Tissue culture independent transformation for Corchorus olitorius

Abstract

In vitro regeneration is difficult for species of Corchorus and several transformation attempts based on tissue culture have failed. We describe a successful transformation protocol for C. olitorius using a technique independent of tissue culture. Primary growth of a plant is brought about from the activities of meristematic region and subsequent division and differentiation of the derivative cells into the tissues and organs of the plant. The principal behind the current study was that if some dividing cells in the meristematic region become transformed by Agrobacterium and the cells retain the capacity for cell division, then the transgene(s) will be transferred to progeny cells, and if some of these cells later differentiate to form floral buds, seeds generated from these buds will inherit the transgene(s) to next generation. In this study young jute plants were transformed at shoot apical meristematic region using Agrobacterium tumefaciens. Heritable transmission of the transgene to progeny from genetically modified plants was confirmed by gus gene expression by histochemical analysis, selection on kanamycin containing medium, RT-PCR, PCR amplification and Southern hybridization. Efficiency of transformation was determined by selection on medium containing kanamycin, and inheritance of transgene to T2 generation plants.