A revised and unified pressure-clamp/relaxation theory for studying plant cell water relations with pressure probes: in-situ determination of cell volume for calculation of volumetric elastic modulus and hydraulic conductivity.

The cell-pressure-probe is a unique tool to study plant water relations in-situ. Inaccuracy in the estimation of cell volume (νo) is the major source of error in the calculation of both cell volumetric elastic modulus (ε) and cell hydraulic conductivity (Lp). Estimates of νo and Lp can be obtained with the pressure-clamp (PC) and pressure-relaxation (PR) methods. In theory, both methods should result in comparable νo and Lp estimates, but this has not been the case. In this study, the existing νo-theories for PC and PR methods were reviewed and clarified. A revised νo-theory was developed that is equally valid for the PC and PR methods. The revised theory was used to determine νo for two extreme scenarios of solute mixing between the experimental cell and sap in the pressure probe microcapillary. Using a fully automated cell-pressure-probe (ACPP) on leaf epidermal cells of Tradescantia virginiana, the validity of the revised theory was tested with experimental data. Calculated νo values from both methods were in the range of optically determined νo (=1.1-5.0nL) for T. virginiana. However, the PC method produced a systematically lower (21%) calculated νo compared to the PR method. Effects of solute mixing could only explain a potential error in calculated νo of <3%. For both methods, this discrepancy in νo was almost identical to the discrepancy in the measured ratio of ΔV/ΔP (total change in microcapillary sap volume versus corresponding change in cell turgor) of 19%, which is a fundamental parameter in calculating νo. It followed from the revised theory that the ratio of ΔV/ΔP was inversely related to the solute reflection coefficient. This highlighted that treating the experimental cell as an ideal osmometer in both methods is potentially not correct. Effects of non-ideal osmotic behavior by transmembrane solute movement may be minimized in the PR as compared to the PC method.

High apoplastic solute concentrations in leaves alter water relations of the halophytic shrub, Sarcobatus vermiculatus.

Predawn plant water potential (Psi(w)) is used to estimate soil moisture available to plants because plants are expected to equilibrate with the root-zone Psi(w). Although this equilibrium assumption provides the basis for interpreting many physiological and ecological parameters, much work suggests predawn plant Psi(w) is often more negative than root-zone soil Psi(w). For many halophytes even when soils are well-watered and night-time shoot and root water loss eliminated, predawn disequilibrium (PDD) between leaf and soil Psi(w) can exceed 0.5 MPa. A model halophyte, Sarcobatus vermiculatus, was used to test the predictions that low predawn solute potential (Psi(s)) in the leaf apoplast is a major mechanism driving PDD and that low Psi(s) is due to high Na+ and K+ concentrations in the leaf apoplast. Measurements of leaf cell turgor (Psi(p)) and solute potential (Psi(s)) of plants grown under a range of soil salinities demonstrated that predawn symplast Psi(w) was 1.7 to 2.1 MPa more negative than predawn xylem Psi(w), indicating a significant negative apoplastic Psi(s). Measurements on isolated apoplastic fluid indicated that Na+ concentrations in the leaf apoplast ranged from 80 to 230 mM, depending on salinity, while apoplastic K+ remained around 50 mM. The water relations measurements suggest that without a low apoplastic Psi(s), predawn Psi(p) may reach pressures that could cause cell damage. It is proposed that low predawn apoplastic Psi(s) may be an efficient way to regulate Psi(p) in plants that accumulate high concentrations of osmotica or when plants are subject to fluctuating patterns of soil water availability.

Seed coat cell turgor in chickpea is independent of changes in plant and pod water potential.

