Evaluation of a Non-Destructive Method for Measuring Turgor Pressure in Helianthus

The portable instrument described by Heathcote, Etherington,and Woodward (1979) for the non-destructive measurement of turgorpressure was evaluated in Helianthus annuus and Helianthus paradoxus.A good correlation was obtained between turgor pressure measuredwith the instrument and turgor pressure estimated by the pressure-volumetechnique for individual leaves allowed to dry after excision;however, variation in both the intercept and slope of the relationshipoccurred between leaves. Consequently, there was no correlationbetween the output of the instrument for individual leaves andthe turgor pressure of the same leaves estimated by conventionalmethods. Moreover, for a given leaf, the instrument had onlya limited ability to detect temporal variation in turgor pressurewhen compared with turgor pressure calculated from measuredvalues of leaf water potential and leaf osmotic potential. Theinstrument's output was influenced by its proximity to majorveins and by leaf thickness. We conclude that variability inleaf thickness and the presence of large veins limits its usefulnessfor measurement of turgor pressure in Helianthus. Key words: Leaf thickness, Turgormeter, Turgor pressure, Helianthus     

Blue Light Pulse-Induced Transient Changes of Electric Potential and Turgor Pressure in the Motor Cells of Phaseolus vulgaris L.

Blue light induces both depolarization of membrane potentialin the motor cell and turgor movement in the laminar pulvinusof bean plant. This paper describes the changes of electricpotential and turgor pressure induced in Phaseolus vulgarisL. by blue light pulses. A transient depolarization of membranepotential as large as 40 mV was induced by a short pulse of15 s blue light in motor cells of the laminar pulvinus. Thischange was not an action potential because of the absence ofa refractory period and threshold. The magnitudes of the responsewere dependent on the fluence of light. The response was long-lived,indicating that continuous input of light energy is not requiredfor a sustained response. The potential change was always followedby a transient turgor movement of the pulvinus. A molecular mechanism similar to a model postulated for theblue light response of stomata may operate in the motor cell.However, the direction of the electrical response to blue light(depolarization) in the motor cell was the opposite of thatin the guard cell (hyperpolarization). Turgor change of themotor cell by blue light was also opposite in direction (decrease). (Received February 19, 1988; Accepted June 28, 1988)     

Direct Measurements of Turgor Pressure Potentials of Guard Cells: II. THE MECHANICAL ADVANTAGE OF SUBSIDIARY CELLS, THE SPANNUNQSPHASE, AND THE OPTIMUM LEAF WATER DEFICIT

Evidence of the mechanical advantage of subsidiary cells wasobtained by simultaneous measurements of turgor pressure potentialsin adjacent subsidiary and guard cells using injection circuitswith two separate needles. In Tradescantia virginiana the mechanicaladvantage approaches two. Using the same technique evidencewas obtained that the Spannungsphase is, in the first place,a turgor relations phenomenon due to the mechanical advantageof epidermal or subsidiary cells. In addition, the evidenceindicated that the elastic properties of guard cell walls mayundergo changes during the Spannungsphase when potassium iontransport commences. During these measurements it was confirmedthat the optimum leaf water deficit for maximum stomatal openingoccurs when the epidermal turgor is near zero. Under these conditionsthe width of the stomatal pore is a function of the turgor pressureof the guard cells, since at zero turgor of the subsidiary cellstheir mechanical advantage has disappeared.     

Turgor Regulation in a Brackish Charophyte, Lamprothamnium succinctum I. Artificial Modification of Intracellular Osmotic Pressure

Internodal cells of Lamprothamnium succinctum cultured in freshwater and brackish water of different salinities maintainedalmost the same turgor pressure at steady state. When the turgorpressure was increased by decreasing the external osmolality,the cells recovered their original turgor pressure within 2h. However, the recovery from decreased turgor pressure required1 day. When salts of the external medium were replaced with sorbitol,the cells still regulated the turgor pressure, indicating thatthe essential factor for the turgor regulation is not the salinitybut the osmolality. Internodal cells with osmotic pressure andion concentrations artificially modified to higher or lowervalues also regained the original turgor pressure by changingtheir intracellular osmotic pressure, whether the cells werecultured in brackish water or fresh water. These results indicate that turgor regulation is intrinsic toLamprothamnium and is initiated by a deviation of turgor pressurefrom the reference value, which is about 0.35 Osm. (Received November 28, 1983; Accepted March 14, 1984)     

