Determination of the pressure in the water-conducting vessels of intactNicotiana rustica L. plants showed that the pressure probe technique gave less-negative values than the Scholander-bomb method. Even though absolute values of the order of -0.1 MPa could be directly recorded in the xylem by means of the pressure probe, pressures between zero and atmospheric were also frequently found. The data obtained by the pressure probe for excised leaves showed that the Scholander bomb apparently did not read the actual tension in the xylem vessles ofNicotiana plants. The possibility that the pressure probe gave false readings was excluded by several experimental controls. In addition, cavitation and leaks either during the insertion of the microcapillary of the pressure probe, or else during the measurements were easily recognized when they occurred because of the sudden increase of the absolute xylem tension to that of water vapour or to atmospheric, respectively. Tension values of the same order could also be measured by means of the pressure probe in the xylem vessels of pieces of stem cut from leaves and roots under water and clamped at both ends. The magnitude of the absolute tension depended on the osmolarity of the bathing solution which was adjusted by addition of appropriate concentrations of polyethylene glycol. Partial and uniform pressurisation of plant tissues or organs, or of entire plants (by means of the Scholander bomb or of a hyperbaric chamber, respectively) and simultaneous recording of the xylem tension using the pressure probe showed that a 1ratio1 response in xylem pressure only occurred under a few circumstances. A 1ratio1 response required that the xylem vessels were in direct contact with an external water reservoir and/or that the tissue was (pre-)infiltrated with water. Corresponding pressure-probe measurements in isolated vascular bundles ofPlantago major L. orP. lanceolata L. plants attached to a Hepp-type osmometer indicated that the magnitude of the tension in the xylem vessels was determined by the external osmotic pressure of the reservoir. These and other experiments, as well as analysis of the data using classical thermodynamics, indicated that the turgor and the internal osmotic pressure of the accessory cells along the xylem vessels play an important role in the maintenance of a constant xylem tension. This conclusion is consistent with the cohesion theory. In agreement with the literature (P.E. Weatherley, 1976, Philos. Trans. R. Soc. London Ser. B23, 435-444; 1982, Encyclopedia of plant physiology, vol. 12B, 79-109), it was found that the tension in the xylem of intact plants under normal and elevated ambient pressure (as measured with the pressure probe) under quasi-stationary conditions was independent of the transpiration rate over a large range, indicating that the conductance of the flow path must be flow-dependent.

Aquaporins are integral membrane proteins of the tonoplast and the plasma membrane that facilitate the passage of water through these membranes. Because of their potentially important role in regulating water flow in plants, studies documenting aquaporin gene expression in specialized tissues involved in water and solute transport are important. We used in situ hybridization to examine the expression pattern of the tonoplast aquaporin ZmTIP1 in different organs of maize (Zea mays L.). This tonoplast water channel is highly expressed in the root epidermis, the root endodermis, the small parenchyma cells surrounding mature xylem vessels in the root and the stem, phloem companion cells and a ring of cells around the phloem strand in the stem and the leaf sheath, and the basal endosperm transfer cells in developing kernels. We postulate that the high level of expression of ZmTIP1 in these tissues facilitates rapid flow of water through the tonoplast to permit osmotic equilibration between the cytosol and the vacuolar content, and to permit rapid transcellular water flow through living cells when required.

A critique of Martin Canny's theory of water transport supported by tissue pressure is given with reference to basic principles of cellular water relations and biomechanics. It is shown that the application of tissue pressure in Canny's theory is neither internally consistent nor compatible with basic biophysics. Canny's translation of tissue pressure into an altered steady-state pressure in the xylem conduits has no defensible mechanism, relying instead on untenable action-at-a-distance and poor definitions. Tissue pressure itself, as defined by Canny and illustrated by the example of a turgid leaf, may well exist. However, it cannot function in whole-plant processes as envisioned by Canny, nor can it exist in the magnitude his theory would require. Rigid outer tissue layers containing internal pressures of the magnitude postulated by Canny would require a tensile strength quite incompatible with the observed biological materials. A simple application of La Place's Law illustrates that this is an issue of scale and that the turgor generated by osmotic potentials must be balanced primarily at the cellular level, and not the tissue level.

