This study aimed to investigate the effects of wind turbulence on CO2 transport within a medium and the extent of measurement errors in a closed chamber. Therefore, in a laboratory with controllable environmental conditions, the measurement performance of the closed chamber at various wind speeds was assessed using a soil respiration calibration apparatus and four types of porous media. The experimental results indicated that the closed chamber under the influence of wind turbulence exhibited varying degrees of underestimation, ranging from −51 to −6%. The effects of wind turbulence were more pronounced in sandy soils. As wind turbulence enhanced gas transport within the medium, the flux measurements of the closed chamber were biased, and this phenomenon was closely related to the medium’s particle size and surface wind speed. To address this issue, it is recommended to conduct long-term monitoring and eliminate errors by averaging repeated measurements, which will improve the accuracy of the ecosystem carbon budget.