A new approach to the investigation on the tonogenic groups of root cell walls

Acid-base properties of wheat, lupin, pea root cell walls were investigated. The roots of etiolated and green plants of different age were analysed by the potentiometric method. The ion exchange capacity of root cell walls (S_i) was estimated at various pH values (pH_i 2 to pH_i 12) and constant ion strength of the solution (10 mM). To analyse polysigmoid curves pH_i =f (S_i), Gregor's equation was used. It was shown that Gregor's model fits fairly well the experimental data. The total quantities of cation-exchange (S_t ^cat) and anion-exchange (S_t ^an) groups were determined in the root cell walls. It was shown that the quantity of anion exchange groups is varied through a small range (60–185 μmol/g dry wt.) in plant species tested, and that the S_t ^cat differs widely from 550 to 1300 μmol/g dry wt. For leguininous plants the quantity of acidic groups (fixed anions) is nearly twice as large as that for cereals. It was found that in seedlings as well as in plants, there are 3 cation-exchange groups and one anion-exchange group in root cell walls. The quantity of functional groups of each type (S^j) was estimated, and the corresponding values of n^j and pK_a ^j were calculated. It can be assumed that the groups with the pK_a ¹ ≈ 3.2 are amine groups, the ones with PK_a ² ≈ 5 are groups of galacturonic acid, the ones with pK_a ≈ 7.5 are the carboxyl groups of the second species, and the ones with pK_a ⁴ ≈ 40 are the phenolic groups. The values of dissociation constants (pK_a ^j) and S^j indicate that the root cell walls of wheat, lupin and pea are identical in qualitative structure of ionogenic groups but vary in the quantity of each ionogenic group. It was demonstrated that the summarized quantity of carboxyl groups (S² + S³) should be connected directly with the pH gradient in the extracellar space at the membrane surface. The gradient arises from ion-exchange reactions between cations of an outer medium and protons of the ionized carboxyl groups of the cell walls. The results suggest that, S_t ^cat and S_t ^an allow the quantitative estimation of ion exchange properties of the cell walls. The resulting parameters (S^j, pK_a ^j and n^j) allow prediction of changes in an ionic composition of a medium that bathes the cell membrane, during the first step of mineral nutrition uptake. This revised version was published online in June 2006 with corrections to the Cover Date.     

Swelling of Root Cell Walls as an Indicator of Their Functional State

The swelling capacity of cell walls isolated from different parts of lupine root was investigated. The water content in fragments of intact roots (Q) and swelling coefficient of standardized samples of cell walls (K^cw) were determined, and the dependences of Q and Kcw on the distance from the root tip (L) were plotted. It was shown that the change in Q value along the stretch of the lupine root reaches its maximum at distances of 1.5-6 cm or 7-12 cm from the root tip in 7-day-old and 14-day-old seedlings, respectively, whereas the K^cw value distribution over the root length is virtually invariable. In the radial direction, both the Q and K^cw values in cortex tissues are about twice higher than in the central cylinder. In our opinion, the changes of both Q and K^cw in the radial direction are associated with different degrees of cross-linking between polymer chains in cell wall structures of root cortex and central cylinder. The results of measurement of the K^cw value are consistent with the widely accepted mechanisms of water transport in roots in the radial direction. These data show that water transport through apoplast to the border between the cortex and central cylinder is accompanied by an increase in the resistance to water flow. Among other factors, this increase is due to a greater degree of cross-linking between cell wall polymers in the central cylinder. The results of measurement of the swelling coefficient of standardized cell wall samples in water and in 10 mM KCl at different pH values show that the swelling capacity of root cell walls varies according to the physicochemical properties of synthetic ion exchangers. Cell walls shrink (cell wall volume decreases) as ion concentration in solution increases and pH decreases. This causes an increase in the hydraulic resistance (or a decrease in the hydraulic conductivity) of apoplast. It was concluded that swelling is determined by the physicochemical properties of the cell wall, whereas the change in the swelling capacity induced by variation of external or internal conditions is an element of the mechanism of regulation of volume water flow in roots.     

