Genetic engineering of Oryza sativa by particle bombardment

Genetic engineering of rice (Oryza sativa L. cv. Pusa basmati 1) using synthetic Cry1Ac gene has been achieved by “particle bombardment”. Scutellar tissues excised after 5 – 6 d from mature seeds cultured on induction medium were bombarded using gold particles coated with a mixture of Cry1Ac and marker genes on medium with osmoticum. Bombarded tissues were subjected to 30 mg dm⁻³ hygromycin selection for two cycles. The selected calli after GUS assay were transferred to shoot regeneration medium. Regenerated shoots were rooted and plantlets (T₀) were grown to full maturity. Polymerase chain reaction (PCR) analysis of T₀ plants using Cry1Ac specific primers revealed the presence of Cry1Ac gene in 65 % plants. Phenotypic assay, β-glucuronidase assay and PCR during T₁ generation revealed the inheritance of the Cry1Ac and marker genes along with the native plant genes.