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.     

Transport constraints on water use by the Great Basin shrub, Artemisia tridentata

As soil and plant water status decline, decreases in hydraulic conductance can limit a plant's ability to maintain gas exchange. We investigated hydraulic limitations for Artemisia tridentata during summer drought. Water use was quantified by measurements of soil and plant water potential ( Ψ ), transpiration and leaf area. Hydraulic transport capacity was quantified by vulnerability to water stress-induced cavitation for root and stem xylem, and moisture release characteristics for soil. These data were used to predict the maximum possible steady-state transpiration rate ( E _crit) and minimum leaf xylem pressure ( Ψ _crit). Transpiration and leaf area declined by ~ 80 and 50%, respectively, as soil Ψ decreased to –2·6 MPa during drought. Leaf-specific hydraulic conductance also decreased by 70%, with most of the decline predicted in the rhizosphere and root system. Root conductance was projected to be the most limiting, decreasing to zero to cause hydraulic failure if E _crit was exceeded. The basis for this prediction was that roots were more vulnerable to xylem cavitation than stems (99% cavitation at –4·0 versus –7·8 MPa, respectively). The decline in water use during drought was necessary to maintain E and Ψ within the limits defined by E _crit and Ψ _crit.     

Theoretical and experimental errors for in situ measurements of plant water potential

Errors in psychrometrically determined values of leaf water potential caused by tissue resistance to water vapor exchange and by lack of thermal equilibrium were evaluated using commercial in situ psychrometers (Wescor Inc., Logan, UT) on leaves of Tradescantia virginiana (L.). Theoretical errors in the dewpoint method of operation for these sensors were demonstrated. After correction for these errors, in situ measurements of leaf water potential indicated substantial errors caused by tissue resistance to water vapor exchange (4 to 6% reduction in apparent water potential per second of cooling time used) resulting from humidity depletions in the psychrometer chamber during the Peltier condensation process. These errors were avoided by use of a modified procedure for dewpoint measurement. Large changes in apparent water potential were caused by leaf and psychrometer exposure to moderate levels of irradiance. These changes were correlated with relatively small shifts in psychrometer zero offsets (−0.6 to −1.0 megapascals per microvolt), indicating substantial errors caused by nonisothermal conditions between the leaf and the psychrometer. Explicit correction for these errors is not possible with the current psychrometer design.     

A temperature-controlled plant psychrometer

When measuring plant water potential in conditions of fluctuating temperature, commercial psychrometers exhibit large errors which are caused by a temperature difference between the reference junctions and the chamber air. A novel psychrometer capable of field use is described which incorporates an electronic device for controlling reference junction temperatures. It consists of a chamber with a thermocouple mounted in it for measuring dewpoints and another thermocouple for sensing the temperature gradients. A thermo-electric heat-pump keeps the temperature of the reference junctions the same as that of the chamber air. Good agreement was found between measurements made with this psychrometer and independent measurements made with the destructive pressure chamber method in the glasshouse and in the field under conditions of widely fluctuating temperature. Corresponding measurements with a commercial psychrometer without temperature control failed to show such agreement. Heavy thermal insulation is necessary, despite the temperature control, which can restrict the psychrometers used. Possibilities for further improvements are discussed.     

A rapid leaf-disc sampler for psychrometric water potential measurements

An instrument was designed which facilitates faster and more accurate sampling of leaf discs for psychrometric water potential measurements. The instrument consists of an aluminum housing, a spring-loaded plunger, and a modified brass-plated cork borer. The leaf-disc sampler was compared with the conventional method of sampling discs for measurement of leaf water potential with thermocouple psychrometers on a range of plant material including Gossypium hirsutum L., Zea mays L., and Begonia rex-cultorum L. The new sampler permitted a leaf disc to be excised and inserted into the psychrometer sample chamber in less than 7 seconds, which was more than twice as fast as the conventional method. This resulted in more accurate determinations of leaf water potential due to reduced evaporative water losses. The leaf-disc sampler also significantly reduced sample variability between individual measurements. This instrument can be used for many other laboratory and field measurements that necessitate leaf disc sampling.     

The origin and theoretical correction of thermal gradients in thermocouple psychrometers

Temperature-compensated psychrometers are explained and a theoretical correction for temperature gradients, with its boundaries, is given. The correction was elaborated for in situ determination of water potential on soybean and tomato stems where it was indispensable. The origin of the thermal gradients in the psychrometric chamber is then discussed. Cooling of the conducting tissue by the circulation of xylem sap seemed to be the major cause for these gradients, while the heating of the psychrometer (thermally insulated) by the surroundings through radiative or conductive transfer was negligible.     

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     

Chloroplast response to low leaf water potentials: I. Role of turgor

The effect of decreases in turgor on chloroplast activity was studied by measuring the photochemical activity of intact sunflower (Helianthus annuus L. cv. Russian Mammoth) leaves having low water potentials. Leaf turgor, calculated from leaf water potential and osmotic potential, was found to be affected by the dilution of cell contents by water in the cell walls, when osmotic potentials were measured with a thermocouple psychrometer. After the correction of measurements of leaf osmotic potential, both the thermocouple psychrometer and a pressure chamber indicated that turgor became zero in sunflower leaves at leaf water potentials of −10 bars. Since most of the loss in photochemical activity occurred at water potentials below −10 bars, it was concluded that turgor had little effect on the photochemical activity of the leaves.     

