Novel plant transformation system by gene-coated gold particle introduction into specific cell using ArF excimer laser

Abstract

Novel methods for foreign-gene introduction into specific cells are required for the functional analysis of specific cells. We report a new transfection method for intact plant cells that uses ArF excimer laser-induced shock waves to introduce foreign gene-coated gold particles into plant cells through the cell wall. Foreign gene (sGFP)-coated gold particles were randomly scattered on the surface of Torenia tissue samples; the laser was irradiated toward the gold particles using a hollow optical fiber and a quartz light-condensing tip to introduce the particles into the target cells. Treated plant tissue samples were then cultured for several weeks on the selection medium, and transformation was assessed by fluorescence microscopy and PCR. Irradiation-induced damage to the cells and coated DNA was insignificant, and the transformation efficiency was similar to that of biolistic transformation based on the particle number. Although this method requires a suitable laser setup and is time-consuming, it could be applied in transformation experiments on specific target cells. Moreover, this method can possibly be used to introduce not only DNA but also biologically active substances such as protein regulators or inhibitors, whose introduction into cells is difficult through the cell wall.