Written Paper

Isolation and culture of mesophyll protoplasts in European plum-trees (Prunus domestica L.) cv. greengage  [2001]

Ferreira, H. (Universidade de Evora (Portugal)) Druart, P. (Station des Cultures Fruitieres e Maraicheres, Gembloux (Belgium)) Kondakova, V. (Institute of Genetic Engineering, Kostinbrod (Bulgaria)) Estacao Nacional de Melhoramento de Plantas, Elvas (Portugal) [Corporate Author]

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Protoplast culture, fusion and somatic hybrids production, in european plum (Prunus domestica L.) cv. 'Rainha Claudia Verde', will provide the variability increase in this specie bypassing self-incompatibility barriers characteristic in this specie. Therefore agronomical and economical cultivars will be created due the utilisation of these breeding technologies. This work developed an efficient protocol to mesophyll protoplast isolation and culture to this particular cultivar of european plum. We obtained 95% viability isolating mesophyll protoplasts, from plants cultivated in Murashige and Skoog medium, (1962) (MS) (0.05 mg/l NAA, 0.75 mg/l BAP, 0.05 mg/l GA3, 0.5% Past-Agar, 2% Sucrose, pH 5.6), using the enzymatic solution, Cellulase Onozuca RS-1%, Hemicellulase-1%, Pectolyase Y 23-0.1% and using one purification method that recovers protoplasts from ML 0.5 solution, layered on the top of the suspension (protoplasts and enzymatic solution) after the first centrifugation at 120 g. The Kao e Michayluk medium, (1975) (KM8p) supplemented with 0.01 mg/l de NAA, 1 mg/l de BAP e 0.5 mg/l de zeatin and the vitamin complex, 0.4 mg/l Thiamine HCl, 2 mg/l Pyridoxine, 2 mg/l Nicotinic Acid, promoted the cell wall regeneration after the 3rd day and the first mitotic division after the 5th day in culture

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Melhoramento (Portugal)

ISSN : 0368-9433