Gene manipulation of Siberian wildrye grass through transgenic technology can modify grass feature forfurther improvement in the forage production. In order to optimize transformation conditions of Siberianwildrye grass, the effects of transformation efficiency was investigated with GUS gene. Seed-derived calli wereinfected and co-cultured with Agrobacterium tumefaciens carrying the binary vector pCAMBIA3301 encodingthe GUS gene in the T-DNA region. The effects of several factors on transformation and the expression of theGUS gene were investigated. The highest transformation efficiency was obtained when embryogenic calli wereinoculated with Agrobacterium strain EHA101 in the presence of 200 μM acetosyringone and coculture for 5days. The present study was framed and materialized to standardize an efficient, high frequency andreproducible genetic transformation protocol for aromatic Siberian wildrye grass using Agrobacteriumtumefaciens.