Author(s):

NN Tiwari, RK Singh and SP Singh

Abstract: Complete elimination of Agrobacterium from plant regeneration media is essential for the successful recovery of putative transgenic sugarcane plantlets. The present investigation determines the eco-friendly cefotaxime concentration for the inhibition of Agrobacterium during co cultivation with sugarcane leaf disc explants. The disk diffusion assay with eight different concentration of cefotaxime viz; 0 (Control), 50, 100, 150, 200, 250, 300 and 350 mg /l were used in Yeast Extract Manitol (YEM) medium. Agrobacterium tumefaciens LBA 4404 strain harboring plasmid pBINAR, carried the selectable marker gene (npt II) for kanamycin. Out of these concentrations, cefotaxime @ 150 mg/l was found eco-friendly dose to suppress the growth of Agrobacterium tumefaciens and produced a vigorous shoots. Plant allows non-phytotoxic regeneration and survival rate was also good on this concentration. Consequently, the efficient protocol established in this study could be applicable for suppressing Agrobacterium tumefactions in further genetic transformation works of sugarcane.

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