Psychrometric Field Measurement of Water Potential Changes following Leaf Excision

In situ measurement of sudden leaf water potential changes has not been performed under field conditions. A laboratory investigation involving the measurement of leaf water potential prior to and 2 to 200 minutes after excision of citrus leaves (Citrus jambhiri) showed good linear correlation (r = 0.99) between in situ leaf psychrometer and Scholander pressure chamber measurements. Following this, a field investigation was conducted which involved psychrometric measurement prior to petiole excision and 1 minute after excision. Simultaneous pressure chamber measurements were performed on neighboring leaves prior to the time of excision and then on the psychrometer leaf about 2 minutes after excision. These data indicate that within the first 2 minutes after excision, psychrometer and pressure chamber measurements were linearly correlated (r = 0.97). Under high evaporative demand conditions, the rate of water potential decrease was between 250 and 700 kilopascals in the first minute after excision. These results show that the thermocouple psychrometer can be used as a dynamic and nondestructive field technique for monitoring leaf water potential.     

Pressure chamber measurements using two wheat leaves

Summary The most widely used technique of leaf water potential measurements is with the Scholander pressure chamber. Representative leaf water potential values require many determinations on individual leaves and this can be time consuming in large fields or experiments with multiple treatments. This paper describes a method of obtaining a mean value more rapidly, by using two leaves in the pressure chamber at the same time, but recording the end point of each leaf separately.     

In situ field measurement of leaf water potential using thermocouple psychrometers

Thermocouple psychrometers are the only instruments which can measure the in situ water potential of intact leaves, and which can possibly be used to monitor leaf water potential. Unfortunately, their usefulness is limited by a number of difficulties, among them fluctuating temperatures and temperature gradients within the psychrometer, sealing of the psychrometer chamber to the leaf, shading of the leaf by the psychrometer, and resistance to water vapor diffusion by the cuticle when the stomates are closed. Using Citrus jambhiri, we have tested several psychrometer design and operational modifications and showed that in situ psychrometric measurements compared favorably with simultaneous Scholander pressure chamber measurements on neighboring leaves when the latter were corrected for the osmotic potential.     

Short communication. Comparative measurements of xylem pressure in transpiring and non-transpiring leaves by means of the pressure chamber and the xylem pressure probe

Simultaneous measurements were made with the xylem pressure probe on exposed, transpiring leaves and with the Scholander pressure chamber on both transpiring and covered, non-transpiring leaves of sugarcane and maize plants. Xylem tensions inferred from pressure chamber balancing pressures on non-transpiring leaves were similar to those measured directly with the xylem pressure probe in transpiring leaves. However, tensions inferred with the pressure chamber on transpiring leaves that were placed in plastics bags just prior to excision were up to 0.6 MPa greater than those measured concurrently with the xylem pressure probe. These findings suggest that relatively large differences in water potential between the xylem and bulk leaf tissue can exist during periods of rapid transpiration, and they confirm that the balance pressure of an excised, previously transpiring leaf is only a measure of the bulk average equilibrium leaf water potential and not of the true xylem pressure that existed prior to excision.Key words: Cohesion-Tension theory, xylem pressure probe, pressure chamber, xylem tension.      

Effect of Cuticular Abrasion on Thermocouple Psychrometric In Situ Measurement of Leaf Water Potential

Cuticular resistance to water vapour diffusion between the substomatalcavity and the sensing psychrometer junction is a problem uniqueto leaf hygrometry. This resistance is not encountered in soilor solution hygrometry. The cuticular resistance may introduceerror in the measurement of leaf water potential. Using in situleaf hygrometers, we studied the effect of abrading the cuticleof Citrus jambhiri Lushington leaves, to reduce the diffusiveresistance. Field measurements of psychrometer water potentialwere compared with Scholander pressure chamber values for adjacentleaves. Different treatments were compared by sealing pairsof psychrometers on either side of the midrib. The time forwater vapour equilibration between the leaf and the psychrometerchamber was greater than 5 h for no abrasion. For abraded leaves,the true water potential value was obtained within an hour.After equilibration, psychrometer values compared favourablywith pressure chamber values for adjacent leaves (r > 0.97).Measured water potential for unabraded leaves did not correlatewell with corresponding pressure chamber measurements. Scanning electron micrographs indicated that the damage causedby abrading leaves for 60 s using carborundum powder (60 µmdiameter) was surface localized, with numerous scratchings ofthe leaf cuticle. The coarse abrasion treatment (aluminium oxide,75 µm diameter) resulted in fewer but larger cavitiesin the epidermis, which may explain the observed variabilityin the corresponding psychrometric measurements. Key words: Leaf water potential, Cuticular resistance, Leaf abrasion, Thermocouple psychrometer     