It is supposed that oscillations in stomatal conductance are associated with the dynamic properties of the loop in which rate of evaporation affects, through physiological processes, the aperture of stomata and stomatal aperture in turn affects rate of evaporation. It is therefore predicted that their occurrence must be influenced by the magnitude of what is termed environmental gain: the sensitivity of rate of evaporation to change in leaf conductance to vapor transfer. Two methods of manipulating gain, and their effects on stomatal behavior in cotton (Gossypium hirsutum L. cv. Deltapine Smooth Leaf), are described. In the first, gain was increased by decreasing ambient humidity; in the second, it was made zero by regulating ambient humidity to keep rate of evaporation constant despite changes in conductance. The results are in accord with the supposition.

Trees grow tall where resources are abundant, stresses are minor, and competition for light places a premium on height growth. The height to which trees can grow and the biophysical determinants of maximum height are poorly understood. Some models predict heights of up to 120 m in the absence of mechanical damage, but there are historical accounts of taller trees. Current hypotheses of height limitation focus on increasing water transport constraints in taller trees and the resulting reductions in leaf photosynthesis. We studied redwoods (Sequoia sempervirens), including the tallest known tree on Earth (112.7 m), in wet temperate forests of northern California. Our regression analyses of height gradients in leaf functional characteristics estimate a maximum tree height of 122-130 m barring mechanical damage, similar to the tallest recorded trees of the past. As trees grow taller, increasing leaf water stress due to gravity and path length resistance may ultimately limit leaf expansion and photosynthesis for further height growth, even with ample soil moisture.

An apparatus has been constructed for the detection of vibrations generated within plant tissue. This can detect vibrations due to water cavitation within fern sporangia. Using this detector 'clicks' have been obtained from Ricinus petioles and leaves in a condition when, from previous work, water cavitation might be expected.Evidence is provided to show: 1. That 'click' production is correlated with the water status of the plant tissue. 2. There are reasons for opposing the possibility that 'clicks' result from tissue fracture alone. 3. The 'clicks' may be detected in a wide range of plants. The implications of the apparent vulnerability of water conducting systems in plants are discussed.

The osmotic pressure of the cell sap of stalk storage parenchyma of sugarcane (Saccharum spp. hybrids) increases by an order of magnitude during ontogeny to reach molar concentrations of sucrose at maturity. Stalk parenchyma cells must either experience very high turgor at maturation or have an ability to regulate turgor. We tested this hypothesis by using pressure probe techniques to quantify parameters of cell and tissue water relations of sugarcane storage parenchyma during ontogeny. The largest developmental change was in the volumetric elastic modulus, which increased from 6 bars in immature tissue to 43 bars in mature tissue. Turgor was maintained relatively low during sucrose accumulation by the partitioning of solutes between the cell and wall compartments. Membrane hydraulic conductivity decreased from about 12 x 10(-7) centimeters per second per bar down to 4.4 x 10(-7) centimeters per second per bar. The 2.7-fold decrease in membrane hydraulic conductivity during tissue maturation was accompanied by a 7.8-fold increase in wall elasticity. Integration of the cell wall and membrane properties appears to be by the opposing effects of turgor on hydraulic conductivity and elastic modulus. The changes in these properties during development of sugarcane stalk tissue may be a way for parenchyma cells to develop a capacity for expansive growth and still serve as a strong sink for storing high concentrations of sucrose.

We proposed the hydraulic limitation hypothesis (HLH) as a mechanism to explain universal patterns in tree height, and tree and stand biomass growth: height growth slows down as trees grow taller, maximum height is lower for trees of the same species on resource-poor sites and annual wood production declines after canopy closure for even-aged forests. Our review of 51 studies that measured one or more of the components necessary for testing the hypothesis showed that taller trees differ physiologically from shorter, younger trees. Stomatal conductance to water vapour (g(s)), photosynthesis (A) and leaf-specific hydraulic conductance (K L) are often, but not always, lower in taller trees. Additionally, leaf mass per area is often greater in taller trees, and leaf area:sapwood area ratio changes with tree height. We conclude that hydraulic limitation of gas exchange with increasing tree size is common, but not universal. Where hydraulic limitations to A do occur, no evidence supports the original expectation that hydraulic limitation of carbon assimilation is sufficient to explain observed declines in wood production. Any limit to height or height growth does not appear to be related to the so-called age-related decline in wood production of forests after canopy closure. Future work on this problem should explicitly link leaf or canopy gas exchange with tree and stand growth, and consider a more fundamental assumption: whether tree biomass growth is limited by carbon availability.

