An Efficient Agrobacterium-Mediated Genetic Transformation Using Embryonic Axis in Cotton (Gossypium hirsutum L.)

摘要: Agrobacterium-mediated genetic transformation approach allows for introducing novel genes in cotton (Gossypium hirsutum L.). Development of efficient regeneration and protocol is very important recalcitrant plants like cotton. In the present study, five-day-old germinated mature embryo parts especially embryonic axis, hypocotyl plumule ‘Lashata’ were wounded inoculated with A. tumefaciens strain GV2260 harbouring plasmid p35S-GUS-INT. The binary p35S-GUS-INT contains neomycin phosphotransferase II (NPTII) gene driven by nopaline synthase (NOS) promoter β-glucuronidase (GUS) controlled cauliflower mosaic virus (CaMV) 35S promoter. After inoculation, explants co-cultivated liquid agar-solidified MS medium 48 h dark condition. Agar-solidified co-cultivation increased frequency as compared to medium. putative primary transformants confirmed histochemical GUS assay, PCR RT-PCR. However, comparing three culture explant, axis had significant difference terms number percentage (%) GUS, RT-PCR positive plant. total efficiency was recorded 3% varying expression levels.