Transformation of callus sugarcane (Saccharum officinarum L.) using Agrobacterium tumefaciens – Mediated method and regeneration of transgenic shoot

Abstract

Sugarcane (Saccharum officinarum L.) is a major industrial crop in the world. The creation of transgenic sugacane varieties contributed to overcome the disadvantages of  traditional farming methods. Immature cutting cane stems after 10 days of putting on MS medium (Murashige and Skoog) supplemented 2,4-dichlorophenoxyacetic acid (2,4-D) 3  mg/L triggered the highest rate of explants inducing callus (91.11%). Calli were cultured on MS medium supplemented 6- Benzyladenin (BA) with concentrations of 0, 0.5, 1.0, 1.5, 2.0, 2.5 mg/L for inducing shoot. All six concentrations of BA could induce shoot, but the concentration of BA 1 mg/L was the most suitable. The 4-to-5-week old calli were introduced desired gene by using Agrobacterium tumefaciens, which was affected by acetosyringone concentration, inoculation and co-culture time. Consequently, transformation with acetosyringone 100 μM or 150 μM could be successful and regenerate transgenic shoots with assumed rate 25%.