Ion-Exchange Properties of Cell Walls Isolated from Lupine Roots

Acid–base properties of cell walls isolated from various root tissues of 7-day-old lupine seedlings and 14-day-old lupine plants grown in various media were studied. The ion-exchange capacity of root cell walls was estimated at various pH values (from 2 to 12) and constant ionic strength (10 mM). The parameters determining the qualitative and quantitative composition of cell wall ionogenic groups along the root length and in its radial direction were estimated using Gregor's model. This model fits the experimental data reasonably well. Four types of ionogenic groups were found in the cell walls: an amino group (pK _a 3), two types of carboxylic groups (pK _a 5 and 7.3, the first being the carboxylic group of galacturonic acid), and a phenolic group (pK _a 10). The number of functional groups of each type was estimated, and the corresponding ionization constant values were calculated. It is shown that the chemical composition of the ionogenic groups was constant along the root length as well as in its radial direction and did not depend on either physiological state or root nutrition, while the number of different groups varied. The content of carboxylic groups of -D-polygalacturonic acid in the root cell walls of 14-day-old plants was shown to depend on the distance from the root tip, being maximal in the zone of lateral roots. The number of these groups was 10- and 2-fold less in the central cylinder compared to that of cortex for 14-day-old plants and 7-day-old seedlings, respectively.     

A model for water transport in the stele of plant roots

Summary The mechanism of water movement across roots is, as yet, not well understood. Some workable black box theories have already been proposed. They, however, assumed unrealistic cell membranes with low values of , or were based on a poor anatomical knowledge of roots. The role of root stele in solute and water transport seems to be especially uncertain. An attempted explanation of the nature of root exudation and root pressure by applying the apoplast canal theory (Katou andFurumoto 1986 a, b) to transport in the root stele is given. The canal equations are solved for boundary conditions based on anatomical and physiological knowledge of the root stele. It is found that the symplast cell membrane, cell wall and net solute transport into the wall apoplast are the essential constituents of the canal system. Numerical analysis shows that the canal system enables the coupled transport of solutes and water into a xylem vessel, and the development of root pressure beyond the level predicted by the osmotic potential difference between the ambient medium and the exudate. Observations on root exudation and root pressure previously reported seem to be explained quite well. It is concluded that the movement of water in the root stele although apparently active is essentially osmotic.Abbreviations J _v ^ex volume exudation per root surface - J₀ non-osmotic exudation - L^r overall radial hydraulic conductivity of an excised root - reflection coefficient - C_s difference in the osmotic concentration between the bathing medium and the exudate - R gas constant - T absolute temperature - C_K molar concentration of K⁺ - C_Cl molar concentration of Cl^– - C_j molar concentration of ion species j - P_j membrane permeability of ion j - z_j valence of ion j - F Faraday constant - V_ix intracellular electric potential with reference to the canal     

Occurrence of the suberized lamella in leaves of grasses of different photosynthetic types. I. In parenchymatous bundle sheaths and PCR (“Kranz”) sheaths

Summary Leaf blades of 42 grasses (Poaceae) have been examined ultrastructurally for the occurrence of a suberized lamella in walls of parenchymatous bundle sheaths and PCR (= Kranz) sheaths in both large and small vascular bundles. The sample includes species from a range of major grass taxa, and represents all photosynthetic types found in the grasses. Three grasses with unusual C₄ leaf anatomy were also included:Alloteropsis semialata, Aristida biglandulosa, Arundinella nepalensis. The presence of a suberized lamella in PCR cell walls was perfectly correlated with photosynthetic type. All PEP-carboxykinase type and NADP-malic enzyme type C₄ species examined possessed a suberized lamella in outer tangential and radial walls, but with variable presence in inner tangential walls. PCR cells of bothAlloteropsis semialata andArundinella nepalensis also possessed a suberized lamella. A lamella was totally absent from parenchymatous bundle sheath cells of the C₃ species examined (5 spp.) and ofPanicum milioides, a C₃-C₄ intermediate. It was also absent from PCR cells of NAD-malic enzyme type C₄ species (14 spp.) andAristida biglandulosa. The results are discussed in relation to the leakage of CO₂ from PCR cells, and to differences between C₄ types in ¹³C values, chloroplast position in PCR cells, and other anatomical characteristics.     

