Environmental Influences on Carbon Recycling in a Terrestrial CAM Bromeliad, Bromelia humilis Jacq.

The effects of night-time temperature, leaf-to-air vapour pressuredeficit (VPD) and water stress on CO₂ recycling in Bromeliahumilis Jacq. grown under two light and nitrogen regimes wereinvestigated. At night-time temperatures above 30°C, integratednet dark CO₂ uptake was severely reduced and CO₂ for malatesynthesis was mainly derived from dark respiration. At 35°C,up to 84% of the CO₂ liberated by dark respiration was refixedinto malic acid. Below 30 °C only nitrogen deficient plantsshowed significant recycling. No significant differences wereobserved between high and low light grown plants in CO₂ recycling.A doubling of leaf-to-air VPD from 7-46 Pa kPa^–1 to 15.49Pa kPa^–1 resulted in a 2- to 20-fold decrease in leafconductance and about 50 to 65% reduction in integrated darkCO₂ uptake. However, about twice as much CO₂ was recycled atthe higher VPD as in the lower. Ten days of water stress resultedin 80 to 100% recycling of respiratory CO₂. Under high VPD andwater stress treatments, the amount of water potentially savedthrough recycling of CO₂ reached 2- to 6-fold of the actualtranspiration. In general, nitrogen deficient plants had higherper cent recycling of respiratory CO₂ in response to high night-timetemperature, increased VPD or water stress. The results emphasizethe ecological relevance of carbon recycling in CAM plants. Key words: Bromelia humilis, CAM, PPFD, dark respiration, temperature, VPD, water stress     

The Effect of Vapour Pressure Deficit on Carbon Isotope Discrimination in the Desert Shrub Larrea tridentata(Creosote Bush)

Larrea tridentata (creosote bush) seedlings were subjected tothree regimens of atmospheric humidity in a growth chamber experiment.Relative humidity was varied to achieve daytime vapour pressuredeficits (VPD) during growth of 29, 48 and 77 kPa. Photosyntheticgas exchange, carbon isotope composition and biomass productionwere measured after 8–10 weeks of treatment. Whereas stomatalconductance (g) declined linearly with increasing ambient VPD,CO₂ assimilation rate (A) was not measurably affected by changesin ambient VPD. This resulted in a decrease in intrinsic wateruse efficiency (ratio of CO₂ assimilation to stomatal conductance;A/_g) with increasing VPD. Leaf carbon isotope discrimination(A) was negatively correlated (r² = 088) with A/g ratios. Carbonisotope discrimination also correlated positively with ratiosof internal (C₁) to ambient (c_a) CO₂ levels determined by gasexchange measurements (c₁/c). The ratio of c₁ to c_a was lowerat higher VPD levels. Leaf biomass decreased with increasingambient VPD and correlated positively with. Root to leaf biomassratio increased at higher VPD levels and correlated negativelywith. Key words: Larrea tridentata, vapour pressure deficit, carbon isotope discrimination, intrinsic water-use efficiency     

The Response of the C3-CAM Tree, Clusia rosea, to Light and Water Stress: I. GAS EXCHANGE CHARACTERISTICS

Gas exchange measurements were undertaken on 2-year-old plantsof Clusia rosea. The plants were shown to have the ability toswitch from C₃-photosynthesis to CAM and vice versa regardlessof leaf age and, under some conditions, CO₂ was taken up continuously,throughout the day and night. The light response was saturatedby 120 µmol m^–2 s^–1 typical of a shade plant. Gas exchange patterns in response to light, water and VPD wereexamined. All combinations of daytime and night-time CO₂ uptakewere observed, with rates of CO₂ uptake ranging from 2 to 11µmol m^–2 s^–1 depending upon water status andlight. Categorization of this plant asC₃, CAM or an intermediateis impossible. Differing VPD affected the magnitude of changesfrom CAM to C₃-photosynthesis (0 to 0.5 and 0 to 6.0 µmolm^–2 s^–1 CO₂, respectively) when plants were watered.Under well-watered conditions, but not under water stress, highPPFD elicited changes from CAM to C₃ gas exchange. This is unusualnot only for a shade plant but also for a plant with CAM. Itis of ecological importance for C. rosea, which may spend theearly years of its life as an epiphyte or in the forest understorey,to be able to maximize photosynthesis with minimal water loss. Key words: Clusia rosea, CAM, C₃, stress     

