Abstract: This study aims to explore the sequence characteristics of Tilia species through the analysis of the internal transcribed spacer(ITS) sequences of ribosomal DNA and apply this to the species identification of the "Bodhi" King tree in Tiantai Mountain. Samples of T. miqueliana were collected from Tiantai County,Taizhou City,Zhejiang Province,and total DNA was extracted using the CTAB method. The ITS sequences were amplified and sequenced via PCR. Sequence alignment and contig assembly were performed using DNAStar software,and a phylogenetic tree was constructed with MEGA-X software to analyze the relationships among T. miqueliana and its close relatives. The results indicated that the ITS sequences of the "Bodhi" King tree from Tiantai showed significant differences from other Tilia species,confirming its identity as T. miqueliana. Additionally,a BLAST comparison revealed sequence similarities ranging from 95.81% to 97.97% between T. miqueliana and other Tilia species,highlighting their phylogenetic relationships. This study not only provides new insights into Tilia taxonomy but also offers significant clues for understanding the adaptation and evolution of Buddhist culture in different geographical environments.