We report on the in-plane anisotropy of the electronic response in the spin-, charge-, and orbital-ordered phase of a half-doped layered-structure manganite. The optical conductivity spectra for a single domain of Eu1/2Ca3/2MnO4 unambiguously show the anisotropic charge dynamics which well corresponds to the theoretical calculation: the optical conductivity with the polarization along the zigzag ferromagnetic chain direction exhibits a smaller gap and a larger intensity at lower energies than that of the perpendicular polarization mostly due to the charge and orbital ordering and the associated quantum interference effect.