A Simple Instrument for Measuring Leaf Extension in Grasses, and its Application in the Study of the Effects of Water Stress on Maize and Sorghum

The design of a simple instrument to monitor leaf expansionin grasses is described. The instrument was used to comparethe effects of water stress on leaf extension of two cultivarsof maize and sorghum. The effect of withholding water for 3days was an appreciable reduction in the rate of leaf expansionin both plants, particularly during the light period. In well-wateredplants of both species, leaf extension continued at a steadyrate even when leaf turgor fell to around 0.1 MPa. In water-stressedmaize plants, leaf turgor during the light period fell to zeroand leaf growth ceased. When turgor was restored, followingstomatal closure, leaf extension resumed at a slow rate. Inunwatered sorghum plants, leaf turgor remained at a value greaterthan 0.1 MPa but the rate of leaf extension was significantlyreduced. The reduction in leaf turgor in the unwanted plantsresulted partly from an increase in solute potential. Zea mays L, maize, Sorghum bicolor L, leaf expansion, leaf turgor, water stress     

Turgor Maintenance by Osmoregulation inBrassica napusandB. junceaunder Field Conditions

Indian mustard (Brassica juncea(L) Czernjacw) maintains higherleaf turgor than canola (B. napusL.) under water deficits andthis is related to the greater yield of mustard under theseconditions. The work reported in this paper was designed tostudy the way mustard maintains this turgor advantage. It wasbased on three field experiments that each used at least twocultivars or lines of each species. The leaf water potentialat which leaves reached zero turgor was consistently lower inmustard than in canola (up to 1.1 MPa lower). This differencearose from a greater rate of decline in leaf osmotic potentialwith declining water potential in mustard rather than from anydifference in the osmotic potential at full turgor. Calculationsof solute accumulation showed that mustard had a greater capacityto osmoregulate than canola, with this capacity being the basisfor its advantage in turgor maintenance. Other differences inplant water relations were consistent with the differences inosmoregulation, with the predicted relative water content ofleaves at an osmotic potential of -2.5 MPa being 0.43 for canolaand 0.61 for mustard. Mustard's greater capacity to accumulatesolutes is concluded to be a major factor in its greater yieldunder water deficits. Brassica napusL.; Brassica juncea(L) Czernjacw; Indian mustard; canola; water deficit; plant water relations; osmoregulation; osmotic adjustment; turgor     

Action Spectrum of Light Pulse-Induced Membrane Depolarization in Pulvinar Motor Cells of Phaseolus

In addition to circadian changes in the membrane potential andleaf movement, light applied to the pulvinus causes changesin both the membrane potential and the pulvinar movement inPhaseolus vulgaris L. Even after a short pulse of light, a transientdepolarization of the membrane occurs and leaf movement is observed.Decreases of turgor pressure of the motor cells are always precededby the depolarization. The direction of the leaf movement canbe explained by the decrease of turgor pressure in the motorcells on the irradiated side of the pulvinus. Using the OkazakiLarge Spectrograph at the National Institute for Basic Biology,we determined the action spectrum of the membrane depolarizationinduced by light pulses (30 s) in motor cells of Phaseolus.The pulvinus was left exposed to air during measurement of themembrane potential with microelectrodes. The action spectrumobtained was in the range of 300 to 730 nm. It had the highestpeak at 460 nm with lower peaks at 380 nm and 420 nm. Almostno sensitivity was observed at wavelengths shorter than 360nm and longer than 520 nm. Red and far-red light had no effecton the depolarization of the motor cell. The features of theaction spectrum are almost the same as those of the Blue-Typeresponse in plants. (Received January 9, 1997; Accepted February 14, 1997)     

Cessation of cell elongation in rye coleoptiles is accompanied by a loss of cell-wall plasticity

