The role of solute accumulation, osmotic adjustment and changes in cell wall elasticity in drought tolerance in Ziziphus mauritiana (Lamk.)

Ber (Ziziphus mauritiana Lamk.) is a major fruit tree crop of the north-west Indian arid zone. In a study of the physiological basis of drought tolerance in this species, two glasshouse experiments were conducted in which trees were droughted during single stress-cycles. In the first experiment, during a 13 d drying cycle, pre-dawn leaf water (leaf) and osmotic () potentials in droughted trees declined from -0.5 and -1.4 MPa to -1.7 and -2.2 MPa, respectively, for a decrease in relative water content () of 14%. During drought stress, changes in sugar metabolism were associated with significant increases in concentrations of hexose sugars (3.8-fold), cyclitol (scyllo-inositol; 1.5-fold), and proline (35-fold; expressed per unit dry weight), suggesting that altered solute partitioning may be an important factor in drought tolerance of Ziziphus. On rewatering pre-dawn leaf and recovered fully, but remained depressed by 0.4 MPa relative to control values, indicating that solute concentration per unit water content had changed during the drought cycle.Evidence for osmotic adjustment was provided from a second study during which a gradual drought was imposed. Pressure-volume analysis revealed a 0.7 MPa reduction in osmotic potential at full turgor, with leaf at turgor loss depressed by 1 MPa in drought-stressed leaves. Coupled with osmotic adjustment, during gradual drought, was a 65% increase in bulk tissue elastic modulus (wall rigidity) which resulted in turgor loss at the same in both stressed and unstressed leaves. The possible ecological significance of maintenance of turgor potential and cell volume at low water potentials for drought tolerance in Ziziphus is discussed.Keywords: Ziziphus mauritiana, drought, solute accumulation, osmotic adjustment, proline.      

Optimization theory of stomatal behaviour II. Stomatal responses of several tree species of north Australia to changes in light, soil and atomospheric water content and temperature

In a companion paper several methods of calculating the marginal unit water cost of plant carbon gain (E/A) were tested to determine whether stomata were behaving optimally in relation to regulating leaf gas exchange. In this paper one method is applied to several tropical tree species when leaf-to-air vapour pressure difference (D), photosynthetic photon flux density, leaf temperature, and atmospheric soil water availability were manipulated. The response of leaves that had expanded during the dry season were also compared to that of leaves that had expanded in the wet season. Few differences in absolute value of E/A, or the form of the relationship, were observed between species or between seasons. In the majority of species, E/A increased significantly as either leaf-to-air vapour pressure difference increased, at a leaf temperature of either 33C or 38C, or as in photosynthetic photon flux density increased. In contrast, as leaf temperature increased at constant D, E/A was generally constant. As pre-dawn water potential declined, E/A declined. The relationship between E/A and D did not differ whether internal or ambient carbon dioxide concentration were kept constant. It is concluded that stomata are only behaving optimally over a very small range of D. If a larger range of D is used, to incorporate values that more closely reflect those experienced by tropical trees in a savanna environment optimization is incomplete.Key words: Stomatal optimization theory, marginal unit water cost.      

Alterations in the Components of Peanut Leaf Water Potential during Desiccation

Changes in components of leaf water potential during soil waterdeficits influence many physiological processes. Research resultsfocusing on these changes during desiccation of peanut (Arachishypogeae L.) leaves are apparently not available. The presentstudy was conducted to examine the relationships of leaf water_l, solute _s and turgor _p potentials, and percent relative watercontent (RWC) of peanut leaves during desiccation of detachedleaves and also during naturally occurring soil moisture deficitsin the field. The relationship of _p to _l and RWC was evaluated by calculating_p from differences in _l and _s determined by thermocouple psychrometryand by constructing pressure-volume (P-V) curves from the _land RWC measurements. Turgor potentials of ‘Early Bunch’and ‘Florunner’ leaves decreased to zero at _l of–1.2 to –1.3 MPa and RWC of 87%. There were no cultivardifferences in the _l at which _p became zero. P-V curves indicatedthat the error of measuring _s after freezing due to dilutionof the cellular constituents was small but resulted in artefactualnegative _p values. Random measurements on two dates of _l, _s, and calculation of_p from well-watered and water-stressed field plots consistingof several genotypes indicated that zero _p occurred at _l of–1.6 MPa. It was concluded that the relationships of _p,_l, _s, and RWC of peanut leaves were similar to leaves of othercrops and that these relationships conferred no unique droughtresistance mechanism to peanut.     