Cell walls as reservoirs of potassium ions for reversible volume changes of pulvinar motor cells during rhythmic leaf movements

The laminar pulvinus of primary leaves of Phaseolus coccineus L. was investigated with respect to the total K⁺ content, the apoplastic K⁺ content, and the water potential of extensor and flexor sections in relation to the leaf positions in a circadian leaf-movement cycle, as well as the cation-exchange properties of isolated extensor- and flexor-cell walls. Turgid tissue showed a high total but low apoplastic K⁺ content, shrunken tissue a low total but high apoplastic K⁺ content. Thus, part of the K⁺ transported into and out of the swelling or shrinking protoplasts is shuttled between the protoplasts and the surrounding walls, another part between different regions of the pulvinus. The K⁺ fraction shuttled between protoplasts and walls was found to be 30–40% of the total transported K⁺ fraction. Furthermore, 15–20% of the total K⁺ content of the tissue is located in the apoplast when the apoplastic reservoir is filled, 5–10% when the apoplastic reservoir is depleted. The ion-exchange properties of walls of extensor and flexor cells appear identical in situ and in isolated preparations. The walls behave as cation exchangers of hhe weak-acid type with a strong dependence of the activity of fixed negative charges as well as of the K⁺-storing capacity on pH and [K⁺] of the equilibration solution. The high apoplastic K⁺ contents of freshly cut tissues reflect the cation-storing capacity of the isolated walls. We suggest that K⁺ ions of the Donnan free space are used for the reversible volume changes (mediating the leaf movement) mainly by an electrogenic proton pump which changes the pH and-or the [K⁺] in the water free space of the apoplast.Abbreviations and symbols DFS Donnan free space - DW dry weight - pK negative logarithm of the equilibrium constant K of the acidic group - WFS water free space - water potential; Indices - cw cell wall - t tissue     

Modulation of a plant mitochondrial K+ATP channel and its involvement in cytochrome c release

Pea stem mitochondria, resuspended in a KCl medium (de-energized mitochondria), underwent a swelling, as a consequence of K⁺ entry, that was inhibited by ATP. This inhibition was partially restored by GTP and diazoxide (K⁺ _ATP channel openers). In addition, glyburide and 5-hydroxydecanate (K⁺ _ATP channel blockers) induced an inhibition of the GTP-stimulated swelling. Mitochondrial swelling was inhibited by H₂O₂, but stimulated by NO. The same type of responses was also obtained in succinate-energized mitochondria. When the succinate-dependent transmembrane electrical potential () had reached a steady state, the addition of KCl induced a dissipation that was inhibited by H₂O₂ and stimulated by NO. The latter stimulation was prevented by carboxy-PTIO, a NO scavenger. Phenylarsine oxide (a thiol oxidant) and NEM (a thiol blocker) stimulated the KCl-induced dissipation of , while DTE prevented this effect in both cases. In addition, DTE transiently inhibited the NO-induced dissipation of , but then it caused a more rapid collapse. These results, therefore, show that the plant mitochondrial K⁺ _ATP channel resembles that present in mammalian mitochondria and that it appears to be modulated by dithiol–disulfide interconversion, NO and H₂O₂. The aperture of this channel was linked to the partial rupture of the outer membrane. The latter effect led to a release of cytochrome c, thus suggesting that this release may be involved in the manifestation of programmed cell death.     

Continuous penicillin G hydrolysis in an electro-membrane reactor with immobilized penicillin G acylase

Penicillin G (2%, w/v in phosphate buffer, pH 8) was hydrolysed in a flow-through, miniature electro-membrane reactor with the penicillin G acylase immobilized in 5% (w/v) polyacrylamide (diam. 10 mm, thickness 2.6 mm, enzyme activity 24 U ml^–1). The conversion of penicillin G increased from 0.15 to almost 0.5 when the electric current applied to the reactor was changed from –600 to +600 A/m² with a substrate residency of 1 h. Symbols and abbreviations c _j ^p & concentration of component j in product stream (M) c _j ^s & concentration of component j in substrate stream (M) c _s ^o & substrate concentration at reactor inlet (M) C _j ^p=c _j ^p/c _S ⁰ & scaled concentration of component j in product stream C _j ^s=c _j ^s/c _S ⁰ & scaled concentration of component j in substrate stream i & electric current density (A/m²) j & reaction component, j P, Q or S P & main reaction product (6-aminopenicillanic acid) PGA & penicillin G acylase Q & side reaction product (phenylacetic acid) S & substrate (penicillin G) Y _s=C _P ^s+C _P ^p & substrate conversion & mean residence time of substrate and product streams in reactor (h) =C _Q ^s+C _Q ^p+C _S ^s+C _S ^s & check-sum of scaled concentrations =C _P ^p/(C _P ^s+C _P ^p) & separation factor of 6-aminopenicillanic acid (0 1)     

