A comparison of the composite and repeat pressurization methods of pressure–volume analysis for shoots of four North American conifer species

Significantly different water relations attributes were derived for temperate conifers measured using the repeat pressurization (RP) and composite (CM) pressure–volume (PV) procedures. In the RP method, single shoots were measured 10–20 times for xylem water potential and mass during air-drying to produce each PV curve. In contrast, for CM PV curves 25–30 shoots were air-dried to relative water contents (R) ranging from 1.0 to 0.5 before being pressurized once. Aggregation of these 25–30 paired values produced single PV curves. Pinus banksiana, P. resinosa and Picea mariana, but not Pinus strobus, had lower full turgor osmotic potential, shallower slope of the linear segment of the PV curves and higher symplast fraction with the CM method. Data points along the linear segment of PV curves were obtained to lower R using the CM method. Reanalysis using similar R ranges eliminated differences between PV methods for Picea mariana but not Pinus banksiana and P. resinosa.     

Discrepancies between water potential isotherm measurements on Pinus ponderosa seedling shoots: xylem hysteresis and apoplasmic osmotic potentials *

Abstract. Sap expression, air drying and a combined technique were used to measure the water potential isotherm of Pinus ponderosa Laws, seedling shoots with the pressure chamber. Discrepancies between water relations parameters derived from these techniques can be partially explained by air entry into air drying tissues, hysteresis in the xylem water potential isotherm and dilution of apoplasmic solutes during sap expression.     

Does Xylem Sap ABA Control the Stomatal Behaviour of Water-Stressed Sycamore (Acer pseudoplatanus L.) Seedlings?

Sycamore seedlings were grown with their root systems dividedequally between two containers. Water was withheld from onecontainer while the other container was kept well-watered. Effectsof soil drying on stomatal behaviour, shoot water status, andabscisic acid (ABA) concentration in roots, xylem sap and leaveswere evaluated. At 3 d, root ABA in the drying container increased significantly,while the root ABA in the unstressed container of the same plantsdid not differ from that of the control. The increase in rootABA was associated with the increase in xylem sap ABA and withthe decrease in stomatal conductance without any significantperturbation in shoot water status. At 7 d, despite the continuous increase in root ABA concentration,xylem sap ABA showed a marked decline when soil water contentwas depleted below 013 g g^–1. This reduction in xylemsap ABA coincided with a partial recovery of stomatal conductance.The results indicate that xylem sap ABA is a function of rootABA as well as the flow rate of water from roots to shoots,and that this ABA can be a sensitive indicator to the shootof the effect of soil drying. Key words: Acer pseudoplatanus L., soil drying, stomatal behaviour, xylem sap ABA     

The Influence of Resaturation Method and Tissue 3Quercus alba L. Seedlings

Parker, W. C. and Pallardy, S. G. 1987. The influence of resaturationmethod and tissue type on pressure-volume analysis of Quercusalba L. seedlings.—J. exp. Bot. 38: 535–549. The effect of resaturation method and amount of woody tissueon pressure-volume analysis was investigated using materialcollected from Quercus alba L. seedlings. Leaves excised fromwell-irrigated, intact plants had lower initial xylem pressurepotentials than did leaves resaturated by two artificial methods.Differential capacity for tissue rehydration among the threemethods was linked to shifts in the relative position of pressure-volumecurves, and differences in the osmotic potential and relativewater content at which turgor loss occurred were observed. Pressure-volumecurves from leaves resaturated by all three methods contained‘plateaus’ near full turgor, where xylem pressurepotential declined only slightly with relative water content.These plateaus were apparently associated with apoplastic waterthat accumulated in intercellular spaces of the leaf near fullturgidity, and acted to buffer changes in leaf xylem pressurepotential as tissues dehydrated. The presence of this waterhas implications for derived water relations parameter estimates.Pressure-volume curves for excised shoots also exhibited plateaus,but the relationship between xylem pressure potential and relativewater content over this region was steeper than was found forleaves. Shoot osmotic potentials were somewhat lower than thosefor leaves. The slope of the linear portion of shoot pressure-volumecurves was more shallow than for single leaves, a response associatedwith comparatively lower values of the symplastic water fractionin shoots. Key words: Pressure-volume curve, tissue-water relations, elasticity     