The mechanism by which endogenous cessation of coleoptile elongationafter emergence of the primary leaf is brought about was investigatedin rye seedlings (Secale cereale L.) that were either grownin darkness or irradiated with continuous white light. In 3-d-oldetiolated (growing) coleoptiles a turgor pressure of 0.59 MPawas measured. In 6-d-old coleoptiles, which had ceased to elongate,cell turgor was 0.51 MPa and thus only 13% lower than in therapidly growing organ. Hence, the driving force for growth (turgor)is largely maintained. Cell-wall plasticity (E_pl) and elasticity(E_Ql were determined with a constant load extensiometer bothin vivo (turgid coleoptile segments) and in vitro (frozen-thawedsamples). Cessation of coleoptile elongation was correlatedwith a 95% reduction in E_pl9 whereas E_Ql was only slightly affected.Extension kinetics were measured with living and frozen-thawedsegments cut from growing and non-growing coleoptiles. The correspondingstress-strain (load-extension) curves indicate that the cellwall of the growing coleoptile behaves like an elastic-plasticmaterial whereas that of the non-growing organ shows the behaviourof an elastic solid. These data demonstate that E_pl representsa true plastic (irreversible) deformation of the cell wall.It is concluded that cessation of coleoptile growth after emergenceof the primary leaf is attributable to a loss of cell-wall plasticity.Hence, a mechanical stiffening of the cell wall and not a lossof turgor pressure may be responsible for the deceleration ofcell elongation in the rye coleoptile. Key words: Extension growth, rye coleoptile, cell-wall extensibility, turgor pressure     

Action Potentials in Lupinus angustifolius L. Shoots: II. DETERMINATION OF THE STRENGTH-DURATION RELATION AND THE ALL-OR-NOTHING LAW

In Lupinus shoots an electrical stimulus (d.c.) produces a potentialwave analogous to the action potential wave (AP) in stimulatedsimple plant cells or in nerves. The method used (Paszewskiand Zawadzki, 1973a, b) is similar to those applied in neurophysiologyin research on the excitability of nerves. In the present paper the strength-duration relation (Eqn. (1))of the excitation in Lupinusitalic has been used to calculatethe values of the rheobase, chronaxie, and useful time of pulse.The rheobase, as a threshold value of the stimulus, is examinedin terms of the applicability of the all-or-nothing law. The results suggest that the origin and propagation of AP inplants and in nerves may occur in a similar way. The analogybetween the impulse propagation in nerve trunks and in Lupinusshoots is discussed.     

Ionic Currents across the Plasmalemma of Chara inflata Cells: III. WATER-RELATIONS PARAMETERS AND THEIR CORRELATION WITH MEMBRANE ELECTRICAL PROPERTIES

The water-relations parameters of Chara inflata cells were determineddirectly using the micro pressure probe technique. The turgorpressure of cells in artificial pond water (₀ = 0.06 MPa) wasabout 0.65 MPa and the half-time (T_1/2) for water exchange wasabout 6.5 s. The calculated values of the hydraulic conductivity(L_P) were in the range 1–2 ? 10^–6m s^–1 (MPa)^–1.The volumetric elastic modulus () was 32.8 MPa for turgor rangingfrom 0.77 to 0.82 MPa. Large changes in the water-relations parameters and the electricalproperties of the membrane occurred when the turgor was decreasedto low values. These changes included: (i) a decrease in theT_1/2 for water exchange, (ii) an increase in L_P and (iii) depolarizationof the membrane potential difference (V_m). The micro pressure probe, which enabled the turgor pressureof the cell to be altered, was used in combination with thevoltage-clamp technique to determine the relationship betweenK⁺ and Cl^– conductances of the plasmalemma and the cellturgor. The K⁺ conductance increased reversibly as the turgorwas reduced in the range 0 to 0.6 MPa and the Cl^– -conductanceincreased as the turgor was reduced in the range 0.1 to 0.5MPa. It is suggested that these pressure-dependent K⁺ and Cl^–conductances may have a dual role in electrical events and thenon-electrical responses such as changes in the cell volume. Key words: Chara inflata, membrane conductances, ion channels, water-relations parameters     

