Chara buckellii, a Euryhaline Charophyte from an Unusual Saline Environment : III. Time Course of Turgor Regulation

The time course of turgor regulation of the euryhaline giant-celled alga, Chara buckellii, is presented. Isolated intermodal cells were challenged by increasing or decreasing the external osmotic pressure by 150 milliosmoles per kilogram with all ions in the media or by dilution, respectively. Regulation following hypotonic stress was complete within 48 hours whereas regulation following hypertonic stress required between 96 and 144 hours. The change in internal osmotic pressure could be entirely accounted for by changes in vacuolar KCl in response to hypotonic stress, but this ion pair only accounted for 45% of the change in response to hypertonic stress. The membrane potential of C. buckellii is normally hyperpolarized with respect to the equilibrium potential for K⁺ (E_K). The membrane depolarized to a level close to E_K in response to hypotonic treatment and this was accompanied by a transient increase in membrane conductance. In response to hypertonic stress, the membrane hyperpolarized transiently, then repolarized to a level close to the control. This was accompanied by a temporary decrease in membrane conductance. The data are discussed with respect to the ecological significance of the time course and ion transport mechanisms during turgor regulation.     

Turgor pressure changes trigger characteristic changes in the electrical conductance of the tonoplast and the plasmalemma of the marine alga Valonia utricularis

The giant marine alga Valonia utricularis is capable of regulating its turgor pressure in response to changes in the osmotic pressure of the sea water. The turgor pressure response comprises two phases, a fast, exponential phase arising exclusively from water shifting between the vacuole and the external medium (time constant about 10 min) and a second very slow, almost exponential phase adjusting (but not always) the turgor pressure near to the original value by release or uptake of KCl (time constant about 5 h). The changes in the vacuolar membrane potential as well as in the individual conductances of the tonoplast and plasmalemma accompanying turgor pressure regulation were measured by using the vacuolar perfusion assembly (with integrated microelectrodes, pressure transducers and pressure‐regulating valves) as described by Wang et al. (J. Membrane Biology 157, 311–321, 1997). Measurements on pressure‐clamped cells gave strong evidence that the turgor pressure, but not effects related to water flow (i.e. electro‐osmosis or streaming potential) or changes in the internal osmotic pressure and in the osmotic gradients, triggers the cascade of osmotic and electrical events recorded after disturbance of the osmotic equilibrium. The findings definitely exclude the existence of osmosensors as postulated for other plant cells and bacteria. There was also evidence that turgor pressure signals were primarily sensed by ion transporters in the vacuolar membrane because conductance changes were first recorded in the many‐folded tonoplast and then significantly delayed in the plasmalemma independent of the direction of the osmotic challenge. Consistently, turgor pressure up‐regulation (but not down‐regulation) could be inhibited reversibly by external addition of the K⁺ transport inhibitor Ba²⁺ and/or by the Cl^– transport inhibitor 4,4′‐diisothiocyanatostilbene‐2,2′‐disulfonic acid (DIDS). Extensive studies under iso‐, hyper‐ and hypo‐osmotic conditions revealed that K⁺ and Cl^– contribute predominantly to the plasmalemma conductance. Addition of 0.3 mm NaCN showed further that part of the K⁺ and Cl^– transporters depended on ATP. These transporters are apparently up‐regulated upon hyper‐osmotic, but not hypo‐osmotic challenge. These findings explain the strong increase of the K⁺ influx upon lowering turgor pressure and the less pronounced pressure‐dependence of the Cl^– influx of V. utricularis reported in the literature. The data derived from the blockage experiments under hypo‐osmotic conditions were also equally consistent with the experimental findings that the K⁺ efflux is solely passive and progressively increases with increasing turgor pressure due to an increase of the volumetric elastic modulus of the cell wall. However, despite unravelling some of the sequences and other components involved in turgor pressure regulation of V. utricularis the co‐ordination between the ion transporters in the tonoplast and plasmalemma remains unresolved because of the failure to block the tonoplast transporters by addition of Ba²⁺ and DIDS from the vacuolar side.     

