Efficient Regeneration and Genetic Transformation of Sugarcane with AVP1 Gene

Agrobacterium mediated genetic transformation is considered to be more efficient and reliable method than direct gene bombardment in sugarcane. In the present study an efficient regeneration from callus and genetic transformation was established by using Arabidopsis Vacuolar Pump (AVP1) gene conferring tolerance against drought and salinity in local cultivar of sugarcane CP77-400. Agrobacterium strain EHA105 harboring pGreen0029 vector containing AVP1 and kanamycin resistant gene Npt II driven by 35S CaMV promoter was employed. Three weeks older calli of CP77-400 were infected with 0.2 optical density of Agrobacterium, 100 uM acetosyringone and co-cultivated for 4 days. The transformed plantlets were selected on 150 mg L of kanamycin and it was found that cefotaxime concentration 250 mg L was effective in 11 controlling Agrobacterium overgrowth as well as to enhances regeneration and transformation efficiency in sugarcane. The transgenic plants were acclimatized in glass house and both AVP1 and Npt II genes in transgenic plants of sugarcane were confirmed at genomic level through DNA amplification using Polymerase Chain Reaction (PCR). The evaluation of transgenic and non transgenic plants showed that transgenic plants more resistance to high salt concentration while the non transgenic plants were died at high salt stress.