Expression of a Na+/H+ antiporter RtNHX1 from a recretohalophyte Reaumuria trigyna improved salt tolerance of transgenic Arabidopsis thaliana

Reaumuria trigyna is an endangered recretohalophyte and a small xeric shrub that is endemic to the eastern Alxa and western Ordos areas of Inner Mongolia, China. Using transcriptome data, we identified a 1662-bp open reading frame encoding a 553-amino-acid protein corresponding to a Na+/H+ antiporter (RtNHX1) from R. trigyna. RtNHX1 was rapidly up-regulated by NaCl and exogenous abscisic acid treatment and had different tissue-specific expression patterns before and after salt-stress treatment. Overexpression of RtNHX1 enhanced seed germination, biomass accumulation, chlorophyll content, and root elongation in transgenic Arabidopsis plants under salt stress and rescued the salt-sensitive deficiencies of the nhx1 mutant. POD and CAT enzyme activities, proline content, and RWC all increased significantly in salt-stressed transgenic Arabidopsis plants, whereas MDA content did not. Additionally, there was a corresponding upregulation of some antioxidant-enzyme, proline biosynthesis and other stress responsive genes (AtPOD1, AtCAT1, AtP5CS1, AtP5CS2, AtRD29A, AtRD29B, AtKIN1, and AtABI2). The transgenic Arabidopsis plants accumulated more K+ and less Na+ in their leaves and had lower Na+/K+ ratios than WT plants. This was reflected in the upregulation of some ion transport-related genes (AtAVP1, AtSOS1, AtKUP6, and AtKUP8). When RtNHX1 was expressed in the AXT3 yeast strain, the accumulation of Na+ and K+ in the vacuole increased and the Na+/K+ ratio decreased. These results reveal that R. trigyna RtNHX1 is a functional antiporter that sequesters Na+ and K+ in the vacuole and could confer salt tolerance on transgenic Arabidopsis plants by maintaining Na+/K+ homeostasis and enhancing osmotic and antioxidant regulatory capacity. These results suggest that RtNHX1 may be a good target for improving salt tolerance in plants.