Effect of irradiance and vapour pressure deficit On stomatal response to CO2 enrichment of four tree species

The stomatal response of seedlings grown in 360 or 720 µmolmol^–1 to irradiance and leaf-to-air vapour pressure deficit(VPD) at both 360 and 720 µmol mol^–1 to CO₂ wasmeasured to determine how environmental factors interact withCO₂ enrichment to affect stomatal conductance. Seedlings offour species with different conductances and life histories,Cercis canadensis (L.), Quercus rubra (L.), Populus deltoides(Bartr. ex Marsh.) P. nigra (L.), and Pinus taeda (L.), weremeasured in hopes of identifying general responses. Conductanceof seedlings grown at 360 and 720 µmol mol^–1 CO₂were similar and responded in the same manner to measurementCO₂ concentration, irradiance and VPD. Conductance was lowerfor all species when measured at 720 than when measured at 360µmol mol^–1 CO₂ at both VPDs ({small tilde}1.5 and{small tilde}2.5 kPa) and all measured irradiances greater thanzero (100, 300, 600,>1600 µmol m^–2 S^–2)The average decrease in conductance due to measurement in elevatedCO₂ concentration was 32% for Cercis, 29% for Quercus, 26% forPopulus, and 11% for Pinus. For alt species, the absolute decreasein conductance due to measurement in CO₂ enrichment decreasedas irradiance decreased or VPD increased. The proportional decreasedue to measurement in CO₂ enrichment decreased in three of eightcases: from 0.46 to 0.10 in Populus and from 0.18 to 0.07 inPinus as irradiance decreased from>1600 to 100 µmolm^–2 s^–1 and from 0.35 to 0.24 in Cercis as VPD increasedfrom 1.3 to 2.6 kPa. Key words: Stomatal conductance, CO₂ enrichment, irradiance, vapour pressure deficit     

Inhibition of nitrogenase activity and nodule oxygen permeability by water deficit

Short-term effects of water deficit on nitrogenase activitywere investigated with hydroponically grown soybean plants (Glycinemax L. Merr. cv. Biloxi) by adding polyethylene glycol (PEG)to the hydroponic solution and measuring nitrogenase activity,nodule respiration, and permeability to oxygen diffusion (P_o).These experiments showed a rapid decrease in acetylene reductionactivity (ARA) and nodule respiration. A consequence of thedecreased respiration rate was that P_o calculated by Fick'sLaw also decreased. However, these results following PEG treatmentwere in direct conflict with a previous report of stabilityin P_o determined by using an alternative technique. To resolvethis conflict, an hypothesis describing a sequence of responsesto the initial PEG treatment is presented. An important findingof this study was that the response to water deficit inducedby PEG occurred in two stages. The first stage of decreasednodule activity was O₂-limited and could be reversed by exposingthe nodules to elevated pO₂. The second stage which developedafter 24 h of exposure to PEG resulted in substantial loss innodule activity and this activity could not be recovered withincreased pO₂. Severe water deficit treatments disrupt noduleactivity to such a degree that O₂ is no longer the major limitation. Key words: Glycine max, N₂ fixation, soybean, oxygen permeability, water deficit     

The Influence of Leaf Water Status and ABA on Leaf Growth and Stomata of Phaseolus Seedlings with Hypoxic Roots

