Abstract
Plastids are surrounded by an envelope consisting of a double membrane. This barrier has to be penetrated or overcome by the DNA when transforming the plastome. Both the biolistic and polyethylene glycol-mediated transformation techniques accomplish this task, albeit by different mechanisms. We were the first laboratory to successfully use the polyethylene glycol (PEG)-method for plastid transformation, yet we use the particle gun when appropriate. In this report we compare the two methods and discuss their shortcomings and advantages. Plastid transformations with various constructs, mainly using theaadA gene as a selective marker, were performed. We point to potential problems likely to be encountered during the transformation and selection processes and offer possibilities for improvement. We give further examples of the successful application of plastome transformation and show its merits in addressing biological questions concerning the elucidation of plastid sequences of unknown function and the control of plastid gene expression.
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