Callose deposition at plasmodesmata

Summary The transport of ions and metabolites through plasmodesmata has been thought to be controlled at the neck region where the cytoplasmic annulus is constricted and where callose has also been localised. In order to determine the possible structural and functional effects of callose, its deposition was inhibited through incubation of the plant tissue with 2-deoxy-D-glucose (DDG) for 1 h prior to fixation in 2.5% glutaraldehyde. The inhibition of callose formation was monitored through aniline blue-induced fluorescence of callose. The neck region of the plasmodesmata fromAllium cepa L. roots treated with DDG exhibited a funnel-shaped configuration. This is in contrast to the plasmodesmata from tissue not incubated with DDG, which exhibited constricted necks similar to those previously reported. Both initial dissection and glutaraldehyde fixation induced neck constriction in plasmodesmata, however, dissection of tissue increased the frequency of constrictions. The inhibition of callose formation by chemical means showed that the neck constrictions and raised collars in this area are artefacts due to physical wounding and glutaraldehyde fixation. The external electron-dense material observed when tannic acid is included in the primary fixative appears to be unrelated to the deposition of callose at the neck region.Abbreviations DDG 2-deoxy-D-glucose     

Coupling of Solute and Solvent Flows in Porous Lipid Bilayer Membranes

The present experiments were designed to evaluate coupling of water and nonelectrolyte flows in porous lipid bilayer membranes (i.e., in the presence of amphotericin B) in series with unstirred layers. Alterations in solute flux during osmosis, with respect to the flux in the absence of net water flow, could be related to two factors: first, changes in the diffusional component of solute flux referable to variations in solute concentrations at the membrane interfaces produced by osmotic flow through the unstirred layers; and second, coupling of solute and solvent flows within the membrane phase. Osmotic water flow in the same direction as solute flow increased substantially the net fluxes of glycerol and erythritol through the membranes, while osmotic flow in the opposite direction to glycerol flow reduced the net flux of that solute. The observed effects of osmotic water flow on the fluxes of these solutes were in reasonable agreement with predictions based on a model for coupling of solute and solvent flows within the membrane phase, and considerably in excess of the prediction for a diffusion process alone.     

Osmotic water flow in leaky epithelia

I review three currently unsolved and controversial problems in understanding solute-linked water transport in epithelia. 1. Values of osmotic water permeability (Posm) calculated from steady-state osmotic flow in response to a gradient of a probe molecule tend to be underestimates, because of three unstirred-layer (USL) effects. These are: dissipation of the probe's gradient by diffusion in USL's; reduction of the probe's gradient, due to the sweeping-away effect of water flow generated by the probe itself; and solute polarization (creation of an opposing gradient of an initially symmetrically distributed solute by the sweeping-away effect). These errors increase with probe permeability, USL thickness, Posm, and concentration ratio of symmetrically distributed solute to probe, and vary inversely as the fractional area available for water flow (e.g., lateral intercellular space width). The form of an osmotic transient, and the possibility of extracting a true Posm value from the transient, depend on the relative values of three time constants: those for solute diffusion in USL's, for solute polarization by water flow in USL's and for measuring water flow. Sweeping-away effects cause major underestimates (by one or more orders of magnitude) in epithelial Posm determinations, as shown by apparent streaming potentials during osmotic flow and by transiently reversed flows after removal of the proble. True Posm values for leaky epithelia probably exceed 10(-3) or 10(-2) cm/sec.osm. The necessary conditions for resolving osmotic transients are set out. 2. I illustrate the difficulties in deciding what fraction of transepithelial water flow is via the cells, and what fraction via the junctions. There is no existing method for answering this question. 3. Controversies about the validity, or need for modification, of the standing-gradient theory are discussed. Progress in this field requires new methods: to resolve osmotic transients; to separate transcellular and transjunctional water flows; and to measure solute concentrations in lateral intercellular spaces directly.     