Cut-shoot vs root rehydration: Effects on pressure-volume analysis of shoots and fascicles of Pinus resinosa seedlings

Two methods of rehydrating red pine ( Pinus resinosa Ait.) shoots for pressure-volume (PV) analysis were compared to clarify the effects of rehydration on estimated tissue water relations of shoots and fascicles. The commonly employed cut-shoot method was compared to rehydration by means of water uptake through the roots of intact plants. Cut-shoot rehydration and increased duration of rehydration significantly decreased estimates of tissue elasticity and relative water content at zero turgor for both shoots and fascicles. Rehydration of cut shoots for 2 days significantly increased the slope of the linear region of shoot pressure-volume (PV) curves and decreased estimates of the apoplastic water fraction. Changes in these and estimates of other water relations attributes were correlated with increased initial water content during rehydration. Estimated apoplastic water fraction was higher for needle fascicles than shoots despite the large amount of woody stem tissue contained in shoots. Fascicle water status strongly influenced shoot water relations, in part due to apparent apoplastic loading with water of fascicles during rehydration.     

The Distribution of Nitrate Assimilation Between Root and Shoot in Vicia faba L.

Nitrate assimilation was examined in two cultivars (Banner Winterand Herz Freya) of Vicia faba L. supplied with a range of nitrateconcentrations. The distribution between root and shoot wasassessed. The cultivars showed responses to increased applied nitrateconcentration. Total plant dry weight and carbon content remainedconstant while shoot: root dry weight ratio, total plant nitrogen,total plant leaf area and specific leaf area (SLA) all increased.The proportion of total plant nitrate and nitrate reductase(NR) activity found in the shoot of both cultivars increasedwith applied nitrate concentrations as did NO₃^–: Kjeldahl-Nratios of xylem sap. The cultivars differed in that a greaterproportion of total plant NR activity occurred in the shootof cv. Herz Freya at all applied nitrate concentrations, andits xylem sap NO₃^–: Kjeldahl-N ratio and SLA were consistentlygreater. It is concluded that the distribution of nitrate assimilationbetween root and shoot of V. faba varies both with cultivarand with external nitrate concentration. Vicia faba L., field bean, nitrate assimilation, nitrate reductase, xylem sap analysis     

Delivery rates of abscisic acid in xylem sap of Ricinus communis L. plants subjected to part-drying of the soil

Abscisic acid (ABA) moving from roots to shoots in the transpirationstream is a potential hormonal message integrating perceptionof a root stress with adaptive changes in the shoot. A twinroot system was used to study ways of estimating effects ofdroughting the upper roots of Ricinus communis L. on abscisicacid (ABA) transport to the shoot in the transpiration stream.Droughted plants transpired more slowly than controls. Droughtingalso increased concentrations of ABA up to I I-fold in sap inducedto flow from the roots of freshly decapitated plants at ratesof whole plant transpiration. However, because of dilution effectsarising from the different sap flows in control and droughtedplants, these changes in ABA concentration in the xylem sapdid not accurately reflect amounts of ABA transported. To overcomethis problem, delivery rates were calculated by multiplyingconcentration with sap flow rate to generate ABA delivery interms of µmol s–1 per plant. Droughting for 24 hor more increased ABA delivery from roots to shoots by 5-fold.Since droughting can alter the relative sizes of the roots andshoots and also the root:shoot ratio these delivery rates wererefined in several ways to reflect both the amount of root generatingthe ABA message and the size of the recipient shoot system. Key words: Abscisic acid, Ricinus communis L., soil drying, xylem sap     

Do increases in xylem sap pH and/or ABA concentration mediate stomatal closure following nitrate deprivation?

