The Diffusive Conductivity of the Stomata of Wheat Leaves

A leaf chamber (described in detail) was used alternately witha resistance porometer to measure resistance to viscous flowof air through the leaf, and with a diffusion porometer to measurethe differential diffusive flow of hydrogen and air (V_H–V_A)through the leaf and the component of hydrogen flow (V'_H) movingstraight across the leaf. The resistance of the mesophyll isneeded for interpretation: estimates by three different methodsfor viscous flow did not agree very well, but two differentmethods for diffusive flow gave good agreement. For wheat leaves,only very large errors are important. Formal analysis is in three appendixes: I. Interpretation ofviscous and diffusive flow in small pores involves some problemsin molecular physics, complicated by the particular geometryof the wheat stoma. With some uncertainty, formal expressionsare derived for the viscous resistance of a single stoma, rv,and for the resistances to diffusion of hydrogen and air, andof water vapour and carbon dioxide, all expressed as r_s persquare centimetre of leaf surface. The analysis for hydrogen/airis the most uncertain; that for water vapour and carbon dioxideis more reliable. II. An indication is given of the flow characteristicsof the leaf-chamber system, from which rv can be derived, andof the basis for estimating mesophyll resistance. III. The methodof converting estimates of r_s into estimates of V_H–V_Aand V'_H is given. The results presented are expressed as nearly as possible interms of the quantities which were measured. For five leavesthe dependence of V_H–V_A on V'_H agrees well with theoreticalpredictions; the dependence of V_H–V_A (and V'_H) on rv,on average, agrees well with prediction, but involves the assumptionthat the stomata get shorter as they close. The agreement isgood enough to suggest that the formal expressions for r_s interms of stomatal dimensions and molecular gas constants arereliable enough to be carried forward into future transpirationand assimilation studies. The minimum value of ra for watervapour (c. 3 sec cm⁺¹) is close to values found elsewhere bydifferent techniques. At very small stomatal openings there was a large deviationfrom predicted behaviour, such as would occur if the imposedexcess air pressure further closed the stomata during viscousflow experiments.     

A Null Balance Carbon Dioxide and Water Vapour Porometer

The Binos IRGA is suitable for portable use in the field aftermodification and can be added to the null-balance water vapourdiffusion porometer in a closed loop with the porometer chamberand used as a null-balance indicator for measurements of CO²flux, without the need for substantial structural or electricalmodification. The null-balance concept works satisfactorily in the measurementof both CO² influx and efflux and enables the CO² flux to bemeasured at either the ambient CO² concentration or at otherknown CO² concentrations. The combined CO² and water vapour porometer enables measurementsof the rate of photosynthesis, and estimates of stomatal andmesophyll conductance to be made extensively in the field withoutthe need for an elaborate mobile laboratory.     

Microanalytical Studies of Potassium and Chloride Fluxes and Stomatal Movements of Two Species: Vicia faba and Pelargonium x hortorum

Potassium and chloride exchange kinetics in guard cells werefound to be related to stomatal opening and closing kineticsin Pelargonium x hortorum and Vicia faba by simultaneously measuringion contents and degrees of opening with an electron probe anda hydrogen diffusion porometer. Large shifts between both kinetics occur during opening andclosing. These results show that maximal potassium content isdelayed, which seems to question the primary role of potassiumin stomatal opening     

A Radioactive Tracer Technique for the Direct Measurement of the Diffusion Resistance of Stomata

A new method is described for measuring the stomatal resistanceto diffusion in leaves. It consists of measuring the radioactivityof air samples which have diffused through a leaf from a porometercontainer enriched with radioactive argon(⁴¹A). Details of thecalibration are given together with the methods adopted to determinethe accuracy of measurement and the optimum time of sampling.Results suggest that in the material investigated the measurementsobtained are essentially those of stomatal resistance.In onesmall experiment inter and intra plant variability in stomatalresistance was investigated; in another, the diffusion porometerwas used to compare the daily march of stomatal resistance withthe rate of transpiration in plants growing under four differentsoil-moisture regimes.     

