Sodium sensing induces different changes in free cytosolic calcium concentration and pH in salt‐tolerant and ‐sensitive rice (Oryza sativa) cultivars

Abstract

Perception of salt stress in plant cells induces a change in the free cytosolic Ca2+, [Ca2+]cyt, which transfers downstream reactions toward salt tolerance. Changes in cytosolic H+ concentration, [H+]cyt, are closely linked to the [Ca2+]cyt dynamics under various stress signals. In this study, salt‐induced changes in [Ca2+]cyt, and [H+]cyt and vacuolar [H+] concentrations were monitored in single protoplasts of rice (Oryza sativa L. indica cvs. Pokkali and BRRI Dhan29) by fluorescence microscopy. Changes in cytosolic [Ca2+] and [H+] were detected by use of the fluorescent dyes acetoxy methyl ester of calcium‐binding benzofuran and acetoxy methyl ester of 2′, 7′‐bis‐(2‐carboxyethyl)‐5‐(and‐6) carboxyfluorescein, respectively, and for vacuolar pH, fluorescent 6‐carboxyfluorescein and confocal microscopy were used. Addition of NaCl induced a higher increase in [Ca2+]cyt in the salt‐tolerant cv. Pokkali than in the salt‐sensitive cv. BRRI Dhan29. From inhibitor studies, we conclude that the internal stores appear to be the major source for [Ca2+]cyt increase in Pokkali, although the apoplast is more important in BRRI Dhan29. The [Ca2+]cyt measurements in rice also suggest that Na+ should be sensed inside the cytosol, before any increase in [Ca2+]cyt occurs. Moreover, our results with individual mesophyll protoplasts suggest that ionic stress causes an increase in [Ca2+]cyt and that osmotic stress sharply decreases [Ca2+]cyt in rice. The [pH]cyt was differently shifted in the two rice cultivars in response to salt stress and may be coupled to different activities of the H+‐ATPases. The changes in vacuolar pH were correlated with the expressional analysis of rice vacuolar H+‐ATPase in these two rice cultivars.