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 theory of cell hydration governed by adsorption of water on cell proteins rather than by osmotic pressure

It is postulated that cell hydration is governed by adsorption of water on cell proteins in accord with the Bradley adsorption isotherm, and that the action of a solute in the surrounding solution is to lower the vapor pressure of the solution so that cell water adsorption is decreased by moving down the Bradley isotherm. From these concepts, it is derived that cell volume (V) should be related to solute concentration (x) by the equationV=−E log₁₀ x+F whereE andF are constants which are independent of type of solute. For a non-adsorbed solute this agrees well with experimental data. For solutes which are adsorbed by cell proteins, a correction in the above equation may be necessary at higher solute concentrations, which is shown to be compatible with various experimental data. The types of experiments which are generally used to support the osmotic pressure theory of cell hydration agree equally well with the adsorption theory. The virtue of the adsorption theory is that, unlike the osmotic pressure theory of cell swelling, it is compatible with permeability of the cell membrane to solutes, which has been experimentally observed for various solutes. The opinions and conclusions contained in this report are those of the author. They are not to be construed as necessarily reflecting the views or the endorsement of the Navy Department.     

Diurnal and Seasonal Changes in Leaf Water Potential Components and Elastic Properties in Response to Water Stress in Apple Trees

Apple trees are very drought tolerant, having the capability to grow and carry on photosynthesis even at low water potentials. Much of the tolerance is due to the ability of apple leaves to maintain turgor potentials at levels conducive to growth and stomatal opening. Diurnally, leaf turgor is maintained through decreases in osmotic potentials (due to active solute accumulation), osmotic adjustment, or to concentration of solutes via tissue water loss. These two processes combined may decrease osmotic potentials by as much as 1.65 MPn during the day. Seasonally, osmotic potentials remain fairly constant, but leaf elasticity increases, allowing growth to continue and stomata to remain open us water and turgor potentials become progressively lower. Release of stored water from plant tissues to the transpiration stream is another means of preventing water potentials from reaching critical values for stomatal closure. A combination of a number of these physiological adaptations may account for much of the drought tolerance in apple trees.     

The control of single-celled cotton fiber elongation by developmentally reversible gating of plasmodesmata and coordinated expression of sucrose and K+ transporters and expansin

Each cotton fiber is a single cell that elongates to 2.5 to 3.0 cm from the seed coat epidermis within approximately 16 days after anthesis (DAA). To elucidate the mechanisms controlling this rapid elongation, we studied the gating of fiber plasmodesmata and the expression of the cell wall-loosening gene expansin and plasma membrane transporters for sucrose and K(+), the major osmotic solutes imported into fibers. Confocal imaging of the membrane-impermeant fluorescent solute carboxyfluorescein (CF) revealed that the fiber plasmodesmata were initially permeable to CF (0 to 9 DAA), but closed at approximately 10 DAA and re-opened at 16 DAA. A developmental switch from simple to branched plasmodesmata was also observed in fibers at 10 DAA. Coincident with the transient closure of the plasmodesmata, the sucrose and K(+) transporter genes were expressed maximally in fibers at 10 DAA with sucrose transporter proteins predominately localized at the fiber base. Consequently, fiber osmotic and turgor potentials were elevated, driving the rapid phase of elongation. The level of expansin mRNA, however, was high at the early phase of elongation (6 to 8 DAA) and decreased rapidly afterwards. The fiber turgor was similar to the underlying seed coat cells at 6 to 10 DAA and after 16 DAA. These results suggest that fiber elongation is initially achieved largely by cell wall loosening and finally terminated by increased wall rigidity and loss of higher turgor. To our knowledge, this study provides an unprecedented demonstration that the gating of plasmodesmata in a given cell is developmentally reversible and is coordinated with the expression of solute transporters and the cell wall-loosening gene. This integration of plasmodesmatal gating and gene expression appears to control fiber cell elongation.     