Turgor regulation in a brackish water charophyte,Lamprothamnium succinctum

Internodal cells of a brackish water charophyte,Lamprothamnium succinctum (A. Br. in Ash.) R.D.W. regulate the turgor pressure in response to changes in both the cellular and the external osmotic pressures. During turgor regulation upon hypotonic treatment, net effluxes of K⁺ and Cl⁻ from the vacuole, membrane depolarization, a transient increase in the electrical membrane conductance and a transient increase in concentration of cytoplasmic Ca²⁺ are induced. Activation of the plasmalemma Ca²⁺ channels and the Ca²⁺-controlled passive effluxes of K⁺ and Cl⁻ through the plasmalemma ion channels are postulated.     

Turgor regulation inValonia macrophysa following acute osmotic shock

Summary The marine algaValonia macrophysa an inhabitant of shallow subtropical waters, is subjected to sudden dilutions of external seawater during rain showers. This study describes the mechanisms involved in turgor pressure regulation following acute hyposmotic shock. Turgor regulation is 88% effective and complete within 4 hr following hyposmotic shocks of up to –10 bar. Loss of vacuolar K⁺, Na⁺ and Cl^– accounts for the decrease in vacuolar osmotic pressure associated with turgor regulation. A novel mechanism of turgor regulation is exhibited byValonia macrophysa given hyposmotic shocks greater than about –4 bar. Such an osmotic shock causes cell wall tension to increase above a critical value of about 6×10⁵ dyne/cm, whereupon the protoplasm ruptures and the cell wall stretches irreversibly at a localized site. The protoplasm rupture is suggested by (1) a large abrupt increase in K⁺ efflux (as measured by⁸⁶Rb⁺), (2) a rapid decrease in turgor pressure as measured with a pressure probe, and (3) sudden depolarization of the vacuole potential. Evidence for an increase in cell wall permeability includes efflux from the vacuole of dextran (mol wt 70,000), which normally has a very low cell wall permeability, and scanning electron micrographs which show a trabeculated scar area in the cell wall. This mechanism of turgor regulation is physiologically important because 98% of the cells regained normal growth rate and turgor following acute osmotic shock.     

Differential effects of turgor on sucrose and potassium transport at the tonoplast and plasma membrane of sugar beet storage root tissue

When turgor was increased, by decreasing the concentration of mannitol bathing discs of sugar beet storage root tissue, the rates of sucrose and potassium uptake into the vacuole were decreased. At all external mannitol concentrations the rate of sucrose and potassium uptake across the plasma membrane was an order of magnitude greater than the rate of quasi-steady uptake into the vacuole, implying a very large efflux. Efflux of both sucrose and potassium was increased at high turgor. However, while increasing turgor decreased the rate of K⁺ uptake, the rate of sucrose uptake at the plasma membrane increased with time. Compartmental analysis of tracer exchange kinetics was used to determine unidirectional K⁺ fluxes. From these results, it was estimated that the increase in K⁺ efflux accompanying a 1.5 MPa increase in turgor could lead to a net increase of 140mol^?3h^?1 in the external potassium concentration. It is suggested that the turgor-imposed increase in solute efflux is a means of regulating intracellular osmotic pressure and/or turgor in sugar beet storage roots, but that sucrose is preferentially retrieved from the apoplast, even under conditions of excessively high turgor. However, much of this sucrose is probably lost from the cell, implying a ‘futile’ sucrose transport cycle at the plasma membrane. The turgor-stimulated leak of potassium could play a major role in the regulation of turgor pressure in sugar beet storage root tissue.     

Turgor regulation in Lamprothamnium papulosum. I. I/V analysis and pharmacological dissection of the hypotonic effect

Current-voltage (I/V) analysis and pharmacological dissection were applied to membranes of Lamprothamnium at the time of hypotonic stress. At least three types of process were found to be involved in the response to this stress.

• 1 The first 10min of exposure to hypotonic medium resulted in a depolarization of about 50mV accompanied by a decrease or no change in conductance. This depolarization occurred with either K⁺ or Ca²⁺ (and consequently C^? channels inactivated.
• 2 The CI^? channels opened mainly in the first 15min of the hypotonic stress, increasing the membrane conductance by about an order of magnitude.
• 3 The K⁺ conductance rose as the Cl^? conductance started to diminish and reached a maximum after about 40 min.

Both types of channel were strongly potential-dependent with a conductance peak between -150 and 0mV. An inactivation of K⁺or CI^? channels resulted in moving the membrane potential away from the conductance maximum toward either E_K or E_CI, diminishing the ion efflux (and turgor regulation). The time courses of the conductance increases remained the same, suggesting that the conductance changes are not driven by feedback to some preset turgor level. The electrophysiology of the Lamprothamnium transporters is compared to that of salt-sensitive charophytes.     