Osmotic Adjustment to Water Stress in Pearl Millet (Pennisetum americanum (L. ) Leeke) in a Controlled Environment

A pressure-volume (P-V) and an expressed sap (cryoscopic) techniquewere compared for assessing osmotic adjustment to water stressby pearl millet (Pennisetum americanum (L. ) Leeke) plants grownin a controlled environment cabinet. For leaf water potentials( ) above the point of zero turgor, there was good agreementbetween estimates of solute potential ( _s)and turgor ( _p) obtainedby the two methods. Reductions in pre-dawn leaf to –1.8 MPa over 5–6d resulted in net solute accumulation as indicated by a fallin s at full hydration of about 0.3 MPa. The degree of osmoticadjustment increased linearly with the decrease in pre-dawn. Adjustment in cv. BJ 104 was significantly (P < 0.05) lessduring a second drought than during a first, and cv. Serere39 was significantly (P < 0.05) less able to adjust osmoticallythan BJ 104. Adjustment was greater in leaves which were undergoing extensiongrowth during the drought than in leaves already fully extendedbefore drought started. Much of the adjustment was lost within24 h following rewatering, the loss being most complete in theolder, fully extended leaves.     

An Error in the Calibration of Xylem-water Potential against Leaf-water Potential

An error occurs in the calibration of xylem pressure potential() against leaf-water potential () when the calibration is madeusing plant material in which the water stress has been inducedartificially after excision. The impostion of water stress afterexcision affects the determination more than it affects , consequentlythe relationship between these two indices of water stress isaltered. Care should be exercised to ensure that identical proceduresare adopted during . calibrations and during susbsequent fieldmeasurements of with the pressure-chamber apparatus.     

Abscisic Acid Accumulation and Water Relations of Four Cultivars of Phaseolus vulgaris L. under Drought

Larqué-Saavedra, A., Rodriguez, M. T., Trejo, C. andNava, T. 1985. Abscisic acid accumulation and water relationsof four cultivars of Phaseolus vulgaris L. under drought.—J.exp. Bot 36: 1787–1792. Plants of four cultivars of Phaseolus vulgaris L. differingin drought resistance were grown in pots under greenhouse conditionsand prior to flowering water was withheld from the pots untilthe mid-day transpiration rate reached values below 1.0 µgH₂O cm^–2 s^–1 (designated the ‘drought’stage). At this point leaves were harvested on 3 or 4 occasionsover 24 h to determine the abscisic acid (ABA) concentration,total water potential (), solute potential (₁) and turgor potential(_p). Results showed that values of , ₁, and _p differed between cultivarswhen they reached the ‘drought’ stage. The stomatalsensitivity to changes in and _p, was as follows: Michoacán12A3 > Negro 150 Cacahuate 72 > Flor de Mayo. These datacorrelated well with the pattern of drought resistance reportedfor the cultivars. ABA accumulation at the ‘drought’ stage differedbetween cultivars at each sampling time, but overall differencesin ABA level between cultivars were not significant. ABA levelsdid not, therefore, correlate with the drought resistance propertiesreported for the cultivars. Results are discussed in relationto and hour of the day when bean samples were taken for ABAanalysis. Key words: Phaseolus vulgaris L., drought resistance, abscisic acid     

Characterization of Spring Wheat Genotypes Differing in Drought-Induced Abscisic Acid Accumulation: II. LEAF WATER RELATIONS

