Diurnal Rb+ Transport from Roots to the Laminar Pulvinus in Phaseolus vulgaris L.

The primary leaves of kidney bean (Phaseolus vulgaris L.) openunder light and close in the dark by the deformation of thepulvinus resulting from diurnal distribution changes of K⁺,Cl^–, organic acid (or H⁺) and NO₃^–. When Rb⁺ was added as a tracer of K⁺ to the seedlings throughtheir roots, it was transported to the pulvinus cells duringthe light period but not during the dark period. Transpirationoccurred vigorously in the light but almost stopped in the dark.We concluded that Rb⁺ absorbed by the roots was carried to thepulvinus by the transpiration stream. Phaseolus vulgaris L., pulvinus, Rb⁺, diurnal transport transpiration stream     

K+ Gradients in the Pulvinus of Phaseolus coccineus during Leaf Movement*

We used quantitative histochemistry to investigate the tissue-specific compartmentation of potassium in the laminar pulvinus of Phaseolus coccineus L. at day and night positions of diurnal leaf movement. The assay was based on the potassium-dependent activation of pyruvate kinase. Total potassium levels of pulvini were higher in the light than in the dark [0.88 and 0.57mol (kg dry weight)^?1, respectively]. Transverse compartmentation of potassium was studied on three tissue slices, representing the middle part, petiolar and laminar sides of individual pulvini. These were dissected further into 10 distinct subsamples (bundle; motor tissues: extensor, flexor; flanks). In the day position the amount of potassium in the extensor was higher than in the night position [1.92 and 1.50 mol (kg dry weight)^?1, respectively]. Flexor changes were opposite [1.13 and 1.65 mol (kg dry weight)^?1, respectively]. In the day position there was a steep and consistent increase in potassium content from the innermost to the outermost zones of the extensor. In the night position this was much more variable. Comparable gradients were not detected in flexor samples. Here highest amounts of potassium were recovered from the middle of the motor tissue. The data specify distinct tissue regions involved in osmotic adjustment during leaf movement in Phaseolus coccineus.     

Changes of the Apoplastic pH and K+ Concentration in the Phaseolus Pulvinus in situ in Relation to Rhythmic Leaf Movements

The apoplastic pH and K⁺ concentration of the extensor of thePhaseolus primary-leaf pulvinus in relation to rhythmic leafmovements have been investigated with double-barrelled ion-sensitivemicro-electrodes. Simultaneous measurements of leaf movementand ion activities in a fine hole of the extensor in situ showedco-existence of ultradian and circadian leaf movements as wellas of ultradian and circadian pH changes in the Water Free Space(WFS) of the extensor apoplast in situ. During circadian leafmovement the H⁺ and K⁺ activities in the WFS of the extensorchange in an antagonistic manner. When extensor cells swell(upward movement of the lamina) the H⁺ activity increases fromapproximately pH 6.7 to 5.9 and the K⁺ concentration decreasesfrom approximately 50 to 10 mol m^–3 and vice versa whenextensor cells shrink. These changes in the ionic activitiesin the WFS must be correlated with large changes in the ioncontent of the DFS and thus support the hypothesis that thecell walls of pulvinar cells serve as reservoirs for K⁺ andH⁺. Key words: Phaseolus pulvinus, apoplastic ionic activities, rhythmic leaf movements, ion-sensitive micro-electrodes (double-barrelled)     

Passive Ca2+ Transport and Ca2+-Dependent K+ Transport in Plasmodium falciparum-Infected Red Cells