The objective of this study was to assess the relative rolesof leaf water status and root-sourced signals in mediating beanleaf responses to root hypoxia. To do so, the roots of beanplants under varied VPD (0.95 kPa to 0.25 KPa) were made hypoxic.Under all conditions, leaf growth rates and stomatal conductanceswere reduced. There was a transitory decline in leaf water potentialat high VPD which accounted for the initial reduction in leafgrowth rates and stomatal conductance. At low VPD, no waterdeficits were detected. Leaf growth inhibition and reduced stomatalconductance under low VPD treatments were unrelated to leafwater status and must be induced by some other factor. In vitrogrowth of leaf discs was reduced by xylem sap collected fromhypoxic roots. Exogenously applied ABA, at high concentrationsin KCl and sucrose, or at low concentrations diluted in xylemsap from aerated plants, inhibited in vitro growth of leaf discs.Applications of ABA in the transpiration stream reduced stomatalconductance.     

Differential effects of phorbol ester (PMA) on blocker-sensitive ENaCs of frog skin and A6 epithelia

Activation ofprotein kinase C with phorbol 12-myristate 13-acetate (PMA) causedcomplex transient perturbations of amiloride-sensitive short-circuitNa⁺ currents(I_Na) in A6epithelia and frog skins that were tissue and concentration dependent.A noninvasive channel blocker pulse method of noise analysis (18) wasused to investigate how PMA caused time-dependent changes of apicalmembrane epithelial Na⁺ channel(ENaC) single-channel currents, channel open probabilities (P_o), andchannel densities(N_T). In A6epithelia, 5 and 50 nM PMA caused within 7 min concentration-dependentsustained decreases ofP_o (~55% belowcontrol, 50 nM) and rapid compensatory transient increases ofN_T within 7 min(~220% above control, 50 nM), resulting in either small transientincreases of I_Naat 5 nM PMA or small biphasic decreases ofI_Na at 50 nM PMA.In contrast to A6 epithelia, 50 and 500 nM PMA in frog skin causedafter a delay of at least 10 min transient increases ofN_T to~60-70% above control at 30-60 min. Unlike A6 epithelia,P_o was increased~15% above control within 7 min and remained within±10-15% of control for the duration of the 2-h experiments.Despite differences in the time courses of secondary inhibition oftransport in A6 epithelia and frog skin, the delayed downregulation oftransport was due to time-dependent decreases ofN_T from theirpreelevated levels in both tissues. WhereasP_o is decreasedwithin minutes in A6 epithelia as measured by noise analysis or bypatch clamp (8), the discrepancy in regulation ofN_T in A6epithelia as measured by noise analysis and patch clamp is most likelyexplained by the inability of on-cell patches formed before treatmentof tissues with PMA to respond to regulation of their channeldensities.

     

LY-294002-inhibitable PI 3-kinase and regulation of baseline rates of Na(+) transport in A6 epithelia

Blocker-inducednoise analysis of epithelial Na⁺ channels (ENaCs) was usedto investigate how inhibition of an LY-294002-sensitive phosphatidylinositol 3-kinase (PI 3-kinase) alters Na⁺transport in unstimulated and aldosterone-prestimulated A6 epithelia. From baseline Na⁺ transport rates(I_Na) of 4.0 ± 0.1 (unstimulated) and9.1 ± 0.9 µA/cm² (aldosterone), 10 µM LY-294002caused, following a relatively small initial increase of transport, acompletely reversible inhibition of transport within 90 min to 33 ± 6% and 38 ± 2% of respective baseline values. Initialincreases of transport could be attributed to increases of channel openprobability (P_o) within 5 min to 143 ± 17% (unstimulated) and 142 ± 10% of control (aldosterone) frombaseline P_o averaging near 0.5. Inhibition oftransport was due to much slower decreases of functional channeldensities (N_T) to 28 ± 4% (unstimulated)and 35 ± 3% (aldosterone) of control at 90 min. LY-294002 (50 µM) caused larger but completely reversible increases ofP_o (215 ± 38% of control at 5 min) andmore rapid but only slightly larger decreases ofN_T. Basolateral exposure to LY-294002 induced nodetectable effect on transport, P_o or N_T. We conclude that an LY-294002-sensitive PI3-kinase plays an important role in regulation of transport bymodulating N_T and P_o ofENaCs, but only when presented to apical surfaces of the cells.