Aspects of the water relations of spring wheat (Triticum aestivumL.) are described for cultivars Highbury (low ABA) and TW269/9(high ABA), and low and high ABA accumulating F⁶selections derivedfrom a cross between them. In a pot experiment, pressure-volume (P-V) curves were constructedfor main stem leaf four (MSL4) of well-watered plants of Highburyand TW269/9. Estimates of solute potential (²) from these curveswere similar for the two cultivars, but varied with the timeof sampling and the time allowed for hydration in dim light. In a field experiment with four low and four high ABA F⁶lines,P-V curves for flag leaves from both droughted and irrigatedplants gave at both zero turgor (^p) and zero water potential(¹) which differed with degree of stress, sampling time andgenotype. ¹was strongly dependent on the initial_L of the leafand was reduced on average by c. 0.4 MPa per MPa decline ininitial L.5, was lower (more negative) by c. 0.1-MPa in theafternoon than in the morning. Overall, was also 0.1 MPa lowerin low ABA lines than in high ABA lines. In another field experiment, flag leaves of five low and fivehigh ABA F⁶lines were sampled over a 4 week period from droughtedplots and _L and 5, measured (the latter by osmometry with expressedsap). For these leaves 5, at zero p or zero _L was consistentlylower by 0.3–0.5 MPa than estimates of 5, from the P-Vcurves with flag leaves. However, data for the low ABA lineswere again lower (by c. 0.1 MPa) than those for high ABA lines. The consequences of these differences in 1 are discussed inrelation to the stimulation of ABA accumulation in low and highABA selections. Key words: Water potential, Solute potential, P-V curves, Wheat (Triticum aestivum), Drought stress     

Alternative Methods of Analysing Water Potential Isotherms: Some Cautions and Clarifications: I. THE IMPACT OF NON-IDEALITY AND OF SOME EXPERIMENTAL ERRORS

In recent years alternative ways have been proposed to transformmeasurements of leaf water potential, , and relative water content,R^*, in order to derive values of osmotic pressure at full turgidityin leaves and shoots, ^o(when 0). Two types of transformationsare usually considered: 1/ versus R^* and versus 1/R^*, and linearregression is used to fit the data in the region where turgoris thought to be zero. It appears that when ^o is estimated bylinear extrapolation of 1/Psi; versus R^* then apoplastic watermight not influence the accuracy of ^o but when the versus \/R^*transformation is used apoplastic water causes an underestimateof ^o. We examine the accuracy of the estimate of ^o obtainedfrom the two transformations when there are random errors in, systematic errors in , and when the osmotic solutions arenon-ideal. The 1/ versus R^* transformation generally producesthe best estimate of ⁰ by linear extrapolation.     

A Simple Thermocouple Psychrometer for Determining Tissue 6

An apparatus for determining water potential of leaves () isdescribed. It is a modification of one previously developedby van Andel for determining osmotic potentials of sap (), forwhich purpose it may still be employed. A constantan—minalphathermocouple is used for direct determination of wet-bulb temperaturedepression of air in moisture equilibrium with leaves. The system has advantages over others in simplicity of manufactureand electrical circuitry, and in adaptability to a range ofsignal reading and recording methods. It is capable of handlingrelatively large samples, e.g. 20 cereal leaves of total area(single side) of 300 cm². Tests of leaf maturity and position effects in two potato cropsare described. Differences between corresponding values of and , considered indicative of cell wall pressures, were lessin lower leaves. In one crop, was lower in relation to leafrelative water content with lower than with upper leaves and,in the other crop, with leaves from the more mature stages ofgrowth. These findings are in accord with a higher rigidityof cell walls in lower and more mature leaves. Mean differences of and involved in the above conclusionswere small, usually less than 2 bar, indicating the potentialusefulness of the apparatus.     

Developmental Responses to Drought and Abscisic Acid in Sunflower Roots: I. ROOT GROWTH, APICAL ANATOMY, OSMOTIC ADJUSTMENT