Manometric Measurement of Turgor Pressures in Laticiferous Phloem Tissues2

A manometric technique for the determination of turgor pressuresin laticiferous phloem tissues has given reproducible resultsin Hevea brasiliensis and a few other tree species. In Hevea,early morning pressures are in the range 7.9–15.0 atmospheres,falling during the day and recovering at night. These diurnalpressure changes are positively correlated with atmosphericrelative humidity and negatively correlated with changes intemperature, evaporation, leaf water deficit, and stomatal opening.They are not displayed by trees devoid of leaves. Thus the lossin turgor is probably the result of withdrawal of water fromthe phloem tissues under transpirational stress. Pressures at the base of the trunk normally exceed those atthe top, the gradient usually approximating to 1 atmosphere/10metres at night and rising up to six times this figure duringthe day. This increase probably reflects the development oftension gradients in the xylem during transpiration. A generalturgor gradient from base to crown does not preclude mass flowin sieve-tubes in the opposite direction provided that ratesof loading and unloading are such that a sufficient osmoticgradient is maintained in them in the required direction. No marked long-term effects of regular tapping on turgor pressurehave been noted in Hevea trees and there is no evidence forseasonal changes in turgor under our conditions.     

Involvement of a Calcium-Dependent Protein Kinase in Hypoosmotic Turgor Regulation in a Brackish Water Characeae Lamprothamnium succinctum

Calcium ion is a key messenger in turgor regulation of internodalcells of Lamprothamnium succinctum in response to hypoosmotictreatment. An increase in the concentration of cytosolic freecalcium ion ([Ca²⁺]_c) is prerequisite for the turgor regulation[Okazaki and Tazawa (1990) J. Membr. Biol. 114: 189], We examinedwhether or not a calcium-dependent protein kinase (CDPK) isinvolved in the Ca²⁺-mediated turgor regulation of Lamprothamniumcells. A 53-kDa CDPK which phosphorylated preferentially histoneH1 but poorly myelin basic protein or casein, was detected inthe cell extract of Lamprothamnium by an in-gel protein kinaseassay. This protein kinase was detected by Western blottingand was immunoprecipitated using an anti-Dunaliella tertiolectaCDPK antibody which can neutralize the Dunaliella CDPK activity[Yuasa et al. (1995) Plant Cell Physiol. 36: 699]. The 53-kDaCDPK was partially purified from Lamprothamnium and its activitywas shown to be inhibited by the antibody and K-252a, a proteinkinase inhibitor. Microinjection of the antibody into the cytosblof Lamprothamnium cells inhibited the decrease in turgor pressurein response to hypoosmotic treatment. However, a transient increasein [Ca²⁺]_c, which was suggested by a transient reduction ofthe velocity of cytoplasmic streaming, was induced in antibody-injectedcells after hypoosmotic treatment. Turgor regulation upon hypoosmotictreatment was inhibited when the cells were treated with K-252a.These results imply that CDPK of Lamprothamnium functions ata down-stream position of Ca²⁺-mobilization in processing turgorregulation in response to hypoosmotic treatment. ² These authors contributed equally to the work.     

The Contribution of Photosynthate to Turgor Pressure Rise in the Planktonic Blue-green Alga Anabaena flos-aquae

The turgor pressure of the cells of the planktonic blue-greenalga Anabaena flos-aquae increases following transfer of thealga from low light intensity to high light intensity. Up tohalf of this rise in turgor pressure may be due to the accumulationof soluble photosynthate. This estimate is based on direct measurementof cell water, extraction and fractionation of ¹⁴C-labelledsoluble products of photosynthesis, and simultaneous measurementof turgor pressure rise.     

