It is well known that nitric oxide (NO) enhances salt tolerance of glycophytes. However, the effect of NO on modulating ionic balance in halophytes is not very clear. This study focuses on the role of NO in mediating K+/Na+ balance in a mangrove species, Kandelia obovata Sheue, Liu and Yong. We first analyzed the effects of sodium nitroprusside (SNP), an NO donor, on ion content and ion flux in the roots of K. obovata under high salinity. The results showed that 100 μM SNP significantly increased K+ content and Na+ efflux, but decreased Na+ content and K+ efflux. These effects of NO were reversed by specific NO synthesis inhibitor and scavenger, which confirmed the role of NO in retaining K+ and reducing Na+ in K. obovata roots. Using western-blot analysis, we found that NO increased the protein expression of plasma membrane (PM) H+-ATPase and vacuolar Na+/H+ antiporter, which were crucial proteins for ionic balance. To further clarify the molecular mechanism of NO-modulated K+/Na+ balance, partial cDNA fragments of inward-rectifying K+ channel, PM Na+/H+ antiporter, PM H+-ATPase, vacuolar Na+/H+ antiporter and vacuolar H+-ATPase subunit c were isolated. Results of quantitative real-time PCR showed that NO increased the relative expression levels of these genes, while this increase was blocked by NO synthesis inhibitors and scavenger. Above results indicate that NO greatly contribute to K+/Na+ balance in high salinity-treated K. obovata roots, by activating AKT1-type K+ channel and Na+/H+ antiporter, which are the critical components in K+/Na+ transport system.