Is the Voltage Gate of Connexins CO<Subscript>2</Subscript>-sensitive? Cx45 Channels and Inhibition of Calmodulin Expression

The sensitivity of Cx45 channels to CO₂, transjunctional voltage (V _j) and inhibition of calmodulin (CaM) expression was tested in oocytes by dual voltage clamp. Cx45 channels are very sensitive to V _j and close with V _j preferentially by the slow gate, likely to be the same as the chemical gate. With a CO₂-induced drop in junctional conductance (G _j), both the speed of V _j-dependent inactivation of junctional current (I _j) and V _j sensitivity increased. With 40-mV V _j-pulses, the of single exponential I _j decay reversibly decreased by 40% during CO₂ application, and G_{j steady state}/G_{j peak} decreased multiphasically, indicating that both kinetics and V _j sensitivity of chemical/slow V _j gating are altered by changes in [H⁺]_i and/or [Ca²⁺]_i. CaM expression was inhibited with oligonucleotides antisense to CaM mRNA. With 15 min CO₂, relative junctional conductance (G _jt/G _jt0) dropped to 0% in controls, but only by 17% in CaM-antisense oocytes. Similarly, V _j sensitivity was significantly lessened in CaM-antisense oocytes. The data indicate that both the speed and sensitivity of V _j-dependent inactivation of the junctional current of Cx45 channels are affected by CO₂ application, and that CaM plays a key role in channel gating.     

How Common Are Intragene Windows with <Emphasis Type="Italic">K</Emphasis><Subscript>A</Subscript> > <Emphasis Type="Italic">K</Emphasis><Subscript>S</Subscript> Owing to Purifying Selection on Synonymous Mutations?

One method for diagnosing the mode of sequence evolution considers the ratio of nonsynonymous substitutions per nonsynonymous site (K _A) to the corresponding figure for synonymous substitutions (K _S). A ratio (K _A/K _S) greater than unity is taken as evidence for positive selection. This, however, need not necessarily be the case. Notably, there is one instance of a high intragenic K _A/K _S peak, revealed by sliding window analysis and observed in two pairwise comparisons, better accounted for by localised purifying selection on synonymous mutations that affect splicing. Is this example exceptional? To address this we isolate intragenic domains with K _A/K _S > 1 from more than 1000 long mouse-rat orthologues. Approximately one K _A/K _S > 1 peak is found per 12–15 kb of coding sequence. Surprisingly, low synonymous substitution rates underpin more incidences than do high nonsynonymous rates. Several reasons, however, prevent us from supposing that the low synonymous rates reflect purifying selection on synonymous mutations. First, for many peaks, the null that the peak is no higher than expected given the underlying rates of evolution, cannot be rejected. Second, of 18 statistically significant incidences with unusually low K _S values, only 3 are repeatable across independent comparisons. At least two of these are within alternatively spliced exons. We conclude that repeatable statistically significant intragenic domains of low intragenic K _S are rare. As so few K _A/K _S peaks reflect increased rates of protein evolution and so few hold statistical support, we additionally conclude that sliding window analysis to infer domains of positive selection is highly error-prone.     