Aeroponically grown sunflower seedlings (Helianthus annuus L.cv. Russian Giant) were droughted or treated with abscisic acid(ABA) for 7 d. Drought stress prompted a three-phase growthresponse in sunflower roots: an initial phase of increased rootelongation was followed by a period of almost complete inhibitionbetween about 6 h and 72 h; this was followed, in turn, by aphase of partial recovery in the rate of root elongation. Droughtdecreased the size of the apical meristem as cells in the proximalregion of the meristem vacuolated and elongated. Root-to-shootbiomass ratios (R:S) increased initially but declined after72 h. Drought stress decreased water potential () and osmoticpotential ( and increased turgor pressure _p in the apical 30mm of the roots. These initial changes were transitory, lastingabout 3 h. Thereafter, and began to rise; _p fell back to controllevels. In the later stages of treatment, fell as the stressgrew more severe, but fp was maintained by osmotic adjustment.Desiccation for 1 h increased turgor pressures in excised 30mm apical segments. The transitory increase in root elongationwas contemporary with the initial rise in _p in the root apices,while the periods of greatest inhibition and partial recoveryin root elongation were contemporary with the periods of declineand partial recovery in the length of the apical meristem respectively.The inhibition of root elongation and the anatomical changesin the root apices were not determined by loss of turgor orlack of photosynthate, but rather appeared to be an active responseby the meristem to a drop in external . Treatment with ABA triggeredmany of the same changes as drought stress: ABA promoted a three-phasegrowth response, increased R:S, triggered the same initial changesin , , and _p, increased _p in excised 3.0 mm apical segments,and induced the same pattern of anatomical changes in the rootapices as drought stress. It is proposed that ABA mediates drought-inducedchanges in the primary development of sunflower roots. Key words: Abscisic acid, apical meristem, drought, osmotic adjustmen     

The Meaning of Matric Potential

The commonly used equation, = P - + , which describes thepartitioning of plant water potential, , into components ofhydrostatic pressure, P, osmotic pressure, , and matric potential,, is misleading. The term , which is supposed to show the influenceof a solid phase on , is zero if a consistent definition ofpressure is used in the standard thermodynamic derivation. However,it can be usefully defined by = + _D, where _D is the osmoticpressure of the equilibrium dialysate of the system. The practicaland theoretical significance of this definition is discussed.     

Photorespiratory glycine enhances glutathione accumulation in both the chloroplastic and cytosolic compartments

Transformed poplars overexpressing -glutamylcysteine synthetase (-ECS) in the chloroplast (Lggs) were used to investigate chloroplastic biosynthesis of glutathione (GSH). In Lggs leaves, GSH contents were enhanced by up to 3.7 fold. In general, the highest GSH contents were observed in lines with highest -glutamylcysteine (-EC) contents. These lines had relatively low glycine. In darkness, foliar GSH decreased and -EC increased. Illumination of pre-darkened Lggs in air resulted in a 5-fold decrease in the -EC : GSH ratio. This light-induced decrease was largely abolished if leaves were illuminated at high CO2. Consequently, the -EC : GSH ratio of illuminated leaves was much higher at high CO2 than in air. At high CO2 total foliar amino acids were higher, but glycine and serine were lower, than in air. These results suggest that photorespiratory glycine is used in chloroplastic GSH synthesis. Despite this net CO2 fixation was similar in Lggs to untransformed poplars. Pre-illuminated leaf discs from Lggs, and poplars overexpression -ECS in the cytosol (ggs), were incubated in darkness with a range of metabolites. After 15 h, discs for both types of transformant incubated on water had accumulated high levels of -EC and showed marked increases in the -EC : GSH ratio. Feeding glycine, serine, glycollate or phosphoserine, attenuated the dark-induced changes in the -EC : GSH ratio, whereas 3-phosphoglycerate (PGA), phosphoenolpyruvate, glycerate, and hydroxypyruvate did not. Glycine produced from glycollate was therefore required for maximal GSH accumulation in both the chloroplastic and cytosolic compartment. Production of glycine from PGA failed to meet the demand of increased GSH synthetic capacity.     

Physiological Responses to Drought of Lolium perenne L.: Measurement of, and Genetic Variation in, Water Potential, Solute Potential, Elasticity and Cell Hydration

Thomas, H. 1987. Physiological responses to drought of Loliumperenne L.: Measurement of, and genetic variation in, waterpotential, solute potential, elasticity and cell hydration.—J.exp. Bot. 38: 115–125. Clonally-replicated genotypes of Loiium perenne L. were grownin a controlled environment. Leaf water potential (_w) osmoticpotential (_s), turgor potential (_p = _w – _s), elasticity(E), leaf hydration (g water per g dry matter, H) and numberof green leaves per tiller (N_GL) were measured before and duringa 42 d drought treatment. A simplified method of estimating E (at _w < 1?0 MPa) usingonly six measurements was developed to permit a measurementrate of 8 leaves per hour. Measurement errors in all characterswere 3% or less. During drought, _w and _s (at _w = 0?5 MPa) decreased significantly,_p and E increased significantly, and H decreased slightly. Plantsize during drought was negatively correlated with _s, and ^Hand positively correlated with _p, osmotic adjustment, E andN_GL. Measurements made on the genotypes before draughting didnot give a reliable indication of their physiological conditionafter adaptation to drought. Genetically controlled variation (‘broad sense heritability’)of drought-adapted plants for E was 15%, _w 23%, _s, 34%, _p, 35%,H 34% and N_GL 64%. The possibilities for, and effectivenessof, divergent selection of genotypes with high and low expressionof the characters are discussed. Key words: Water relations, Lolium, genetic variation     

