Ti plasmid homologous sequences present in tissues from Agrobacterium plasmid-transformed Petunia protoplasts

• Published in: Plant and cell physiology 1982-04, Vol.23 (3), p.451-458
• Main Authors: Draper, J, Davey, M.R, Freeman, J.P, Cocking, E.C. (Nottingham Univ. (UK)), Cox, B.J
• Format: Article
• Language: English
• Subjects: ACIDE; ACIDE AMINE; ACIDE NUCLEIQUE; ACIDE ORGANIQUE; ACIDOS; ACIDOS NUCLEICOS; ACIDOS ORGANICOS; ACIDS; ADN; Agrobacterium tumefaciens; AMELIORATION GENETIQUE; AMINO ACIDS; AMINOACIDOS; BACTERIA; BACTERIA GRAM NEGATIVA; BACTERIE; BACTERIE GRAM NEGATIF; BREEDING; BREEDING METHODS; COMPOSE NUCLEIQUE; COMPOSE ORGANIQUE; COMPOSE ORGANOAZOTE; COMPUESTO ORGANICO DEL NITROGENO; COMPUESTOS NUCLEICOS; COMPUESTOS ORGANICOS; CYTOPLASMIC INHERITANCE; DICOTILEDONEAS; DICOTYLEDONE; DICOTYLEDONS; DNA; DNA-DNA hybridisation; ECONOMIC PLANTS; GENETICA; GENETICS; GENETIQUE; GRAM-NEGATIVE BACTERIA; HERBACEOUS ORNAMENTALS; HEREDITE; HEREDITE CYTOPLASMIQUE; HERENCIA; HERENCIA CITOPLASMATICA; HIBRIDACION; HORMONAS; HORMONE; Hormone autotrophy; HORMONES; HYBRIDATION; HYBRIDIZING; INHERITANCE; MEJORAMIENTO; METHODE; METHODE D'AMELIORATION; METHODS; METODOS; METODOS DE MEJORAMIENTO; MONOCARPOS ORNAMENTALES; NUCLEIC ACIDS; NUCLEIC COMPOUNDS; Octopine; ORGANIC ACIDS; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; ORNAMENTAL MONOCARPS; ORNAMENTAL PLANTS; ORNITHINE; ORNITINA; PETUNIA; Petunia hybrida; Petunia protoplasts; PLANTAS; PLANTAS DE VALOR ECONOMICO; PLANTAS HERBACEAS ORNAMENTALES; PLANTAS ORNAMENTALES; PLANTE; PLANTE D'INTERET ECONOMIQUE; PLANTE ORNEMENTALE; PLANTE ORNEMENTALE HERBACEE; PLANTE ORNEMENTALE MONOCARPIQUE; PLANTS; Poly-L-ornithine; Polyethylene glycol; PROTOPLASM; PROTOPLASMA; PROTOPLASME; SOLANACEAE; Ti plasmid; Transformation; TUBIFLORAE
• ISSN: 0032-0781; 1471-9053
• DOI: 10.1093/oxfordjournals.pcp.a076368

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.