Light-promoted changes in apoplastic K+ activity in the Samanea saman pulvinus,monitored with liquid membrane microelectrodes

The movement of Samanea saman (Jacq.) Merrill leaflets is a consequence of the re-distribution of K⁺ and anions between motor cells on opposite sides of the pulvinus. We used a K⁺-sensitive microelectrode to study dynamic changes in K⁺ transport through motor-cell membranes during and immediately after change in illumination. Potassium-ion-sensitive and reference microelectrodes were inserted into extensor or flexor tissue of a whole pulvinus in white light (WL). A brief pulse of red light (RL) followed by darkness (D) (a) increased K⁺ activity in the extensor apoplast, indicating K⁺ release by the protoplast; and (b) decreased K⁺ activity in the flexor apoplast, indicating K⁺ uptake by the protoplast. White light after 35–40 min D reversed K⁺ activity in the extensor apoplast to approximately its original value. Blue light substituted partially for WL in this regard. Potassium-ion activity in the flexor apoplast reverted to approximately its original value after 2 h, with or without white illumination. Our data support the hypothesis that K⁺ efflux from extensor cells and K⁺ uptake by flexor cells following a WLRLD transition occurs by way of K⁺ channels.Abbreviations L light - WL white light - RL red light - BL blue light - D darkness     

Incorporation of 14CO2 into C4 acids by leaves of C3-C4 intermediate and C3 species of Moricandia and Panicum at the CO2 compensation concentration

Comparative ¹⁴CO₂ pulse-¹²CO₂ chase studies performed at CO₂ compensation ()-versus air-concentrations of CO₂ demonstrated a four-to eightfold increase in assimilation of ¹⁴CO₂ into the C₄ acids malate and aspartate by leaves of the C₃-C₄ intermediate species Panicum milioides Nees ex Trin., P. decipiens Nees ex Trin., Moricandia arvensis (L.) DC., and M. spinosa Pomel at . Specifically, the distribution of ¹⁴C in malate and aspartate following a 10-s pulse with ¹⁴CO₂ increases from 2% to 17% (P. milioides) and 4% to 16% (M. arvensis) when leaves are illuminated at the CO₂ compensation concentration (20 l CO₂/l, 21% O₂) versus air (340 l CO₂/l, 21% O₂). Chasing recently incorporated ¹⁴C for up to 5 min with ¹²CO₂ failed to show any substantial turnover of label in the C₄ acids or in carbon-4 of malate. The C₄-acid labeling patterns of leaves of the closely related C₃ species, P. laxum Sw. and M. moricandioides (Boiss.) Heywood, were found to be relatively unresponsive to changes in pCO₂ from air to . These data demonstrate that the C₃-C₄ intermediate species of Panicum and Moricandia possess an inherently greater capacity for CO₂ assimilation via phosphoenolpyruvate (PEP) carboxylase (EC 4.1.1.31) at the CO₂ compensation concentration than closely related C₃ species. However, even at , CO₂ fixation by PEP carboxylase is minor compared to that via ribulosebisphosphate carboxylase (EC 4.1.1.39) and the C₃ cycle, and it is, therefore, unlikely to contribute in a major way to the mechanism(s) facilitating reduced photorespiration in the C₃-C₄ intermediate species of Panicum and Moricandia.Abbreviations Rubisco ribulose-1,5-bisphosphate carboxylase/oxygenase - PEP phosphoenolpyruvate - CO₂ compensation concentration - 3PGA 3-phosphoglycerate - SuP sugar monophosphates - SuP₂ sugar bisphosphates Published as Paper No. 8249, Journal Series, Nebraska Agricultural Research Division     

Effects of temperature and CO2 enrichment on kinetic properties of NADP+-malate dehydrogenase in two ecotypes of Barnyard grass (Echinochloa crus-galli (L.) Beauv.) from contrasting climates

Summary The apparent energy of activation (E _a), Michaelis-Menten constant (K _mfor oxaloacetate), V _max/K _mratios and specific activities of NADP⁺-malate dehydrogenase (NADP⁺-MDH; EC 1.1.1.82) were analyzed in plants of Barnyard grass from Québec (QUE) and Mississippi (MISS) acclimated to two thermoperiods 28/22°C, 21/15°C, and grown under two CO₂ concentrations, 350 l l^-1 and 675 l l^-1. E _avalues of NADP⁺-MDH extracted from QUE plants were significantly lower than those of MISS plants. K _mvalues and V _max/K _mratios of the enzyme from both ecotypes were similar over the range of 10–30°C but reduced V _max/K _mratios were found for the enzyme of QUE plants at 30 and 40°C assays. MISS plants had higher enzyme activities when measured on a chlorophyll basis but this trend was reversed when activities were expressed per fresh weight leaf or per leaf surface area. Activities were significantly higher in plants of both populations acclimated to 22/28°C. CO₂ enrichment did not modify appreciably the catalytic properties of NADP⁺-MDH and did not have a compensatory effect upon catalysis or enzyme activity under cool acclimatory conditions. NADP⁺-MDH activities were always in excess of the amount required to support observed rates of CO₂ assimilation and these two parameters were significantly correlated. The enhanced photosynthetic performance of QUE plants under cold temperature conditions, as compared to that of MISS plants, cannot be attributed to kinetic differences of NADP⁺-malate dehydrogenase among these ecotypes.     