Turgor and Longitudinal Tissue 12Helianthus annuus

The quantitative relationship between turgor and the pressureexerted by the inner tissues (cortex, vascular tissue, and pith)on the peripheral cell walls (longitudinal tissue pressure)was investigated in hypocotyls of sunflower seedlings (Helianthusannuus L.) In etiolated hypocotyls cell turgor pressures, asmeasured with the pressure probe, were in the range 0·38to 0·55 MPa with an average of 0·48 MPa. In irradiatedhypocotyls turgor pressures varied from 0·40 to 0·57MPa with a, mean at 0·49 MPa. The pressure exerted bythe inner tissues on the outer walls was estimated by incubatingpeeled sections in a series of osmotic test solutions (polyethyleneglycol 8000). The length change was measured with a transducer.In both etiolated and irradiated hypocotyls an external osmoticpressure of 0·5 MPa was required to inhibit elongationof the inner tissues, i.e. the average cell turgor and the longitudinaltissue pressure are very similar quantities. The results indicatethat the turgor of the inner tissues is displaced to and borneby the thick, growth-limiting peripheral cell walls of the hypocotyl. Key words: Helianthus annuus, hypocotyl growth, tissue pressure, turgor pressure, wall stress     

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

Previous single-cell studies on the upper epidermis of barleyleaves have shown that cells differ systematically in theirsolute concentrations depending on their location relative tostomatal pores and veins and that during NaCl stress, gradientsin osmotic pressure () develop (Fricke et al., 1995, 1996; Hinde,1994). The objective of the present study was to address thequestion to which degree these intercellular differences insolute concentrations and it are associated with intercellulardifferences in turgor or water potential (). Epidermal cellsanalysed were located at various positions within the ridgeregions overlying large lateral or intermediate veins, in thetrough regions between those veins or in between stomata (i.e.interstomatal cells). Turgor pressure of cells was measuredusing a cell pressure probe, and of extracted cell sap wasdetermined by picolitre osmometry. For both large and intermediatelateral veins, there were no systematic differences in turgorbetween cells located at the base, mid or top of ridges, regardlessof whether plants were analysed at low or high PAR (10 or 300–400µmol photons m^–2 s^–1). However, turgor withina ridge region was not necessarily uniform, but could vary byup to 0.14 MPa (1.4 bar) between adjacent cells. In 60 out of63 plants, turgor of ridge cells was either slightly or significantlyhigher than turgor of trough (lowest turgor) or interstomatalcells (intermediate turgor). The significance and magnitudeof turgor differences was higher in plants analysed under highPAR or local air flow than in plants analysed under low PAR.The largest (up to 0.41 MPa) and consistently significant differencesin turgor were found in plants treated for 3–9 d priorto analysis with 100 mM NaCl. For both NaCl-treated and non-treated(control) plants, differences in turgor between cell types weremainly due to differences in since differences in were negligible(0.01–0.04 MPa). Epidermal cell , in NaCl-treated plantswas about 0.38 MPa more negative than in control plants dueto higher . Turgor pressures were similar. Following a suddenchange in rooting-medium or air humidity, turgor of both ridgeand trough cells responded within seconds and followed the sametime-course of relaxation. The half time (T_1/2) of turgor relaxationwas not limited by the cell's T_1/2 for water exchange. Key words: Barley leaf epidermis, cell turgor, heterogeneity, NaCl stress, osmotic pressure, water potential     

Cell elongation, turgor and osmotic pressure in developing sunflower hypocotyls

The relationship between cell elongation, change in turgor andcell osmotic pressure was investigated in the sub-apical regionof hypocotyls of developing sunflower seedlings (Helianthusannuus L.) that were grown in continuous white light. Cell turgorwas measured with the pressure probe. The same hypocotyl sectionswere used for determination of osmotic pressure of the tissuesap. Acceleration of cell elongation during the early phaseof growth was accompanied by a 25% decrease in both turgor andosmotic pressure. During the linear phase of growth both pressuresremained largely constant. The difference between turgor andosmotic pressure (water potential) was –0.10 to –0.13MPa. Excision of one cotyledon had no effect on growth, turgorand osmotic pressure. However, after removal of both cotyledonscell elongation ceased and a substantial decrease in both pressureswas measured. In addition, we determined the longitudinal tissuepressure in seedlings from which one or both cotyledons hadbeen removed. Tissue pressure and turgor were very similar quantitiesunder all experimental conditions. Our results demonstrate thatturgor and cell osmotic pressure show a parallel change duringdevelopment of the stem. Cessation of cell elongation afterremoval of the cotyledons is attributable to a decrease in turgor(tissue) pressure, which provides the driving force for growthin the hypocotyl of the intact plant. Key words: Cell elongation, Helianthus annuus, osmotic pressure, tissue pressure, turgor     