The Use of the Pressure Chamber Technique for Measurement of the Water Potential of Transpiring Plant Organs

The existence of water potential gradients in flowering shoots and leaves of roses (Rosa sp., cv. Baccara) and along flag leaves of wheat (Triticum aestivum L.) were studied by means of the Scholander pressure chamber. In roses grown in greenhouse, the water potential measured in transpiring shoots was higher than in leaves detached from these shoots, whereas the potential differences between leaf and shoot after equilibration in the dark were small or negligible. A progressive decrease in water potential was found upon repeated measurement on the same organ; this decline was steeper in leaves than in shoots. Extrapolating this decline to excision time resulted in water potential values which, in transpiring shoots, were 3 to 5 bars higher than in leaves. Detopping the flower bud did not alter this pattern, indicating that the highest water potential in the shoot was in the stem. In field-grown wheat, the water potential measured in a whole flag leaf was about 6 bars higher than that measured in the apical one-third of the leaf, and this difference disappeared after equilibrating the detached leaf for 1 h in the dark. These potential differences indicate the presence of resistances along the water path in the organ. The results obtained by the pressure chamber represent the highest water potential in the organ, rather than the average water potential.     

Differences between tree species in hydraulic press calibration of leaf water potential are correlated with specific leaf area

Abstract To determine the usefulness of the J-14 Hydraulic Press (Campbell Scientific, Inc., Logan, Utah, U.S.A.) in estimating leaf water potential, we calibrated the J-14 Press against a Scholander-type pressure chamber for leaves of various tree species. The species tested were: Acer saccharum, Acer negundo, Acer rubrum. Populus tremuloides, Populus grandidentata, Quercus rubra, and Brassaia actinophylla (Schefflera). The regression calibrations were linear with standard errors about the regression less than 0.1 MPa. The regression equations for the four genera were significantly different, with the y- intercept increasing and the slope decreasing in order of decreasing specific leaf area (SLA). There were no significant differences between species of the calibration lines within the genera Acer and Populus. These data may indicate that leaves with lower SLA resist mechanical compression by the hydraulic press, causing the J-14 Press to be less sensitive to differences of leaf water potential. Therefore the J-14 Press is only a relative measure of leaf water status and does not measure leaf water potential.     

Stomatal Control of Photosynthesis of Eucalyptus globulus Labill. Trees under Field Conditions in Portugal

Pereira, J. S., Tenhunen, J. D. and Lange, O. L. 1987. Stomatalcontrol of photosynthesis of Eucalyptus globulus Labill. treesunder field conditions in Portugal.—J. exp. Bot. 38: 1678–1688. Stomatal behaviour of adult leaves of Eucalyptus globulus treeswas studied under field conditions in Portugal. In the absenceof severe plant water stress stomata were open when the summedtotal of photosynthetically active photon flux density incidenton both leaf surfaces was above 100 µmol m²s¹ and leafconductance to water vapour reached 245 mmol m ² s¹ on a total(both epidermes) leaf area basis. The stomata of both leaf epidermesresponded similarly to changes in solar radiation and waterstress. Water stress resulted in decreasing daily maxima inleaf conductance as predawn leaf water potential decreased.Maximal leaf conductance decreased to less than 50 mmol m ²s ¹ when predawn leaf water potential decreased below —1·0MPa. At similar values of predawn leaf water potential stomatawere more closed as the leaf to air water vapour partial pressuredifference increased. The effect of increasing air dryness onstomata was greatest at high predawn leaf water potential. Dailymaxima in photosynthetic rates and in leaf conductance werelinearly related to one another in spring and summer. Both decreasedwith increase in leaf water stress. In autumn and winter, increasesin leaf conductance occurring under natural conditions duringthe course of the day were not necessarily accompanied by increasesin net photosynthesis. Stomata were more closed in the afternoonthan in the morning at the same rates of net photosynthesis,temperature or leaf to air water vapour partial pressure difference. Key words: Eucalyptus globulus,, photosynthesis, stomata, water stress.     