Previous reports have indicated that Plasmodium falciparum-infected red cells (pRBC) have an increased Ca²⁺ permeability. The magnitude of the increase is greater than that normally required to activate the Ca²⁺-dependent K⁺ channel (K _Ca channel) of the red cell membrane. However, there is evidence that this channel remains inactive in pRBC. To clarify this discrepancy, we have reassessed both the functional status of the K _Ca channel and the Ca²⁺ permeability properties of pRBC. For pRBC suspended in media containing Ca²⁺, K _Ca channel activation was elicited by treatment with the Ca²⁺ ionophore A23187. In the absence of ionophore the channel remained inactive. In contrast to previous claims, the unidirectional influx of Ca²⁺ into pRBC in which the Ca²⁺ pump was inhibited by vanadate was found to be within the normal range (30–55 μmol (10¹³ cells · hr)⁻¹), provided the cells were suspended in glucose-containing media. However, for pRBC in glucose-free media the Ca²⁺ influx increased to over 1 mmol (10¹³ cells · hr)⁻¹, almost an order of magnitude higher than that seen in uninfected erythrocytes under equivalent conditions. The pathway responsible for the enhanced influx of Ca²⁺ into glucose-deprived pRBC was expressed at approximately 30 hr post-invasion, and was inhibited by Ni²⁺. Possible roles for this pathway in pRBC are considered. Received: 12 May 1999/Revised: 8 July 1999     

The Phototropic Pulvinus of Bean Phaseolus vulgaris L. - Functional Features

Abstract: The laminar pulvinus of bean ( Phaseolus vulgaris L.) is a highly specialized organ for reorienting the lamina, and exhibits positive phototropic curvature. Structural and ultrastructural features of the pulvinus were studied to determine their possible role in its phototropic response. The vascular tissue forms a flexible, relatively inextensible central core enclosed by a starch sheath and surrounded by a multi-layered motor tissue. Phototropic curvature is a result of opposite anisotropic changes in volume of the cells in the exposed and opposite sectors of the motor tissue. Radial inflexibility of the epidermis and axial plasticity constrain these changes to the pulvinar axis. Anisotropic changes in volume of motor cells reduce the osmotic work involved. Motor cells exhibit features that are associated with high synthetic activity: thick cytoplasm with numerous ribosomes, polysomes, RER and SER, well-developed mitochondria and a large nucleus. Numerous, well-developed chloroplasts, with little starch, are increasingly abundant toward the periphery. The intercellular system is limited and partially filled with matrix. Stomata are absent and the motor tissue lacks vascularization. These features support the suggestion that the primary role of the chloroplasts in the photonastic response is photophosphorylation and photosynthetic electron transport (Koller et al., 1995^[339]).     

The Rhythmic Leaf Movements after Regeneration of Partially Excised Pulvinus in Phaseolus vulgaris L.

Unlike the petiole or stem, the laminar pulvinus of the primaryleaves of Phaseolus vulgaris L. regenerated after a partialexcision. The histological and physiological aspects of theseregeneration processes have been studied. On the third day afterthe excision of the flexor (or extensor) region, the pulvinuswas regenerated. When the major part of the extensor was cutaway, the period and phase of the circadian leaf movements wereunchanged whereas the amplitude was greatly reduced. When theflexor region was excised, period, phase and amplitude weremaintained. Some changes could be seen in the ultradian movementsafter excision of flexor as well extensor regions. (Received August 31, 1988; Accepted March 30, 1989)     

Rhythmic and Blue Light-Induced Turgor Movements and Electrical Potential in the Laminar Pulvinus of Phaseolus vulgaris L.

The well-known circadian movement in Phaseolus leaves is usuallysuperimposed with small and somewhat irregular rhythmic changesof pulvinar movement. This study examines the relationship betweenthese small pulvinar movements and the membrane potential ofthe motor cell as well as the effect of light on them. Bluelight affected both the movements and potential while red andgreen lights of the same photon flux had little effect. Also,the difference in the membrane potential between the extensorand flexor cells was found to be closely related to the rhythmicturgor movement of the pulvinus. Changes in the potential differencealways preceded the movement. Sequential changes of the potentialdifference and turgor pressure in the motor cells, includingthe light-induced ones, are discussed in relation to the leafmovement. (Received August 8, 1985; Accepted November 11, 1985)     

