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Vacuolar sodium/proton (Na + /H + ) antiporters (NHXs) can stabilize ion contents to improve the salt tolerance of plants. Here,  GhNHX3D  was cloned and characterized from upland cotton ( Gossypium hirsutum ). Phylogenetic and sequence analyses showed that  GhNHX3D  belongs to the vacuolar-type NHXs. The GhNHX3D-enhanced green fluorescent protein (eGFP) fusion protein localized on the vacuolar membrane when transiently expressed in  Arabidopsis  protoplasts. The quantitative real-time PCR (qRT-PCR) analysis showed that  GhNHX3D  was induced rapidly in response to salt stress in cotton leaves, and its transcript levels increased with the aggravation of salt stress. The introduction of  GhNHX3D  into the salt-sensitive yeast mutant ATX3 improved its salt tolerance. Furthermore, silencing of  GhNHX3D  in cotton plants by virus-induced gene silencing (VIGS) increased the Na +  levels in the leaves, stems, and roots and decreased the K +  content in the roots, leading to greater salt sensitivity. Our results indicate that  GhNHX3D  is a member of the vacuolar NHX family and can confer salt tolerance by adjusting the steady-state balance of cellular Na +  and K +  ions.

GhNHX3D, a Vacuolar-Localized Na+/H+ Antiporter, Positively Regulates Salt Response in Upland Cotton