Control of thickness of collenchyma cell walls by pectins

Near-isotropic stresses were generated within collenchyma cell walls of celery (Apium graveolens L.) by exchanging K⁺ for Ca²⁺ ions, varying the ionic strength and de-esterifying the pectic carboxyl groups, treatments that changed the free-charge density of the pectic polysaccharides. The collenchyma strands swelled radially with increasing free-charge density but there was very little longitudinal swelling. Depolymerising the pectins by -elimination also induced much more radial than longitudinal swelling. Supported by earlier work on Nitella, these results indicate that pectins control the interlamellar spacing in cell walls and hold them together across their thickness, particularly against turgor stresses tending to delaminate the walls at the cell corners.The author thanks J.S.G. Reid (Department of Biological Sciences, University of Stirling, UK) and M. Demarty (SCUEOR, University of Rouen, France) for critical comments.     

Physiological responses of lupin roots to high pH

High pH seems to be a major constraint limiting the production of narrow-leafed lupin (Lupinus angustifolius L.) on alkaline soils. Whereas there has been much interest in soil acidity, relatively little is known about the effect of high pH on the growth of roots of higher plants.Elongation of roots of L. angustifolius was particularly sensitive to pH6.0 compared with other species. The effect of high pH in decreasing root elongation in L. angustifolius occurred within one hour. It was via an effect on cell elongation and not cell division and the effect was readily reversible. The mechanisms of the adverse effect of high pH are unknown. The permeability ratio of K⁺ to Na⁺ in the plasma-membrane of the root cortical cells was similar in solutions of both low pH and high pH. Reduced cell growth at high pH was not associated with an inefficiency of proton extrusion to the bulk solution by roots of this species. Nevertheless, increasing buffer concentration in the external solution decreased root elongation more in L. angustifolius than in Lupinus pilosus and Pisum sativum.     

The active species of “CO2” utilized in ferredoxin-linked carboxylation reactions

The active species of CO₂ , i.e. CO₂ or HCO ₃ ^– –(H₂CO₃) utilized by enzymes catalyzing ferredoxin-linked carboxylation reactions was determined. The enzyme investigated was pyruvate synthase from Clostridium pasteurianum (EC 1.2.7.1; Pyruvate: ferredoxin oxidoreductase). Data were obtained which were compatible with those expected if CO₂ is the active species.The dissociation constant (K _S) of the enzyme-CO₂ complex was measured. At pH 7.2 K _Sfor CO₂ of pyruvate synthase was found to be approximately 5 mM.Abbreviations Fd ferredoxin No distinctions are made between CO₂, H₂CO₃, HCO ₃ ^– and CO ₃ ⁼ when the symbol CO₂ is used.     

K+ current stimulation by Cl- in the midgut epithelium of tobacco hornworm (Manduca sexta)

