Biolistic transformation of sugarcane (Saccharum officinarum L.) with the Oryza sativa L. H+-PPase gene for improved salt stress tolerance

Several biotic and abiotic stresses adversely affect sugarcane crop. Amongst the stresses, salinity causes substantial losses to yield. Lack of conducive environment for flowering unstable genome are main constraints in crop improvement via conventional breeding. Therefore, genetic engineering using Oryza sativa L. H+-PPase gene (OVP1) confer salt tolerance is preferred alternative strategy. In present research, tissue culture biolistic transformation protocols were optimized. For callus induction, Dichlorophenoxyacetic acid (2,4-D) was most effective. Maximum induction (50.22%) achieved on CM-3 augmented with 2.5 mgL-1 2,4-D. The induced calli subjected transformation. transformation, gold particles (0.6 1.0 µm) plasmid DNA (100 50 ng) used 1X 2X bombardment. Significant differences (P≤0.05) observed amongst concentrations. frequencies when bombarded 0.6 µm coated 100 ng (23.60%). Selection regeneration transgenic performed MS media BAP (2.5 mgL-), NAA (1 mgL-1) hygromycin (25 2% coconut water. Regenerated plants after root containing (2 water transferred loam clay (1:1) acclimatized greenhouse condition. confirmed through PCR RT-PCR.