Abstract
Suspension cell protoplasts of albino Petunia hybrida have been transformed by isolated Agrobacterium tumefaciens Ti plasmid. Uptake of octopine Ti plasmid (pTiACH5) into protoplasts was stimulated by poly-L-ornithine and polyethylene glycol (PEG). The frequency and efficiency of transformation of protoplasts to phytohormone autotrophy was compared using the two uptake agents with various concentrations of plasmid. Transformation was most efficient with PEG-mediated uptake, 5 μg of Ti plasmid per 106 protoplasts giving a frequency of 6×10−5. Octopine was not synthesised in any of the transformants after the second subculture on hormone-free medium. DNA-DNA hybridisation showed the presence of DNA homologous to the T-DNA region of pTiACH5 in all clones analysed.
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