Developments in Mass Flow Porometry

A ventilated leaf attachment clamp is described with which lateralair flow can be measured separately from through flow. Stomatalconductances are expressed with dimensions: mm s^–1. Basedon parallel measurements with a diffusion porometer the relationbetween mass flow and diffusive flow conductances has been experimentallydetermined for three species. Records of stomatal movementsin air of different compositions are presented. Key words: Stomata, porometry, diffusive and mass flow conductance     

Design calibration and field use of a stomatal diffusion porometer

Modifications of the design and calibration procedure of a diffusion porometer permit determinations of stomatal resistance which agree well with results obtained by leaf energy balance. The energy balance and the diffusion porometer measurements indicate that the boundary layer resistances of leaves in the field are substantially less than those predicted from heat transport formulas based on wind flow and leaf size.     

Ineffectiveness of Abscisic Acid in Stomatal Closure of Yellow Lupin, Lupinus luteus var. Weiko III

Stomata of yellow lupin leaves are remarkably insensitive toabscisic acid (ABA). Stomatal resistance was monitored usingboth a viscous now porometer and a diffusion porometer. Resultswere confirmed with scanning electron microscopy. When exogenousABA solutions were supplied via petioles, 10^–6 M solutionshad no effect on stomatal resistance. Upper (adaxial) stomatawere not affected by 10^–5 M ABA but lower stomata showed3-fold more resistance after 2 h. Stomata of both surfaces closedafter 30 min in 10^–4 M ABA. Isolated epidermal peels of lupin leaves were floated on ABAsolutions yet upper surface peels showed no stomatal closingin 10^–4 M ABA, while lower surface stomata closed to abarely significant extent. Stomata of intact leaves were not very sensitive to darkness,showing at most a doubling in resistance after 6 h darkness.Complete stomatal closure, however, was readily produced bywilting leaves. Hence, lupin stomata are physically capableof closing. Endogenous ABA levels of water-stressed leaves increased approximately10-fold, which corresponds to concentrations below 10 µMABA. It is concluded that ABA is unlikely to play a role incontrolling short-term stomatal response of lupins.     

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.     

Stomatal Response of some Cultivated and Wild Tuber-bearing Potatoes in Warm Tropics as Influenced by Water Deficits

Leaf resistances of 14 cultivated potato genotypes (Solanumspp) and three tuber-bearing wild Solanum species were comparedwhen plants were grown under water stress at two tropical sitesFactors investigated were diurnal changes in leaf resistance,the effect of plant age, transient drought versus well-wateredconditions of potted and field-grown plants These measurementswere carried out in order to determine the stomatal behaviourof tuber-bearing genotypes and species Significant genotypic differences in leaf resistances were notedwithin the cultivated genotypes All genotypes had higher resistanceswhen water-stressed, but LT-7 appeared to have the lowest leafresistances Genetic differences in stomatal behaviour of tuber-bearingSolanum species were confirmed Abaxial stomatal resistancesof water-stressed plants of the species ranged between 1 74and 13 8 s cm^–1 Stomata of S chacoense were less affectedby drought (three-fold) than S tuberosum (four-fold) The greatesteffect was on S jungasense (five-fold) and on S raphanifoliumThese data show that stomata behaviour among tuber-bearing Solanumspecies is sufficiently different to warrant investigationsof drought-resistance in potato species under dry hot conditions Solanum tuberosum L., Solanum raphanifolium, Solanum chacoense, Solanum jungasense, leaf resistance     

Effect of Elevated CO2 on Stomatal Density and Distribution in a C4 Grass and a C3 Forb under Field Conditions

