S-glutathionylation, the formation of disulphide of glutathione and target protein cysteine residues, is a post-translational modification that modulates the function of target protein. Besides its formation, the deglutathionylation of protein can also be reversibly catalyzed by glutaredoxin (Grx). The S-glutathionylation modification is also considered to be a protective mechanism for preventing cysteine residues of protein from irreversible oxidation. Since it changes the structure and function of the redox-sensitive thiol-containing protein, S-glutathionylation therefore is a mechanism responsible for the regulation of protein function. The changes in the levels of S-glutathionylation in mammalian cells are associated with many pathologic mechanisms. However, the research of S-glutathionylation in plants is just the beginning. In this paper, the research progress in mechanism, detecting method, and physiological action of S-glutathionylation were reviewed. Finally, the important problems in the future research were also put forward.