Abstract:The plant expression vector with Imtl (inositol O-methyltransferase) gene, pDH5, was constructed. The Imtl gene was introduced to tobacco (Nicotiana tabacum cv. SR1) via Agrobacterium-mediated transformation. Thirty kanamycin-resistant shoots in MSS (MSO + 1 mg/L 6-BA + 30 g/L sucrose) selective medium was transferred to MSr (MSO + 3 g/L sucrose + 7 g/L glucose) medium supplemented with 1.2%~1.5% NaC1 for rooting. Eight plantlets rooted among the thirty shoots. PCR and Southern blot assay indicated that the Irntl gene was integrated into tobacco genome in six plantlets. HPLC analysis showed that the product of Imtl gene, D-ononitol accumulated in tobacco fresh leaf cells in a scope of 100 - 654 nmol/g FW. Western blot demonstrated the expression of Imtl gene in tobacco cells.