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.     

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.     

Measuring water potential (activity) from free water to oven dryness

Water activities (potentials) in plant materials were measured over the range from free water to oven dryness with a Spanner thermocouple psychrometer. In a two-step procedure, water was first condensed on the thermocouple junction for several minutes. The sample was then inserted under the wet thermocouple and the maximum psychrometric cooling was measured in about 10 seconds. Calibration was with saturated salt slurries of known water activities. Psychrometric cooling was a nearly linear function of the water activity and of the negative log of the water potential. The psychrometric cooling to water activity relationship agreed with wetbulb temperature depression to relative humidity relationships given in tables. Water activities of wheat grains and leaves decreased sharply in a curvilinear fashion as their water contents decreased. Some problems of the procedure are discussed.     

Comparison of Isoprene Emission, Intercellular Isoprene Concentration and Photosynthetic Performance in Water-Limited Oak (Quercus pubescens Willd. and Quercus robur L.) Saplings

Abstract: The influence of prolonged water limitation on leaf gas exchange, isoprene emission, isoprene synthase activities and intercellular isoprene concentrations was investigated under standard conditions (30 °C leaf temperature and 1000 μmol photons m^-2 s^-1 PPFD) in greenhouse experiments with five-year-old pubescent oak ( Quercus pubescens Willd.) and four-year-old pedunculate oak ( Quercus robur L.) saplings. Net assimilation rates proved to be highly sensitive to moderate drought in both oak species, and were virtually zero at water potentials (Ψ_pd) below - 1.3 MPa in Q. robur and below - 2.5 MPa in Q. pubescens . The response of stomatal conductance to water stress was slightly less distinct. Isoprene emission was much more resistant to drought and declined significantly only at Ψ_pd below - 2 MPa in Q. robur and below - 3.5 MPa in Q. pubescens . Even during the most severe water stress, isoprene emission of drought-stressed saplings was still approximately one-third of the control in Q. robur and one-fifth in Q. pubescens . Isoprene synthase activities were virtually unaffected by drought stress. Re-watering led to partial recovery of leaf gas exchange and isoprene emission. Intercellular isoprene concentrations were remarkably enhanced in water-limited saplings of both oak species during the first half of the respective drought periods with maximum mean values up to ca. 16 μl l^-1 isoprene for Q. pubescens and ca. 11 μl l^-1 isoprene for pedunculate oak, supporting the hypothesis that isoprene serves as a short-term thermoprotective agent in isoprene-emitting plant species.     

Effects of salt secretion on psychrometric determinations of water potential of cotton leaves

Thermocouple psychrometers gave lower estimates of water potential of cotton leaves than did a pressure chamber. This difference was considerable for turgid leaves, but progressively decreased for leaves with lower water potentials and fell to zero at water potentials below about −10 bars. The conductivity of washings from cotton leaves removed from the psychrometric equilibration chambers was related to the magnitude of this discrepancy in water potential, indicating that the discrepancy is due to salts on the leaf surface which make the psychrometric estimates too low. This error, which may be as great as 400 to 500%, cannot be eliminated by washing the leaves because salts may be secreted during the equilibration period. Therefore, a thermocouple psychrometer is not suitable for measuring the water potential of cotton leaves when it is above about −10 bars.     

Errors in measuring water potentials of small samples resulting from water adsorption by thermocouple psychrometer chambers

The adsorption of water by thermocouple psychrometer assemblies is known to cause errors in the determination of water potential. Experiments were conducted to evaluate the effect of sample size and psychrometer chamber volume on measured water potentials of leaf discs, leaf segments, and sodium chloride solutions. Reasonable agreement was found between soybean (Glycine max L. Merr.) leaf water potentials measured on 5-millimeter radius leaf discs and large leaf segments. Results indicated that while errors due to adsorption may be significant when using small volumes of tissue, if sufficient tissue is used the errors are negligible. Because of the relationship between water potential and volume in plant tissue, the errors due to adsorption were larger with turgid tissue. Large psychrometers which were sealed into the sample chamber with latex tubing appeared to adsorb more water than those sealed with flexible plastic tubing. Estimates are provided of the amounts of water adsorbed by two different psychrometer assemblies and the amount of tissue sufficient for accurate measurements of leaf water potential with these assemblies. It is also demonstrated that water adsorption problems may have generated low water potential values which in prior studies have been attributed to large cut surface area to volume ratios.     