Osmotic acclimation of the brackish water xanthophyceae,Vaucheria dichotoma (L.) MARTIUS: Inorganic ion composition and amino acids

The salinity tolerance ofVaucheria dichotoma, a siphonous Xanthophycean alga was investigated. The alga survived an external osmotic potential range between 74 and 1, 176 mOsmol (ca. 2.5 and 40.0 ppt. (parts per thousand]). Turgor pressure was regulated in salinities ranging from 74 to 441 mOsmol. With further increase of the salinity, turgor pressure decreased from 153 to 9 mOsmol (0.44 to 0.08 MPa). At 441 mOsmol salinity the major intracellular ions were present in the following concentrations (mM/l cell water): K⁺, 145; Na⁺; 90; sulphate, 91; Cl⁻, 91. Under the most severe salinity stress (1,176 mOsmol) the ionic concentration increased to (mM/l cell water): K⁺, 250; Na⁺, 75; sulphate, 35; Cl⁻, 351. The content of amino acids: alanine (Ala), threonine (Thr and glutamic acid (Glu) was lower, nerver exceeding 5–11 mM, however; the concentrations were positively correlated with salinity.     

Partial Turgor Pressure Regulation in Chara canescens and its Implications for a Generalized Hypothesis of Salinity Response in Charophytes

Concentrations of ions and sucrose in the vacuolar sap of Chara canescens growing in an oligohaline lake (1.5 ‰) were estimated over the main growth period of the plants. During fructification vacuolar sap contained a mean of 41 mol m^?3 (range 10.2–61.8) sucrose. The mean turgor pressure was 239 mosmol kg^?1 (range 219–264). In long- and short-term experiments these plants were subjected to increasing salinities up to 22 ‰. When salinity was increased from 1.5 to 4.4 ‰ turgor pressure was restored to only 80 % of the initial value. This reduced level of turgor pressure was maintained up to a salinity of 22 ‰. The increase in vacuolar osmotic potential was due to the monovalent ions Na⁺, K⁺ and Cl^?. The relative amounts of Na⁺ and K⁺ participating in the regulation process were dependent on external salinity. The regulatory mechanisms observed in the brackish water species Ch. canescens are compared with those reported from freshwater and euryhaline species.     

Turgor regulation in the salt-tolerant alga Chara longifolia

Chara longifolia is a salt‐tolerant Charophyte which regulates its turgor inresponse to osmotic stress. Membrane depolarization, in creased membrane conductance, and cessation of cytoplasmic streaming (due to increase in cytoplasmic Ca^{2 +} ) precede regulation in response to hypotonic stress. Measurements of these three parameters are presented here with simultaneous turgor measurements. Variability in the occurrence, rate and extent of turgor regulation in individual cells was correlated with magnitude of the stress. Hypertonic stress showed the same slow time course as was found previously, requiring several days for complete regulation. Fifty μ M nifedipine, a Ca^{2 +} channel blocker, inhibited turgor regulation. In the presence of 5 μ M nifedipine, turgor regulation was delayed. An increase in conductance preceded regulation, but membrane depolarization was less and no detectable change in cytoplasmic streaming was observed, requiring modifications to a previously presented model for turgor regulation. There was no significant difference in ⁴⁵Ca^{2 +} influx under control and stress conditions. However, the control flux was insensitive to nifedipine, whereas under stress the flux is inhibited 54% by nifedipine. We suggest that osmotic stress results in a rapid increase in a nifedipine‐sensitive Ca^{2 +} entry mechanism, followed very quickly by a decrease in the control entry mechanism.     

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

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.     