Stomatal conductance (g(s)) of pepper (Capsicum annuum L.) plants decreased during the second photoperiod (day 2) after withholding nitrate (N). Stomatal closure of N-deprived plants was not associated with a decreased shoot water potential (Psi(shoot)); conversely Psi(shoot) was lower in N-supplied plants. N deprivation transiently (days 2 and 3) alkalized (0.2-0.3 pH units) xylem sap exuded from de-topped root systems under root pressure, and xylem sap expressed from excised shoots by pressurization. The ABA concentration of expressed sap increased 3-4-fold when measured on days 2 and 4. On day 2, leaves detached from N-deprived and N-supplied plants showed decreased transpiration rates when fed an alkaline (pH 7) artificial xylem (AX) solution, independent of the ABA concentration (10-100 nM) supplied. Thus changes in xylem sap composition following N deprivation can potentially close stomata. However, the lower transpiration rate of detached N-deprived leaves relative to N-supplied leaves shows that factors residing within N-deprived leaves also mediate stomatal closure, and that these factors assume greater importance as the duration of N deprivation increases.     

Relative water content and water potential of tissue 1

Relative water content (RWC) and water potential as measuredwith the pressure chamber were evaluated as indicators of waterstatus of tissue-cultured apple shoots and plantlets (shootswith roots). During the hydration required for RWC measurement,both water content and water potential exhibited the same hydrationkinetics, indicating that 10 h were required for full hydration.Once full hydration was reached, shoot mass remained relativelyconstant. Moisture release characteristics were also constructedand the associated shoot and plantlet water relations parameterswere estimated. Underin vitroconditions, both shoot and plantletwater potential were similar to the water potential of the culturemedium in which they were grown. The moisture release characteristicof shoots and plantlets was consistent with that expected fortypical plant tissues, and gave estimates of maximum modulusof elasticity (6.201.14 MPa), osmotic potential at saturation(–0.85 0.10 MPa), osmotic potential at zero turgor (–1.16 0.14 MPa) and RWC at zero turgor (78 2%) which were similarto values in the literature. Higher values of leaf conductanceand RWC were found in shoots and plantlets placed at 95% RH(21 C) compared to those at 90% RH. Plantlets had higher valuesof both conductance and RWC compared to shoots, suggesting thatinvitroroots are functional in water uptake. Relative water contentwas related to measures of physiological activity such as leafconductance, and it was also easier to measure than water potential.Relative water content is suggested as a sound index of waterstatus in tissue culture plants. Key words: Conductance, microculture, water status, water stress.     

Does ABA in the Xylem Control the Rate of Leaf Growth in Soil-Dried Maize and Sunflower Plants?

Maize (Zea mays L.) and sunflower (Helianthus annuus L.) plantswere grown in large volumes of soil and leaf growth rate wasmonitored on a daily basis. Half the plants were given a soildrying treatment and when they showed a significant restrictionof growth rate (compared to both their daily growth rate beforedrying and the average growth rate of well-watered plants onthe same day), leaf water relations were measured and xylemsap was extracted using several techniques. There was a significant negative log-linear relationship betweenthe rate of leaf growth and the concentration of ABA in thexylem for both species. There was no clear relationship betweenleaf growth rate and leaf water potential or turgor for eitherspecies. Assessment of different methods for sampling xylemsap suggests that exudates collected from stem stumps or samplescollected by pressurizing the whole root system are suitablefor estimating ABA concentration in xylem, at least with largeplants of maize or sunflower, provided the first few hundredcubic millimetres of collected sap are used for the assay. Centrifugationof sections of stems resulted in dilution of ABA in the xylemsap with sap squeezed from parenchyma tissue. This is because,at least in plants subjected to mild soil drying, the concentrationof the ABA in the xylem is far higher than that in the cellsap of stem tissue. Results support the proposal that ABA plays a major role asa chemical signal involved in the root-to-shoot communicationof the effects of soil drying. The non-hydraulic restrictionof leaf growth by a chemical signal can be explained by theextra root-sourced ABA in the xylem and may be an importantcomponent of the modification of growth and development whichresults from prolonged soil drought. Key words: Soil drying, ABA, leaf growth, Zea mays L., Helianthus annuus L.     

