Objective Chemotaxis and electrotaxis are the primary mechanisms underlying directed cell migration, and they play important roles in the physiology and pathology of organisms. However, there are differences between the two. This paper presents a comparative study of the roles of Dictyostelium discoideum genes gbpC and gbpD in cell electrotaxis and chemotaxis in order to gain further insight into the differences between these two mechanisms of migration.Methods The gbpC/gefT- mutant strain and gbpD/gefU- mutant strain were placed in a 12 V/cm direct current (DC) electric field to investigate the direction and velocity of cell movement and the changes in cell electrotaxis; Lifeact-GFP (F-actin) was electroporated into cells and the distribution of F-actin during cell movement was observed under a fluorescence microscope; Western blot was used to quantify the phosphorylation of myosin regulatory light chain (RLC) in cells stimulated by DC electric fields.Results The gefT- mutant cells lost polarization but retained electrotaxis at a level similar to the wild-type cells; the gefU- mutant cells showed hyperpolarization but significantly reduced electrotaxis; in a DC field, F-actin was predominantly distributed in the pseudopods in both mutant and wild-type cells; there were differences in myosin RLC phosphorylation between the cell lines in electric fields. Phosphorylation was time-dependent in wild-type cells, whereas phosphorylation first decreased rapidly and then increased in gefT- mutant cells. Time-dependent dephosphorylation occurred in gefU- mutant cells.Conclusion Our findings indicate that gbpC and gbpD play differntal roles between the chemotaxis and electrotaxis of Dictyostelium discoideum and provide further evidence that electrical and chemical signals drive the directed migration of cells through different mechanisms.