Tick-borne encephalitis virus (TBEV) has been classified into three subtypes, namely the European (Eu-TBEV), Far Eastern (FE-TBEV), and Siberian (Sib-TBEV). In this study, we discovered a new subtype of TBEV in wild rodent Marmota himalayana in Qinghai-Tibet Plateau in China, proposed as subtype Himalayan (Him-TBEV). Two complete genomes of TBEV were obtained from respiratory samples of 200 marmots. The phylogenetic analysis using the E protein and polyprotein demonstrated that the two strains of Him-TBEV formed an independent branch, separated from Eu-TBEV, Sib-TBEV, and FE-TBEV. The nomenclature of Him-TBEV as a new subtype was also supported by comparative analysis using nucleotide and amino acid sequences of E protein and polyprotein. For E protein, The Him-TBEV showed 82.6-84.6% nucleotide identities and 92.7-95.0% amino acid identities with other three subtypes. For polyprotein, the Him-TBEV showed 83.5-85.2% nucleotide identities and 92.6-94.2% amino acids identities with other three subtypes. Furthermore, of 69 amino acid substitutions profiles detected in complete polyprotein of 112 strains of TBEV, Him-TBEV subtype displayed unique amino acids in the 36 positions. Notably, for the subtype-specific amino acid position 206 of E protein, Him-TBEV shared the Val with Eu-TBEV, but differed from FE-TBEV and Sib-TBEV. The evolutionary analysis with BEAST suggested that Him-TBEV diverged from other subtypes of eastern TBEV group about 2469 years ago. It should be mentioned that Qinghai-Tibet Plateau in China is the plague endemic region where Marmota himalayana is the primary host. The public health significance of discovery of Him-TBEV in Marmota himalayana must be carefully evaluated.