Role of the Plasma Membrane H+-ATPase in K+ Transport

The role of the plant plasma membrane H+-ATPase in K+ uptake was examined using red beet (Beta vulgaris L.) plasma membrane vesicles and a partially purified preparation of the red beet plasma membrane H+-ATPase reconstituted in proteoliposomes and planar bilayers. For plasma membrane vesicles, ATP-dependent K+ efflux was only partially inhibited by 100 [mu]M vanadate or 10 [mu]M carbonyl cyanide-p-trifluoromethoxyphenylhydrazone. However, full inhibition of ATP-dependent K+ efflux by these reagents occurred when the red beet plasma membrane H+-ATPase was partially purified and reconstituted in proteoliposomes. When reconstituted in a planar bilayer membrane, the current/voltage relationship for the plasma membrane H+-ATPase showed little effect of K+ gradients imposed across the bilayer membrane. When taken together, the results of this study demonstrate that the plant plasma membrane H+-ATPase does not mediate direct K+ transport chemically linked to ATP hydrolysis. Rather, this enzyme provides a driving force for cellular K+ uptake by secondary mechanisms, such as K+ channels or H+/K+ symporters. Although the presence of a small, protonophore-insensitive component of ATP-dependent K+ transport in a plasma membrane fraction might be mediated by an ATP-activated K+ channel, the possibility of direct K+ transport by other ATPases (i.e. K+-ATPases) associated with either the plasma membrane or other cellular membranes cannot be ruled out.     

影响玉米根中K+、Na+离子向地上部输送的因素（简报）

培养液中含有 1mmol·L-1K 、Na 离子的玉米根运往木质部渗出液中的K /Na 比最大 ;低温不影响渗出液中的K /Na 比 ,但极大地减少了根部向地上部输送的K 、Na 总量 ;钙的加入可以大大提高渗出液中的K /Na 比 ,减少由根部向地上部输送的Na 量。     

Transport ratios of reconstituted (H+ + K+)-ATPase

Gastric (H+ + K+)-ATPase was reconstituted into artificial phosphatidylcholine/cholesterol vesicles by means of a freeze-thaw-sonication procedure. The passive and active transport mediated by these vesicles were measured (Skrabanja, A.T.P., Asty, P., Soumarmon, A., De Pont, J.J.H.H.M. and Lewin, M.J.M. (1986) Biochim. Biophys. Acta 860, 131-136). To determine real initial velocities, the proteoliposomes were separated from non-incorporated enzyme, by means of centrifugation on a sucrose gradient. The purified proteoliposomes were used to measure active H+ and Rb+ transport, giving at room-temperature velocities of 46.3 and 42.5 mumol per mg per h, respectively. A transport ratio of two cations per ATP hydrolyzed was also measured. These figures indicate that the enzyme catalyzes an electroneutral H+-Rb+ exchange.     

Methylammonium Transport in Phaseolus vulgaris Leaf Slices

Methylammonium (as a nonmetabolized analog of ammonium) transport was studied in leaf slices of Phaseolus vulgaris L. var. `Hawkesbury Wonder.' The relationship of influx to external pH (6.0-10.5) shows that the influx at low external pH is a larger fraction of that at high external pH than would be expected from the pK_α of methylammonium and the assumption that only CH₃NH₂ is entering the cells. The relationship between methylammonium influx and external methylammonium concentration shows some evidence of saturation; this is a function of the transport system rather than of the (limited) methylammonium metabolism in the cells. The “equilibrium” concentration ratio for methylammonium between leaf slices and bathing medium is far higher than can be explained by the transport of CH₃NH₂ alone and the pH of the compartments involved. These three lines of evidence strongly suggest that there is an influx of CH₃NH₃⁺, possibly by a uniporter driven by the electrical potential of the cytoplasm with respect to the medium, as has been shown for other plant cells. Competitive inhibition of methylammonium influx by ammonium suggests that there is also an ammonium transport system. The significance of this for the recycling of N within the plant and for exchange of gaseous NH₃ between leaves and the atmosphere is discussed.     

