Abstract: In order to obtain the efficient expression vectors for new lily antiviral germplasm, CMV 2b gene 271 bp and LMoV cp gene 428 bp fragments were chosen as the target sequence to construct the vectors by homologous sequence. The required virus was confirmed by RT-PCR method from infected leaves. And then PCR method was used to obtain the cDNA from the total RNA. Using the cDNA as template, the target fragments were obtained under specific primers with different restriction enzyme cutting sites. The two fragments were inserted into both sides of intron of pFGC5941 in forwards and reverse after recombinant clone. The RNAi vector was introduced into Agrobacterium tumefaciens EHA105. Restriction enzyme digestion showed that the RNAi vectors were constructed successfully. The results of PCR confirmed that both of the two vectors pFGC5941-C2 and pFGC5941-L2 were introduced into Agrobacterium tumefaciens EHA105. The results of PCR and sequencing showed that the constructed vectors and the engineering bacteria were completely consistent with the expected, the RNAi vectors were constructed successfully, which could be applied in transgenic breeding engineering for lily viral resistance.