Optimization of Agrobacterium Mediated Genetic Transformation in Paspalum scrobiculatum L. (Kodo Millet)

An efficient and reproducible protocol for Agrobacterium tumefaciens mediated genetic transformation was developed kodo millet (Paspalum scrobiculatum L.) by optimizing various parameters. strains EHA 105 LBA 4404 harboring plasmids pCNL 56 pCAMBIA 2300, respectively, provided the highest efficiency. Addition of acetosyringone (AS) in infection medium (200 µM-EHA 105, 250 µM-LBA 4404) co-cultivation (50 µM) increased Transient stable expression gus gene confirmed with histochemical assay infected embryos leaves transformed plants, respectively. The best GUS response obtained pretreatment callus an antinecrotic mixture (10 mg/L Cys + 5 Ag 2.5 As) at time 20 min followed 3 days (EHA 105) (LBA dark. Regenerated transgenic plants were after 8 to 10 weeks selection on induction (NAA 0.5 mg/L, BAP 1 mg/L) containing 50 Kan Cef rooted 2 MS PAA (1 phytagel. plantlets established greenhouse showed normal growth. Therefore, present study can be used development improved varieties millet.