Endogenous electrical current leaves the limb and prelimb region of the Xenopus embryo

The electrical current pattern around the developing Xenopus laevis embryo was mapped with a vibrating probe. Current (taken as the movement of positive charge) was found to leave the emerging hind limb bud and to enter the gill region of the stage 47 embryo. The magnitude of the current leaving the limb was about 7 μA/cm² and the current entering the gill was about 60 μA/cm². Other regions of smaller outward current were found between the limb and gill. At stage 43, prior to the appearance of the limb bud, a highly localized region of outward current existed in the general area from which the bud would later emerge. The inward current was localized to the gill bud, as in the older embryo. The ionic basis of the currents could not be determined. In about one-third of the cases studied, the inward current was sodium sensitive since the removal of external sodium or the addition of amiloride reversably blocked the current. In the remaining cases, however, removal of sodium did not change (or else increased) the current. No other external ion (Ca²⁺, Mg²⁺, K⁺) could be identified as the current-carrying ion; the possibility of an outward movement of some anion such as Cl^? or HCO^?₃ remains.     

Correlation between Root-Generated Ionic Currents, pH, Fusicoccin, Indoleacetic Acid, and Growth of the Primary Root of Zea mays

Correlations between root-generated ionic currents, extracellular pH, indoleacetic acid, fusicoccin, and growth were investigated. Current consistently entered the meristematic and elongating tissues of intact growing roots of Zea mays cv Golden Bantam. Mature root regions generated the outward limb of the current loop. Ion-substitution and pH-profile experiments suggested that the bulk of the ionic current was carried by H⁺. Calcium ions did not carry current, but calcium may regulate the proton circulation since the proton current density was slightly larger in calcium-depleted media. Increased root elongation at low pH was associated with increased current density and an extended zone of inward current. Conversely decreased elongation at high pH was associated with a reduced current density and a more restricted zone of inward current. The effect of the fungal toxin fusicoccin was to increase the current density of the inward limb of the ion current and to increase root extension. Concentrations of indoleacetic acid that reduced root growth, also reduced the density of the inward current and shortened the inward current zone. The results emphasize the point that roots are electrically contiguous over many millimeters and that the electrophysiology of root growth is best studied in intact root systems.     

Demonstration of Voltage-Dependent and TTX-Sensitive Na+-Channels in Human Melanocytes

The electrophysiological properties of cultured human melanocytes were investigated using the whole-cell configuration of the patch-clamp technique. Depolarizations to membrane potentials more positive than -30 mV resulted in the rapid development (<1 ms to peak) of an inward current. The maximum peak current was observed at +10 mV and reached an average amplitude of about 270 pA. During the depolarizations, the current inactivated with a time constant of about 2 ms. The current was abolished by the addition of 0.3 μM tetrodotoxin, a blocker of voltage-gated Na⁺-channels, and disappeared when Na⁺ was omitted from the extracellular medium. In addition, the melanocytes contain at least two types of outward K⁺-current. The first type, observed in every cell, was highly sensitive (K_i 1 mM) to the K⁺-channel blocker TEA, required depolarizations beyond zero to be activated and did not inactivate. The second type was less regularly observed (10% of the cells). This current activated at more negative voltages (–20 mV), was resistant to TEA (20 mM) but was blocked by 2 mM 4-aminopyridine and inactivated rapidly during depolarizations. We conclude that human melanocytes are equipped with voltage-dependent Na⁺-channels, a delayed rectifying K⁺-current and a K⁺-current similar to the A-current in neurones.     

Growth, Gravitropism, and Endogenous Ion Currents of Cress Roots (Lepidium sativum L.) : Measurements Using a Novel Three-Dimensional Recording Probe

