Impacts of Ashe juniper (Juniperus ashei J. Buchholz) and karst geology on the regional water cycle in the Edwards plateau region of Texas are complex and not well understood. The objective of our study was to gain a comprehensive understanding of the subsurface flow processes occurring at a juniper woodland site on the Edwards Plateau near Honey Creek State Natural Area. A 2.3-m-deep, 7-m-long trench was excavated at the downslope end of a 7 by 14 m experimental plot, and time domain reflectometry (TDR) probes were installed at various locations within the trench face to measure volumetric water contents. A multi-port telescopic boom type rainfall simulator was used to provide artificial rainfall on the plot. Six rainfall simulations and two dye-tracer tests were conducted on the plot during a 7-mo period. Subsurface flow was visually inspected at various locations on the trench face during artificial rainfall experiments and water content was monitored near slow and fast flow regions using TDR probes. The total volume of subsurface flow was also recorded after each rainfall simulation event. Results demonstrated that subsurface flow occurred in a bimodal manner, consisting of preferential/macropore flow around juniper root channels and planar fractures in the limestone, and pseudo-matrix flow through the soil matrix (water flowing primarily through the intermediate layers and lenses of soil between the rock layers). Preferential/macropore flow at the trench face depended on imposed boundary conditions and was independent of antecedent moisture content in the soil matrix. Pseudo-matrix flow response time decreased with high rainfall. During large rainfall events (>200 mm), water exchange was observed between the fractures and soil matrix. No apparent water exchange occurred between fractures and the soil matrix during small rainfall events. The dye studies indicated that fractures and juniper root channels are primary pathways for preferential/macropore flow occurring within the plot.