The Drosophila melanogaster white-eyed w1118 line serves as a blank control, allowing genetic recombination of any gene of interest along with a readily recognizable marker. w1118 flies display behavioral susceptibility to environmental stimulation such as light. It is of great importance to characterize the behavioral performance of w1118 flies because this would provide a baseline from which the effect of the gene of interest could be differentiated. Little work has been performed to characterize the walking behavior in adult w1118 flies. Here we show that pulsed light stimulation increased the regularity of walking trajectories of w1118 flies in circular arenas. We statistically modeled the distribution of distances to center and extracted the walking structures of w1118 flies. Pulsed light stimulation redistributed the time proportions for individual walking structures. Specifically, pulsed light stimulation reduced the episodes of crossing over the central region of the arena. An addition of four genomic copies of mini-white, a common marker gene for eye color, mimicked the effect of pulsed light stimulation in reducing crossing in a circular arena. The reducing effect of mini-white was copy-number-dependent. These findings highlight the rhythmic light stimulation-evoked modifications of walking behavior in w1118 flies and an unexpected behavioral consequence of mini-white in transgenic flies carrying w1118 isogenic background.