A novel, three-dimensional recording, vibrating probe was used for measuring the density and direction of the endogenous ionic current of cress roots (Lepidium sativum L.) bathed in low salt media (artificial pond water, APW). Roots submerged in regular APW and growing vertically show the following current pattern. Current of 0.7 microampere/square centimeter density enters or leaves the root cap; the current changes direction frequently. Current of 1.6 microamperes/square centimeter enters the meristem zone most of the time. Maximum current with a density of 2.2 microamperes/square centimeter enters the apical elongating zone, i.e. between 0.8 and 1.2 millimeters behind the root tip. The current density decreases to 1.4 microamperes/square centimeter at 2 millimeters, i.e. in the central elongating zone, and to 1.0 microampere/square centimeter at 3 millimeters, i.e. in the basal elongating zone. The current direction changes from inward to predominantly outward between 1.2 and 3 millimeters behind the tip. Measurements on opposite flanks of the roots indicate that the current pattern is fairly symmetrical. After placing the roots horizontally, the density of the endogenous current remains stable, but the current direction changes at the root cap and in the meristem zone. The current leaves the root on the upper side and enters on the lower side, causing a highly asymmetrical current pattern at the very tip. The current pattern at the upper and lower side further away from the tip remains the same as in vertical roots. Roots submerged in low Ca²⁺ APW show a very different current pattern, no gravitropism, and no change of the current pattern after horizontal orientation. In these roots current enters the root cap and the basal elongating zone and leaves the apical elongating zone. Three conclusions are drawn from these results: First, plant roots elongate by two different modes of growth that are correlated with different current directions. They grow by cytoplasmic enlargement at sites of inward current and by turgor-driven elongation at sites of outward current. Second, a change in the current pattern at the root cap and in the meristem zone is a clear indicator of later gravitropism. Third, Ca²⁺ ions are involved in the gravistimulated change in the current pattern, probably affecting the activity of plasmalemma H⁺-ATPases.     

MEMBRANE POTENTIAL AND ENDOGENOUS ION CURRENT OF PHYCOMYCES SPORANGIOPHORES

Glass microelectrodes were inserted into the growing zone of sporangiophores of Phycomyces blakesleeanus that had been submersed in artificial pond water. The membrane potential (inside negative) increased with increasing pH of the bathing solution from an average of ?98 mV at pH 5 up to ?131 mV at pH 7. Removal of Ca²⁺ from the medium hyperpolarized the membrane potential in the wild type, but caused a significant depolarization in the blue-light-insensitive madC mutant. KCN, diethylstilbestrol, and N,N′-dicyclohexylcarbodiimide depolarized the membrane potential in both the wild type and the madC mutant, while fusicoccin had no effect. Endogenous ion current of up to 2 μA cm^?2 was measured in the growing zone of sporangiophores with an extracellular vibrating electrode. The current density and current pattern varied with the pH of the medium. At pH 5 most sporangiophores had weak inward current along the growing zone, whereas at pH 7 most sporangiophores had strong outward current. The response of the membrane potential to specific inhibitors and the presence of an endogenous ion current indicate an electrogenic H⁺-ATPase in the plasma membrane. The results show a negative correlation between growth rate of sporangiophores growing in buffered aqueous medium and magnitude of membrane potential, as well as density of outward current. They also indicate an important role of protons in controlling the growth of Phycomyces sporangiophores.     

Ion channels of intact young root hairs from Medicago sativa

Root hairs are a primary site for nutrient absorption and for initiation of signalling processes linked to variations of the root environment:plant-microbe interactions or abiotic changes. In many of these cases, the earliest detectable response is the modification of plasma membrane transports, detected through alteration of the electrical membrane potential. In spite of this, root hairs have not been extensively used in electrophysiological research so far. Problems with cell shape and current coupling are often prohibitive for microelectrode voltage-clamp on intact root hairs. In the present study, these difficulties have been overcome and the ion channel currents are described for young root hairs from alfalfa seedlings (Medicago sativa cv Sitel). Electrophysiological and pharmacological studies indicated an inward rectifying K⁺ time-dependent current. This current was sensitive to tetraethylammonium and Cs⁺ (10 mM each). Two other currents never shown in root hairs were described: an outward rectifying time-dependent K⁺ current, inhibited by tetraethylammonium and Cs⁺ (10 mM each) allowing K⁺ efflux under strong depolarizations and an instantaneous inward current identified as an anion current, inhibited by 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid and anthracene-9-carboxylic acid (100 μM each). These results should contribute to the understanding of root hair development and of signalling processes in M. sativa root hairs.     

