Agrobacterium mediated rice transformation

Immature embryo (12-15 days after pollination ) —->

Precultured on ND2 medium for 4 days at 26C in darkness —->

Immersed in the bacterial suspension for 20 minutes —->

Incubated on ND2-As medium for 3 days at 26C in darkness —->

Cultured on ND2CCH25 medium for 2 weeks for selection —->

Subcultured on ND2CCH50 medium for 4 weeks ( 2 generations ) for
selection —->

Resistant calli were put on 3M filter paper overnight for osmotic
treatment —->

Resistant calli were transferred to NN.1B2H medium for regeneration

Regenerated seedlings were transferred to MS0 medium for further growth

Plantlets about 10cm high were grown to maturity in greenhouse in soil