vessels of roots of Zea mays measured by pressure-probe techniques

Turgor pressure was measured in cortical cells and in xylem elements of excised roots and roots of intact plants of Zea mays L. by means of a cell pressure probe. Turgor of living and hence not fully differentiated late metaxylem (range 0.6–0.8 MPa) was consistently higher than turgor of cortical cells (range 0.4–0.6 MPa) at positions between 40 and 180 mm behind the root tip. Closer to the tip, no turgor difference between the cortex and the stele was measured. The turgor difference indicated that late-metaxylem elements may function as nutrient-storage compartments within the stele. Excised roots were attached to the root pressure probe to precisely manipulate the xylem water potential. Root excision did not affect turgor of cortical cells for at least 8 h. Using the cell pressure probe, the propagation of a hydrostatic pressure change effected by the root pressure probe was recorded in mature and immature xylem elements at various positions along the root. Within seconds, the pressure change propagated along both early and late metaxylems. The half-times of the kinetics, however, were about five times smaller for the early metaxylem, indicating they are likely the major pathway of longitudinal water flow. The hydraulic signal dissipated from the source of the pressure application (cut end of the root) to the tip of the root, presumably because of radial water movement along the root axis. The results demonstrate that the water status of the growth zone and other positions apical to 20 mm is mainly uncoupled from changes of the xylem water potential in the rest of the plant.

Effects of NaCl and CaCl2 on Water Transport across Root Cells of Maize (Zea mays L.) Seedlings

The effect of salinity and calcium levels on water flows and on hydraulic parameters of individual cortical cells of excised roots of young maize (Zea mays L. cv Halamish) plants have been measured using the cell pressure probe. Maize seedlings were grown in one-third strength Hoagland solution modified by additions of NaCl and/or extra calcium so that the seedlings received one of four treatments: control; +100 millimolar NaCl; +10 millimolar CaCl(2); +100 millimolar NaCl + 10 millimolar CaCl(2). From the hydrostatic and osmotic relaxations of turgor, the hydraulic conductivity (Lp) and the reflection coefficient (sigma(s)) of cortical cells of different root layers were determined. Mean Lp values in the different layers (first to third, fourth to sixth, seventh to ninth) of the four different treatments ranged from 11.8 to 14.5 (Control), 2.5 to 3.8 (+NaCl), 6.9 to 8.7 (+CaCl(2)), and 6.6 to 7.2 . 10(-7) meter per second per megapascal (+NaCl + CaCl(2)). These results indicate that salinization of the growth media at regular calcium levels (0.5 millimolar) decreased Lp significantly (three to six times). The addition of extra calcium (10 millimolar) to the salinized media produced compensating effects. Mean cell sigma(s) values of NaCl ranged from 1.08 to 1.16, 1.15 to 1.22, 0.94 to 1.00, and 1.32 to 1.46 in different root cell layers of the four different treatments, respectively. Some of these sigma(s) values were probably overestimated due to an underestimation of the elastic modulus of cells, sigma(s) values of close to unity were in line with the fact that root cell membranes were practically not permeable to NaCl. However, the root cylinder exhibited some permeability to NaCl as was demonstrated by the root pressure probe measurements that resulted in sigma(sr) of less than unity. Compared with the controls, salinity and calcium increased the root cell diameter. Salinized seedlings grown at regular calcium levels resulted in shorter cell length compared with control (by a factor of 2). The results demonstrate that NaCl has adverse effects on water transport parameters of root cells. Extra calcium could, in part, compensate for these effects. The data suggest a considerable apoplasmic water flow in the root cortex. However, the cell-to-cell path also contributed to the overall water transport in maize roots and appeared to be responsible for the decrease in root hydraulic conductivity reported earlier (Azaizeh H, Steudle E [1991] Plant Physiol 97: 1136-1145). Accordingly, the effect of high salinity on the cell Lp was much larger than that on root Lp(r).