     

Time, Temperature and Germination of Pearl Millet (Pennisetum typhoides S. & H.): I. CONSTANT TEMPERATURE

The germination of pearl millet (Pennisetum typhoides S. &H.) seeds was investigated at constant temperatures between12 ?C and 47 ?C on a thermal gradient plate. The rate of germination increased linearly with temperaturefrom a base T_b to a sharply defined optimum T_o beyond whichthe rate decreased linearly with temperature, reaching zeroat T_m. The linearity of the response both above and below T_oallowed time and temperature to be combined in a thermal timeat which a specified fraction of the seeds germinated. Withinthe population T_b and T_m were constant.     

Regulation of the epithelial Na(+) channel by extracellular acidification

The effect of extracellular acidification wastested on the native epithelial Na⁺ channel (ENaC) in A6epithelia and on the cloned ENaC expressed in Xenopusoocytes. Channel activity was determined utilizing blocker-inducedfluctuation analysis in A6 epithelia and dual electrode voltage clampin oocytes. In A6 cells, a decrease of extracellular pH(pH_o) from 7.4 to 6.4 caused a slow stimulation of theamiloride-sensitive short-circuit current (I_Na)by 68.4 ± 11% (n = 9) at 60 min. This increaseof I_Na was attributed to an increase of openchannel and total channel (N_T) densities. Similar changes were observed with pH_o 5.4. The effects ofpH_o were blocked by buffering intracellularCa²⁺ with 5 µM1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid. Inoocytes, pH_o 6.4 elicited a small transient increase of theslope conductance of the cloned ENaC (11.4 ± 2.2% at 2 min)followed by a decrease to 83.7 ± 11.7% of control at 60 min (n = 6). Thus small decreases of pH_ostimulate the native ENaC by increasing N_T butdo not appreciably affect ENaC expressed in Xenopus oocytes.These effects are distinct from those observed with decreasingintracellular pH with permeant buffers that are known to inhibit ENaC.

     

The Influence of Temperature on Seed Germination in Cultivars of Common Bean

Common bean (Phaseolus vulgaris L.) is grown over a wide rangeof environments, including sites with low or high soil temperaturesat sowing time. To describe the temperature responseof seedgermination, 20 bean genotypes were evaluated using a rolledpaper towel system with 11 constant temperatures ranging from12 to 34 °C. Germination response was characterized by fittingcumulative counts using a maximum-likelihood analysis. Rateof germination increased from abase temperature (T_b) typicallynear 8 °C to an optimal development temperature (T_o) of29 to 34 °C. T_b did not differ among common bean genotypes.Mesoamerican germplasm showed slightlyhigher T_o than Andeangermplasm, but there was large variation in T_o within each ofthe two gene pools. The single accession of tepary bean (P.acutifolius) evaluated appeared to be the mosttolerant to highgermination temperatures. Key words: Common bean, seed germination rate, temperature     

Respiration and germination rates of tomato seeds at suboptimal temperatures and reduced water potentials

