In the current research an efficient transformation system for sugarcane was established. Shoot tip of variety HSF-240, excised from a six months old field grown plants were used as explant. For transformation, Agrobacterium tumefaciens strain EHA101 with vector pIG121 Hm, harboring GUS, HPTII and NPTII genes were used. HPTII is a hygromycin resistant while NPTII is a kanamycin resistant gene. Effects of Acetosyringone, duration of co-cultivation and pre-selection, concentration of cefotaxime and hygromycin in medium on transformation efficiency were studied. High transformation efficiency and 60% GUS expression was observed when 50 μM acetosyringone was added in the co-cultivation medium. Among different durations of co-cultivation, 48 h produced high (40%) transient GUS positives with an absolute control of bacterial growth. For pre-selection, seven days gave a high transformation efficiency of 10%. Cefotaxime concentration of 1000 mg/L proved optimal for pre-selection of the explants with efficient control of bacterial growth. A high regeneration (31%; P < 0.01) of the transformants was observed at 50 mg/L hygromycin. Presence of GUS gene was confirmed by PCR analysis and only the transgenic plants contained the 430 bp fragment of GUS gene. The new protocol developed in this study could be used for the efficient transformation of sugarcane with desired gene to produce insect/pest resistant, drought tolerant and high yielding sugarcane varieties in future