Phosphate cycles in energy crop systems with emphasis on the availability of different phosphate fractions in the soil

The growing cells of hydroponic maize roots expand at constant turgor pressure (0.48 MPa) both when grown in low-(0.5 mol m^-3 CaCl₂) or full-nutrient (Hoagland's) solution and also when seedlings are stressed osmotically (0.96 MPa mannitol). Cell osmotic pressure decreases by 0.1–0.2 MPa during expansion. Despite this, total solute influx largely matches the continuously-varying volume expansion-rate of each cell. K⁺ in the non-osmotically stressed roots is a significant exception-its concentration dropping by 50% regardless of the presence or absence of K⁺ in the nutrient medium. This corresponds to the drop in osmotic pressure. Nitrate appears to replace Cl^- in the Hoagland-grown cells.Analogous insensitivity of solute gradients to external solutes is observed in the radial distribution of water and solutes in the cortex 12 mm from the tip. Uniform turgor and osmotic pressures are accompanied by opposite gradients of K⁺ and Cl^-, outwards, and hexoses and amino acids, inwards, for plants grown in either 0.5 mol m^-3 CaCl₂ or Hoagland's solution (with negligible Cl^-). K⁺ and Cl^- levels within both gradients were slightly higher when the ions were available in the medium. The gradients themselves are independent of the direction of solute supply. In CaCl₂ solution all other nutrients must come from the stele, in Hoagland's solution inorganic solutes are available in the medium.24 h after osmotic stress, turgor pressure is recovered at all points in each gradient by osmotic adjustment using organic solutes. Remarkably, K⁺ and Cl^- levels hardly change, despite their ready availability. Hexoses are responsible for some 50% of the adjustment with mannitol for a further 30%. Some 20% of the final osmotic pressure remains to be accounted for. Proline and sucrose are not significantly involved. Under all conditions a standing water potential step of 0.2 MPa between the rhizodermis and its hydroponic medium was found. We suggest that this is due to solute leakage.Abbreviations EDX energy dispersive X-ray microanalysis - water potential - 11-1 cell osmotic pressure - P turgor pressure     

Increase in cytoplasmic calcium content in internodal cells of Lamprothamnium upon hypotonic treatment

Abstract Internodal cells of Lamprothamnium succinctum, a brackish water Characeae, regulate turgor pressure in response to changes in external osmotic pressure (turgor regulation). When internodal cells were transferred to a hypotonic medium containing 3.9 mol m^?3 Ca²⁺, the cell osmotic pressure decreased and the original turgor pressure was recovered. During turgor regulation Ca content of the cytoplasm increased significantly. Lowering the external Ca²⁺ concentration from 3.9 to 0.01 mol m^?3 inhibited this increase in cytoplasmic calcium content. In a hypotonic medium containing 0.01 mol m^?3 Ca²⁺, turgor regulation was inhibited as previously reported (Okazaki & Tazawa, 1986a). Thus transient increase in cytoplasmic Ca, probably in the ionized form, induced by hypotonic treatment may play an important role in turgor regulation.     

The role of calcium in turgor regulation in Chara longifolia

The salt-tolerant alga Chara longifolia (Robinson) is capable of regulating its turgor in response to hypotonic stress resulting from a decrease in the osmotic pressure of the medium. This regulatory process takes only 40 min in small cells (length ≤ 10 mm), but requires 3d in large cells (length ≥30mm). Turgor regulation in small cells is comprised of two phases, a fast phase reducing the increased turgor by about 25% in the First 5 min, and a second phase reducing the turgor to near the original value within 40 min. The second phase is inhibited by reducing the concentration of Ca²⁺ in the external medium from 4.6 to 0.01 mol m^?3; the first phase is less affected by the reduction of Ca²⁺. In the first 5 min of stress, the membrane depolarizes in a voltage-dependent fashion, electrical conductance of the membrane increases transiently and cytoplasmic streaming is inhibited. When the external Ca²⁺ concentration is lowered, conductance does not increase and streaming continues unaffected. In a low ionic strength medium, Ca²⁺ is not required in the medium for turgor regulation. To test the hypothesis that there is increased Ca²⁺ entry from the medium during turgor regulation, we measured the influx of ⁴⁵Ca²⁺ into the cell. We found an increased influx of Ca²⁺, from 18 to 36 nmol m^?2 s^?1 during the first 30 to 90 s following osmotic stress. This increase was evident only in cells below about 7 mm in length, and was more marked in smaller cells.     