The role of the lateral intercellular spaces and solute polarization effects in the passive flow of water across the rabbit gallbladder

Summary Osmotic water flows were measured acrossin vitro preparations of the rabbit gallbladder by a gravimetric technique. The bladders exhibited asymmetrical osmotic behavior, in which theL _p (hydraulic conductivity) for water flow from mucosa to serosa was up to four times greater than theL _p for water flow in the opposite direction. This result is similar to the effects of osmotic gradients on ion and nonelectrolyte permeability reported in the first paper. As in the case of solute permeability, these changes inL _p are accounted for by changes in the dimensions of the lateral intercellular spaces of the epithelium. These spaces are thus a final common pathway for the movement of both solutes and water across the epithelium. We also observed osmotic flow transients in which the initialL _p was about an order of magnitude greater than the steady stateL _p . These transients are largely explained by solute polarization in the unstirred layers adjacent to the epithelial membranes. A comparison between streaming potentials and water flows showed that streaming potentials are directly proportional to the rate of flow only over a limited range. These observations are readily explained on the basis of structural changes and solute polarization effects. Finally, the routes of water flow across epithelia are discussed in the light of our observations.     

Water Flow Across the Sieve Tube Boundary: Estimating Turgor and Some Implications for Phloem Loading and Unloading. IV. Root Tips and Seed Coats

In the present paper, the theory developed in Part I of thisseries is applied to seed coats of Phaseolus vulgaris and somecombined data on root tips of Hordeum distichum and Hordeumvulgare. Because of the large back-pressures implied, it isconcluded that phloem transport into these primary sinks wouldbe physiologically impossible in the absence of a symplasticpathway for the unloading of water from sieve elements. In thiscase, unloading of water and sucrose will occur predominantlyas a pressure-driven flow of solution through plasmodesmata,although diffusion can contribute significantly to the plasmodesmatalsucrose flux. At least 20% of the plasmodesmata connecting sieveelements and adjacent cells must be unobstructed if large changesin turgor and osmotic pressure are to be avoided. Dependingon the membrane area available for water fluxes, it is possiblethat the difference in water potential across the sieve-tubeplasmalemma can lead to significant errors when axial turgorgradients are estimated from gradients of osmotic pressure andexternal water potential. The magnitude and even the sign ofthese errors is uncertain, but it is possible that sieve-tubeturgor pressures will be significantly underestimated in primarysinks Phloem, turgor, osmotic pressure, plasmodesmata, Munch hypothesis, Phloem unloading     

Growth of the Maize Primary Root at Low Water Potentials : II. Role of Growth and Deposition of Hexose and Potassium in Osmotic Adjustment

Primary roots of maize (Zea mays L. cv WF9 × Mo17) seedlings growing in vermiculite at various water potentials exhibited substantial osmotic adjustment in the growing region. We have assessed quantitatively whether the osmotic adjustment was attributable to increased net solute deposition rates or to slower rates of water deposition associated with reduced volume expansion. Spatial distributions of total osmotica, soluble carbohydrates, potassium, and water were combined with published growth velocity distributions to calculate deposition rate profiles using the continuity equation. Low water potentials had no effect on the rate of total osmoticum deposition per unit length close to the apex, and caused decreased deposition rates in basal regions. However, rates of water deposition decreased more than osmoticum deposition. Consequently, osmoticum deposition rates per unit water volume were increased near the apex and osmotic potentials were lower throughout the growing region. Because the stressed roots were thinner, osmotic adjustment occurred without osmoticum accumulation per unit length. The effects of low water potential on hexose deposition were similar to those for total osmotica, and hexose made a major contribution to the osmotic adjustment in middle and basal regions. In contrast, potassium deposition decreased at low water potentials in close parallel with water deposition, and increases in potassium concentration were small. The results show that growth of the maize primary root at low water potentials involves a complex pattern of morphogenic and metabolic events. Although osmotic adjustment is largely the result of a greater inhibition of volume expansion and water deposition than solute deposition, the contrasting behavior of hexose and potassium deposition indicates that the adjustment is a highly regulated process.     

