Food And Pharma International

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Abstract: Agrobacterium-mediated genetic transformation was done in BRRI dhan-29 using
Agrobacterium strain LBA4404 harboring the plasmid pBI121. The plasmid carries CaMV35S promoter driving
the expression of GUS and Kanamycin resistant genes. The β-glucuronidase (GUS) gene served as a reporter
gene in the histochemical assay and the neomycin phosphotransferase II (nptII) gene used for the identification
of resistance to kanamycin. The incubation period of 30 minutes was found to be the most effective for infection
and integration of foreign genes into callus. Transformed calli were selected along with a control in the
regeneration medium containing 100 mg/l of kanamycin. Embryogenic calli were found to show the best
transformation ability producing 30 % GUS positive result in histochemical assay after 3 days of co-cultivation.
GUS expression was also observed in the leaf and root of putative plants prior to final confirmation of
transformation through PCR amplification.

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