Abstract: BIOLOG-ECO methed was used for the investigation of the effect of 480 g·L^-1 chlorpyrifos emulsifiable concentrate (EC) on soil microbial functional diversity. The results demonstrated that (1) the average well color development (AWCD) of the lower chloripyrifos concentration treatments (≤12.5 mg·kg^-1) were greater than that of the control soil, and the irritation disappeared after used 20 d. On the contrary, the ability of microbial utilizing carbon source was inhibited in the highest chloripyrifos concentration treatment at 125 mg·kg^-1. (2) Between the chloripyrifos treatments and control, the carbon source utilization characterization of soil microbial exhibited significant differences, as utilization of ester compounds was improved, while that of sugar compounds was not improved significantly. (3) The results of principal component analysis (PCA) show that the carbon source utilization of soil microbial at treatment had significant difference with below 12.5 mg·kg^-1 treatments and the control. (4) The diversity index results showed that the highest concentration of chloripyrifos (at 125 mg·kg^-1) in soil could significantly inhibit soil microbial community richness, while the lower level of chloripyrifos (≤12.5 mg·kg^-1) promoted microbial community richness.