Through the experiment simulated sand columns, the biodegradation characteristics of diesel in sand layers (including fine sand, medium sand and coarse sand) with different depths and moisture contents were studied. The results show that the depth and moisture content of medium are important factors in affecting the efficiency of diesel degradation. In the same medium conditions, the higher moisture content of the medium, the higher biological activity, and biological degradation efficiency of diesel are observed. The nature of medium affects the efficiency of diesel degradation in vadose zone. The finer particles of the medium, the higher ability of diesel degradation is. It is expressed as: fine sand > medium sand > coarse sand. Volatilization and biodegradation are important factors in affecting natural attenuation of diesel in vadose zone.