Effects of mono- and multi-valent cations on the inward-rectifying potassium channel in isolated protoplasts from maize roots

Transport properties mediated by ionic channels were studied by the patch-clamp technique in protoplasts from cortical parenchyma cells of maize roots (CPMR). While outward currents could be seen only occasionally, macroscopic voltage- and time-dependent potassium-selective inward currents (I_K+in) were frequently observed in the whole-cell configuration. These currents increased continuously as a function of K⁺ concentration (in the range 3 – 200 mm) and the slow-saturating macroscopic chord-conductance was fitted by a Michaelis-Menten function with K_m = 195 ± 39 mm. Other ions, like sodium and lithium, did not permeate at all through the maize root inward-channel, or like ammonium (P_NH4+/ P_K+ = 0.16 0.25) and rubidium (P_Rb+/P_K+≈ 0.10) displayed a very low permeability ratio. Up to 5 mm Rb⁺ did not induce any inhibition of the K⁺ inward current, whereas submillimolar concentrations of Cs⁺ were sufficient to block, in a voltage-dependent manner, the inward currents. A decrease of the external potassium concentration favoured Cs⁺ inhibition (K_m = 89 ± 6 μm and 26 ± 2 μm in 200 and 100 mm KCl, respectively). The potassium inward-currents were reversibly and consistently inhibited by submillimolar external concentrations of the metal ions Ni²⁺, Zn²⁺ and Co²⁺, while 1 mm La³⁺ only slightly decreased (≈10%) both the single channel conductance (9.2 ± 1.2 pS in 100 mm potassium) and the macroscopic current. In contrast to the case with Cs⁺, inhibition induced by other metal ions did not show any voltage dependence. These results suggest that, as with animal potassium channels, the inward channel of maize-root cortical cells has a narrow pore of permeation and metal ions decrease the K⁺ current, possibly by acting on binding sites located outside the pore. Received: 21 February 1997 / Accepted: 27 May 1997     

Inward and outward K+-selective currents in the plasma membrane of protoplasts from maize root cortex and stele

In an attempt to understand the processes mediating ion transport within the root, the patch clamp technique was applied to protoplasts isolated from the cortex and stele of maize roots and their plasma membrane conductances investigated. In the whole-cell configuration, membrane hyperpolarization induced a slowly activating inwardly rectifying conductance in most protoplasts isolated from the root cortex. In contrast, most protoplasts isolated from the stele contained a slowly activating outwardly rectifying conductance upon plasma membrane depolarization. The reversal potential of the inward current indicated that it was primarily due to the movement of K⁺; the outwardly rectifying conductance was comparatively less selective for K⁺. Membrane hyperpolarization beyond a threshold of about ?70 mV induced inward currents. When E_K was set negative of this threshold, inward currents activated negative of E_K and no outward currents were observed positive of E_K. Outward currents in the stelar protoplasts activated at potentials positive of ?85 mV. However, when E_K was set positive of ?85 mV a small inward current was also observed at potentials negative (and slightly positive) of the equilibrium potential for K⁺. Inwardly and outwardly rectifying K⁺ channels were observed in outside-out patches from the plasma membrane of cortical and stelar cells, respectively. Characterization of these channels showed that they were likely to be responsible for the macroscopic ‘whole-cell’ currents. Inward and outward currents were affected differently by various K⁺ channel blockers (TEA⁺, Ba²⁺ and Cs⁺). In addition, Ca²⁺ above 1 mM partially blocked the inward current in a voltage-dependent manner but had little effect on the outward current. It is suggested that the inwardly rectifying conductance identified in protoplasts isolated from the cortex probably represents an important component of the low-affinity K⁺ uptake mechanism (mechanism II) identified in intact roots. The outwardly rectifying conductance identified in protoplasts isolated from the stele could play a role in the release of cations into the xylem vessels for transport to the shoot.     

Response of grape rootstocks to salinity: changes in root growth, polyamines and abscisic acid

Effects of salinity (0, 50, 100 and 250 mM NaCl) on growth, root:shoot dry mass ratio, osmotic potential (ψ_x), electrolyte leakage and contents of Na⁺ and K⁺, polyamines and abscisic acid (ABA) were studied in the grape rootstocks Dogridge, 1613, St. George and Salt Creek. In control rootstocks, the root length was highest in Dogridge and contents of K⁺ and ABA in Salt Creek. Salinity treatments increased root Na⁺ and decreased K⁺ content and St. George exhibited highest Na⁺ content and Na⁺:K⁺ ratio. The root:shoot dry mass ratio in all rootstocks increased upto 100 mM NaCl. With increasing NaCl concentration, putrescine, spermine and spermidine contents showed consistent increase and putrescine increase was highest in St. George and spermidine and spermine in the Dogridge and Salt Creek. Under salinity, the ABA content increased in all the rootstocks but more in Salt Creek and Dogridge than in St. George.     