Studies with Composite Membranes: II. Measurement of Asymmetric Properties

Electrical potentials arising across composite membranes when they separate the same concentration of a (1:1) electrolyte or electrolytes have been measured. These potentials have been shown to arise from differences in the transport number of counterions contacting the two faces of the membrane which contained in its body a high concentration of electrolyte and polyelectrolyte. When the concentration of this trapped electrolyte or polyelectrolyte is low, the asymmetry potentials are small. Although measurements of current-voltage relations provided evidence for the existence of asymmetry between the two faces of the membrane, osmotic flow of water in either direction across the membrane and the salt flow in the two directions were symmetrical. These solvent and solute flux measurements lasted more than 30 hr. Short-term (about 4 hr) flux measurements, however, using tritiated water (THO), gave flows which were different in the two directions. Similarly, the salt flows measured using ²²Na isotope were different in the two directions. The usefulness of the present system as a model to use for studies concerned with carrier transport problems in biology has been pointed out.     

Impact of osmotic and matric water stress on germination, growth, mycelial water potentials and endogenous accumulation of sugars and sugar alcohols in Fusarium graminearum

Studies were conducted to determine the effect of osmotic (NaCl, glycerol) and matric (PEG 8000) water stress on temporal germination and growth of two F. graminearum strains over the water potential range of -0.7 to -14.0 MPa at 15 and 25 C. The effect on endogenous water potentials and accumulation of sugars and sugar alcohols also were measured. For both strains, germination occurred rapidly over the same range of osmotic or matric potential of -0.7 to -5.6 MPa after 4-6 h incubation. At lower osmotic and matric potentials (-7.0 to -8.4 MPa), there was a lag of up to 24 h before germination. Optimum germ-tube extension occurred between -0.7 and -1.4 MPa for both strains but varied with the solute used. Growth was optimal at -1.4 MPa and 25 C in response to matric stress, with the minimum being about -8.0 and -11.2 MPa at 15 and 25 C, respectively. In contrast, F. graminearum grew fastest at -0.7 MPa and was more tolerant of solute stress modified with either glycerol or NaCl with a minimum of about -14.0 MPa at 15 and 25 C. A decrease in the osmotic/matric water potential of the media caused a large decrease in the mycelial water potential (Ψ(c)) as measured by thermocouple psychrometry. In general, the concentration of total sugar alcohols in mycelia increased as osmotic and matric potential were reduced to -1.2 MPa. However, this increase was more evident in mycelia from glycerol-amended media. The quality of the major sugar alcohol accumulated depended on the solute used to generate the water stress. The major compounds accumulated were glycerol and arabitol on osmotically modified media and arabitol on matrically modified media. In response to matric stress, the concentration of trehalose in colonies generally was higher in the case of osmotic stress. In each water-stress treatment there was a good correlation between Ψ(c) and total sugar alcohol content.     

Osmotic Flow of Water across Permeable Cellulose Membranes

Direct measurements have been made of the net volume flow through cellulose membranes, due to a difference in concentration of solute across the membrane. The aqueous solutions used included solutes ranging in size from deuterated water to bovine serum albumin. For the semipermeable membrane (impermeable to the solute) the volume flow produced by the osmotic gradient is equal to the flow produced by the hydrostatic pressure RT ΔC, as given by the van't Hoff relationship. In the case in which the membrane is permeable to the solute, the net volume flow is reduced, as predicted by the theory of Staverman, based on the thermodynamics of the steady state. A means of establishing the amount of this reduction is given, depending on the size of the solute molecule and the effective pore radius of the membrane. With the help of these results, a hypothetical biological membrane moving water by osmotic and hydrostatic pressure gradients is discussed.     