Chloride transport and the membrane potential in the marine alga,Halicystis parvula

Summary The relationship between the rate of Cl^– transport and the electrical properties ofHalicystis parvula was investigated. Three metabolic inhibitors-darkness, cyanide (2mm), and low temperature (4°C)-all rapidly and reversibly reduce both the short circuit current (SCC), which is a measure of net Cl^– transport, and the vacuole electrical potential (PD). Plotting thePD vs. SCC for inhibited cells yields a linear regression with ay-intercept of zero. ThePD is also greatly reduced when the [Cl^–] of the external medium is lowered. Raising the external [K⁺] produces an appreciable, but less than Nernstian, depolarization, while increasing the external [H⁺] tenfold has no net effect on thePD. Decreasing the external [Na⁺] by tenfold produces only a slight depolarization. Thus, the outer plasma membrane appears to be moderately selective for K⁺ over Na⁺ or H⁺. The effects of ion substitutions in the vacuolar perfusing solutions on thePD reveal that the vacuolar membrane does not discriminate electrically between Cl^– and the much larger anions, isethionate and benzenesulfonate, or between Na⁺ and K⁺. The data suggest that in internally perfused cells ofH. parvula generation of thePD of –50 to –60 mV by a transport system involving only electroneutral pumps is unlikely and that most of thisPD is generated by an electrogenic Cl^– pump.     

Turgor and net ion flux responses to activation of the osmotic MAP kinase cascade by fludioxonil in the filamentous fungus Neurospora crassa

The internal hydrostatic pressure (turgor) of the filamentous fungus Neurospora crassa is regulated at about 400–500 kiloPascals, primarily by an osmotic MAP kinase cascade which activates ion uptake from the extracellular medium and glycerol synthesis. In the absence of hyperosmotic stress, the phenylpyrrole fungicide fludioxonil activates the osmotic MAP kinase cascade, resulting in cell death. Turgor, the electrical potential and net ion fluxes were measured after treatment with fludioxonil. In wildtype, fludioxonil causes a hyperpolarization of the plasma membrane and net H⁺ efflux from the cell, consistent with activation of the H⁺-ATPase. At the same time, net K⁺ uptake occurs, and turgor increases (about 2-fold above normal levels). None of these changes are observed in the os–2 mutant (which lacks a functional MAP kinase, the last of the three kinases in the osmotic MAP kinase cascade). Tip growth ceases as hyperpolarization, net ion flux changes, and turgor increases begin. The inappropriate turgor increase is the probable cause of eventual lysis and death. The results corroborate a multi-pathway response to hyperosmotic stress that includes activation of plasma membrane transport. The relation to cell expansion (tip growth) is not direct. Increases in turgor due to ion transport might be expected to increase growth rate, but this does not occur. Instead, there must be a complex regulatory interplay between the growth and the turgor driving force, possibly mediated by regulation of cell wall extensibility.     

The Role of Potassium in the Control of Turgor Pressure in a Gas-Vacuolate Blue-Green Alga

The contribution of K⁺ accumulation to cell turgor pressurewas investigated in the gas-vacuolate blue-green alga Anabaenaflos-aquae. The cell turgor pressure, measured by the gas vesiclemethod, drops in cells suspended in culture medium depletedof K⁺ but rapidly rises again, by 100 kPa or more, when K⁺ isresupplied. A similar though rather slower rise in turgor pressureis supported by an equivalent concentration of Rb⁺. The internalK⁺ concentration rose from 66 to 91 mM when K⁺ was suppliedat an external concentration of 0.4 mM. This rise was light-dependent.Greater increases in internal K⁺ concentration and turgor pressureoccurred when K⁺ was supplied at a higher concentration, 3.6mM. In both cases over 60% of the observed turgor pressure risecould be accounted for by accumulation of K⁺. The turgor pressurerise supported by light-stimulated K⁺ uptake can cause collapseof enough of the alga's gas vesicles to destroy its buoyancy.The effect of K⁺ availability on buoyancy regulation by planktonicblue-green algae is discussed.     

Involvement of 14-3-3 proteins in the osmotic regulation of H+-ATPase in plant plasma membranes

 Taking the binding of fusicoccin to plasma membranes as an indicator of complex formation between the 14-3-3 dimer and H⁺-ATPase, we assessed the effect of osmotic stress on the interaction of these proteins in suspension-cultured cells of sugar beet (Beta vulgaris L.). An increase in osmolarity of the cell incubation medium, accompanied by a decrease in turgor, was found to activate the H⁺ efflux 5-fold. The same increment was observed in the number of high-affinity fusicoccin-binding sites in isolated plasma membranes; the 14-3-3 content in the membranes increased 2- to 3-fold, while the H⁺-ATPase activity changed only slightly. The data obtained indicate that osmotic regulation of H⁺-ATPase in the plant plasma membrane is achieved via modulation of the coupling between H⁺ transport and ATP hydrolysis, and that such regulation involves 14-3-3 proteins. Received: 10 February 2000 / Accepted: 31 March 2000     