An analysis of resistance to water flow through wheat and tall fescue leaves during pressure chamber efflux experiments

Abstract. This is a physical analysis of water movement in wheat ( Triticum ) and tall fescue ( Festuca arundinacea ) leaves placed in the Scholander pressure chamber. It takes into account the efflux resistances of water movement through the xylem and water flow across the cell membranes. Xylem resistance was estimated using Poiseuille's law.
Leaves which had been pressurized in the chamber were embedded, sectioned, examined under a light microscope and photographed. Cells were intact but distorted and xylem vessels were intact. Cells in portions of the blade squeezed by the chamber sealing grommet were crushed, but xylem vessels remained intact.
By applying pressure several tenths of a megapascal in excess of the balance pressure, water was forced from each leaf through the severed end which protruded from the chamber. Efflux curves were drawn by plotting the total water expressed as a function of time after the pressure increase. Water efflux from the shortest wheat leaf lasted only 10 min while efflux from the longest continued for up to 40 min. The efflux from a tall fescue leaf which was rehydrated and cut to a shorter length was much more rapid than efflux from the original leaf.
Experiments combined with mathematical analysis suggested that the effect of leaf length on efflux is related to a high resistance to water flow through vascular bundles. Xylem resistance would be sufficient to produce this effect if it were 10 times greater than that predicted by Poiseuille's law. Both the observations of water flow from the cut end of the leaf and the mathematical model suggested very little water flows from bundles with vessels of diameter less than 12 μm. The apparent explanation is high resistance to water flow through these small diameter vessels.     

The temperature dependence of shoot hydraulic resistance: implications for stomatal behaviour and hydraulic limitation

Recent soil pressurization experiments have shown that stomatal closure in response to high leaf–air humidity gradients can be explained by direct feedback from leaf water potential. The more complex temperature‐by‐humidity interactive effects on stomatal conductance have not yet been explained fully. Measurements of the change in shoot conductance with temperature were made on Phaseolus vulgaris (common bean) to test whether temperature‐induced changes in the liquid‐phase transport capacity could explain these temperature‐ by‐humidity effects. In addition, shoot hydraulic resistances were partitioned within the stem and leaves to determine whether or not leaves exhibit a greater resistance. Changes in hydraulic conductance were calculated based on an Ohm’s law analogy. Whole‐plant gas exchange was used to determine steady‐ state transpiration rates. A combination of in situ psychrometer measurements, Scholander pressure chamber measurements and psychrometric measurements of leaf punches was used to determine water potential differences within the shoot. Hydraulic conductance for each portion of the pathway was estimated as the total flow divided by the water potential difference. Temperature‐induced changes in stomatal conductance were correlated linearly with temperature‐induced changes in hydraulic conductance. The magnitude of the temperature‐induced changes in whole‐plant hydraulic conductance was sufficient to account for the interactive effects of temperature and humidity on stomatal conductance.     

The Use of 14C-Ethane Diol as a Quantitative Tracer for the Transpirational Volume Flow of Water and an Investigation of the Effects of Salinity upon Transpiration, Net Sodium Accumulation and Endogenous ABA in Individual Leaves of Oryza sativa L.

Yeo, A. R., Yeo, M. E., Caporn, S. J. M., Lachno, D. R. andFlowers, T. J. 1985. The use of ¹⁴C-ethane diol as a quantitativetracer for the transpirational volume flow of water and an investigationof the effects of salinity upon transpiration, net sodium accumulationand endogenous ABA in individual leaves of Oryza sativa L.—J.exp. Bot. 36: 1099–1109. Oryza sativa L. (rice) seedlings growing in saline conditionsexhibit pronounced gradients in leaf sodium concentration whichis always higher in the older leaves than the younger ones.Individual leaf transpiration rates have been investigated todiscover whether movement of sodium in the transpiration streamis able to explain these profiles from leaf to leaf. The useof ¹⁴C labelled ethane diol to estimate transpiration was evaluatedby direct comparison with values obtained by gas exchange measurements.Ethane diol uptake was linearly related to the transpirationalvolume flow and accurately predicted leaf to leaf gradientsin transpiration rate in saline and non-saline conditions. ¹⁴C-ethanediol and ²²NaCl were used to compare the fluxes of water andsodium into different leaves. The youngest leaf showed the highesttranspiration rate but the lowest Na accumulation in salineconditions; conversely, the older leaves showed the lower transpirationrates but the greater accumulation of Na. The apparent concentrationof Na in the xylem stream was 44 times lower into the youngerleaf 4 than into the older leaf 1. Exposure to NaCl (50 molm^–3) for 24 h elicited an increase in endogenous ABA inthe oldest leaf only, but no significant changes occurred inthe younger leaves. Key words: —Salinity, rice, Oryza sativa L., transpiration, volume flow, abscisic acid     

An Impasse in Plant Water Relations?

