A simple and environment-friendly procedure for isolating high-quality plant DNA was presented in this paper. The extraction buffer contained 2%CTAB, 1% SDS, 2%PVP, 100 mmol/L Tris-Cl (pH8.0), 20 mmol/L EDTA (pH8.0) and 1.4 mol/L NaCl. In this way, DNA of 12 various plants was extracted and successfully used for PCR amplification by ITS4/ITS5 eukaryotic universal primers. This DNA extract method left out phenol-chloroform extraction, which was safe and time-effective and made it attractive in large-scale DNA extraction．