Stomatal response to drying soil in relation to changes in the xylem sap composition of Helianthus annuus. I. The concentration of cations, anions, amino acids in, and pH of, the xylem sap

Sunflower plants (Helianthus annuus L.) were subjected to soil drying with their shoots either kept fully turgid using a Passioura-type pressure chamber or allowed to decrease in water potential. Whether the shoots were kept turgid or not, leaf conductance decreased below a certain soil water content. During the soil drying, xylem sap samples were taken from individual intact and transpiring plants. Xylem sap concentrations of nitrate and phosphate decreased with soil water content, whereas the concentrations of the other anions (SO₄² and Cl^?) remained unaltered. Calcium concentrations also decreased. Potassium, magnesium, manganese and sodium concentrations stayed constant during soil drying. In contrast, the pH, the buffering capacity at a pH below 5 and the cation/anion ratio increased after soil water content was lowered below a certain threshold. Amino acid concentration of the xylem sap increased with decreasing soil water content. The effect of changes in ion concentrations in the xylem sap on leaf conductance is discussed.     

Water relations in silver birch during springtime: How is sap pressurised?

• Positive sap pressures are produced in the xylem of birch trees in boreal conditions during the time between the thawing of the soil and bud break. During this period, xylem embolisms accumulated during wintertime are refilled with water. The mechanism for xylem sap pressurization and its environmental drivers are not well known.
• We measured xylem sap flow, xylem sap pressure, xylem sap osmotic concentration, xylem and whole stem diameter changes, and stem and root non‐structural carbohydrate concentrations, along with meteorological conditions at two sites in Finland during and after the sap pressurisation period.
• The diurnal dynamics of xylem sap pressure and sap flow during the sap pressurisation period varied, but were more often opposite to the diurnal pattern after bud burst, i.e. sap pressure increased and sap flow rate mostly decreased when temperature increased. Net conversion of soluble sugars to starch in the stem and roots occurred during the sap pressurisation period. Xylem sap osmotic pressure was small in comparison to total sap pressure, and it did not follow changes in environmental conditions or tree water relations.
• Based on these findings, we suggest that xylem sap pressurisation and embolism refilling occur gradually over a few weeks through water transfer from parenchyma cells to xylem vessels during daytime, and then the parenchyma are refilled mostly during nighttime by water uptake from soil. Possible drivers for water transfer from parenchyma cells to vessels are discussed. Also the functioning of thermal dissipation probes in conditions of changing stem water content is discussed.

     

Influence of Xylem Water Potential on Leaf Elongation and Osmotic Adjustment of Wheat and Lupin

The leaf elongation rate and osmotic pressure at full turgorof wheat (Triticum aestivum L.) and lupin (Lupinus cosentiniiGuss.) were measured in well watered plants, in plants thatwere allowed to dry the soil slowly over 7 d, and in plantsin which the water potential of the leaf xylem was maintainedhigh by applying pressure to the roots during the drying cycle.Maintenance of high xylem water potentials failed to preventa reduction in the rate of leaf elongation as the soil dried,while the osmotic pressure at full turgor and the degree ofosmotic adjustment increased as the soil water content decreased.The rate of leaf elongation was reduced more and the degreeof osmotic adjustment was higher in leaves with high xylem waterpotentials than in those in which leaf xylem potentials wereallowed to decrease as soil water content decreased. Osmoticadjustment was linearly correlated with the reduction in leafelongation rate in both wheat and lupin. Key words: Osmotic adjustment, leaf elongation, turgor regulation     

Characterization of Cd translocation and identification of the Cd form in xylem sap of the Cd-hyperaccumulator Arabidopsis halleri