Diurnal Changes in the Sugar and Organic Anion Concentrations in Red Beet Leaves

Marked diurnal fluctuations in the concentrations of ethanolsoluble sugars and organic anions occurred in red beet leaves.These fluctuations were related to solar radiation and air temperature.Leaves from four plots which had received different manurialtreatments contained significantly different mean sugar andanion concentrations, but the patterns of the diurnal fluctuationswere not affected by the manurial treatments. The results indicated a daily production of photosynthate equalto about 19 per cent per day (leaf-weight basis), compared tothe net assimilation rate of 17 per cent per day as estimatedfor similar plants from dry-weight increase. From the results no evidence can be adduced to support the hypothesisthat sugar accumulated in the leaves to an extent which inhibitedphotosynthesis.     

Wirkung von K+ auf die Fluxe und den Transport von Na+ in Gerstenwurzeln,K+-stimulierter Na+-Efflux in der Wurzelrinde

Summary Barley roots grown on a nutrient solution containing 1 mM Na⁺ but no K⁺ are capable of a considerable Na⁺ transport via the symplasm of the root and the xylem vessels. K⁺ added to the medium surrounding the root cortex severely inhibits this transport after a lag period at a high rate constant (Fig. 3).It is likely that the fluxes of Na⁺ are changed drastically during this transition from low to high K⁺ status. Although originally limited to steady state fluxes, the extended method of efflux analysis for excised roots (Pitman, 1971) has been applied to the non-steady fluxes which occur upon the addition of K⁺ to the roots. It is shown that besides other changes the efflux of ²²Na⁺ through the cortex of barley roots is stimulated instantaneously (Fig. 5) by the addition of K⁺ and presumably by an influx of K⁺ ions. From this a transient, K⁺-stimulated Na⁺ efflux at the plasmalemma of the cortical cells can be estimated. It amounts to 10.9 moles/g fw · h compared to the control efflux of 3.3 moles/g fw · h without K⁺.The stimulated efflux is attributed to a Na⁺ efflux pump at the plasmalemma and is thus related to the K-Na-selectivity of barley plants. The inhibition of the Na⁺ transport by K⁺ is probably a consequence of this increased efflux of Na⁺ from the symplasm through the root cortex.     

Prestin's Anion Transport and Voltage-Sensing Capabilities Are Independent

The integral membrane protein prestin, a member of the SLC26 anion transporter family, is responsible for the voltage-driven electromotility of mammalian outer hair cells. It was argued that the evolution of prestin's motor function required a loss of the protein's transport capabilities. Instead, it was proposed that prestin manages only an abortive hemicycle that results in the trapped anion acting as a voltage sensor, to generate the motor's signature gating charge movement or nonlinear capacitance. We demonstrate, using classical radioactive anion ([¹⁴C]formate and [¹⁴C]oxalate) uptake studies, that in contrast to previous observations, prestin is able to transport anions. The prestin-dependent uptake of both these anions was twofold that of cells transfected with vector alone, and comparable to SLC26a6, prestin's closest phylogenetic relative. Furthermore, we identify a potential chloride-binding site in which the mutations of two residues (P328A and L326A) preserve nonlinear capacitance, yet negate anion transport. Finally, we distinguish 12 charged residues out of 22, residing within prestin's transmembrane regions, that contribute to unitary charge movement, i.e., voltage sensing. These data redefine our mechanistic concept of prestin.     