Two common tallgrass prairie species, Andropogon gerardii, thedominant C₄ grass in this North American grassland, and Salviapitcheri, a C₃ forb, were exposed to ambient and elevated (twiceambient) CO₂ within open-top chambers throughout the 1993 growingseason. After full canopy development, stomatal density on abaxialand adaxial surfaces, guard cell length and specific leaf mass(SLM; mg cm^-2) were determined for plants in the chambers aswell as in adjacent unchambered plots. Record high rainfallamounts during the 1993 growing season minimized water stressin these plants (leaf xylem pressure potential was usually >-1·5 MPa in A. gerardii) and also minimized differencesin water status among treatments. In A. gerardii, stomatal densitywas significantly higher (190 ± 7 mm^-2; mean ±s.e.) in plants grown outside of the chambers compared to plantsthat developed inside the ambient CO₂ chambers (161 ±5 mm^-2). Thus, there was a significant 'chamber effect' on stomataldensity. At elevated levels of CO₂, stomatal density was evenlower (P < 0·05; 121 ± 5 mm^-2). Most stomatawere on abaxial leaf surfaces in this grass, but the ratio ofadaxial to abaxial stomatal density was greater at elevatedlevels of CO₂. In S. pitcheri, stomatal density was also significantlylower when plants were grown in the open-top chambers (235 ±10 mm^-2 outside vs. 140 ± 6 mm^-2 in the ambient CO₂ chamber).However, stomatal density was greater at elevated CO₂ (218 ±12 mm^-2) compared to plants from the ambient CO₂ chamber. Theratio of stomata on adaxial vs. abaxial surfaces did not varysignificantly in this herb. Guard cell lengths were not significantlyaffected by growth in the chambers or by elevated CO₂ for eitherspecies. Growth within the chambers resulted in lower SLM inS. pitcheri, but CO₂ concentration had no effect. In A. gerardii,SLM was lower at elevated CO₂. These results indicate that stomataland leaf responses to elevated CO₂ are species specific, andreinforce the need to assess chamber effects along with treatmenteffects (CO₂) when using open-top chambers.Copyright 1994, 1999Academic Press Andropogon gerardii, elevated CO₂, Salvia pitcheri, stomatal density, tallgrass prairie     

The Influence of Prior Water Stress and Abscisic Acid Foliar Spraying on Stomatal Responses to CO2, IAA, ABA, and Calcium in Leaves of Solanum melongena

The influence of a water stress or foliar ABA spraying pretreatmenton stomatal responses to water loss, exogenous ABA, IAA, Ca²⁺,and CO₂ were studied using excised leaves of Solanum melongena.Both pretreatments increased stomatal sensitivity of water loss,in the presence and absence of CO₂, but decreased stomatal sensitivityto exogenous ABA. CO₂ greatly reduced the effect of exogenouslyapplied ABA. IAA decreased leaf diffusion resistance for controland ABA sprayed leaves, but did not influence the LDR of previouslywater-stressed leaves. CA²⁺ did not influence LDR of any leavesof any treatments. Key words: Water stress, stomatal response, pretreatments     

Studies on the Movement of Water Through Apple Trees

Resistances to the flow of water through young potted appletrees were estimated by measuring the transpiration rate oftrees with and without root systems. Root system resistanceswere obtained by difference. Whole-plant resistances were ofthe order 10 x 10¹³ Pa s m^–3 and there was some evidencethat root resistances (R_r) varied with transpiration rate; theratio R_r:R_x (where R_x is resistance to water flow in the stemsystem) altered from 2:1 at relatively high transpiration ratesto 1:1 at lower rates. The trunk of a 9-year-old orchard tree (trunk diameter {smalltilde}7 cm, height {small tilde}2.5 m) was cut under water andestimates of the flow resistances in this tree were obtained.These were much lower than the resistances to flow in the pottedtrees. Capacitance (defined as the change in stored water content perunit change in plant water potential) values were calculatedfor the small trees and the large tree from measurements ofweight and water potential changes after the trees were removedfrom water. They were very similar on a weight basis (approx.2.0 x 10^–8 kg kg^–1 Pa^–1). Leaf capacitancevalues ({small tilde}1 x 10^–8 kg Pa^–1 m^–2)were also obtained. Stomatal conductances decreased with water potential and increasedwith short-wave radiation, but the relationships were not definitive.Estimates of boundary layer conductance in a greenhouse (verylow wind speeds) were of the same order ({small tilde}5 mm s^–1)as values obtained previously.     

The Effect of Wind on Grasses: 1. CUTICULAR AND STOMATAL TRANSPIRATION

Transpiration of Festuca arundinacea Schreb, strain S170, wasmeasured at two different wind speeds in a controlled-environmentwind tunnel. As a result of a wind increase from 1 m s^–1to 3.5 m s^–1 above the sward, transpiration graduallyincreased, especially at night-time. Similar effects were notedin three other grass species. The transpiration increase couldbe attributed to decreases in stomatal and cuticular resistances,as a result of leaf buffeting.     