Solute Leakage from the Isolated Parenchyma of Allium cepa and Kalanchoe daigremontiana

Efflux of different solutes from leaf slices of Kalanchoëdaigremontiana and from slices of the onion bulb scale was reinvestigatedwith respect to (1) dependence on turgor, (2) selectivity, (3)integrity of protoplasts and cellular changes. In both materials efflux of solutes (electrolytes or sugars)is non-selective and strongly dependent on turgor. Treatmentof tissue slices with hypotonic solutions (below a criticalosmotic pressure) resulted in high leakage rates, an increasein free space and an increased number of damaged cells. Lowconcentrations of calcium did not prevent this loss of retentionand cell stability. Part of the surviving cells were found to have a strongly decreasedosmotic pressure of cell sap. Leakage did not occur simultaneouslyat all cells of the tissue slice. It can be concluded that effluxfrom parenchyma cells in hypotonic solutions results from irreversibleosmotic breakdown and reversible membrane defects both favouredby high turgor. Key words: Parenchyma cells, Allium cepa, Kalanchoé daigremontiana, Solute efflux, Viability, Permeability, Plasmoptysis     

A critical re-examination of pressure-volume analysis of conifer shoots: comparison of three procedures for generating PV curves on shoots of Pinus resinosa Ait. seedlings

Prediction of water relations attributes for red pine (Pinusresinosa Ait.) derived from pressure-volume (PV) curves varieddepending on which of three methods was used. The sap expressionmethod entailed the enclosure of a shoot in a pressure chamberand expression of xylem sap by applying a constant selectedpressure until sap flow ceased, at which point xylem water potentialand shoot weight were measured. A sap expression PV curve wasformed by aggregating pairs of water potential-weight measurements,each pair supplied by one of 25 shoots. The repeat pressurizationmethod involved repeatedly measuring xylem water potential andshoot weight on a single shoot drying on a laboratory bench.Repeat pressurization PV curves were constructed from data providedby a single shoot. The composite method utilized single measurementsof xylem water potential and shoot weight on 25-30 differentshoots ranging in relative water content from about 1.0 to 0.5achieved by bench drying. Composite PV curves were constructedfrom aggregate data supplied by a population of shoots. Therewas close agreement in all PV attributes generated using repeatpressurization and sap expression methods. In contrast, withthe composite PV method, there was a fundamental differencein the slope of the linear region of the PV curves, causingosmotic potentials at full turgor and turgor loss to be morenegative, and relative water content at turgor loss to be lowerand symplast fraction to be higher. Comparison of compositeand repeat pressurization PV curves over the same ranges inwater content did not eliminate differences in derived waterrelations attributes. Differences in water potential isothermsrelated to the PV procedures used suggest that prolongedor repeatedexposure to gas at high pressure may introduce errors in theestimation of water relations attributes. Key words: Pinus resinosa, pressure chamber, pressure volume, tissue water relations     

The Use of Coleoptile Responses to Water Stress to Differentiate Wheat Genotypes for Osmoregulation, Growth and Yield

Differences in osmoregulation between genotypes of wheat (Triticumaestivum L.) were identified by measuring responses of coleoptilesto water stress. Seeds were germinated in Petri dishes on germinationpads. Water stress was applied when coleoptiles were 1 cm longby either adding polyethylene glycol 6000 (20% w/w) or by removingthe lids of the dishes and allowing the pads to dry by evaporationfor a period of 2 d, or from the beginning of germination byvarying the amount of water in the germination pads. Measurementsof the water and osmotic potentials of shoots and roots (usingthermocouple psychrometers) were used to estimate turgor pressures.Wheat genotypes showed the same segregation for shoot and rootlengths and turgor pressures as they had done in previous studiesfor flag-leaf osmoregulation and yield. It was concluded thatobservations of the responses of coleoptile length to waterstress could be used in plant-breeding applications to identifydifferences in osmoregulation between genotypes of wheat. Triticum aestivum, osmoregulation, turgor maintenance, coleoptile growth, root, selection criterion