A low-cost psychrometer for field measurements of atmospheric humidity

A design for an aspirated wet and dry bulb psychrometer, suitable for continuous field measurement of atmospheric humidity, is described. The psychrometer is simple and cheap to make, with a material cost of UK £40 per unit. It is mainly constructed from ready-made plastic plumbing parts, thus avoiding the need for expensive workshop facilities. The wet and dry bulb temperature sensors are suitable for direct connection to a solid-state data-logger. During extensive field use, it has proved reliable and easy to maintain. The accuracy of the device was assessed by intercomparison with an Assman psychrometer, a standard instrument for measuring atmospheric humidity. The test was conducted under high ambient saturation deficits (4.08 ± 1.22 kPa) and shortwave radiation fluxes in Niger, West Africa. Reasonable agreement between the two instruments was observed, with the low-cost psychrometer giving a mean saturation deficit 0.12 ± 0.25 kPa higher than the Assman.     

An evaluation of techniques for measuring yield turgor in excised Salix leaves

Abstract. The use of three techniques for determining yield turgor in excised Salix leaves was investigated. These were the osmotic-solutions technique, the psychrometer technique, and the pressure-chamber technique. The application of the osmotic-solutions technique to a range of leaf types was discussed and the appropriate corrections for volume changes and the contribution of apoplastic water were detailed. It was concluded that the osmotic-solutions technique is not satisfactory for use with slowly growing and/or very elastic leaves. The psychrometer and pressure-chamber techniques were both simple compared with the osmotic-solutions technique, and gave values for yield turgor in the range of 0·3–0·5 MPa. A disadvantage of the psychrometer technique for field applications is that it requires one psychrometer chamber per sample. The pressure-chamber technique was modified for use as a field technique where multiple sampling could be easily and inexpensively achieved. Particular care was required with this technique to prevent water loss from the leaf during stress relaxation, but simple and effective procedures for doing so were found. The modified pressure-chamber technique described here, is recommended as the preferred technique for measuring the yield turgor of leaves in experiments where many simultaneous estimates of yield turgor are to be made.     

A developmental study of Glycine max cell and protoplast isolation in relation to leaf age and photosynthetic competence

Leaf mesophyll cells were isolated from developing first trifoliate leaves of Glycine max (L.) Merr cv. Fiskeby V using a mechanical isolation procedure combined with low speed centrifugation. Cell yields of 17 ± 1.7% were routinely obtained with 55–75% intactness, as assessed by staining techniques, fluorescence transients and the ability of cells to convert to protoplasts after enzyme treatment. Rates of leaf photosynthesis were maximal in 27-day-old plants [280 μmol O₂ evolved (mg chlorophyll)^-1h^-1], from which isolated cells and protoplasts gave rates of up to 140 μmol O₂ evolved (mg chlorophyll)^-1 h^-1. Results are discussed in relation to leaf development and cell status during the attainment of photosynthetic competence.     

A new stem hygrometer, corrected for temperature gradients and calibrated against the pressure bomb

Abstract A simple stem hygrometer for attachment to a bared section of sapwood or a cross-sectional cut end of a shoot is described. Two welded chromelconstantan thermocouples inside the chamber, one touching the sample and the other in the chamber air, allowed measurement of and correction for the temperature gradient between the sample and the dewpoint measuring junction. The instrument was attached to the cut end of an apical shoot of Thuja occidentalis L. protuding from a Scholander-Hammel pressure bomb. Cut-end water potential (ψ_hyg), measured using the stem hygrometer, was compared to xylem pressure potential (ψ_xp) while the latter was manipulated in the pressure bomb. After an initial equilibration time of 3–4 h, hygrometer equilibrium values were achieved within 1.5–4.0 min of changing ψ_xp in the pressure bomb. The half-time (ψ_1/2) for vapour pressure equilibration was 15–40 s. Stable temperature gradients between the sample and dewpoint measuring junction of 0.01–0.1°C were measured. Correcting ψ_hyg for the temperature gradient resulted in excellent agreement with ψ_xp.     

Changes in leaf hydraulics and stomatal conductance following drought stress and irrigation in Ceratonia siliqua (Carob tree)

Changes in leaf hydraulic conductance (K) were measured using the vacuum chamber technique during dehydration and rehydration of potted plants of Ceratonia siliqua . K of whole, compound leaves as well as that of rachides and leaflets decreased by 20–30% at leaf water potentials (Ψ_L) of −1.5 and −2.0 MPa, i.e. at Ψ_L values commonly recorded in field-growing plants of the species. Higher K losses (up to 50%) were measured for leaves at Ψ_L of −2.5 and −3.0 MPa, i.e. near or beyond the leaf turgor loss point. Leaves of plants rehydrated while in the dark for 30 min, 90 min and 12 h recovered from K loss with characteristic times and to extents inversely proportional to the initial water stress applied. Leaf conductance to water vapour of plants dehydrated to decreasing Ψ_L and rehydrated at low transpiration was inversely related to loss of K, thus suggesting that leaf vein embolism and refilling (and related changes in leaf hydraulics) may play a significant role in the stomatal response.     

Correction of thermal gradient errors in stem thermocouple hygrometers

Stem thermocouple hygrometers were subjected to transient and stable thermal gradients while in contact with reference solutions of NaCl. Both dew point and psychrometric voltages were directly related to zero offset voltages, the latter reflecting the size of the thermal gradient. Although slopes were affected by absolute temperature, they were not affected by water potential. One hygrometer required a correction of 1.75 bars water potential per microvolt of zero offset, a value that was constant from 20 to 30 C.