The effects of exogenous ABA applied to maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) roots on plant responses to chilling stress

This work aimed to discuss the effects of exogenous abscisic acid (ABA) on the root growth regulation of maize seedlings under chilling stress. The roots of the maize cultivar Zhengdan 958 were irrigated with ABA (10^?7, 10^?6, 10^?5 and 10^?4 M) at the third true leaf stage under chilling duration (0, 2, 4, 6, and 8 days). The biomass, the phenylalanine ammonia lyase (PAL), and polyphenol oxidase (PPO) enzyme activities, total phenolic and flavonoid contents, the ferric reducing ability of plasma (FRAP) antioxidant capacity, and 2,2-azinobis (3-ethlbenzothiazo-line-6-sulfonic acid) diammonium salt radical (ABTS^·+) scavenging capacity of the roots of maize seedlings were measured after the treatment. The results showed that appropriate concentrations of exogenous ABA effectively enhanced root biomass, increased PAL and PPO enzyme activities, and significantly increased total phenolic contents and flavonoid contents. Moreover, the ABA markedly improved the FRAP antioxidant capacity and ABTS^·+ scavenging capacity under low-temperature stress. These results indicate that ABA-treated maize seedlings are resistant to chilling stress and that the optimum concentration of ABA is 10^?5 M. Exogenous applications of ABA have a concentration effect in alleviating chilling stress, in which low concentrations have a promoting effect and high concentrations have an inhibiting effect.     

Time-dependent Outward Currents through the Inward Rectifier Potassium Channel IRK1 : The Role of Weak Blocking Molecules

Outward currents through the inward rectifier K⁺ channel contribute to repolarization of the cardiac action potential. The properties of the IRK1 channel expressed in murine fibroblast (L) cells closely resemble those of the native cardiac inward rectifier. In this study, we added Mg²⁺ (0.44–1.1 mM) or putrescine (∼0.4 mM) to the intracellular milieu where endogenous polyamines remained, and then examined outward IRK1 currents using the whole-cell patch-clamp method at 5.4 mM external K⁺. Without internal Mg²⁺, small outward currents flowed only at potentials between −80 (the reversal potential) and ∼−40 mV during voltage steps applied from −110 mV. The strong inward rectification was mainly caused by the closed state of the activation gating, which was recently reinterpreted as the endogenous-spermine blocked state. With internal Mg²⁺, small outward currents flowed over a wider range of potentials during the voltage steps. The outward currents at potentials between −40 and 0 mV were concurrent with the contribution of Mg²⁺ to blocking channels at these potentials, judging from instantaneous inward currents in the following hyperpolarization. Furthermore, when the membrane was repolarized to −50 mV after short depolarizing steps (>0 mV), a transient increase appeared in outward currents at −50 mV. Since the peak amplitude depended on the fraction of Mg²⁺-blocked channels in the preceding depolarization, the transient increase was attributed to the relief of Mg²⁺ block, followed by a re-block of channels by spermine. Shift in the holding potential (−110 to −80 mV), or prolongation of depolarization, increased the number of spermine-blocked channels and decreased that of Mg²⁺-blocked channels in depolarization, which in turn decreased outward currents in the subsequent repolarization. Putrescine caused the same effects as Mg²⁺. When both spermine (1 μM, an estimated free spermine level during whole-cell recordings) and putrescine (300 μM) were applied to the inside-out patch membrane, the findings in whole-cell IRK1 were reproduced. Our study indicates that blockage of IRK1 by molecules with distinct affinities, spermine and Mg²⁺ (putrescine), elicits a transient increase in the outward IRK1, which may contribute to repolarization of the cardiac action potential.     

Ca2+ and cell signalling in guard cells

Abscisic acid (ABA)-induced stomatal closure involves two different signalling chains, only one of which is Ca²⁺-dependent. ABA induces deactivation of the inward K⁺ channel and activation of an inward 'background' current, changes also produced by high cytoplasmic Ca²⁺ or injection of inositol 1,4,5-trisphosphate. It is argued that ABA produces local increases in Ca²⁺, which are obligatory for the response, even where global increases are not observed with present methodology. Deactivation of the inward K⁺ channel is abolished in the presence of internal Ca²⁺ chelator, but not by external Ca²⁺ chelator, arguing for release from internal stores. ABA-induced turnover in the polyphosphoinositide cycle occurs within 30 s, and may precede the electrical changes. Activation of the outward K⁺ channel is Ca²⁺-independent; changes in cytoplasmic pH, of unknown origin, may be responsible.     

