Agrobacterium -Mediated Genetic Transformation in Soybean (Glycine max (L.) Merr.)
B.D. Ranjitha Kumari *
Department of Plant Science, Bharathidasan University, Tiruchirappalli-620 024, TN, India
A. Settu
Department of Plant Science, Bharathidasan University, Tiruchirappalli-620 024, TN, India
G. Sujatha
Department of Plant Science, Bharathidasan University, Tiruchirappalli-620 024, TN, India
*Author to whom correspondence should be addressed.
Abstract
An efficient and reproducible protocol for Agrobacterium-mediated transformation has been standardization in soybean (Glycine max (L.) Merr.) with a binary vector pBI 121. Putatively transgenic shoots were selected on kanamycin medium. GUS assay was performed to confirm the presence of GUS gene from putatively transgenic shoots. These three confirmed transgenic were multiplied followed by successful rooting in the media containing 1.5 mg/l IBA. Plants with well-developed roots were successfully hardened in plastic cups containing sand, soil and sawdust (1:1:1). Stable integration of the transgene expression (uid A and npt II) has been confirmed by PCR analysis. Additional molecular analysis by Southern blot hybridization confirmed the presence of npt II gene in putatively transformed plants.
Keywords: Agrobacterium tumefaciens, Glycine max, GUS gene, Soybean, Transgenic shoots