Control:Article Title: Equipment and protocol for measurement of extracellular electrical signals, gas exchange and turgor pressure in plants.
Article Snippet: This is an open access article under the CC BY-NC-ND license http://creativecommons.org/licenses/by-nc-nd/4.0/ ) Specifications table Subject Area Agricultural and Biological Sciences More specific subject area Plant Electrophysiology Protocol name Extracellular measurement of electrical signals in plants Reagents/tools Equipment and software Data acquisition interface (Lab-Trax 4/24T) - World Precision Instruments LabScribe version 3 - iWorx Systems Inc. Infra-red gas analyzer (IRGA) - IRGA, Model Li-6400, Li-Cor Patch clamp pressure probe (ZIM-probe) – YARA ZIM-plant Technology Computer with USB interface. .. Electronic components Faraday cage with LED lamps that may be linked to a timer to control the photoperiod; Silver wire (0.5 or 0.25 mm diameter) - World Precision Instruments, e.g., cat. No. AGW 2010 and AGT 1010, respectively; Copper wire of 0.2 mm diameter - from local electronics supplier Gold pin - World Precision Instruments, e.g., cat. No. 5482 Gold socket - World Precision Instruments, e.g., cat. No. 5483 BNC cable - from local electronics supplier Male DIN8 connector - from local electronics supplier Reagent 3 M KCl solution Experimental design The experiments were carried out in the Laboratory of Plant Stress Study (LEPSE) of University of São Paulo, Brazil. ..
Article Title: Equipment and protocol for measurement of extracellular electrical signals, gas exchange and turgor pressure in plants
Article Snippet: .. Reagents/tools , Equipment and software Data acquisition interface (Lab-Trax 4/24T) - World Precision Instruments LabScribe version 3 - iWorx Systems Inc. Infra-red gas analyzer (IRGA) - IRGA, Model Li-6400, Li-Cor Patch clamp pressure probe (ZIM-probe) – YARA ZIM-plant Technology Computer with USB interface. Electronic components Faraday cage with LED lamps that may be linked to a timer to control the photoperiod; Silver wire (0.5 or 0.25 mm diameter) - World Precision Instruments, e.g., cat. No. AGW 2010 and AGT 1010, respectively; Copper wire of 0.2 mm diameter - from local electronics supplier Gold pin - World Precision Instruments, e.g., cat. No. 5482 Gold socket - World Precision Instruments, e.g., cat. No. 5483 BNC cable - from local electronics supplier Male DIN8 connector - from local electronics supplier Reagent 3 M KCl solution. .. Experimental design , The experiments were carried out in the Laboratory of Plant Stress Study (LEPSE) of University of São Paulo, Brazil. The tomato and sunflower plants grew in a greenhouse equipped with evaporative coolers. We transferred 30- to 60-day-old plants from the greenhouse to the laboratory where the measurements of gas exchange, turgor pressure and electrical signals were performed simultaneously. Leaf turgor pressure was measured by using the leaf patch clamp pressure probe which enables real-time recording of the turgor variation. It is a non-destructive or invasive method, so the measurements can be performed for several days without interruption. To ensure a perfect stabilization of the measures, the probe was installed in plants, even in the greenhouse at least two days before the plants are transferred to the laboratory. In the laboratory, the plants were placed in the faraday cage and the electrical system installed as described below. Only after the electrical measurement is stabilized (around 3 h after inserting the electrodes), the leaf was closed in the IRGA camera to start gas exchange measurements. CO 2 assimilation rate ( A , μmol m −2 s −1 ), transpiration ( E , mmol m −2 s −1 ), stomatal conductance ( g s , mol m −2 s −1 ) were measured continuously for eight hours, from 9 a.m. to 5 p.m. The equipment (IRGA) was programmed using the 'autolog' function to record measures in time intervals from 10 to 300 s. We analyzed the electrical activity of the plants as the number of signals generated, amplitude, speed and duration of the signals. In order to record action potentials (AP) evoked by electrical stimulation, an electric current was supplied extracellularly through Ag/AgCl electrodes. Different voltages were used to determine the excitation threshold. For post-irrigation records, the plants were subjected to water deficit and after eight days (determined in preliminary tests) re-irrigated. To record variation potentials (PVs), a burning stimulus was applied by placing the flame approximately 3 cm from the leaf for 10 s..