{alpha}-Amylase Production by Pre-mature Wheat (Triticum aestivum L.) Embryos

Embryo/scutellar tissue of pre-mature wheat grains usually containslittle -amylase but readily produces the enzyme upon removalfrom the caryopsis. Enzyme production is accompanied by cytologicalchanges in the scutellar epithelium cells characteristic ofgerminating mature embryos, although -amylase production bypre-mature tissue is not always associated with germinativegrowth of the embryonic axis. Production of -amylase is influencedby embryo age, stimulated by GA₃ and overall is inhibited byABA. Examination by isoelectric focussing and rocket-line immuno-electrophoresisreveals the presence of both -AMYl and -AMY2 isoenzymes, thelatter being the major constituent. In the presence of ABA certain-AMY2 isoenzymes not detected previously are observed. Key words: a-Amylase, wheat, embryo     

Mycorrhizal influence on hydraulic and hormonal factors implicated in the control of stomatal conductance during drought

During drying, mycorrhizal plants often maintain higher stomatalconductance (g_s) than similarly-sized and -nourished non-mycorrhizalplants, but the mechanism of mycorrhizal influence remains unclear.Several hydraulic and non-hydraulic factors previously implicatedin control of stomatal behavior during drought were measured,to learn which are affected when roots of cowpea (Vigna unguiculata[L.] Walp. cv. White Acre) are extensively colonized by Glomusintraradices Schenck and Smith isolate UT143. At low soil watercontents (), mycorrhizal plants maintained higher g_s, transpirationand shoot water potential () than non-mycorrhizal plants. Thesehigher foliar water status characters were associated with lowerxylemsap abscisic acid concentrations ([ABA]) and lower ABAfluxes to leaves in mycorrhizal plants at low soil . Stomatalconductance was most closely correlated with xylem-sap [ABA],ABA flux to leaves and shoot . Stomatal conductance was notcorrelated with xylemsap concentrations of calcium or zeatinriboside equivalents, or with xylem-sap pH, nor were these xylem-sapconstituents affected by mycorrhizal symbiosis. Stomata of mycorrhizaland non-mycorhizal leaves showed similar sensitivities to ABA,whether leaves were intact or detached. It is concluded thatmycorrhizal fungi probably increased the capability of rootsystems to scavenge water in drier soil, resulting in less strainto foliage and hence higher g_s, and shoot at particular soil. Key words: Abscisic acid, cytokinins, Glomus intraradices     

Reduced aleurone {alpha}-amylase production in aged wheat seeds is accompanied by lower levels of high-pI [IMAGE]-amylase transcripts and reduced response to gibberellic acid

With the aim of determining the level at which ageing exerts its effect on the expression of -amylase, GA3 regulation of -amylase production was studied in isolated aleurone layers from aged wheat seeds. GA3-induced -amylase activity was lower in the tissue from aged seeds than in controls. However, the proportion of ³⁵S-methionine incorporated into -amylase was higher in the aged than in control tissue. The pattern of -amylase isoforms was resolved by isoelectric focusing and showed that two isogroups were present with the activity of the high-pI isogroup being higher in the control than in the aged lot. These apparently contradictory results may be explained in terms of differences in isozyme expression. Studies on the expression of -amylase genes indicated a reduction in the level of high-pI mRNA in aged tissue. Dose-response curves showed lower GA3-responsiveness of aleurone layers from aged seeds as compared to the controls. From these results, it is proposed that the diminished capacity of -amylase production in aleurone from aged seeds is apparently due to a decrease in the expression of the high-pI -amylase genes, and this reduction is associated with a decrease in the response to GA3.Key words: Seed ageing, wheat aleurone, gibberellic acid, -amylase isozymes, gene expression.      

