Effect of trimethyltin chloride on slow vacuolar (SV) channels in vacuoles from red beet (Beta vulgaris L.) taproots

In the present study, patch-clamp techniques have been used to investigate the effect of trimethyltin chloride (Met3SnCl) on the slow vacuolar (SV) channels in vacuoles from red beet (Beta vulgaris L.) taproots. Activity of SV channels has been measured in whole-vacuole and cytosolic side-out patch configurations. It was found that addition of trimethyltin chloride to the bath solution suppressed, in a concentration-dependent manner, SV currents in red beet vacuoles. The time constant, τ, increased significantly in the presence of the organotin. When single channel activity was analyzed, only little channel activity could be recorded at 100 μM Met3SnCl. Trimethyltin chloride added to the bath medium significantly decreased (by ca. threefold at 100 μM Met3SnCl and at 100 mV voltage, as compared to the control medium) the open probability of single channels. Single channel recordings obtained in the presence and absence of trimethyltin chloride showed that the organotin only slightly (by <10%) decreased the unitary conductance of single channels. It was also found that Met3SnCl significantly diminished the number of SV channel openings, whereas it did not change the opening times of the channels. Taking into account the above and the fact that under the here applied experimental conditions (pH = 7.5) Met3SnCl is a non-dissociated (more lipophilic) compound, we suggest that the suppression of SV currents observed in the presence of the organotin results probably from its hydrophobic properties allowing this compound to translocate near the selectivity filter of the channel

Role of auxin (IAA) in the regulation of slow vacuolar (SV) channels and the volume of red beet taproot vacuoles

Background: Auxin (IAA) is a central player in plant cell growth. In contrast to the well-established function of the plasma membrane in plant cell expansion, little is known about the role of the vacuolar membrane (tonoplast) in this process. Results: It was found that under symmetrical 100 mM K+ and 100 μM cytoplasmic Ca2+ the macroscopic currents showed a typical slow activation and a strong outward rectification of the steady-state currents. The addition of IAA at a final concentration of 1 μM to the bath medium stimulated the SV currents, whereas at 0.1 and 10 μM slight inhibition of SV currents was observed. The time constant, decreased in the presence of this hormone. When single channels were analyzed, an increase in their activity was recorded with IAA compared to the control. The single-channel recordings that were obtained in the presence of IAA showed that auxin increased the amplitude of the single-channel currents. Interestingly, the addition of IAA to the bath medium with the same composition as the one that was used in the patch-clamp experiments showed that auxin decreased the volume of the vacuoles. Conclusions: It is suggested that the SV channels and the volume of red beet taproot vacuoles are modulated by auxin (IAA)

Effect of Auxin (IAA) on the Fast Vacuolar (FV) Channels in Red Beet (Beta vulgaris L.) Taproot Vacuoles

In contrast to the well-studied e ect of auxin on the plasma membrane K+ channel activity, little is known about the role of this hormone in regulating the vacuolar K+ channels. Here, the patch-clamp technique was used to investigate the e ect of auxin (IAA) on the fast-activating vacuolar (FV) channels. It was found that the macroscopic currents displayed instantaneous currents, which at the positive potentials were about three-fold greater compared to the one at the negative potentials. When auxin was added to the bath solution at a final concentration of 1 M, it increased the outward currents by about 60%, but did not change the inward currents. The imposition of a ten-fold vacuole-to-cytosol KCl gradient stimulated the e ux of K+ from the vacuole into the cytosol and reduced the K+ current in the opposite direction. The addition of IAA to the bath solution with the 10/100 KCl gradient decreased the outward current and increased the inward current. Luminal auxin reduced both the outward and inward current by approximately 25% compared to the control. The single channel recordings demonstrated that cytosolic auxin changed the open probability of the FV channels at the positive voltages to a moderate extent, while it significantly increased the amplitudes of the single channel outward currents and the number of open channels. At the positive voltages, auxin did not change the unitary conductance of the single channels. We suggest that auxin regulates the activity of the fast-activating vacuolar (FV) channels, thereby causing changes of the K+ fluxes across the vacuolar membrane. This mechanism might serve to tightly adjust the volume of the vacuole during plant cell expansion

A Comparison of the Effect of Lead (Pb) on the Slow Vacuolar (SV) and Fast Vacuolar (FV) Channels in Red Beet (Beta vulgaris L.) Taproot Vacuoles

