In halophytic plants, the high-affinity potassium transporter HKT gene family can selectively uptake K＋ in the presence of toxic concentrations of Na＋. This has so far not been well examined in glycophytic crops. Here, we report the characterization of SbHKTI;4, a member of the HKT gene family from Sorghum bicolor. Upon Na＋ stress, SbHKT1;4 expression was more strongly upregulated in salt-tolerant sorghum accession, correlating with a better balanced Na＋/ K＋ ratio and enhanced plant growth. Heterogeneous expression analyses in mutants of Saccharomyces cerevisiae and Arabidopsis thaliana indicated that overexpressing SbHKT1;4 resulted in hypersensitivity to Na＋ stress, and such hypersensitivity could be alleviated with the supply of elevated levels of K＋, implicating that SbHKT1;4 may mediate K＋ uptake in the presence of excessive Na＋. Further electrophysiological evidence demonstrated that SbHKT1;4 could transport Na＋ and K＋ when expressed in Xenopus laevis oocytes. The relevance of the finding that SbHKTI;4 functions to maintain optimal Na＋/K＋ balance under Na＋ stress to the breeding of salt-tolerant glycophytic crops is discussed.