In perennial plants, freeze-thaw cycles during the winter months can induce the formation of air bubbles in xylem vessels, leading to changes in their hydraulic conductivity. Refilling of embolized xylem vessels requires an osmotic force that is created by the accumulation of soluble sugars in the vessels. Low water potential leads to water movement from the parenchyma cells into the xylem vessels. The water flux gives rise to a positive pressure essential for the recovery of xylem hydraulic conductivity. We investigated the possible role of plasma membrane aquaporins in winter embolism recovery in walnut (Juglans regia). First, we established that xylem parenchyma starch is converted to sucrose in the winter months. Then, from a xylem-derived cDNA library, we isolated two PIP2 aquaporin genes (JrPIP2,1 and JrPIP2,2) that encode nearly identical proteins. The water channel activity of the JrPIP2,1 protein was demonstrated by its expression in Xenopus laevis oocytes. The expression of the two PIP2 isoforms was investigated throughout the autumn-winter period. In the winter period, high levels of PIP2 mRNA and corresponding protein occurred simultaneously with the rise in sucrose. Furthermore, immunolocalization studies in the winter period show that PIP2 aquaporins were mainly localized in vessel-associated cells, which play a major role in controlling solute flux between parenchyma cells and xylem vessels. Taken together, our data suggest that PIP2 aquaporins could play a role in water transport between xylem parenchyma cells and embolized vessels.

A method is described which permits measurement of sap pressure in the xylem of vascular plants. As long predicted, sap pressures during transpiration are normally negative, ranging from -4 or -5 atmospheres in a damp forest to -80 atmospheres in the desert. Mangroves and other halophytes maintain at all times a sap pressure of -35 to -60 atmospheres. Mistletoes have greater suction than their hosts, usually by 10 to 20 atmospheres. Diurnal cycles of 10 to 20 atmospheres are common. In tall conifers there is a hydrostatic pressure gradient that closely corresponds to the height and seems surprisingly little influenced by the intensity of transpiration. Sap extruded from the xylem by gas pressure on the leaves is practically pure water. At zero turgor this procedure gives a linear relation between the intracellular concentration and the tension of the xylem.

10 MPa) air-seed pressure primarily because of decreasing pit membrane conductivity. Vessel conductivity (per length and wall area) increased with vessel length as higher lumen conductivity overcame low pit conductivity. At the]]>

3 megapascals. This same pressure difference was found to be sufficient to force air across intervessel pits from air injection experiments of hydrated stem segments. This suggests air entry at pits is causing embolism in dehydrating stems. (b) Treatments that increased the permeability of intervessel pits to air injection also caused xylem to embolize at less negative xylem pressures. Permeability was increased either by perfusing stems with solutions of surface tension below that of water or by perfusion with a solution of oxalic acid and calcium. The mechanism of oxalic-calcium action on permeability is unknown, but may relate to the ability of oxalate to chelate calcium from the pectate fraction of the pit membrane. (c) Diameter of pores in pit membranes measured with the scanning electron microscope were within the range predicted by hypothesis (

Ultrasonic acoustic emissions (AE) in the frequency range of 0.1 to 1 megahertz appear to originate in the sapwood of Thuja occidentalis L. The AE are vibrations of an impulsive nature. The vibrations can be transduced to a voltage waveform and amplified. The vibrations of each AE event begin at a large amplitude which decays over 20 to 100 microseconds. Strong circumstantial evidence indicates that the ultrasonic AE result from cavitation events because: (a) they occur only when the xylem pressure potential Psi(xp) is more negative than a threshold level of about -1 megapascal; (b) the rate of AE events increases as Psi(xp) decreases and when the net rate of water loss increases; (c) the AE can be stopped by raising Psi(xp) above -1 megapascal. Ultrasonic AE have been measured in whole terminal shoots allowed to dry in the laboratory, in isolated pieces of sapwood as they dried in the laboratory, and in whole terminal shoots in a pressure bomb when Psi(xp) was decreased by lowering the gas pressure in the pressure bomb.

