Potassium Efflux and Cytosol Acidification as Primary Anoxia-Induced Events in Wheat and Rice Seedlings

Both ion fluxes and changes of cytosolic pH take an active part in the signal transduction of different environmental stimuli. Here we studied the anoxia-induced alteration of cytosolic K⁺ concentration, [K⁺]_cyt, and cytosolic pH, pH_cyt, in rice and wheat, plants with different tolerances to hypoxia. The [K⁺]_cyt and pH_cyt were measured by fluorescence microscopy in single leaf mesophyll protoplasts loaded with the fluorescent potassium-binding dye PBFI-AM and the pH-sensitive probe BCECF-AM, respectively. Anoxic treatment caused an efflux of K⁺ from protoplasts of both plants after a lag-period of 300-450 s. The [K⁺]_cyt decrease was blocked by tetraethylammonium (1 mM, 30 min pre-treatment) suggesting the involvement of plasma membrane voltage-gated K⁺ channels. The protoplasts of rice (a hypoxia-tolerant plant) reacted upon anoxia with a higher amplitude of the [K⁺]_cyt drop. There was a simultaneous anoxia-dependent cytosolic acidification of protoplasts of both plants. The decrease of pH_cyt was slower in wheat (a hypoxia-sensitive plant) while in rice protoplasts it was rapid and partially reversible. Ion fluxes between the roots of intact seedlings and nutrient solutions were monitored by ion-selective electrodes and revealed significant anoxia-induced acidification and potassium leakage that were inhibited by tetraethylammonium. The K⁺ efflux from rice was more distinct and reversible upon reoxygenation when compared with wheat seedlings.

Keywords: Oryza sativa; Triticum aestivum; acidification; anoxic signaling; fluorescence microscopy; pH; potassium.