The Derivation of the Cell Wall Elasticity Function from the Cell Turgor Potential

An equation is derived expressing average turgor pressure ofa leaf (_p) as a function of relative water content (RWC). Basedon this derivation, the relationships of the bulk elastic modulus(_v) and both RWC and _p, are formulated and discussed. The bulkelastic modulus (_v) becomes zero for _p = 0, that is at the turgorloss point for the leaf. At full water saturation the valueof ev is proportional to the water saturation turgor potential_p(max). The factor relating _P and _v (structure coefficient ,Burstrom, Uhrstr?m and Olausson, 1970) changes only very littlefor values of _p, which are not too close to zero. An exampleis given for the calculation from experimental data of the turgorpressure function, the structure coefficient function, and the_v function. Key words: Cell wall, Turgor pressure, Bulk elastic modulus     

Stomatal Response to Water Stress and its Relationship to Bulk Leaf Water Status and Osmotic Adjustment in Pearl Millet (Pennisetum americanum [L.] Leeke)

The water potential () at which stomata completed closure (_8Lmin)was determined for pearl millet (Pennisetum americanum [L.]Leeke) at two growth stages by monitoring changes in leaf conductance(g_L) and following shoot detachment. Leaf water status wasevaluated concurrently using a pressure-volume (P-V) technique. In a pot experiment with young vegetative plants, _8Lmin closelyapproximated to the estimated at zero turgor (_u) both for controland for drought-conditioned plants which had osmotically adjusted.However, for penultimate leaves of field-grown flowering plants,_8Lmin was found to be 0.61 (irrigated plants) and 0.87 (droughtedplants) MPa below _u. In drought-stressed field-grown plants,osmotic adjustment (characterized by a decrease in solute (osmotic)potential (_s ) at both full hydration and zero turgor) was insufficientto maintain a positive bulk leaf turgor potential (_p) once had declined to below about -1.5 MPa. It is suggested that localizedadjustment by the stomatal complex in response to environmentaldifferences, leaf ageing and/or ontogenetic change, is responsiblefor the uncoupling of stomatal from bulk leaf water status. Key words: Stomata, Water stress, Pennisetum americanum     

Adaptation of the Euryhaline Charophyte Lamprothamnium papulosum to Brackish and Freshwater: Turgor Pressure and Vacuolar Solute Concentrations During Steady-State Culture and After Hypo-osmotic Treatment

The euryhaline charophyte Lamprothamnium papulosum (Wallr.)J. Gr. was adapted to media with decreasing salinities rangingfrom 550 to 0 mosmol kg^–1. Vegetative plants grown inmedia with osmotic pressures (⁰) in the range of 550 to 130mosmol kg^–1 maintained a constant turgor pressure () at309 + 7 mosmol kg^–1. The ions K+, Na+ and Cl–, werethe predominant solutes in the vacuole. Changes in their concentrationsaccount for the variation in internal osmotic pressure (¹) with,⁰. The divalent ions Mg²⁺, Ca²⁺ and were also present in significant amounts, but their concentrationsdid not alter with changes in, ⁰. In cells subjected to hypo-osmotic shock the regulation of was incomplete. The turgor pressure increased from 302 to 383mosmol kg^–1. The first rapid response to the sudden decreasein ⁰ was a loss of K⁺ and Cl^–. In contrast to the decreasein ionic concentrations an accumulation of sucrose occurredwhich could account for the increase of . The increase in sucroseconcentration started 24 to 48 h after the downshock and reachedits highest value after 3 to 4 weeks. The sucrose concentrationin the vacuole was up to 320 mol m^–3. During this timethe ionic content continued to decrease but did not counterbalancethe sucrose concentration sufficiently to regain the original. High sucrose levels accompanied by an enhanced were also observedduring the period of fructification (sexual reproduction: formationof antheridia and oogonia) in Lamprothamnium kept under conditionsof constant salinity. It is concluded that high sucrose content and elevated arecharacteristic of sexual reproduction in this charophyte. Lamprothamniumis able to tolerate different during various developmentalstages (e.g. vegetative and reproductive phases). Key words: Lamprothamnium papulosum, sucrose, turgor pressure