Suboptimal temperature (T) affects germination rates (reciprocalof time to radicle emergence) on a thermal time basis; thatis, the T in excess of a base or minimum temperature multipliedby the time to a given per cent germination [t_g) is a constant.Respiration rates are also sensitive to T, and proportionalrelationships are often found between respiration rates andgermination rates. Reduced water potential () delays seed germinationon a hydrotime basis (i.e. the in excess of a base water potentialmultiplied by t_g is a constant). It was tested whether respirationrates prior to radicle emergence vary in proportion to T and as expected from the thermal and hydrotime models. Respirationrates (C0₂ evolution) of cold-tolerant, rapidly germinating(PI 341988) and cold-sensitive, more slowly germinating (T5)tomato [Lycopersicon esculentum Mill.) seeds were evaluatedover a range of T and conditions. For both genotypes, respirationrates until the beginning of radicle emergence were relatedto T on a thermal time basis and increased approximately linearlywith above -2.0 MPa, consistent with the hydrotime model. Respirationrates were uniquely related to germination rates, regardlessof whether germination timing was affected by T, , or genotype.However, germination timing was unaffected when respirationrates were manipulated by varying 0₂ partial pressure. Thus,while both germination and respiration rates vary with T and consistent with thermal and hydrotime models of biologicaltime, respiration rates per se were not the limiting factorin germination timing of tomato seeds. Key words: Lycopersicon esculentum Mill., tomato, germination, respiration, temperature, water potential     

The Influence of Temperature on Seed Germination Rate in Grain Legumes: II. INTRASPECIFIC VARIATION IN CHICKPEA (Cicer arietinum L.) AT CONSTANT TEMPERATURES

Positive linear relationships were shown between constant temperaturesand the rates of progress of germination to different percentiles,G, for single populations of each of five genotypes of chickpea(Cicer anetinum L.). The base temperature, T_b, at which therate of germination is zero, was 0·0°C for all germinationpercentiles of all genotypes. The optimum temperature, T_o(G),at which rate of germination is most rapid, varied between thefive genotypes and also between percentiles within at leastone population. Over the sub-optimal temperature range, i.e.from T_b to T_o(G), the distribution of thermal times within eachpopulation was normal. Consequently a single equation was appliedto describe the influence of sub-optimal temperatures on rateof germination of all seeds within each population of each genotype.The precision with which optimum temperature, T_b(G), could bedefined varied between populations. In each of three genotypesthere was a negative linear relationship between temperatureabove T_b(G) and rate of germination. For all seeds within anyof these three populations thermal time at supra-optimal temperatureswas constant. Variation in the time taken to germinate at supra-optimaltemperatures was a consequence of normal variation in the ceilingtemperature, T_o(G)—the temperature at or above which rateof progress to germination percentile G is zero. A new approachto defining the response of seed germination rate to temperatureis proposed for use in germplasm screening programmes. In two populations final percentage germination was influencedby temperature. The optimum constant temperature for maximumfinal germination was between 10°C and 15°C in thesepopulations; approximately 15°C cooler than the optimumtemperature for rate of germination. It is suggested that laboratorytests of chickpea germination should be carried out at temperaturesbetween 10°C and 15°C. Key words: Chickpea, seed germination rate, temperature     

The Influence of Temperature on Seed Germination Rate in Grain Legumes: I. A COMPARISON OF CHICKPEA, LENTIL, SOYABEAN AND COWPEA AT CONSTANT TEMPERATURES

For a single seed population of each of four species of grainlegume positive linear relationships were shown between temperatureand rate of germination for different fractions (G) of eachpopulation, from a base temperature, T_b(G), at which germinationrate is zero, to an optimum temperature, T_o(G) at which germinationrate is maximal. At constant temperatures warmer than T_o(G)there were negative relations (probably linear) between temperatureand rate of germination to the maximum temperature for germination,T_m(G), Within each population T_b(G) did not differ, but it didvary between species, viz.0.0?C, 0.25?C, 4.and 8.5?C for chickpea(Cicer arietinum L.), lentil (Lens culinaris Medic.), soyabean(Glycine max [ Merr.) and cowpea (Vigna unguiculata [L.] Walp.),respectively. In contrast, T_o(G) varied both within each populationand also between the four species: 80% of seeds in each populationhad T_o(G) values within the range 31.8?C to 33.8 ?C, 24.0?Cto 24.4?C, 34.0?C to 34.5?C and 33.2?C to >40?C, respectively.Values of T_m(G) were much more vanable: the 80% population rangewas 48 .0?C to 60.8?C for chickpea, 31.8?C to 34.4?C for lentiland 46.8?C to 55.2?C for soyabean; reliable estimates couldnot be made for cowpea, but the results suggest higher and morevariable values of T_m(G) than in the other three species. Atsub-optimal temperatures the distribution of thermal time forthe different fractions of each population was normal, exceptfor lentil where it was log-normal. A single equation is proposedto describe the influence of sub-optimal temperatures on ratesof germination for whole seed populations. At supra-optimaltemperatures, variation in thermal time for the different fractionsof each population was only slight. The implications of thesefindings for the adaptation of grain legume crops to differentenvironments, and for the screening of germplasm, are discussed. Key words: Seed germination rate, temperature, grain legumes     