Software:Article Title: Equipment and protocol for measurement of extracellular electrical signals, gas exchange and turgor pressure in plants
Article Snippet: .. Reagents/tools , Equipment and software Data acquisition interface (Lab-Trax 4/24T) - World Precision Instruments LabScribe version 3 - iWorx Systems Inc. Infra-red gas analyzer (IRGA) - IRGA, Model Li-6400, Li-Cor Patch clamp pressure probe (ZIM-probe) – YARA ZIM-plant Technology Computer with USB interface. Electronic components Faraday cage with LED lamps that may be linked to a timer to control the photoperiod; Silver wire (0.5 or 0.25 mm diameter) - World Precision Instruments, e.g., cat. No. AGW 2010 and AGT 1010, respectively; Copper wire of 0.2 mm diameter - from local electronics supplier Gold pin - World Precision Instruments, e.g., cat. No. 5482 Gold socket - World Precision Instruments, e.g., cat. No. 5483 BNC cable - from local electronics supplier Male DIN8 connector - from local electronics supplier Reagent 3 M KCl solution. .. Experimental design , The experiments were carried out in the Laboratory of Plant Stress Study (LEPSE) of University of São Paulo, Brazil. The tomato and sunflower plants grew in a greenhouse equipped with evaporative coolers. We transferred 30- to 60-day-old plants from the greenhouse to the laboratory where the measurements of gas exchange, turgor pressure and electrical signals were performed simultaneously. Leaf turgor pressure was measured by using the leaf patch clamp pressure probe which enables real-time recording of the turgor variation. It is a non-destructive or invasive method, so the measurements can be performed for several days without interruption. To ensure a perfect stabilization of the measures, the probe was installed in plants, even in the greenhouse at least two days before the plants are transferred to the laboratory. In the laboratory, the plants were placed in the faraday cage and the electrical system installed as described below. Only after the electrical measurement is stabilized (around 3 h after inserting the electrodes), the leaf was closed in the IRGA camera to start gas exchange measurements. CO 2 assimilation rate ( A , μmol m −2 s −1 ), transpiration ( E , mmol m −2 s −1 ), stomatal conductance ( g s , mol m −2 s −1 ) were measured continuously for eight hours, from 9 a.m. to 5 p.m. The equipment (IRGA) was programmed using the 'autolog' function to record measures in time intervals from 10 to 300 s. We analyzed the electrical activity of the plants as the number of signals generated, amplitude, speed and duration of the signals. In order to record action potentials (AP) evoked by electrical stimulation, an electric current was supplied extracellularly through Ag/AgCl electrodes. Different voltages were used to determine the excitation threshold. For post-irrigation records, the plants were subjected to water deficit and after eight days (determined in preliminary tests) re-irrigated. To record variation potentials (PVs), a burning stimulus was applied by placing the flame approximately 3 cm from the leaf for 10 s..
Patch Clamp:Article Title: Equipment and protocol for measurement of extracellular electrical signals, gas exchange and turgor pressure in plants
Article Snippet: .. Reagents/tools , Equipment and software Data acquisition interface (Lab-Trax 4/24T) - World Precision Instruments LabScribe version 3 - iWorx Systems Inc. Infra-red gas analyzer (IRGA) - IRGA, Model Li-6400, Li-Cor Patch clamp pressure probe (ZIM-probe) – YARA ZIM-plant Technology Computer with USB interface. Electronic components Faraday cage with LED lamps that may be linked to a timer to control the photoperiod; Silver wire (0.5 or 0.25 mm diameter) - World Precision Instruments, e.g., cat. No. AGW 2010 and AGT 1010, respectively; Copper wire of 0.2 mm diameter - from local electronics supplier Gold pin - World Precision Instruments, e.g., cat. No. 5482 Gold socket - World Precision Instruments, e.g., cat. No. 5483 BNC cable - from local electronics supplier Male DIN8 connector - from local electronics supplier Reagent 3 M KCl solution. .. Experimental design , The experiments were carried out in the Laboratory of Plant Stress Study (LEPSE) of University of São Paulo, Brazil. The tomato and sunflower plants grew in a greenhouse equipped with evaporative coolers. We transferred 30- to 60-day-old plants from the greenhouse to the laboratory where the measurements of gas exchange, turgor pressure and electrical signals were performed simultaneously. Leaf turgor pressure was measured by using the leaf patch clamp pressure probe which enables real-time recording of the turgor variation. It is a non-destructive or invasive method, so the measurements can be performed for several days without interruption. To ensure a perfect stabilization of the measures, the probe was installed in plants, even in the greenhouse at least two days before the plants are transferred to the laboratory. In the laboratory, the plants were placed in the faraday cage and the electrical system installed as described below. Only after the electrical measurement is stabilized (around 3 h after inserting the electrodes), the leaf was closed in the IRGA camera to start gas exchange measurements. CO 2 assimilation rate ( A , μmol m −2 s −1 ), transpiration ( E , mmol m −2 s −1 ), stomatal conductance ( g s , mol m −2 s −1 ) were measured continuously for eight hours, from 9 a.m. to 5 p.m. The equipment (IRGA) was programmed using the 'autolog' function to record measures in time intervals from 10 to 300 s. We analyzed the electrical activity of the plants as the number of signals generated, amplitude, speed and duration of the signals. In order to record action potentials (AP) evoked by electrical stimulation, an electric current was supplied extracellularly through Ag/AgCl electrodes. Different voltages were used to determine the excitation threshold. For post-irrigation records, the plants were subjected to water deficit and after eight days (determined in preliminary tests) re-irrigated. To record variation potentials (PVs), a burning stimulus was applied by placing the flame approximately 3 cm from the leaf for 10 s..
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