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Plant and Cell Physiology, 1982, Vol. 23, No. 3 451-458
© 1982

Article

Ti Plasmid Homologous Sequences Present in Tissues from Agrobacterium Plasmid-transformed Petunia Protoplasts

J. Draper1, M. R. Davey1, J. P. Freeman1, E. C. Cocking1 and B. J. Cox2

1Plant Genetic Manipulation Group, Department of Botany, University of Nottingham, University Park Nottingham NG7 2RD, U.K.
2Bio Sciences Group, ICI Corporate Laboratory Runcorn, Cheshire WA7 4QE, U.K.

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.

(Received November 9, 1981; Accepted January 29, 1982)
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