The effect of material surface morphology on the periodic subwavelength of nano-structures induced by a femtosecond (fs) laser was investigated systematically from the initial surface roughness, the different scratches, the pre-formed ripples, and the “layer-carving” technology experiments. The results of the comparative experiments indicate that the initial surface conditions of the target surface have no obvious effects on the spatial structured periods (SSPs) and the ripple orientation of the periodic nano-structures induced by a fs laser, which agreed well with the foretold present surface two-plasmon resonance (STPR) model. Furthermore, different shapes of nano-grids with high regularity and uniformity were obtained by fs-laser fabrication.