Abscisic Acid Response of Corn (<Emphasis Type="Italic">Zea mays</Emphasis> L.) Roots and Protoplasts to Lanthanum

Lanthanum ions antagonize calcium and are used as a Ca²⁺ channel blocker but their direct effects are unknown. We investigated lanthanum effects on endogenous abscisic acid (ABA) levels in protoplasts and intact primary roots of Zea mays L. Application of 1 mM La³⁺ reduced primary root elongation, caused swelling of root tips, and essentially doubled the ABA content in intact roots but decreased ABA in root protoplasts in a concentration-dependent manner. Osmotic stress increased ABA level in protoplasts more than in intact roots. Temporal ABA changes in response to La³⁺ treatment indicate that La³⁺ affects root growth at least partially via ABA pathway.     

Whole-cell K+ current activation in response to voltages and carbachol in gastric parietal cells isolated from guinea pig

Summary Patch-clamp studies of whole-cell ionic currents were carried out in parietal cells obtained by collagenase digestion of the gastric fundus of the guinea pig stomach. Applications of positive command pulses induced outward currents. The conductance became progressively augmented with increasing command voltages, exhibiting an outwardly rectifying current-voltage relation. The current displayed a slow time course for activation. In contrast, inward currents were activated upon hyperpolarizing voltage applications at more negative potentials than the equilibrium potential to K⁺ (E _K). The inward currents showed time-dependent inactivation and an inwardly rectifying current-voltage relation. Tail currents elicited by voltage steps which had activated either outward or inward currents reversed at nearE _K, indicating that both time-dependent and voltagegated currents were due to K⁺ conductances. Both outward and inward K⁺ currents were suppressed by extracellular application of Ba²⁺, but little affected by quinine. Tetraethylammonium inhibited the outward current without impairing the inward current, whereas Cs⁺ blocked the inward current but not the outward current. The conductance of inward K⁺ currents, but not outward K⁺ currents, became larger with increasing extracellular K⁺ concentration. A Ca²⁺-mobilizing acid secretagogue, carbachol, and a Ca²⁺ ionophore, ionomycin, brought about activation of another type of outward K⁺ currents and voltage-independent cation currents. Both currents were abolished by cytosolic Ca²⁺ chelation. Quinine preferentially inhibited this K⁺ current. It is concluded that resting parietal cells of the guinea pig have two distinct types of voltage-dependent K⁺ channels, inward rectifier and outward rectifier, and that the cells have Ca²⁺-activated K⁺ channels which might be involved in acid secretion under stimulation by Ca²⁺-mobilizing secretagogues.     

Biodiversity and extracellular enzymatic activity of heterotrophic bacterial communities in Bardawil Lagoon,Egypt

The biodiversity of heterotrophic viable bacteria (209 isolates) in the hypersaline Bardawil Lagoon, Egypt, was studied. Composition and extracellular activities of viable culturable heterotrophic bacteria (VCHB) in the water and in non-colonised and seagrass-colonised sediments of Bardawil Lagoon were determined bimonthly during 1997 and 1998. The average ± SD total Kjeldhal nitrogen was 1.69 ± 0.44 mg l^?1 in water, 335.95 ± 19.22 mg kg^?1 in colonised sediments, and 215.5 ± 16.0 mg kg^?1 in non-colonised sediments. Exoenzymatic bacterial activity (glycosidase) presented a seasonal trend with average values of 1.02 ±0.16 μM cm^?3 min^?1 in colonised sediment samples and was 0.36 ± 0.27 μM cm^?3 min^?1 in non-colonised sediments. Mean of VCHB was 4 017 ± 565 cfu g^?1 and 1 195 ± 242 cfu g^?1 for colonised and non-colonised sediments, respectively. Bacterial isolates from Bardawil Lagoon water and sediments yielded a wide diversity of VCHB: a total of 209 different species, belonging to 13 genera from the water and 12 genera from the sediments.     