Hydrostatic pressures developed by osmotically swelling vesicles bound to planar membranes

When phospholipid vesicles bound to a planar membrane are osmotically swollen, they develop a hydrostatic pressure (delta P) and fuse with the membrane. We have calculated the steady-state delta P, from the equations of irreversible thermodynamics governing water and solute flows, for two general methods of osmotic swelling. In the first method, vesicles are swollen by adding a solute to the vesicle-containing compartment to make it hyperosmotic. delta P is determined by the vesicle membrane's permeabilities to solute and water. If the vesicle membrane is devoid of open channels, then delta P is zero. When the vesicle membrane contains open channels, then delta P peaks at a channel density unique to the solute permeability properties of both the channel and the membrane. The solute enters the vesicle through the channels but leaks out through the region of vesicle-planar membrane contact. delta P is largest for channels having high permeabilities to the solute and for solutes with low membrane permeabilities in the contact region. The model predicts the following order of solutes producing pressures of decreasing magnitude: KCl greater than urea greater than formamide greater than or equal to ethylene glycol. Differences between osmoticants quantitatively depend on the solute permeability of the channel and the density of channels in the vesicle membrane. The order of effectiveness is the same as that experimentally observed for solutes promoting fusion. Therefore, delta P drives fusion. When channels with small permeabilities are used, coupling between solute and water flows within the channel has a significant effect on delta P. In the second method, an impermeant solute bathing the vesicles is isosmotically replaced by a solute which permeates the channels in the vesicle membrane. delta P resulting from this method is much less sensitive to the permeabilities of the channel and membrane to the solute. delta P approaches the theoretical limit set by the concentration of the impermeant solute.     

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.     

Measurement of grouped intracellular solute osmotic virial coefficients

Models of cellular osmotic behaviour depend on thermodynamic solution theories to calculate chemical potentials in the solutions inside and outside the cell. These solutions are generally thermodynamically non-ideal under cryobiological conditions. The molality-based Elliott et al. form of the multi-solute osmotic virial equation is a solution theory which has been demonstrated to provide accurate predictions in cryobiological solutions, accounting for the non-ideality of these solutions using solute-specific thermodynamic parameters called osmotic virial coefficients. However, this solution theory requires as inputs the exact concentration of every solute in the solution being modeled, which poses a problem for the cytoplasm, where such detailed information is rarely available. This problem can be overcome by using a grouped solute approach for modeling the cytoplasm, where all the non-permeating intracellular solutes are treated as a single non-permeating “grouped” intracellular solute. We have recently shown (Zielinski et al., J Physical Chemistry B, 2017) that such a grouped solute approach is theoretically valid when used with the Elliott et al. model, and Ross-Rodriguez et al. (Biopreservation and Biobanking, 2012) have previously developed a method for measuring the cell type-specific osmotic virial coefficients of the grouped intracellular solute. However, the Ross-Rodriguez et al. method suffers from a lack of precision, which—as we demonstrate in this work—can severely impact the accuracy of osmotic model predictions under certain conditions. Thus, we herein develop a novel method for measuring grouped intracellular solute osmotic virial coefficients which yields more precise values than the existing method and then apply this new method to measure these coefficients for human umbilical vein endothelial cells.     

Osmoregulation,solute distribution,and growth in soybean seedlings having low water potentials