The Dielectric Relaxation of Water in Plant Tissue

Thermally stimulated depolarization current (TSDC) measurementson plant leaves and stems of six different species in the temperaturerange of 77–300 K revealed the existence of three differentdispersions. The first dispersion at low temperatures, whichis attributed to the relaxation of loosely bound water moleculeswas studied in detail in an attempt to obtain information onthe possible structures of water in plant tissue. Its characteristicsdiffer for various plant tissues, indicating a different organizationof water in those plant tissues. The dispersion can be describedby a continuous distribution of relaxation times t with boththe activation energy W and the pre-exponential factor To inthe Arrhenius equation being distributed parameters. The spectrumof W and T_o was determined for E. globulus and O. europaea leafsamples. The mean values of T and W are larger and that of T_osmaller than the corresponding values for free (bulk) water.The results favour a model of the organization of water in clustersrather than in multilayers and indicate a stronger binding ofwater in living systems. Key words: Dielectric relaxation, distribution of relaxation times, free and bound water     

Time-dependent stimulation by aldosterone of blocker-sensitive ENaCs in A6 epithelia

To study and define the early time-dependent response (6 h) ofblocker-sensitive epithelial Na⁺channels (ENaCs) to stimulation ofNa⁺ transport by aldosterone, weused a new modified method of blocker-induced noise analysis todetermine the changes of single-channel current (i_Na) channel open probability(P_o), andchannel density(N_T) undertransient conditions of transport as measured by macroscopic short-circuit currents(I_sc). In threegroups of experiments in which spontaneous baseline rates of transportaveraged 1.06, 5.40, and 15.14 µA/cm², stimulation of transportoccurred due to increase of blocker-sensitive channels.N_T variedlinearly over a 70-fold range of transport (0.5-35µA/cm²). Relatively small andslow time-dependent but aldosterone-independent decreases ofP_o occurredduring control (10-20% over 2 h) and aldosterone experimentalperiods (10-30% over 6 h). When theP_o of control andaldosterone-treated tissues was examined over the 70-fold extendedrange of Na⁺ transport,P_o was observedto vary inversely withI_sc, falling from~0.5 to ~0.15 at the highest rates ofNa⁺ transport or ~25% per3-fold increase of transport. Because decreases ofP_o from anysource cannot explain stimulation of transport by aldosterone, it isconcluded that the early time-dependent stimulation ofNa⁺ transport in A6 epithelia isdue exclusively to increase of apical membraneN_T.

     

