Genetic Augmentation of Sugarcane Through Direct Gene Transformation with Osgly II Gene Construct

Owing to bottlenecks like complex genetic architecture, high ploidy level, specific climatic requirements for flowering coupled with longer life cycle in traditional sugarcane breeding, genetic augmentation of elite sugarcane cultivars for traits like abiotic and biotic tolerance are realistic goals for biotechnology endeavours. Glyoxalase I and glyoxalase II have been proved to confer improved salinity tolerance in tobacco and rice. Direct genetic transformation in sugarcane with salinity conferring Osgly gene construct in commercial sugarcane variety CoJ 83 has been reported. Embryogeneic calli lines were established on MS + 2,4-D (4.0 mgL −1 ) + Kinetin (0.5 mgL −1 ) + Sucrose (30 gL −1 ) medium. Selection of transformants was carried on callus stage on MS + 2,4-D (2.5 mgL −1 ) + Kinetin (0.5 mgL −1 ) + Hygromycin (35 ppm) medium for three cycles of 2 weeks each and regenerated on selection medium, i.e. MS + IAA (5 mgL −1 ) + Kinetin (0.5 mgL −1 ) + BAP (0.5 mgL −1 ) + Sucrose (30 gL −1 ) + Hygromycin (35 ppm). Putative transformants were pre-screened for expression of GUS reporter gene. PCR based molecular analysis of GUS positive transformants showed that 1.0 kb Osgly II DNA has been integrated into corresponding sugarcane variety and after further confirmation, this may lay the foundation for salinity tolerant transgenic breeding in sugarcane.