Soybean (Glycine max (L.) Merr.) seedlings osmoregulate when the supply of water is limited around the roots. The osmoregulation involves solute accumulation (osmotic adjustment) by the elongating region of the hypocotyls. We investigated the relationship between growth, solute accumulation, and the partitioning of solutes during osmoregulation. Darkgrown seedlings were transplanted to vermiculite containing 1/8 (0.13 x) the water of the controls. Within 12–15 h, the osmotic potential of the elongating region had decreased to-12 bar, but it was-7 bar in the controls. This osmoregulation involved a true solute accumulation by the hypocotyls, since cell volume and turgor were virtually the same regardless of the water regime. The hypocotyls having low water potentials elongated slowly but, when deprived of their cotyledons, did not elongate or accumulate solute. This result indicated a cotyledonary origin for the solutes and a dependence of slow growth on osmotic adjustment. The translocation of nonrespired dry matter from the cotyledons to the seedling axis was unaffected by the availability of water, but partitioning was altered. In the first 12 h, dry matter accumulated in the elongating region of the 0.13 x hypocotyls, and osmotic adjustment occurred. The solutes involved were mostly free amino acids, glucose, fructose, and sucrose, and these accounted for most of the increased dry weight. After osmotic adjustment was complete, dry matter ceased to accumulate in the hypocotyls and bypassed them to accumulate in the roots, which grew faster than the control roots. The proliferation of the roots resulted in an increased root/shoot ratio, a common response of plants to dry conditions.Osmotic adjustment occurred in the elongating region of the hypocotyls because solute utilization for growth decreased while solute uptake continued. Adjustment was completed when solute uptake subsequently decreased, and uptake then balanced utilization. The control of osmotic adjustment was therefore the rate of solute utilization and, secondarily, the rate of solute uptake. Elongation was inhibited by unknown factors(s) despite the turgor and substrates associated with osmotic adjustment. The remaining slow elongation depended on osmotic adjustment and represented some optimum between the necessary inhibition for solute accumulation and the necessary growth for seedling establishment.     

Symplasmic transport of water along the root depends on pressure

The gradient NMR method was applied to study intercellular water flows in root segments of maize (Zea mays L.) under disturbance of root hydrodynamic system by the increase in external pressure up to 4 MPa. The rate of intercellular water flows along the root symplast was found to depend on the magnitude and dynamics of pressure changes. Based on the previously predicted cupola-shaped dependency of water flow on the aperture of plasmodesmal neck constrictions, we assume that the external pressure stimulates (via cytosolic calcium) the activity of contractile structures localized in the neck regions of plasmodesmata. Cells of Chlorella vulgaris were taken for comparison since their water relations and cell structure differ strongly from the root cells of maize. The results show that the diffusional water flow in Chlorella is independent of external pressure both in intact cells and in algae, whose plasma membrane was artificially permeabilized.     

Osmosis and solute—Solvent drag

In 1903, George Hulett explained how solute alters water in an aqueous solution to lower the vapor pressure of its water. Hulett also explained how the same altered water causes osmosis and osmotic pressure when the solution is separated from liquid water by a membrane permeable to the water only. Hulett recognized that the solute molecules diffuse toward all boundaries of the solution containing the solute. Solute diffusion is stopped at all boundaries, at an open-unopposed surface of the solution, at a semipermeable membrane, at a container wall, or at the boundary of a solid or gaseous inclusion surrounded by solution but not dissolved in it. At each boundary of the solution, the solute molecules are reflected, they change momentum, and the change of momentum of all reflected molecules is a pressure, a solute pressure (i.e., a force on a unit area of reflecting boundary). When a boundary of the solution is open and unopposed, the solute pressure alters the internal tension in the force bonding the water in its liquid phase, namely, the hydrogen bond. All altered properties of the water in the solution are explained by the altered internal tension of the water in the solution. We acclaim Hulett's explanation of osmosis, osmotic pressure, and lowering of the vapor pressure of water in an aqueous solution. His explanation is self-evident. It is the necessary, sufficient, and inescapable explanation of all altered properties of the water in the solution relative to the same property of pure liquid water at the same externally applied pressure and the same temperature. We extend Hulett's explanation of osmosis to include the osmotic effects of solute diffusing through solvent and dragging on the solvent through which it diffuses. Therein lies the explanations of (1) the extravasation from and return of interstitial fluid to capillaries, (2) the return of luminal fluid in the proximal and distal convoluted tubules of a kidney nephron to their peritubular capillaries, (3) the return of interstitial fluid to the vasa recta, (4) return of aqueous humor to the episcleral veins, and (5) flow of phloem from source to sink in higher plants and many more examples of fluid transport and fluid exchange in animal and plant physiology. When a membrane is permeable to water only and when it separates differing aqueous solutions, the flow of water is from the solution with the lower osmotic pressure to the solution with the higher osmotic pressure.     

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

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