Recovery of hydraulic conductivity after the induction of embolisms was studied in woody stems of laurel (Laurus nobilis). Previous experiments confirming the recovery of hydraulic conductivity when xylem pressure potential was less than -1 MPa were repeated, and new experiments were done to investigate the changes in solute composition in xylem vessels during refilling. Xylem sap collected by perfusion of excised stem segments showed elevated levels of several ions during refilling. Stem segments were frozen in liquid N2 to view refilling vessels using cryoscanning electron microscopy. Vessels could be found in all three states of presumed refilling: (a) mostly water with a little air, (b) mostly air with a little water, or (c) water droplets extruding from vessel pits adjacent to living cells. Radiographic probe microanalysis of refilling vessels revealed nondetectable levels of dissolved solutes. Results are discussed in terms of proposed mechanisms of refilling in vessels while surrounding vessels were at a xylem pressure potential of less than -1 MPa. We have concluded that none of the existing paradigms explains the results.

Hydraulic properties (half-time of water exchange, T1/2, and hydraulic conductivity, Lp; T1/2 approximately 1/Lp) of individual cells in the cortex of young corn roots were measured using a cell pressure probe for up to 6 h to avoid variations between cells. When pulses of turgor pressure of different size were imposed, T1/2 (Lp) responded differently depending on the size. Pulses of smaller than 0.1 MPa, which induced a small proportional water flow, caused no changes in T1/2 (Lp). Medium-sized pulses of between 0.1 and 0.2 MPa caused an increase in T1/2 (decrease in Lp) by a factor of 4 to 23. The effects caused by medium-sized pulses were reversible within 5-20 min. When larger pulses of more than 0.2 MPa were employed, changes were not reversible within 1-3 h, but could be reversed within 30 min in the presence of 500 nM of the stress hormone ABA. Cells with a short T1/2 responded to the aquaporin blocker mercuric chloride (HgCl2). The treatment had no effect on cells which exhibited long T1/2 following a mechanical inhibition by the large-pulse treatment. Step decreases in pressure resulted in the same inhibition as step increases. Hence, the treatment did not cause a stretch-inhibition of water channels and was independent of the directions of both pressure changes and water flows induced by them. It is concluded that inhibition is caused by the absolute value of intensities of water flow within the channels, which increased in proportion to the size of step changes in pressure. Probable mechanisms by which the mechanical stimuli are perceived are (i) the input of kinetic energy to the channel constriction (NPA motif of aquaporin) which may cause a conformational change of the channel protein (energy-input model) or (ii) the creation of tensions at the constriction analogous to Bernoulli's principle for macroscopic pores (cohesion-tension model). Estimated rates of water flow within the pores were a few hundred micro m s-1, which is too small to create sufficient tension. They were much smaller than those proposed for AQP1. Based on literature data of single-channel permeability of AQP1, a per channel energy input of 200 kBxT (kB=Boltzmann constant) was estimated for the energy-input model. This should be sufficient to initiate changes of protein conformation and an inactivation of channels. The data indicate different closed states which differ in the amount of distortion and the rates at which they relax back to the open state.

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.

After Renner had shown convincingly in 1925 that the transpirational water loss generates tensions larger than 0.1 MPa (i.e. negative pressures) in the xylem of cut leafy twigs the Cohesion Theory proposed by Bohm, Askenasy, Dixon and Joly at the end of the 19th century was immediately accepted by plant physiologists. Introduction of the pressure chamber technique by Scholander et al. in 1965 enforced the general belief that tension is the only driving force for water lifting although substantial criticism regarding the technique and/or the Cohesion Theory was published by several authors. As typical for scientific disciplines, the advent of minimal- and non-invasive techniques in the last decade as well as the development of a new, reliable method for xylem sap sampling have challenged this view. Today, xylem pressure gradients, potentials, ion concentrations and volume flows as well as cell turgor pressure gradients can be monitored online in intact transpiring higher plants, and within a given physiological context by using the pressure probe technique and high-resolution NMR imaging techniques, respectively. Application of the pressure probe technique to transpiring plants has shown that negative absolute pressures (down to - 0.6 MPa) and pressure gradients can exist temporarily in the xylem conduit, but that the magnitude and (occasionally) direction of gradients contrasts frequently the belief that tension is the only driving force. This seems to be particularly the case for plants faced with problems of height, drought, freezing and salinity as well as with cavitation of the tensile water. Reviewing the current data base shows that other forces come into operation when exclusively tension fails to lift water against gravity due to environmental conditions. Possible candidates are longitudinal cellular and xylem osmotic pressure gradients, axial potential gradients in the vessels as well as gel- and gas bubble-supported interfacial gradients. The multiforce theory overcomes the problem of the Cohesion Theory that life on earth depends on water being in a highly metastable state.