Arabidopsis halleri is a Cd hyperaccumulator; however, the mechanismsinvolved in the root to shoot translocation of Cd are not wellunderstood. In this study, we characterized Cd transfer fromthe root medium to xylem in this species. Arabidopsis halleriaccumulated 1,500 mg kg^–1 Cd in the shoot without growthinhibition. A time-course experiment showed that the releaseof Cd into the xylem was very rapid; by 2 h exposure to Cd,Cd concentration in the xylem sap was 5-fold higher than thatin the external solution. The concentration of Cd in the xylemsap increased linearly with increasing Cd concentration in theexternal solution. Cd transfer to the xylem was completely inhibitedby the metabolic inhibitor carbonyl cyanide 3-chlorophenylhydrazone(CCCP). Cd concentration in the xylem sap was decreased by increasingthe concentration of external Zn, but enhanced by Fe deficiencytreatment. Analysis with ¹¹³Cd-nuclear magnetic resonance (NMR)showed that the chemical shift of ¹¹³Cd in the xylem sap wasthe same as that of Cd(NO₃)₂. Metal speciation with Geochem-PCalso showed that Cd occurred mainly in the free ionic form inthe xylem sap. These results suggest that Cd transfer from theroot medium to the xylem in A. halleri is an energy-dependentprocess that is partly shared with Zn and/or Fe transport. Furthermore,Cd is translocated from roots to shoots in inorganic forms.     

Hydraulic control of stomatal conductance in Douglas fir [Pseudotsuga menziesii (Mirb.) Franco] and alder [Alnus rubra (Bong)] seedlings

Experiments were conducted on 1-year-old Douglas fir [Pseudotsuga menziesii (Mirb.) Franco] and 2- to 3-month-old alder [Alnus rubra (Bong)] seedlings growing in drying soils to determine the relative influence of root and leaf water status on stomatal conductance (g_c). The water status of shoots was manipulated independently of that of the roots using a pressure chamber that enclosed the root system. Pressurizing the chamber increases the turgor of cells in the shoot but not in the roots. Seedling shoots were enclosed in a whole-plant cuvette and transpiration and net photosynthesis rates measured continuously. In both species, stomatal closure in response to soil drying was progressively reversed with increasing pressurization. Responses occurred within minutes of pressurization and measurements almost immediately returned to pre-pressurization levels when the pressure was released. Even in wet soils there was a significant increase in g_c with pressurization. In Douglas fir, the stomatal response to pressurization was the same for seedlings grown in dry soils for up to 120 d as for those subjected to drought stress over 40 to 60 d. The stomatal conductance of both Douglas fir and alder seedlings was less sensitive to root chamber pressure at higher vapour pressure deficits (D), and stomatal closure in response to increasing D from 1.04 to 2.06 kPa was only partially reversed by pressurization. Our results are in contrast to those of other studies on herbaceous species, even though we followed the same experimental approach. They suggest that it is not always appropriate to invoke a ‘feedforward’ model of short-term stomatal response to soil drying, whereby chemical messengers from the roots bring about stomatal closure.     

Signalling mechanisms involved in the response of two varieties of Humulus lupulus L. to soil drying: I. changes in xylem sap pH and the concentrations of abscisic acid and anions

Background and aims

Soil drying leads to the generation of chemical signals in plants that regulate water use via control of the stomatal aperture. The aim of our work was to identify the presence and identity of potential chemical signals, their dynamics, and their relationship with transpiration rate during soil drying in hop (Humulus lupulus (L.)) plants.

Methods

We used pressure chamber technique for measurement of shoot water potential and collection of shoot xylem sap. We analyzed concentrations of abscisic acid (ABA), nitrate, phosphate, sulphate and malate in sap and also the rate of whole plant transpiration.

Results

Transpiration rate decreased prior to changes in shoot water potential. The concentration of ABA in xylem sap continuously increased from early to later stages of water stress, whereas in leaves it increased only at later stages. Shoot sap pH increased simultaneously with the decrease of transpiration rate. Xylem sap alkalization was in some cases accompanied by a decrease in nitrate concentration and an increase in malate concentration. Concentration of sulphate increased in xylem sap during drying and sulphate in combination with a higher ABA concentration enhanced stomatal closure.

Conclusions

Several early chemical signals appear in sap of hop plants during soil drying and their impact on transpiration may vary according to the stage of soil drying.     