Mechanisms underlying turgor regulation in the estuarine alga Vaucheria erythrospora (Xanthophyceae) exposed to hyperosmotic shock

Aquatic organisms are often exposed to dramatic changes in salinity in the environment. Despite decades of research, many questions related to molecular and physiological mechanisms mediating sensing and adaptation to salinity stress remain unanswered. Here, responses of Vaucheria erythrospora, a turgor‐regulating xanthophycean alga from an estuarine habitat, have been investigated. The role of ion uptake in turgor regulation was studied using a single cell pressure probe, microelectrode ion flux estimation (MIFE) technique and membrane potential (E_m) measurements. Turgor recovery was inhibited by Gd³⁺, tetraethylammonium chloride (TEA), verapamil and orthovanadate. A NaCl‐induced shock rapidly depolarized the plasma membrane while an isotonic sorbitol treatment hyperpolarized it. Turgor recovery was critically dependent on the presence of Na⁺ but not K⁺ and Cl^? in the incubation media. Na⁺ uptake was strongly decreased by amiloride and changes in net Na⁺ and H⁺ fluxes were oppositely directed. This suggests active uptake of Na⁺ in V. erythrospora mediated by an antiport Na⁺/H⁺ system, functioning in the direction opposite to that of the SOS1 exchanger in higher plants. The alga also retains K⁺ efficiently when exposed to high NaCl concentrations. Overall, this study provides insights into mechanisms enabling V. erythrospora to regulate turgor via ion movements during hyperosmotic stress.     

Ionic processes underlying the decrease in osmotic potential of Chara L-cell fragments in dilute electrolyte solutions

Movements of ions are considered to be governed by the electroneutrality rule. Therefore, a cation moving across the cell membrane into the cell either passively or actively should move together with its counterion, an anion, in equal amounts of charge or in exchange for another cation inside the cell. This means that the net influx of the cation in question should be affected by the permeability of its counterion and/or another cation inside the cell. To examine osmotic and ionic regulation in Chara cells, cell fragments of Chara having a lower osmotic pressure than normal (L-cell fragments) were prepared. The L-cell fragments were individually put into various dilute electrolyte solutions and their osmotic potentials were measured with a turgor balance. Concentrations of K⁺, Na⁺, Ca²⁺, Mg²⁺, Cl^?, NO^?₃. and SO^2?₄. in the external electrolyte solutions in which L-cells had been incubated were also analysed by ion chromatography. The results showed that in 0.5 mM KCL + 0.1 mM CaCl₂ solution, Chara L-cell fragments absorbed K⁺ and Cl^? to maintain electroneutrality and then regained their osmotic potential very rapidly. When the anion was Cl, the cation absorbed at the highest rate was K⁺ On the other hand, when the cation was K, the anion absorbed at the highest rate was Cl, Other ions Ca²⁺, SO^2?₄ and NO^?₃ showed much less permeability than K⁺ and Cl ^?for the Chara plasma membrane. The conclusion from these findings was that due to the constraint of electroneutral transport, the uptake rate of a salt into L-cells is limited by the permeability of the least permeable ion.     

Ptk2 contributes to osmoadaptation in the filamentous fungus Neurospora crassa

Hyphal tip-growing organisms often rely upon an internal hydrostatic pressure (turgor) to drive localized expansion of the cell. Regulation of the turgor in response to osmotic shock is mediated primarily by an osmotic MAP kinase cascade which activates osmolyte synthesis and ion uptake to effect turgor recovery. We characterized a Neurospora crassa homolog (PTK2) of ser/thr kinase regulators of ion transport in yeast to determine its role in turgor regulation in a filamentous fungi. The ptk2 mutant is osmosensitive, and has lower turgor poise than wildtype. The cause appears to be lower activity of the plasma membrane H⁺-ATPase. Its role in osmoadaptation is unrelated to the activity of the osmotic MAP kinase cascade. Instead, it acts in an alternative pathway that, like the osmotic MAP kinase cascade, also involves ion transport mediated osmoadaptation.