Abstract: In order to establish a system for molecular identification of tobacco virus diseases, primers for TMV, CMV and PVY detection were used in RT-PCR for TMV samples, and the genetic sequence of the amplified product was analyzed at nucleic acid molecular level. The results of RT-PCR indicated that the pathogen was TMV and the amplification product was 923 bp in length. NCBI BLAST showed that it shared 97% sequence identity with 3 China isolates. Sequence analysis indicated that the fragment located at the 3' terminal of TMV genome and included a part of movement protein(MP) gene, complete coat protein(CP) gene and 3' terminal' s non-coding region. The phylogenetic tree analysis of CP gene was conducted, and the sequence has been logged in GenBank.