Balling and Zimmermann [Planta 182 (1990), 325–338] used a pressure probe to measure directly negative pressures in the xylem of transpiring plants. They obtained data that challenge the standard framework that plant physiologists use when thinking about plant water relations, and, most notably, found a substantial discrepancy between their measurements of xylem pressure and of leaf water potential measured with a Scholander pressure bomb. Their data are critically examined and it is shown that most of them can be accommodated within the established principles of plant water relations. In particular, there are several reasons, consistent with the established principles, why leaf water potential and xylem pressure may differ.     

Field Estimates of Root and Xylem Resistances in Pinus contorta using Root Excision

A root excision technique was used to estimate the proportionof total resistance to water flux residing in the soil, theroot, and the xylem of lodgepole pine (Pinus contorta Douglex. Loud.) trees in the field. Root excision at mid-day alwaysresulted in rapid recovery of leaf water potential when waterwas supplied to the cut stem, suggesting a high soil-root resistance.Transpiration was unaffected if leaf water potential beforecutting was not limiting leaf conductance. By mid-June wateruptake by the excised stem always exceeded calculated crowntranspiration indicating recharge of internal sapwood storage.Predawn leaf water potential before root excision was highlycorrelated with total soil-plant resistance (r² = 0·89)and calculated root water uptake (r² = 0·92).     

Leaf water relations of black alder [Alnus glutinosa (L.) Gaertn.] growing at neighbouring sites with different water regimes

 The tree species black alder [Alnus glutinosa (L.) Gaertn.] typically inhabits wet sites in central Europe but is also successful on well drained soils. To test the physiological adjustment of the species in situ, conductances, transpiration rates and water potentials (Scholander pressure chamber) of black alder leaves were investigated at two neighbouring sites with different water regimes: alder trees at an occasionally water logged alder forest and alder shrubs in a nearby, much drier hedgerow. Additional experiments with alder cuttings in nutrient culture showed that leaf conductances and gas exchange were both strongly influenced by the substrate water potential. In situ however, there was little spatial variability within the different parts of a crown and we found that physiological regulation at leaf level was hardly influenced by different site water regimes or different tree sizes. Diurnal courses of leaf water relations as well as their regulation at the leaf level (e.g. the hyperbolic relationship between conductances and ΔW) were strikingly similar at both sites. Leaf water potential in black alder was shown to be a consequence of immediate transpiration rates, which were high in comparison to other tree species (up to 4 mmol H₂O m^–2 s^–1), rather than the water potentials being a factor that influenced conductance and, therefore, transpiration. The always high leaf conductances and consequent high transpiration rates are interpreted as a strategy to maximise productivity through low stomatal limitation at sites where water supply is usually not limited. However, at the same time this behaviour restricts black alder to sites where at least the deep-going roots can exploit water. Received: 10 September 1998 / Accepted: 12 January 1999     

Leaf water potentials measured with a pressure chamber

Leaf water potentials were estimated from the sum of the balancing pressure measured with a pressure chamber and the osmotic potential of the xylem sap in leafy shoots or leaves. When leaf water potentials in yew, rhododendron, and sunflower were compared with those measured with a thermocouple psychrometer known to indicate accurate values of leaf water potential, determinations were within ± 2 bars of the psychrometer measurements with sunflower and yew. In rhododendron. water potentials measured with the pressure chamber plus xylem sap were 2.5 bars less negative to 4 bars more negative than psychrometer measurements.

The discrepancies in the rhododendron measurements could be attributed, at least in part, to the filling of tissues other than xylem with xylem sap during measurements with the pressure chamber. It was concluded that, although stem characteristics may affect the measurements, pressure chamber determinations were sufficiently close to psychrometer measurements that the pressure chamber may be used for relative measurements of leaf water potentials, especially in sunflower and yew. For accurate determinations of leaf water potential, however, pressure chamber measurements must be calibrated with a thermocouple psychrometer.

     

A Rapid Field Method for Measuring Photosynthesis with Labelled Carbon Dioxide

A portable field apparatus for the measurement of photosynthesisis described. Short pulses (20 s) of air containing ¹⁴CO₂ areapplied to circular leaf areas of 1 cm², enclosed in a transparentminiature chamber; the radioactivity of the leaf areas is measuredby liquid scintillation, using a new in-vial wet combustiontechnique. Typical daily courses of photosynthesis in maizeand cotton are presented.     