Little is known about the effect of lead on the activity of the vacuolar K+ channels. Here, the patch-clamp technique was used to compare the impact of lead (PbCl2) on the slow-activating (SV) and fast-activating (FV) vacuolar channels. It was revealed that, under symmetrical 100-mM K+, the macroscopic currents of the SV channels exhibited a typical slow activation and a strong outward rectification of the steady-state currents, while the macroscopic currents of the FV channels displayed instantaneous currents, which, at the positive potentials, were about three-fold greater compared to the one at the negative potentials. When PbCl2 was added to the bath solution at a final concentration of 100 M, it decreased the macroscopic outward currents of both channels but did not change the inward currents. The single-channel recordings demonstrated that cytosolic lead causes this macroscopic effect by a decrease of the single-channel conductance and decreases the channel open probability. We propose that cytosolic lead reduces the current flowing through the SV and FV channels, which causes a decrease of the K+ fluxes from the cytosol to the vacuole. This finding may, at least in part, explain the mechanism by which cytosolic Pb2+ reduces the growth of plant cells

Long range correlations of the ion current in SV channels. Met3PbCl influence study

The long-range correlations within the current signal time series of the Beta vulgaris vacuolar membrane under the influence of organolead compound (Met3PbCl) are investigated. The current time series is transformed into a dwell time series. Then the rescaled range and detrended fluctuations analyses are used. It is shown that the presence of Met3PbCl in the solution decreases the mean value of the Hurst exponent and therefore influences the longrange correlations in ionic channel current. This observation is statistically significant. An ion channel model is built and the experimental results reconstructed and analysed

The effect of DC electric field on the elongation growth, proton extrusion and membrane potential of Zea mays L. coleoptile cells; a laboratory study

Background:In this study, we investigated the effect of an electric field, with an intensity similar to that of the Earth’s field, on plant cells growth. The molecular mechanism underlying this effect remains unclear. Results: It was found that the electric field, depending on the applied voltage, its duration and the polarization of the maize seedlings, stimulated or inhibited the growth of the seedling organs (root, mesocotyl and coleoptile). Moreover, it was also noticed that the gravitropic response of maize seedlings was inhibited at all voltages studied. Simultaneous measurements of growth and external medium pH show that auxin(IAA, indole-3-acetic acid)- and fusicoccin(FC)-induced elongation growth and proton extrusion of maize coleoptile segments were significantly inhibited at higher voltages. The ionic current flowing through the single coleoptile segment during voltage application was 1.7-fold lower in segments treated with cation channel blocker tetraethylammonium chloride (TEA-Cl) and 1.4-fold higher with IAA compared to the control. The electrophysiological experiments show that the electric field caused the depolarization of the membrane potential of parenchymal coleoptile cells, which was not reversible over 120 min. Conclusion: It is suggested that a DC electric field inhibits the plasma membrane H+ pump activity and K+ uptake through voltage-dependent, inwardly rectifying ZMK1 channels (Zea mays K+ channel 1). The data presented here are discussed, taking into account the “acid growth hypothesis” of the auxin action and the mechanism of gravitropic response induction

The Effects of 1,4-Naphthoquinone (NQ) and Naphthazarin (5,8-Dihydroxy-1,4-naphthoquinone, DHNQ) Individually and in Combination on Growth and Oxidative Stress in Maize (<i>Zea mays</i> L.) Seedlings

This study investigated the effects of 1,4-naphthoquinone (NQ) and naphthazarin (5,8-dihydroxy-1,4-naphthoquinone, DHNQ) individually and in combination, applied at low concentrations (0.1, 1, and 10 nM), on growth, hydrogen peroxide (H2O2) production, catalase activity, and lipid peroxidation in maize seedlings. It was found that NQ at 0.1 and 1 nM and DHNQ at 0.1 nM significantly stimulated the fresh weight of the aboveground parts of the seedlings (APS), while the fresh weight of the underground parts of the seedlings (UPS) was enhanced only at 0.1 nM NQ. Interestingly, DHNQ at higher concentrations (1 and 10 nM) significantly diminished the fresh weight of the APS and UPS. When NQ and DHNQ were applied together, an increase in the fresh weight of the APS at all of the concentrations studied was observed. It was also found that NQ and DHNQ individually and in combination, at all concentrations studied, decreased the H2O2 production in the aboveground and underground parts of maize seedlings. The presence of the DHNQ at higher concentrations (1 and 10 nM) triggered an increase in the catalase (CAT) activity of the UPS and APS compared to the control. However, NQ added at 1 nM decreased the CAT activity of both the UPS and APS, while 10 nM increased the CAT activity of UPS. NQ and DHNQ applied together at 0.1 and 10 nM almost completely inhibited catalase activity in the UPS and APS. The data that were obtained for lipid peroxidation, measured as the malondialdehyde (MDA) concentration, indicated that NQ and DHNQ at all concentrations studied decreased the MDA content of the UPS, while both naphthoquinones increased it in APS. The data presented here are discussed taking into account the mechanisms via which naphthoquinones interact with biological systems