An electrophysiological study of the membrane properties of the immature and mature oocyte of the Batstar,Patiria miniata

Immature oocyte membrane properties of a starfish, Patiria miniata, were investigated by microelectrode techniques. The resting membrane potential in artificial seawater (ASW) was ?78.5 ± 6.7 mV (n = 61, inside negative). This was mainly accounted for by a selective permeability to potassium ions. Potassium ion-selective microelectrodes were used to measure intracellular K⁺ ion activity, which was 350 mM. The sodium to potassium permeability ratio was 0.02 ± 0.01 (n = 4). The current-voltage relation was nonlinear. The I–V curve included both areas of inward and outward rectification. The dependence of inward rectification upon the K⁺ ion electrochemical gradient was demonstrated. The membrane was capable of a regenerative action potential due to permeability changes for Ca²⁺ and Na⁺ ions. The Ca and Na components of the action potential were identified. The Ca component was reversibly suppressed by cobalt and irreversibly blocked by D-600. The Na component was tetrodotoxin (TTX) insensitive. The excitable response of P. miniata oocytes is similar to that described by Miyazaki et al. (1975a) for those of the starfish Asterina pectinifera.Immature oocytes were stimulated to mature with 10^?5M 1-methyladenine (1-MA) during continuous monitoring of the membrane potential. The resting potential in ASW became more inside negative during maturation. This change of the passive membrane property of the oocyte may be accounted for by the increased selectivity to K⁺ ions. The specific membrane resistance near the resting potential increased from 4.2 ± 1.4 to 21 ± 8.7 kΩ·cm² (n = 15) during maturation, while the specific membrane capacitance decreased slightly from 2 ± 0.5 to 1.7 ± 0.6 μF/cm² (n = 5). Maturation had little effect upon the active membrane properties.     

NO 3 − transport across the plasma membrane of Arabidopsis thaliana root hairs: Kinetic control by pH and membrane voltage

High-affinity nitrate transport was examined in intact root hair cells of Arabidopsis thaliana using electrophysiological recordings to characterise the response of the plasma membrane to NO ₃ ^? challenge and to quantify transport activity. The NO ₃ ^? -associated membrane current was determined using a three-electrode voltage clamp to bring membrane voltage under experimental control and to compensate for current dissipation along the longitudinal cell axis. Nitrate transport was evident in the roots of seedlings grown in the absence of a nitrogen source, but only 4–6 days postgermination. In 6-day-old seedlings, additions of 5–100 μm NO ₃ ^? to the bathing medium resulted in membrane depolarizations of 8–43 mV, and membrane voltage (V _m) recovered on washing NO ₃ ^? from the bath. Voltage clamp measurements carried out immediately before and following NO ₃ ^? additions showed that the NO ₃ ^? -evoked depolarizations were the consequence of an inward-directed current that appeared across the entire range of accessible voltages (?300 to +50 mV). Both membrane depolarizations and NO ₃ ^? -evoked currents recorded at the free-running voltage displayed quasi-Michaelian kinetics, with apparent values for K_m of 23 ± 6 and 44 ± 11 μm, respectively and, for the current, a maximum of 5.1 ± 0.9 μA cm^?2. The NO ₃ ^? current showed a pronounced voltage sensitivity within the normal physiological range between ?250 and ?100 mV, as could be demonstrated under voltage clamp, and increasing the bathing pH from 6.1 to 7.4–8.0 reduced the current and the associated membrane depolarizations 3- to 8-fold. Analyses showed a well-defined interaction between the kinetic variables of membrane voltage, pH_o and [NO ₃ ^? ]_o. At a constant pH_o of 6.1, depolarization from ?250 to ?150 mV resulted in an approximate 3-fold reduction in the maximum current but a 10% rise in the apparent affinity for NO ₃ ^? . By contrast, the same depolarization effected an approximate 20% fall in the K_m for transport as a function in [H⁺]_o. These, and additional characteristics of the transport current implicate a carrier cycle in which NO ₃ ^? binding is kinetically isolated from the rate-limiting step of membrane charge transit, and they indicate a charge-coupling stoichiometry of 2(H⁺) per NO ₃ ^? anion transported across the membrane. The results concur with previous studies showing a high-affinity NO ₃ ^? transport system in Arabidopsis that is inducible following a period of nitrogen-limiting growth, but they underline the importance of voltage as a kinetic factor controlling NO ₃ ^? transport at the plant plasma membrane.     