Seasonal variation in the tissue water relations of Picea glauca

Seasonal variation in water relations of 3-yearold white spruce (Picea glauca (Moench) Voss) shoots, monitored with pressure-volume curves over 28 months, was closely related to shoot phenology and was sensitive to environmental fluctuations during both summer growth and winter dormancy. Turgor maintenance capacity was lowest during rapid shoot elongation from late May to early July; this was indicated by the lowest total turgor pressures, the highest (least negative) osmotic potentials at full turgor and the turgor loss point, the smallest differences between osmotic potentials at full turgor and the turgor loss point, the highest relative water contents at turgor loss and a linear decline in cell elasticity with decreasing turgor pressure. This suggests that the high susceptibility of white spruce seedlings to growth check after transplanting is largely attributable to the poor turgor maintenance capacity of this species in early summer.     

Cycling of amino-nitrogen between shoots and roots in wheat seedlings

Summary One part of a split root system of wheat seedlings received full nutrient solution with¹⁵N-nitrate, the other received an identical solution with unlabelled nitrate. Appearance of labelled amino compounds was measured in the xylem sap exuding from roots not supplied directly with¹⁵N-nitrate after removal of the¹⁵N-nitrate-fed roots. This material indicates cycling of nitrogen from the shoots and through the roots. About 60 per cent of the nitrogen in the xylem appears to be cycling in this way.     

The role of abscisic acid and water relations in drought responses of subterranean clover

The role of water relations and abscisic acid (ABA) in the responsesto drought were studied in a mediterranean forage crop, Trifoliumsubterraneum L. under field conditions. Soil and plant waterstatus, leaf gas exchange parameters, and xylem sap ABA contentwere determined at different times during a long-term soil dryingepisode in irrigated and droughted plants. The diurnal time-coursesof these parameters were also measured at the end of a droughtperiod. In response to soil drying stomatal conductance (g) was reducedearly to 50% that of irrigated plants before any substantialchange in water potential was detected. A close logarithmicregression between photosynthesis rate (A) and g was present.For the first weeks of drought the decline in A was less pronouncedthan in g, thus increasing water use efficiency. Stomatal conductanceduring diurnal time-courses showed no consistent relationshipswith respect to etther ABA or leaf water potential. Throughoutthe experimental period dependence of g on leaf water statuswas evident from the tight correlation (r²=0.88, P<0.01)achieved between stomatal conductance and midday water potential,but the correlation was also high when comparing g with respectto ABA content in xylem sap (r=0.83, P<0.001). However, thestomata from drought acclimated plants were apparently moresensitive to xylem ABA content. For similar xylem ABA concentrationsstomatal conductance was significantly higher in irrigated thanin waterstressed plants. Key words: Drought, stomatal conductance, water potential, abscisic acid     

Relationships between sap flow and water potential in woody or perennial plants on islands of the Great Barrier Reef

Abstract. This paper describes studies on trees of Pisonia grandis , bushes of Argusia argentea , and the perennial herb Melanthera biflora , growing on One Tree Island, a coral cay of the Great Barrier Reef with 'soil' of coarse coral rubble. Water potential (Ψ_b, measured on small shoots with a pressure chamber), sap flow, stomatal conductance, vapour pressure deficit and photon flux density were monitored over day/night cycles. Sap flow and Ψ_b responded to changes in light and humidity. From these experiments good linear correlations were found between sap flow in a shoot and Ψ_b of similar adjacent shoots. The linearity suggests that the resistance to sap flow is constant as Ψ_b varies. The correlation, however, does not indicate a causal relationship between Ψ_b of an individual shoot on the plant and its sap flow. Ψ_b was only slightly different in shaded shoots from those in sunshine, although sap flow would be expected to differ between them. Enclosing shoots and so reducing their transpiration and sap flow to very low rates resulted in only small changes in Ψ_b of the enclosed shoots; Tb of such enclosed shoots should closely approximate that of the xylem at the point of shoot attachment. From these results it is suggested that the resistance to water flow in shoot and leaf xylem is small compared to the resistance further down the plant, in the root or at the root/soil interface. Shoot xylem water potential would be similar for all parts of the plant, and in such plants the water potential of shoots in the shade would be determined by the overall water use of the plant.