Summary Chloride-stimulated K⁺ secretion by Manduca sexta midgut (5th-instar larvae) was measured as K⁺-carried short-circuit current of the tissue mounted in an Ussing chamber. Microscopic parameters, such as single-channel current and channel density for the rate-determining passive transport step across the basolateral goblet cell membrane (i.e. K⁺ channels), were estimated by means of current-fluctuation analysis of the K⁺ channel blockade by haemolymph-side Ba²⁺ ions. Ba²⁺ was equally effective with Cl^- or gluconate (Glu^-) as the principal ambient anion. The Ba²⁺-induced K⁺ channel conduction noise is reflected by a Lorentzian, or relaxation, noise component in the power spectrum of the K⁺ current fluctuations. A reduced Lorentzian plateau value, but an unchanged corner frequency, were observed when Cl^- was replaced by Glu^-. The results from the analysis of a two-state model of K⁺ channel block by Ba²⁺, with respect to the anion-replacement effects, suggest that the observed changes in K⁺ current and Lorentzian plateau value mirror a complex change of the underlying parameters: Cl^- omission reduces single channel current but increases channel density so that the product of single channel current and channel density is smaller in Glu^- than in Cl^-. It seems likely that basolateral K⁺ channels (1) are subject to anionic gating ligands, and (2) depend on anions with respect to the rate of K⁺ transfer through and open K⁺ channel.Abbreviations a.c. alternating current - single-channel conductance - E _K K⁺ Nernst potential - f frequency contained in current noise - f _c corner frequency - Glu^- gluconate - G _t transepithelial conductance - I _sc short-circuit current - I _K K⁺ current - I _K(max) maximal K⁺ current - i single-channel current - K _Ba barium inhibition constant - K _m Michaelis constant of saturating K⁺ current - k ₀₁ and k ₁₀ barium association and dissociation rate constant, respectively - M K⁺ channel density - S _f power density - S _o Lorentzian plateau value - P _o channel-open probability - P _K K⁺ permeability - V _sc cellular potential at short-circuit These results have already been presented in part, at the 1989 joint meeting of the German and Israel Physiological Societies in Jerusalem (Zeiske et al. 1990).     

Comparative effects of growth irradiance on photosynthesis and leaf anatomy of Flaveria brownii (C4-like), Flaveria linearis (C3–C4) and their F1 hybrid

Photosynthetic rates and related anatomical characteristics of leaves developed at three levels of irradiance (1200, 300 and 80 umol · m^–2 · s^–1) were determined in the C₄-like species Flaveria brownii A.M. Powell, the C₃–C₄-intermediate species F. linearis Lag., and the F₁ hybrid between them (F. brownii × F. linearis). In the C₃–C₄ and F₁ plants, increases in photosynthetic capacity per unit leaf area were strongly correlated with changes in mesophyll area per unit leaf area. The C₄-like plant F. brownii, however, showed a much lower correlation between photosynthetic capacity and mesophyll area per unit leaf area. Plants of F. brownii developed at high irradiance showed photosynthetic rates per unit of mesophyll cell area 50% higher than those plants developed at medium irradiance. These results along with an increase in water-use efficiency are consistent with an increase of C₄ photosynthesis in high-irradiance-grown F. brownii plants, whereas in the other two genotypes such plasticity seems to be absent. Photosynthetic discrimination against ¹³C in the three genotypes was less at high than at low irradiance, with the greatest change occurring in F. brownii. Discrimination against ¹³C expressed as ¹³C was linearly correlated (r ² = 0.81; P<0.001) with the ratio of bundle-sheath volume to mesophyll cell area when all samples from the three genotypes were combined. This tissue ratio increased for F. brownii and the F₁ hybrid as growth irradiance increased, indicating a greater tendency towards Kranz anatomy. The results indicated that F. brownii had plasticity in its C₄-related anatomical and physiological characteristics as a function of growth irradiance, whereas plasticity was less evident in the F₁ hybrid and absent in F. linearis.Abbreviations A leaf surface area - A_ma, A_mn, A_lm total ma, mn or lm cell surface area - bs vascular bundle sheath - lm large spongy-mesophyll cells - ma mesophyll cells adjacent to bundle sheath - mn mesophyll cells not adjacent to bundle sheath - P_n net photosynthesis - (H, M, L) PPFD (high, medium, low) photosynthetic photon flux density - SLDW specific leaf dry wight - V_bs bs volume - V_{(ma + mn + bs)} total photosynthetic tissue volume - ¹³C ¹³C discrimination We thank Mrs. Lisa Smith for technical assistance in light microscopy and Dr. Ned Friedman (Department of Botany, University of Georgia, Athens, GA, USA) for the use of digitizing equipment. Participation of Dr. J.L. Araus in this work was supported by a grant Beca de Especialización para Doctores y Tecnólogos en el Extranjero, from Ministerio de Educatión y Ciencia, Spain.     