Relative importance of Na+, Cl−, and abscisic acid in NaCl induced inhibition of root growth of rice seedlings

The relative importance of endogenous abscisic acid (ABA), as well as Na⁺ and Cl^– in NaCl-induced responses related to growth in roots of rice seedlings were investigated. The increase in ammonium, proline and H₂O₂ levels, and cell wall peroxidase (POD) activity has been shown to be related to NaCl-inhibited root growth of rice seedlings. Increasing concentrations of NaCl from 50 to 150 mM progressively decreased root growth and increased both Na⁺ and Cl^–. Treatment with NaCl in the presence of 4,4-diisothiocyano-2,2-disulfonic acid (DIDS, a nonpermeating amino-reactive disulfonic acid known to inhibit the uptake of Cl^–) had less Cl^– level in roots than that in the absence of DIDS, but did not affect the levels of Na⁺, and responses related to growth in roots. Treatment with 50 mM Na-gluconate (the anion of which is not permeable to membrane) had similar Na⁺ level in roots as that with 100 mM NaCl. It was found that treatment with 50 mM Na-gluconate effected growth reduction and growth-related responses in roots in the same way as 100 mM NaCl. All these results suggest that Cl^– is not required for NaCl-induced responses in root of rice seedlings. Endogenous ABA level showed no increase in roots of rice seedlings exposed to 150 mM NaCl. It is unlikely that ABA is associated with NaCl-inhibited root growth of rice seedlings.     

Does root-sourced ABA have a role in mediating growth and stomatal responses to soil compaction in tomato (Lycopersicon esculentum)?

Isogenic wild-type (Ailsa Craig) and abscisic acid (ABA)-deficient mutant (flacca) genotypes of tomato were used to examine the role of root-sourced ABA in mediating growth and stomatal responses to compaction. Plants were grown in uniform soil columns providing low to moderate bulk densities (1.1–1.5 g cm^?3), or in a split-pot system, which allowed the roots to divide between soils of the same or differing bulk density (1.1/1.5 g cm^?3). Root and shoot growth and leaf expansion were reduced when plants were grown in compacted soil (1.5 g cm^?3) but leaf water status was not altered. However, stomatal conductance was affected, suggesting that non-hydraulic signal(s) transported in the transpiration stream were responsible for the observed effects. Xylem sap and foliar ABA concentrations increased with bulk density for 10 and 15 days after emergence (DAE), respectively, but were thereafter poorly correlated with the observed growth responses. Growth was reduced to a similar extent in both genotypes in compacted soil (1.5 g cm^?3), suggesting that ABA is not centrally involved in mediating growth in this severely limiting ‘critical’ compaction stress treatment. Growth performance in the 1.1/1.5 g cm^?3 split-pot treatment of Ailsa Craig was intermediate between the uniform 1.1 and 1.5 g cm^?3 treatments, whereas stomatal conductance was comparable to the compacted 1.5 g cm^?3 treatment. In contrast, shoot dry weight and leaf area in the split-pot treatment of flacca were similar to the 1.5 g cm^?3 treatment, but stomatal conductance was comparable to uncompacted control plants. These results suggest a role for root-sourced ABA in regulating growth and stomatal conductance during ‘sub-critical’ compaction stress, when genotypic differences in response are apparent. The observed genotypic differences are comparable to those previously reported for barley, but occurred at a much lower bulk density, reflecting the greater sensitivity of tomato to compaction. By alleviating the severe growth reductions induced when the entire root system encounters compacted soil, the split-pot approach has important applications for studies of the role of root-sourced signals in compaction-sensitive species such as tomato.     

Voltage-sensitive chloride ion channels in Anopheles gambiae Sua-1B cells

In this study, we performed electrophysiological analysis of Anopheles gambiae Sua-1B cells having “neuron-like” morphologies using the patch clamp method. The recorded cells (n = 79) had processes resembling axons/dendrites, with 63 % unipolar, 22 % bipolar, and 15 % multipolar. While no inward currents were observed following step depolarizations (holding potential = ?80 mV), a slowly activating outward current was observed in 96 % of the cells, especially at depolarized potentials. The amplitude of the current was attenuated nearly 70 % by reducing extracellular Cl^? ion concentration, or by incubating with 100 μM DIDS, a known voltage-sensitive chloride channel blocker, suggesting that the current was mediated by chloride ions. No qualitative difference was found between recordings made with Cs⁺ ions in the intracellular pipette solution (inhibits K⁺ currents) and those made with normal physiological solution, indicating a deficiency of potassium channels. Additionally, recordings made with Ca²⁺-free extracellular bath solution eliminated the slowly activating outward current. A subset of cells (n = 3) lacked this current, but had outward currents with voltage-dependent properties similar to those of volume-regulated chloride channels. Taken together, our results suggest that the voltage-sensitive currents observed in the majority of Sua-1B cells are mediated primarily by chloride channels of the calcium-dependent subtype.