A simple,fast and efficient method of constructing dual-target CRISPR/Cas9 vector was established based on the Golden Gate ligation. The two designed small target DNA fragments and one exogenous fragment were constructed together into the CRISPR/Cas9 vector with T4 DNA ligase. 20 dual-target CRISPR/Cas9 vectors against 10 reported rice genes were constructed with this method,and the positive rate was 100% with no mutation in sequences. This vector system provides an easy,rapid and efficient method for constructing dual-target CRISPR/Cas9 vector for rice. It also provides a reference for the construction of dual-target CRISPR/Cas9 vectors in other plants.