Abstract: The identification of excellent resistance germplasm resources to bacterial wilt, broadening the genetic background and improving the disease resistance level of tobacco varieties are the research focuses of current tobacco resistance breeding. In this study, Dixie Bright 101, K326, and Honghuadajinyuan were used as control varieties, and three Ralstonia solanacearum strains FJ-SMNT, FQY-4, and Y45 were used to inoculate tobacco plants. We then recorded the disease index and evaluated the bacteria wilt resistance of Qiongzhongwuzhishan-1, a local sun-cured tobacco germplasm resource. And based on the resequencing data of germplasm resources, the characteristic SNP fingerprint of Qiongzhongwuzhishan-1 were developed. The results showed that Qiongzhongwuzhishan-1 had a slow onset, mild disease occurence, and a mild increase in the disease index, with good resistance to all three strains. The level of resistance was significantly higher than that of the resistant control. The specific fingerprint consisted of 339 SNP loci, and a detection program for this fingerprint was developed based on the cloud computing platform. The reliability of this fingerprint was verified using sequencing technologies, and it was demonstrated that this fingerprint could accurately identify Qiongzhongwuzhishan-1 from randomly sampled specimens. The results from this study indicated that Qiongzhongwuzhishan-1 is one of the germplasm resources with the best resistance to tobacco bacteria wilt found in China.