Turgor pressure in cells of the pod wall and the seed coat of chickpea (Cicer arietinum L.) were measured directly with a pressure probe on intact plants under initially dry soil conditions, and after the plants were irrigated. The turgor pressure in cells of the pod wall was initially 0.25 MPa, and began to increase within a few minutes of irrigation. By 2-4 h after irrigation, pod wall cell turgor had increased to 0.97 MPa. This increase in turgor was matched closely by increases in the total water potential of both the pod and the stem, as measured by a pressure chamber. However, turgor pressure in cells of the seed coat was relatively low (0.10 MPa) and was essentially unchanged up to 24 h after irrigation (0.13 MPa). These data demonstrate that water exchange is relatively efficient throughout most of the plant body, but not between the pod and the seed. Since both the pod and the seed coat are vascularized tissues of maternal origin, this indicates that at least for chickpea, isolation of the water relations of the embryo from the maternal plant does not depend on the absence of vascular or symplastic connections between the embryo and the maternal plant.

Influence of stem diameter, water content, and freezing-thawing on bacterial canker development in excised stems of dormant stone fruit.

ABSTRACT In excised dormant stems of peach (Prunus persica), prune (Prunus domestica), and almond (Prunus dulcis), stem diameter, stem hydration, and freezing-thawing influenced the extent of infection caused by Pseudomonas syringae pv. syringae. Bacterial lesion length increased with increasing stem diameter, demonstrating the need to account for the effects of stem diameter when lesion length data are analyzed. Lesion length increased or decreased with stem hydration or dehydration, respectively. However, tissue water content was not a good indicator of tissue susceptibility to infection by P. syringae pv. syringae, as larger diameter stems had larger lesions and lower water content than did smaller diameter stems. After freezing at -5 degrees C for 12 to 24 h, inoculations made during the thawing process produced significantly larger lesions than did inoculations performed before freezing or after thawing. These results support the hypothesis that the increased susceptibility to bacterial canker that is associated with noninjurious freezing is a result of the increased passive spread of bacteria through water redistribution when inoculation is performed during the thawing process. Plant tissue water relationship characteristics that can influence water movement during freezing and thawing may be an important component of bacterial canker development in stone fruit trees.

Maintenance of turgor by rapid sealing of puncture wounds in leaf epidermal cells.

When leaf epidermal cells are puncture wounded with a glass microcapillary tip, a small droplet of fluid is discharged and then evaporates, leaving a solid residue on the cell surface. For puncture wounds of about 3.5 micrometers in diameter, this process is complete within 2 to 3 seconds. A second puncture wound also exhibits a similar discharge, indicating the persistence of some turgor pressure within the cell, despite damage to the cell wall. Direct measurement of turgor on the large epidermal cells of Tradescantia virginiana L. demonstrated that turgor was substantially maintained (91-96%) after puncture wounding. Anatomical and histochemical evidence suggests that the damaged portion of the cell wall was sealed with an amorphous plug of material comprised of pectinaceous polysaccharides. Rapid sealing of puncture wounds and the maintenance of turgor in epidermal cells may be an important functional component of plant adaptation to physical damage such as that caused by insect feeding.

Cell turgor changes associated with ripening in tomato pericarp tissue.

The pressure microprobe was used to determine whether the turgor pressure in tomato (Lycopersicon esculentum Mill., variety "Castelmart") pericarp cells changed during fruit ripening. The turgor pressure of cells located 200 to 500 micrometers below the fruit epidermis was uniform within the same tissue (typically +/- 0.02 megapascals), and the highest turgors observed (<0.2 megapascals) were much less than expected, based on tissue osmotic potential (-0.6 to -0.7 megapascals). These low turgor values may indicate the presence of apoplastic solutes. In both intact fruit and cultured discs of pericarp tissue, a small increase in turgor preceded the onset of ripening, and a decrease in turgor occurred during ripening. Differences in the turgor of individual intact fruit occurred 2 to 4 days before parallel differences in their ripening behavior were apparent, indicating that changes in turgor may reflect physiological changes at the cell level that precede expression of ripening at the tissue level.

Dynamic Relation between Expansion and Cellular Turgor in Growing Grape (Vitis vinifera L.) Leaves.

