Improvement of bioactive bead-mediated transformation by concomitant application of electroporation
Techniques of exogenous gene introduction in plant cells have been actively applied for producing transgenic cultured cells that produce high levels of useful biological substances and are used for the molecular breeding of plants. A novel gene transfer technique has been developed using calcium alginate micro-beads (bioactive beads) to entrap genetic material. Although this technique has several advantages such as high transformation efficiency and the ability to introduce large DNA molecules, the precise optimal application conditions remain to be determined. Here, the optimal conditions for the concomitant use of bioactive beads and electroporation, such as polyethylene glycol (PEG) concentration, alginate concentration, electric field intensity, pulse duration, and pulse numbers, were determined for improved gene transfer efficiency of bioactive bead-mediated plant transformation. Consequently, highly efficient transient transgene expression (up to 4.7%) was achieved by applying electrical pulses (0.75 kV CM<sup>−1</sup>, 30 μs, 3×) to a protoplast suspension in 12% polyethylene glycol (PEG).
- Plant biotechnology
Plant biotechnology 25(4), 387-390, 2008-09-01
Japanese Society for Plant Cell and Molecular Biology