Action potentials inAcetabularia: Measurement and simulation of voltage-gated fluxes

Summary Amounts and temporal changes of the release of the tracer ions K⁺ (⁸⁶Rb⁺),²²Na⁺, and³⁶Cl^– as well as of H⁺ in the course of action potentials inAcetabularia have been recorded. New results and model calculations confirm in quantitative terms the involvement of three major ion transport systemsX in the plasmalemma: Cl^– pumps, K⁺ channels, and Cl^– channels (which are marked in the following by the prefixes,P, K andC) with their equilibrium voltages _X V _e and voltage/time-dependent conductances, which can be described by the following, first approximation. Let the maximum (ohmic) conductance of each of the three populations of transporter species be about the same (_P L, _KL,_C L=1) but voltage gating be different: the pump ( _{p V e} about –200 mV) being inactivated (open,oclosed,c) at positive going transmembrane voltages,V _m; the K⁺ channels (_K V _e about –100 mV) are inactivated at negative goingV _m; and the Cl^– channels (_C V _e: around 0 mV), which are normally closed (c) at a restingV _m (near_PV_e) go through an intermediate open (o) state at more positiveV _m before they enter a third shut state (s) in series. Model calculations, in which voltage sensitivities are expressed by the factorf=exp(V _mF/(2RT)), simulate, the action potential fairly well with the following parameters (_PK_co10/f ks^–1,_PK_oc1000·f ks^–1,_KK_co200·f ks^–1,_Kk_oc2/f ks^–1,_cK_co500·f ks^–1,_CK_oc5/f ks^–1,_CK_so0.1/f ks^–1,_Ck_os20·f ks^–1). It is also shown that the charge balance for the huge transient Cl^– efflux, which frequently occurs during an action potential, can be accounted for by the observation of a corresponding release of Na⁺.     

Gas-exchange of ears of cereals in response to carbon dioxide and light

Data for the maximum carboxylation velocity of ribulose-1,5-biosphosphate carboxylase, V_m, and the maximum rate of whole-chain electron transport, J_m, were calculated according to a photosynthesis model from the CO₂ response and the light response of CO₂ uptake measured on ears of wheat (Triticum aestivum L. cv. Arkas), oat (Avena sativa L. cv. Lorenz), and barley (Hordeum vulgare L. cv. Aramir). The ratio J_m/V_m is lower in glumes of oat and awns of barley than it is in the bracts of wheat and in the lemmas and paleae of oat and barley. Light-microscopy studies revealed, in glumes and lemmas of wheat and in the lemmas of oat and barley, a second type of photosynthesizing cell which, in analogy to the Kranz anatomy of C₄ plants, can be designated as a bundle-sheath cell. In wheat ears, the CO₂-compensation point (in the absence of dissimilative respiration) is between those that are typical for C₃ and C₄ plants.A model of the CO₂ uptake in C₃–C₄ intermediate plants proposed by Peisker (1986, Plant Cell Environ. 9, 627–635) is applied to recalculate the initial slopes of the A(p^c) curves (net photosynthesis rate versus intercellular partial pressure of CO₂) under the assumptions that the J_m/V_m ratio for all organs investigated equals the value found in glumes of oat and awns of barley, and that ribulose-1,5-bisphosphate carboxylase is redistributed from mesophyll to bundle-sheath cells. The results closely match the measured values. As a consequence, all bracts of wheat ears and the inner bracts of oat and barley ears are likely to represent a C₃–C₄ intermediate type, while glumes of oat and awns of barley represent the C₃ type.Abbreviations A net photosynthesis rate (mol·m^-2·s^-1) - J_m maximum rate of whole-chain electron transport (mol·e^-·m^-2·s^-1) - p^c (bar) intercellular partial pressure of CO₂ - PEP phosphoenolpyruvate - PPFD photosynthetic photon flux density (mol quanta·m^-2·s^-1) - RuBPCase ribulose bisphosphate carboxylase/oxygenase - RuBP ribulose bisphosphate - V_m maximum carboxylation velocity of RuBPCase (mol·m^-2·s^-1) - T^* CO₂ compensation point in the absence of dissimilative respiration (bar)