Measurements of the growth and water relations of expanding grape (Vitis vinifera L.) leaves have been used to determine the relationship between leaf expansion rate and leaf cell turgor. Direct measurement of turgor on the small (approximately 15 micrometer diameter) epidermal cells over the midvein of expanding grape leaves was made possible by improvements in the pressure probe technique. Leaf expansion rate and leaf water status were perturbed by environmentally induced changes in plant transpiration. After establishing a steady state growth rate, a step decrease in plant transpiration resulted in a rapid and large increase in leaf cell turgor (0.25 megapascal in 5 minutes), and leaf expansion rate. Subsequently, leaf expansion rate returned to the original steady state rate with no change in cell turgor. These results indicate that the expansion rate of leaves may not be strongly related to the turgor of the leaf cells, and that substantial control of leaf expansion rate, despite changes in turgor, may be part of normal plant function. It is suggested that a strictly physical interpretation of the parameters most commonly used to describe the relationship between turgor and growth in plant cells (cell wall extensibility and yield threshold) may be inappropriate when considering the process of plant cell expansion.

Direct measurement of turgor and osmotic potential in individual epidermal cells : independent confirmation of leaf water potential as determined by in situ psychrometry.

The pressure probe, which is routinely used to measure the turgor potential (Psi(p)) of individual epidermal cells in Tradescantia virginiana (L.), has also been used to sample small volumes of vacuolar fluid from these same cells (as low as 0.02 nl) for measurement of cellular solute (osmotic) potential (Psi(s)) in a micro freezing point osmometer. The water potential components Psi(p) and Psi(o) have been used to calculate the total water potential of individual epidermal cells (Psi(cell)) which has then been directly compared to the total leaf water potential (Psi(leaf)) measured psychrometrically. The relation of Psi(leaf) and Psi(cell) to leaf transpiration indicates that in T. virginiana, a relatively straightforward relation exists between the level of water flow through the leaf tissue, and the DeltaPsi within the leaf, between two points along the water flow pathway. Substantial agreement was found between the two independent, in situ methods of measuring Psi when extrapolated to zero transpiration conditions. These results are discussed with respect to the thermodynamics of water transport in plant tissues.

In Situ Measurement of Epidermal Cell Turgor, Leaf Water Potential, and Gas Exchange in Tradescantia virginiana L.

A combined system has been developed in which epidermal cell turgor, leaf water potential, and gas exchange were determined for transpiring leaves of Tradescantia virginiana L. Uniform and stable values of turgor were observed in epidermal cells (stomatal complex cells were not studied) under stable environmental conditions for both upper and lower epidermises. The changes in epidermal cell turgor that were associated with changes in leaf transpiration were larger than the changes in leaf water potential, indicating the presence of transpirationally induced within-leaf water potential gradients. Estimates of 3 to 5 millimoles per square meter per second per megapascal were obtained for the value of within-leaf hydraulic conductivity. Step changes in atmospheric humidity caused rapid changes in epidermal cell turgor with little or no initial change in stomatal conductance, indicating little direct relation between stomatal humidity response and epidermal water status. The significance of within-leaf water potential gradients to measurements of plant water potential and to current hypotheses regarding stomatal response to humidity is discussed.

Theoretical and experimental errors for in situ measurements of plant water potential.

Errors in psychrometrically determined values of leaf water potential caused by tissue resistance to water vapor exchange and by lack of thermal equilibrium were evaluated using commercial in situ psychrometers (Wescor Inc., Logan, UT) on leaves of Tradescantia virginiana (L.). Theoretical errors in the dewpoint method of operation for these sensors were demonstrated. After correction for these errors, in situ measurements of leaf water potential indicated substantial errors caused by tissue resistance to water vapor exchange (4 to 6% reduction in apparent water potential per second of cooling time used) resulting from humidity depletions in the psychrometer chamber during the Peltier condensation process. These errors were avoided by use of a modified procedure for dewpoint measurement. Large changes in apparent water potential were caused by leaf and psychrometer exposure to moderate levels of irradiance. These changes were correlated with relatively small shifts in psychrometer zero offsets (-0.6 to -1.0 megapascals per microvolt), indicating substantial errors caused by nonisothermal conditions between the leaf and the psychrometer. Explicit correction for these errors is not possible with the current psychrometer design.