A Na +/H + antiporter-like protein (NHXLP) was isolated from Sorghum bicolor L. ( SbNHXLP) and validated by overexpressing in tomato for salt tolerance. Homozygous T 2 transgenic lines when evaluated for salt tolerance, accumulated low Na + and displayed enhanced salt tolerance compared to wild-type plants (WT). This is consistent with the amiloride binding assay of the protein. Transgenics exhibited higher accumulation of proline, K +, Ca 2+, improved cambial conductivity, higher PSII, and antioxidative enzyme activities than WT. Fluorescence imaging results revealed lower Na + and higher Ca 2+ levels in transgenic roots. Co-immunoprecipitation experiments demonstrate that SbNHXLP interacts with a Solanum lycopersicum cation proton antiporter protein2 ( SlCHX2). qRT-PCR results showed upregulation of SbNHXLP and SlCHX2 upon treatment with 200 mM NaCl and 100 mM potassium nitrate. SlCHX2 is known to be involved in K + acquisition, and the interaction between these two proteins might help to accumulate more K + ions, and thus maintain ion homeostasis. These results strongly suggest that plasma membrane bound SbNHXLP involves in Na + exclusion, maintains ion homeostasis in transgenics in comparison with WT and alleviates NaCl stress.