Semiochemicals play important roles in mate and host recognition of herbivorous insects, such as moths, and flight tunnels have been an effective tool in the identification of these bioactive compounds. However, more work has been carried out on pheromones than on host plant cues, and few examples exist where flight tunnel evaluations of host cues have resulted in a lure that is attractive under field conditions. Our goal was to determine whether the flight tunnel could be used to evaluate the response of a specialist moth, grape berry moth (GBM), to its host plant (grapevines), by incorporating ecological and physiological aspects of GBM biology. We found grape shoot tips and mature leaves were more attractive to female GBM than unripe and ripe berries or flowers. Under optimized flight tunnel conditions, approximately 80% of tested females flew upwind and closely approached or landed on the most preferred target. Mating status, wind speed, the time of day, and the presence/absence of patterns that resemble grape tissues on the top of the flight tunnel all significantly affected the responses of female GBM. Consideration of these factors in flight tunnel assays will aid in the development of a synthetic lure that can be used to monitor female moths in the field.