Enhanced Transformation Efficiency of Saccharum officinarum by Vacuum Infiltration Assisted Agrobacterium-mediated Transformation

Transgenic plants have been extensively produced for years with improved agronomic characteristics, enhanced disease and insect pest resistance. However, improved and reproducible system of gene transformation remains the main focal point in plant genetic engineering. In the present study, an efficient Agrobacterium-mediated stable transformation protocol for sugarcane (Saccharum officinarum L.) was optimized using embryogenic calli and green fluorescent protein (gfp) as explants and reporter gene, respectively. Parameters studied were optical density of bacterial culture and agro-infiltration under different pressure regimes. Results indicated that out of four different treatments, inoculation of calli with bacterial culture of OD(600)0.4 through vacuum infiltration at -50 kPa produced maximum number of transgenic plants. This newly developed protocol of Agrobacterium-mediated transformation of sugarcane has shown significant improvement over conventional procedure in terms of its transformation efficiency. This protocol can be employed to develop transgenic sugarcane plants, having tolerance/resistance against various biotic and abiotic stresses. (C) 2014 Friends Science Publishers