Light Distribution and Photosynthesis in Field Crops

In a new model of light distribution in field crops a parameters is the fraction of light passing through unit leaf layer withoutinterception. Radiation profiles measured with solarimetersand photocells give values of s from 0.7 for grasses to 0.4for species with prostrate leaves. Knowing s, leaf transmissionT and leaf-area index L the light distribution in a field cropmay be described by a binomial expansion of the form {s+(I-s)T)^L.To calculate crop photosynthesis at given light intensity thisexpansion is combined with two parameters describing the shapeof the light-response curve of single leaves. Finally, the assumptionthat solar radiation varies sinusoidally allows daily totalphotosynthesis to be estimated from daylength and insolation. The theory predicts about the same potential photosynthesisin a cloudy temperte climate with long days as in a more sunnyequatorial climate with short days. When L < 3 photosynthesisincreases as s decreases, i.e. as leaves become more prostrate;but when L > 5, photosynthesis increases as s increases,i. e. as leaves become more erect. Assuming that respirationis proportional to leaf area, estimated dry-matter productionagrees well with field measurements on sugar-beet, sugar-cane,kale, and subterranean clover. Estimates of maximum gross photosynthesis(for sugar-cane and maize) range from 60 to 9 g m^–2 day^–1depending on insolation.     

Effects of Low Concentrations of Sulphur Dioxide on Net Photosynthesis and Dark Respiration of Vicia faba

Rates of net photosynthesis, P_N, and dark respiration of Viciafaba plants were measured in the laboratory in clean air andin air containing up to 175 parts 10^–9 (500 µg m^–3)SO₂. At all SO₂ concentrations exceeding 35 parts 10^–9,P_N was inhibited compared with clean air. At light saturation,the magnitude of inhibition depended on SO₂ concentration butat low irradiances the inhibition was independent of concentration.Dark respiration rates increased substantially, independentof concentration. When exposures continued for up to 3 days,P_N returned to clean air values about 1 h after fumigation ceased:dark respiration recovered after one photoperiod. There wereno visible injuries. Reviewing possible mechanisms responsible for the inhibitionof P_N, it is suggested that SO₂ competes with CO₂ for bindingsites in RuBP carboxylase. Analysis of resistance analoguesdemonstrates that SO₂ altered both stomatal and internal (residual)resistances. A model of crop photosynthesis shows the implications of theobserved responses for the growth of field crops in which plantsare assumed to respond like laboratory plants. Photosynthesisof the crop would be less sensitive than that of individualplants to SO₂ concentration. Daily dry matter accumulation ofhypothetical ‘polluted crops’ would be substantiallyless than clean air values but would vary relatively littlewith SO₂ concentration. It is concluded that physiological basesexist to account for observed reductions in growth of plantsat very low SO₂ concentrations, and that thresholds for plantresponses to SO₂ require reassessment.     

Two types of ventilated porometers compared on broadleaf and coniferous species

Two ventilated porometers (diffusion and steady-state) were compared on four broadleaf and five coniferous species. The diffusion porometer gave consistently lower conductance values for both types of species, reflecting a direct stomatal response to low chamber humidity. At high conductance values, the porometers produced a linear and nearly equal response, but the diffusion porometer was less sensitive at low conductance values. This was due to lower air flow (20% of the velocity in the steady-state porometer) and water vapor sorption (by its acrylic plastic chamber). The broadleaf species had less variation (R² = 0.81) than did the coniferous species (R² = 0.61), but, with the latter, there was better correspondence between the two porometers, possibly due to sampling technique. Conductance values were clustered by species.     

