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Plants are subject to stimuli from the environment on which they strongly depend and in contrast to animals, they are unable to escape harmful influences. Therefore, being able to receive stimuli they have developed adequate responses to them. Such a reaction can occur in the area of a stimulus action or cover the whole plant or its parts. In the latter case, it is a systemic reaction. The plant reaction is expressed by various intensity, rate and kind of response. It is interesting to know the character of the signal informing about a stimulus, the routes of its propagation and the transmission mechanism. Three conceptions of excitation are distinguished: 1) propagation of chemical agents formed at the site of a stimulus action with the flow of the phloem sap or through the atmosphere (in the case of volatile substances) to other plant parts, 2) a very fast transmission by the xylem in the wave of hydraulic pressure formed after a plant damage. From combining the "hydraulic" and "chemical" hypothesis a conception of hydraulic dispersion has been formulated which assumes that chemical substances synthetized after an injury can be transferred very fast with the wave of hydraulic pressure changes in the whole plant, 3) a stimulus evokes the action potential (AP), and its transmission along the whole plant, plant organ or specialized tissue, by local circuits from cell to cell. Strong, damaging stimuli can evoke variation potentials (VPs), the character of which differs from APs. It is postulated that transmission of VP occurs by a hydraulic dispersion and electrical changes seem to be secondary phenomena.
The object of our patch-clamp study was the liverwort Marchantia polymorpha. Basic solutions were symmetrical in the bath and in the pipette and contained 100 mM KCl and 0.5 mM CaCl2. Whole vacuole recordings mainly showed slowly activating outward rectifying currents, typical for SV (slowvacuolar) channels. The unitary conductance registered from isolated patches was 74.7±4.0 pS at 100 mV. Replacement of K+ with Na+ caused reduction of the unitary conductance to 59.8±6.87 pS. Tenfold decrease of the KCl shifted the reversal potential close to EK and caused reduction of the SV unitary conductance to 31.5±5.5 pS. Moreover, the gradient of KCl revealed the current at the negative potential; the unitary conductance at −100 mV was 43.3±0.5 pS, whereas the gradient of Na+ did not evoke such an effect. This may suggest that inwardly rectifying K+ channels exist in the tonoplast of the liverwort. Supported by NCN grant 2013/09/B/NZ1/01052
An effect of lithium (Li+) on growth, circumnutation, and glutamate-induced excitation in sunflower (Helianthus annuus L.) seedlings was investigated using time-lapse photography and extracellular electrical potential measurements. The seedlings were treated with a micro and millimolar concentration of lithium chloride (LiCl) both persistently in a hydroponic medium and acutely through Li+ injection. The length of hypocotyls, fresh weight of seedlings, intensity and period of circumnutation, and the number of action potentials (APs) after glutamate(Glu) injection were determined. It was found that the circumnutation intensity and period did not depend on hypocotyl length and fresh weight of seedlings. Under persistent Li+ treatment, the circumnutation intensity was constant at a concentration between 0.2 and 20 mM although the hypocotyls were significantly shorter in relation to the control, whereas at a concentration of 40 mM circumnutation intensity decreased without any changes in the hypocotyl length. Under persistent treatment with 0.5 and 20 mM Li+, the period of circumnutation was significantly prolonged. The number of APs in a Glu-induced series significantly increased in the seedlings exhibiting an Li+-induced decrease in circumnutation intensity (in 40 and 60 mM Li+). Additionally Li+ injection before Glu injection also augmented the series of APs in seedlings growing without Li+ in hydroponic medium. These Li+- sensitive responses demonstrated that circumnutation and growth are partly independent processes and reveal a relationship between circumnutation intensity and excitability in Helianthus annuus seedlings.
In specialised sensitive plants such as Mimosa, Dionaea, and Aldrovanda, rapid organ movement is observed and the excitationturgor loss mechanism is the basis for rapid leaf or trap closure. In non-specialised ubiquitous plants, slow movement named circumnutation is common and it is driven by turgor. We examine whether transmembrane potential changes such as oscillations and long distance signal action and variation potentials are involved in this movement. Video camera recordings combined with extracellular measurement of electrical potential changes are applied. Additionally, intracellular microelectrodes and patchclamp measurements are engaged. Novel software Circumnutation Tracker has been developed to track organ movement and standardisation of circumnutation parameters. Helianthus annuus and Arabidopsis thaliana are studied and a model of the circumnutation mechanism is proposed.
Liverworts are pioneer plants that colonized lands. They had to cope with frequent sea water flooding causing salt stress. The role of vacuoles and in particular slow-activating (SV) channels in the salt stress tolerance was addressed in the present study. A patch-clamp method was used to study sodium fluxes through the tonoplast of the liverwort Conocephalum conicum. The whole-vacuole measurements carried out in a symmetrical Na⁺ concentration allowed recording of slowly activated outward currents typical for SV channels. In a Na⁺ gradient promoting an efflux of Na⁺ from the vacuole, the outward rectifying properties of SV channels were reduced and inward Na⁺ currents with different inactivation rates were recorded. Single channel analysis proved that a decrease in cytoplasmic Na⁺ concentration evoked an increase in the open probability of the channels and shifted the activation voltages towards negative values. The number of SV channels recorded at negative voltages was dependent on the vacuolar calcium and decreased at the high concentration of this ion in the vacuole. In some of the tested patches, the channels exhibited a flickering type of activity and two different conductance levels. The role of SV channels in Na⁺ accumulation during salt stress and its removal after periods of flooding is discussed in the present paper.
Multichannel extracellular measurements of electrical potential changes allowed comparison of the electrical activity in two types of tomato: wild type (MTwt) and ABA-deficient, draught-sensitive mutant (MTsit). After electrical stimulation of the stem, MTsit generated action potentials (APs) with an amplitude of 40 mV and half-time (τ1/2) 19 s compared to MTwt 20 mV and 19 s, respectively. Intracellular microelectrodes were used to detect APs triggered by light in the leaves and cotyledons of MTwt. APs were characterized by long latency and their amplitudes were 77±2 mV and τ1/2 58±2 s in leaves, and 67±14 mV, τ1/2 13±2 s in the cotyledons. Turning off the light evoked 25 mV hyperpolarization in the leaf and 10 mV in the cotyledon.
Action potentials generated spontaneously (SAPs) and evoked by electrical stimulation (APs) in tomato plants (Solanum lycopersicum L.) cv. Micro-Tom ABA-deficient mutants (sitiens—MTsit) and its wild type (MTwt) were characterized by continuous monitoring of electrical activity for 66 h and by application of an electrical stimulation supplied extracellularly. MTsit generated SAPs which spread along the stem, including petioles and roots with an amplitude of 44.6 ± 4.4 mV, half-time (t) of 33.1 ± 2.9 s and velocity of 5.4 ± 1.0 cm min-1. Amplitude and velocity were 43 and 108 % higher in MTsit than in MTwt, respectively. The largest number of SAPs was registered in the early morning in both genotypes. MTsit was less responsive to electrical stimuli. The excitation threshold and the refractory period were greater in MTsit than in MTwt. After current application, APs were generated in the MTwt with 21.2 ± 2.4 mV amplitude and propagated with 5.6 ± 0.5 cm min-1 velocity. Lower intensity stimuli did not trigger APs in these plants. In MTsit APs were measured with amplitude of 26.8 ± 4.8 mV and propagated with velocity of 8.5 ± 0.1 cm min-1.
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