Transport of H+, K+, Na+ and Ca++ in Streptococcus

Summary The streptococci differ from other bacteria in that cation translocations (with the possible exception of one of the K⁺ uptake systems) occur by primary transport systems, i.e., by cation pumps which use directly the free energy released during hydrolysis of chemical bonds to power transport. Transport systems in other bacteria, especially for Na⁺ and Ca⁺⁺, are often secondary, using the free energy of another ion gradient to drive cation transport. In streptococci H⁺ efflux occurs via the F₁F₀-ATPase. This enzyme is composed of eight distinct subunits. Three of the subunits are embedded in the membrane and form a H⁺ channel; this is called the F₀ portion of the enzyme. The other five subunits form the catalytic part of the enzyme, called F₁, which faces the cytoplasm and can easily be stripped from the membrane. Physiologically, this enzyme functions as a H⁺-ATPase, pumping protons out of the cell to form an electrochemical proton gradient, . The F₁F₀-ATPase, however, is fully reversible and if supplied with P_i, ADP and a + of sufficient magnitude (ca –200 mv) catalyzes the synthesis of ATP. Streptococcus faecalis can accumulate K⁺ and establish a gradient of 50 000:1 (in>out) under some conditions. Uptake occurs by two transport systems. The dominant, constitutive system requires both an electrochemical proton gradient and ATP to operate. The minor, inducible K⁺ transport system, which has many similarities to the K⁺-ATPase of the Kdp transport system found in Escherichia coli, requires only ATP to power K⁺ uptake.Sodium extrusion occurs by a Na⁺/H⁺-ATPase. Exchange is electroneutral and there is no requirement for a . The possibility that the Na⁺/H⁺-ATPase may consist of two parts, a catalytic subunit and a Na⁺/H⁺ antiport subunit, is suggested by the finding that damage to the Na⁺ transport system either through mutation or protease action leads to the appearance of -requiring Na⁺/H⁺ antiporter activity.Ca⁺⁺ like Na⁺ is extruded from metabolizing, intact cells. Transport requires no but does require ATP. Reconstitution of Ca⁺⁺ transport activity with accompanying Ca⁺⁺-stimulated ATPase activity into proteoliposomes suggests that Ca⁺⁺ is transported by a Ca⁺⁺-translocating ATPase.Where respiring organelles and bacteria use secondary transport systems the streptococci have developed cation pumps. The streptococci, which are predominantly glycolyzing bacteria, generate a much inferior to that of respiring organisms and organelles. The cation pumps may have developed simply in response to an inadequate .Abbreviations electrochemical potential of protons - membrane potential - pH pH gradient - p proton-motive force - DCCD N,Na¹-dicyclohexlcarbodiimide - TCS tetrachlorosalicylanilide - FCCP carbonylcyanide-p-trifluoromethylphenylhydrazone - CCCP carbonylcyanie-m-chlorophenylhydrazone - TPMP⁺ triphenylmethyl phosphonium ion - DDA⁺ dibenzyldimethylammonium ion - Hepes 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid - EGTA ethyleneglycol-bis (amino-ethyl-ether)-N,N-tetraacetic acid     

O_2~-·与红细胞膜阴离子转运蛋白相互关系的研究

利用４,４′二硫氰２,２′二磺基芪（ＤＩＤＳ）对带３（ｂａｎｄ３）蛋白阴离子转运功能的专一抑制作用,研究了超氧阴离子（Ｏ－２·）进入红细胞对带３蛋白的依赖关系;同时根据红细胞在等渗氯化铵溶液中溶血的ｔ１／２,探讨了细胞外大量产生Ｏ－２·对带３蛋白转运阴离子功能的影响。结果表明：４０μｍｏｌ／ＬＤＩＤＳ完全抑制带３蛋白的阴离子转运功能,但并不影响Ｏ－２·进入细胞,证明Ｏ－２·进入细胞对ｂａｎｄ３蛋白无依赖关系;Ｏ－２·通过氧化修饰红细胞膜蛋白的－ＳＨ,使带３蛋白的阴离子转运能力下降,并可用－ＳＨ还原剂ＤＴＴ与巯基乙醇修复。     

Relation between Anion Transport and Water Flow in Tomato Plants

Lowering the water potential of culture solutions from ?0.4 to ?5.4 atm reduced both phosphorus and bromide transport to the shoot, hut the content in the roots was not affected. Reductions in phosphorus transport to the shoot were measured during the first four hours of treatment and were related to concurrent decreases in water flow and not to an impairment of active phosphorus transport. The effect of low water potential on phosphorus transport to shoots was similar at external phosphorus concentrations between 0.6 and 15 mg/l. Phosphorus transport was greater in the dark at ?0.4 atm than in the light at ?5.4 atm even when these treatments gave the same overall rates of water flow; this is attributed to a different pattern of water flow through the various root zones. The results suggest that the main effect of water flow on anion transport to shoots occurred after the ions had been actively adsorbed by the roots and was not due to mass flow increasing ion delivery to sites of active uptake.