Hormonal regulation of ENaCs: insulin and aldosterone

Although a variety of hormones and other agents modulate renalNa⁺ transport acting by way of theepithelial Na⁺ channel (ENaC), themode(s), pathways, and their interrelationships in regulation of thechannel remain largely unknown. It is likely that several hormones maybe present concurrently in vivo, and it is, therefore, important tounderstand potential interactions among the various regulatory factorsas they interact with the Na⁺transport pathway to effect modulation ofNa⁺ reabsorption in distal tubulesand other native tissues. This study represents specifically adetermination of the interaction between two hormones, namely,aldosterone and insulin, which stimulate Na⁺ transport by entirelydifferent mechanisms. We have used a noninvasive pulse protocol ofblocker-induced noise analysis to determine changes in single-channelcurrent (i_Na),channel open probability (P_o), andfunctional channel density(N_T) ofamiloride-sensitive ENaCs at various time points following treatmentwith insulin for 3 h of unstimulated control and aldosterone-pretreatedA6 epithelia. Independent of threefold differences of baseline values of transport caused by aldosterone, 20 nM insulin increased by threefold and within 10-30 min the density of the pool of apical membrane ENaCs(N_T) involvedin transport. The very early (10 min) increases of channel density wereaccompanied by relatively small decreases ofi_Na(10-20%) and decreases ofP_o (28%) in the aldosterone-pretreated tissues but not the control unstimulated tissues. The early changes ofi_Na,P_o, andN_T weretransient, returning very slowly over 3 h toward their respectivecontrol values at the time of addition of insulin. We conclude thataldosterone and insulin act independently to stimulate apicalNa⁺ entry into the cells of A6epithelia by increase of channel density.

     

Dibasic phosphorylation sites in the R domain of CFTR have stimulatory and inhibitory effects on channel activation

To better understand the mechanisms by which PKA-dependent phosphorylation regulates CFTR channel activity, we have assayed open probabilities (P_o), mean open time, and mean closed time for a series of CFTR constructs with mutations at PKA phosphorylation sites in the regulatory (R) domain. Forskolin-stimulated channel activity was recorded in cell-attached and inside-out excised patches from transiently transfected Chinese hamster ovary cells. Wild-type CFTR and constructs with a single Ser-to-Ala mutation as well as octa (Ser-to-Ala mutations at 8 sites) and constructs with one or two Ala-to-Ser mutations were studied. In cell-attached patches, Ser-to-Ala mutations at amino acids 700, 795, and 813 decreased P_o, whereas Ser-to-Ala mutations at 737 and 768 increased P_o. In general, differences in P_o were due to differences in mean closed time. For selected constructs with either high or low values of P_o, channel activity was measured in excised patches. With 1 mM ATP, P_o was similar to that observed in cell-attached patches, but with 10 mM ATP, all constructs tested showed elevated P_o values. ATP-dependent increases in P_o were due to reductions in mean closed time. These results indicate that R-domain phosphorylation affects ATP binding and not the subsequent steps of hydrolysis and channel opening. A model was developed whereby R-domain phosphorylation, in a site-dependent manner, alters equilibrium between forms of CFTR with low and high affinities for ATP. site-directed mutagenesis; kinase-dependent activation; cell-attached patch clamp; open probability; mean open time     

Intracellular pH shifts in cultured kidney (A6) cells: effects on apical Na+ transport

We report, for the epithelialNa⁺ channel (ENaC) in A6 cells,the modulation by cell pH (pH_c)of the transepithelial Na⁺ current(I_Na), thecurrent through the individual Na⁺channel (i), the openNa⁺ channel density(N_o), and thekinetic parameters of the relationship betweenI_Na and theapical Na⁺ concentration. Thei andN_o were evaluatedfrom the Lorentzian I_Na noise inducedby the apical Na⁺ channel blocker6-chloro-3,5-diaminopyrazine-2-carboxamide.pH_c shifts were induced, understrict and volume-controlled experimental conditions, byapical/basolateral NH₄Cl pulses orbasolateral arrest of theNa⁺/H⁺exchanger (Na⁺ removal; block byethylisopropylamiloride) and were measured with the pH-sensitive probe2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein. Thechanges in pH_c were positivelycorrelated to changes inI_Na and theapically dominated transepithelial conductance. The sole pH_c-sensitive parameter underlyingI_Na wasN_o. Only thesaturation value of theI_Na kinetics wassubject to changes in pH_c.pH_c-dependent changes inN_o may be causedby influencingP_o, the ENaC openprobability, or/and the total channel number,N_T = N_o/P_o.

     

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

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