A Handpiece for the Simultaneous Measurement of Photosynthetic Rate and Leaf Diffusive Conductance: I. DESIGN

The design and construction of a handpiece for the simultaneousmeasurement of photosynthetic rate and diffusive conductanceof cereal leaves in almost natural conditions in the field isdescribed. Photosynthetic rate is found from the assimilationof ¹⁴CO₂ by part of the leaf which is temporarily enclosed ina small hand-held chamber. This chamber also acts as a stirred,steady-state water vapour diffusion porometer, allowing thesimultaneous measurement of relative humidity from which theleaf diffusive conductance is estimated. The instrument alsomeasures the leaf and air temperatures and incident photon fluxdensity. The important criteria of the performance of such ahandpiece are discussed, and the sensors which measure the physicalvariables of humidity, temperature, and photon flux densityare described. An automatic sequencing system built from logiccircuits which displays the measured values of these variablesand times the operations is also described.     

In-Phase Cycling of Photosynthesis and Conductance at Saturating Carbon Dioxide Pressure Induced by Increases in Water Vapour Pressure Deficit

Bunce, J. A. 1987. In-phase cycling of photosynthesis and conductanceat saturating carbon dioxide pressure induced by increases inwater vapour pressure deficit.—J. exp. Bot. 38: 1413–1420. The leaf to air water vapour deficit was increased suddenlyfrom about 1·0 to 2·5 IcPa for single leaves ofsoybean (Glycine max L. Merr.) plants held at 30 °C, 2·0mmol m ^–2 s^–1 photosynthetic photon flux density(PPFD) and carbon dioxide pressures saturating to photosynthesis.After a lag of about 10 min, photosynthetic rate and stomatalconductance to water vapour began to decrease, and then cycledin phase with each other. The period of the cydes was about20 min. During these cycles the substomatal carbon dioxide pressurewas constant in the majority of leaves examined, and was alwaysabove saturation for photosynthesis. Epidermal impressions showedthat most stomata changed in aperture during the cycles, andthat very few were ever fully closed. Water potential measuredon excised discs changed by at most 0·1 MPa from theminima to the maxima in transpiration rate. In contrast, forleaves of sunflower (Helianthus animus L.) grown at low PPFD,the increase in VPD led to leaf wilting and decreased photosynthesis,followed by recovery of turgor and photosynthesis as stomatalconductance began to decrease. In these leaves photosynthesisand conductance then cycled approximately 180° out of phase.It is suggested that in soybeans decreased leaf conductanceinduced by high VPD provided a signal which decreased the rateof photosynthesis at carbon dioxide saturation by a mechanismthat was not related to a water deficit in the mesophyll. Key words: Photosynthesis, stomatal conductance, cycling, vapour pressure deficit     

Xylem pressure potential and chlorophyll fluorescence as indicators of freezing survival in black locust and Western hemlock seedlings

Xylem pressure potential was determined using the Scholander pressure chamber on stems of cold hardened and non-hardened black locust (Robinia pseudoacacia L.) seedlings following freezing to various nonlethal and lethal temperatures and subsequent thawing. Correlation was found between immediate xylem pressure potential and long-term seedling survival. Chlorophyll fluorescence transients were monitored using needles of western hemlock (Tsuga heterophylla (Raf.) Sarg.) seedlings following freezing to various non-lethal and lethal temperatures and subsequent thawing. Immediate and repeatable differences in fluorescence transients correlated with long-term seedling survival. Methodology is described and correlations discussed relative to using either chlorophyll fluorescence or xylem pressure potential as an immediate indicator of long-term freezing survival in woody plant seedlings.     

Apparatus for the Simultaneous Measurement of Water Vapour and Carbon Dioxide Exchanges of Single Leaves

Equipment is described which delivers air with concentrationsof CO₂ and water vapour closely controlled in the ranges 0 to2500 ppm and 5 to 15 mb respectively, at flow rates of up to10 1 min^-1, to each four leaf chambers. The leaf temperatureis controlled to ±0.5 °C and, with a light intensityof 0.3 cal cm^-2 min^-1 visible radiation (0.4 to 0.7 µm)leaf temperature can be maintained at 17.5 °C.The apparatusused to measure the concentration differences between the watervapour and CO₂ entering and leaving the leaf chamber (used tocalculate transpiration, photosynthetic, and respiration rates)is described in detail.Results of tests, which show the necessityfor mounting a fan within the leaf chamber, are reported.Typicallight- and CO₂-response curves are given for kale leaves (Brassicaoleracca var. acephala) and an attempt is made to quantify theerrors in the measurement of photosynthesis and transpiration.