Stomatal Responses Measured Using a Viscous Flow (Liquid) Porometer

The design of a liquid flow porometer for the measurement ofstomatal responses in epidermal peels is described. The systemis computerized and allows automatic measurement of about 1000stomata (of Commelina communis L.) in three parallel experiments.Simultaneous microscopic observations of the stomata is alsopossible. Conditions are described for rapid measurement ofstomatal responses with greater resolution than that achievedby conventional microscopic measurement techniques. To determinewhich feature(s) of the stomatal pore influenced the flow ratethe most precise estimates of pore diameter and area were madeusing a cryo-fixation technique, scanning electron microscopyand image analysis. Flow rate showed a near linear dependenceon pore area. The LFP was evaluated for measuring stomatal responsesby comparison with conventional Petri-dish experiments. Responsesto KCl, CO₂, fusicoccin, and ABA were similar using the twomethods, though there was little or no response to light inthe LFP. Pore widths were also lower in the LFP under similarexperimental conditions. The probable causes of these phenomenaare discussed. Key words: Liquid flow porometer, stomatal responses, Commelina     

Circadian Rhythm of Stomatal Movements in Epidermal Strips

Measurements were made of changes in stomatal pore widths inepidermal strips of leaves ofVicia faba and Commelina communis.Strips were incubated in dilute KCI solutions (1 and 10 molm^–3) flowing through a perfusion chamber on the stageof a microscope and kept for 4 d in continuous light. Circadianrhythms of stomatal apertures were detected in both species.Although the amplitude was small it was statistically significant.It is concluded that at least partof the mechanism for the stomatalrhythm resides in the epidermis, probably in the guard cells. Key words: Cireadian rhythm, epidermal strips, stomata     

The Flux and Distribution of Xylem Sap Calcium to Adaxial and Abaxial Epidermal Tissue 8

Inherent differences in the responses of stomata on abaxialand adaxial epidermal surfaces of leaves of Commelina communishave previously been suggested to be due to differences in theconcentrations of apoplastic Ca. Adaxial stomata have also beenreported to be more sensitive than abaxial stomata to appliedabscisic acid (ABA). The aims of these experiments were to determinethe validity of these conclusions and to see if xylem sap Cahas a role in determining the response of stomata to ABA. It can be shown from measurements of relative stomatal resistance(determined with a viscous flow porometer) and stomatal conductancethat stomata were more open in plants grown on 8-0 mol m^–3Ca, than with those grown on 2-0 mol m^–3 Ca. When attachedleaves were fed with ABA via the transpiration stream neitherthe extent nor the rate with which conductance declined wasdependent on Ca nutrition. The extent of Ca accumulation within both epidermes was relatedto the concentration of Ca in the rhizosphere and in the xylemsap. It did not, however, appear to reflect the apparent differencesin the flux of the transpiration stream between the two epidermes.Plants growing at the lower Ca concentration accumulated proportionallymore epidermal Ca relative to Ca in xylem sap. The evidencepresented suggests that Ca movement from the xylem to the epidermiscannot be simply described by a mass flow model, and that thedistribution of Ca is not an adequate explanation of the differencesin the behaviour of adaxial and abaxial stomata. The potentialrole for changes in xylem sap Ca to act as a regulator of stomatalbehaviour are discussed. Key words: Abscisic acid, calcium, Commelina communis L., stomatal conductance     

New fava bean guard cell signaling mutant impaired in ABA-induced stomatal closure

We isolated a mutant from Vicia faba L. cv. House Ryousai. Itwilts easily under strong light and high temperature conditions,suggesting that its stomatal movement may be disturbed. We determinedresponses of mutant guard cells to some environmental stimuli.Mutant guard cells demonstrated an impaired ability to respondto ABA in 0.1 mM CaCl₂ and stomata did not close in thepresence of up to 1 mM ABA, whereas wild-type stomata closedwhen exposed to 10 µM ABA. Elevating external Ca²⁺caused a similar degree of stomatal closure in the wild typeand the mutant. A high concentration of CO₂ (700 µlliter^–1) induced stomatal closure in the wild type, butnot in the mutant. On the basis of these results, we proposethe working hypothesis that the mutation occurs in the regiondownstream of CO₂ and ABA sensing and in the region upstreamof Ca²⁺ elevation. The mutant is named fia (fava bean impairedin ABA-induced stomatal closure). ³ Corresponding author: E-mail, smoiwai{at}agri.kagoshima-u.ac.jp;Fax, +81-99-285-8556.