PEG-Mediated Protoplast Transformation with Naked DNA

Direct introduction of DNA into plant protoplasts facilitates a rapid analysis of transient gene expression, as well as the generation of stably transformed transgenic plants. Transient gene expression assays performed after DNA transformation permit a comparative analysis of cis–actmg regulatory sequences and then function m transcriptional control of plant genes by signaling pathways mediating cellular responses to different environmental and hormonal stimuli (1 ). There are a number of methods for introducing DNA into plant protoplasts, but the most commonly used technique 1s the polyethylene glycol (PEG)–mediated DNA uptake. The PEG–mediated transformation is simple and efficient, allowing a simultaneous processing of many samples, and yields a transformed cell population with high survival and division rates (2 ). The method utihzes inexpensive supplies and equipments, and helps to overcome a hurdle of host range limitations of Agrobactenum–mediated transformation The PEG–mediated DNA transfer can be readily adapted to a wide range of plant species and tissue sources.