Abstract: In this study, the sequences of 200 bp of tomato spotted wilt orthotospovirus(TSWV) NSm, NSs genes were constructed to pTRV−PTV00 silencing vector and infiltrated agrobacterium GV3101 in Nicotiana benthamiana. About 10 days after injection, the plants with injecting PDS gene became white, the plants with injecting the vectors of pTRV−PTV00−NSm, pTRV−PTV00−NSs and pTRV−PTV00 were inoculated with TSWV. The expression of NSm and NSs genes was detected by qRT−PCR in the systematic leaves(one leaf above the inoculated leaf) after manifestation, and the gene silencing efficiency was obtained. The silencing efficiency of NSm and NSs gene was 86.7% and 92.00%, respectively. The results showed that silencing efficiency of NSm and NSs gene silencing was close to 90.00% constructed by pTRV−PTV00 vector, which was suitable for the construction of viral gene silencing vectors, and NSm and NSs gene silencing vectors could be used for subsequent gene function studies.