Biobarriers, as permeable reactive barriers (PRBs), are a common technology that mainly uses bacteria to remediate groundwater in polluted aquifers. In this study, we propose to use Trametes versicolor, a white-rot fungus, as the reactive element because of its capacity to degrade a wide variety of highly recalcitrantand xenobiotic compounds. A laboratory-scale artificial aquifer was constructed to simulate groundwater flow under real conditions in shallow aquifers. Orange G dye was chosen as a contaminant to visually monitor the hydrodynamic behaviour of the system and any degradation of the dye by the fungus. Batch experiments at different pH values (6 and 7) and several temperatures (15°C, 18°C, 20°C and 25°C) were performed to select the appropriate residence time and glucose consumption rate required for continuous treatment. The maximum Orange G degradation was 97%. Continuous degradation over 85% was achieved for more than 8 days. Experimental results indicate for the first time that this fungus can potentially be used as a permeable reactive barrier in real aquifers.