The present invention discloses a method for improving the ability to resist against an intrusive DNA virus of a plant. The present invention provides a method for making a plant with improved ability to resist against a DNA virus, specifically can comprising the following steps: 1) selecting a sequence comply with 5-NX-NGG-3 or 5-CCN-NX-3 from the genomic sequence of the DNA virus as the target sequence designing a DNA sequence reverse-complementary to said target sequence 2) constructing said DNA sequence to a vector for expressing CRISPR/Cas9 nuclease so as to obtain a recombinant vector capable of transcribing a guide RNA and expressing a Cas9 protein the crRNA in said guide RNA contains a RNA fragment transcribed from said DNA fragment 3) introducing said recombinant vector into a recipient plant so as to obtain a plant with improved ability to resist against said DNA virus. The method of the invention merely depends on the genomic sequence of the virus, without the need of knowing specific functions of the viral genes. Therefore, the method can be widely used for resisting against various known double-stranded DNA virus or single-stranded virus with double-stranded DNA as an intermediate.
