Rice black-streaked dwarf virus (RBSDV) disease has seriously affected rice production in China in recent years, giving rise to an imperative of breeding rice new varieties with high resistance to RBSDV. Rice resources highly resistant to RBSDV play key roles in breeding procedure. In this study, 2000 local varieties were evaluated on the resistance to RBSDV for two consecutive years. 38 varieties without disease symptoms were primarily screened, of which 6 japonica varieties with preferred agronomic traits were selected to perform artificial inoculation identification. One variety,12G1, highly resistance to RBSDV was obtained ultimately. The expression of RBSDV in 12G1 decreased 71.5 times compared to the sensitive control, indicating a notable inhibition of RBSDV replication in the disease-resistant variety. This study laid the material foundation for RBSDV resistance breeding procedure and cloning of resistance genes.