Abstract
Cytosolic pH (pHcyt) regulates a wide range of cellular processes in plants. Changes in pHcyt occurring under the effect of different stressors can participate in signal transmission. The dynamics of pHcyt under the action of external factors, including significant factors for open ground crops such as temperature, remains poorly understood, which is largely due to the difficulty of intracellular pH registration using standard methods. In this work, model plants of potato (one of the essential crops) expressing a fluorescent ratiometric pH sensor Pt-GFP were created. The calibration obtained in vivo allowed for the determination of the pHcyt values of the cells of the leaves, which is 7.03 ± 0.03 pH. Cooling of the whole leaf caused depolarization and rapid acidification of the cytosol, the amplitude of which depended on the cooling strength, amounting to about 0.2 pH units when cooled by 15 °C. When the temperature rises to 35–40 °C, the cytosol was alkalized by 0.2 pH units. Heating above the threshold temperature caused the acidification of cytosol and generation of variation potential. The observed rapid changes in pHcyt can be associated with changes in the activity of H+-ATPases, which was confirmed by inhibitory analysis.
Highlights
Potato is one of the most important crops grown in open ground conditions
Alkalization is observed in cases of penetration of pathogen fungus Alternaria alternata hyphae into the plant cell [21], gravistimulation [22], the external addition of indole-3-acetic acid (IAA) [23] and anoxia, which acidifies cytosol by 0.8–1.2 units [24]
It should be noted that the majority of research on the effect of temperature changes on cytosolic pH has been carried out on cell cultures [26] or in the cells of individual organs [25], and the pH has not been studied in the whole organs or plant
Summary
Potato is one of the most important crops grown in open ground conditions. Cultivation in open ground conditions exposes potato plants to a greater influence of uncontrollable external factors than growing in greenhouses under a controlled level of humidity, degree of irrigation and other conditions [1,2,3]. Temperature changes affect the activity of a number of processes, including photosynthesis and starch metabolism [7,8], which can lead to plant death or crop yield reduction. A decrease in yield is mediated via changes in metabolism, the direct regulation of the activity of key enzymes and the initiation of a plant defense reaction, which occurs with the participation of signaling systems. The importance of cytosolic pH for the regulation of many cellular processes has been the reason for creating of various methods for its measurement, which has different degrees of invasiveness and toxicity for the plant and the duration of observation. This work describes the creation of potato plants with the genetically encoded Pt-GFP sensor and their application for the registration of pH shifts under the local and systemic action of a temperature stimulus
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