Regeneration and Agrobacterium-mediated transformation of a popular indica rice variety, ADT39

Abstract

A simple and efficient protocol for the regeneration and Agrobacterium-mediated transformation of an agronomically useful indica rice variety, ADT39 has been standardized. Initiation of callusing and its subculture were best achieved in MS medium supplemented with 300 mg/l casein enzymatic hydrolysate. The best callusing and regeneration responses were observed at concentrations of 2 mg/l 2,4-D and 1.5 mg/l BAP. Optimal transformation efficiency of 22.2% was obtained using high concentrations of augmentin as a bacteriostatic agent for a short period to inhibit the growth/ persistence of Agrobacterium, without compromising the regeneration potential of the tissue. Using a viral promoter-driven GUS reporter gene, morphologically normal fertile plants were obtained. Molecular analysis of the above transgenic plants indicated several independent, single-copy transgene insertion events. Expression of the reporter gene was detected in the above plants. Mendelian inheritance of the transgene in the progeny was also observed.