ISOLATION AND CULTURE OF OLIVE (OLEA EUROPAEA L.) CULTIVAR PROTOPLASTS.
Viable olive (Olea europaea L.) protoplasts were isolated from in vitro cultured leaf mesophyll tissue of cv Dolce Agogia and petiole-derived callus of cv Dolce Agogia and Canino by an overnight incubation in an enzyme solution containing 1.5 % (w/v) Driselase, 500 mg/l of KCl, 500 mg/l of CaCl2.H2O, 0.6 M Mannitol, 5mM 2-N-morpholinoethane sulfonic acid (MES), 100 mg/l reduced glutathione (GSH) and 100 mg/l ascorbic acid (AA) at pH 5.6. Protoplasts were cultured in MS-basal medium supplemented with 5 mg/l thidiazuron (TDZ), 0.01 mg/l 1-naphtaleneacetic acid (NAA), 0.75 % w/v sucrose and 9 %, w/v mannitol as the osmoticum at pH 5.8. Petiole-derived callus from cv Dolce Agogia and Canino gave high yields of viable protoplasts whereas leaf mesophyll tissue from cv Dolce Agogia gave a poor yield of protoplasts of low viability. Only the cells derived from protoplasts of petiole callus from cv Dolce Agogia and Canino were able to divide themselves.
Perri, E., Parlati, M.V. and Rugini, E. (1994). ISOLATION AND CULTURE OF OLIVE (OLEA EUROPAEA L.) CULTIVAR PROTOPLASTS.. Acta Hortic. 356, 51-53
In vitro culture, petiole, leaf, shoot, 'Dolce Agogia', 'Canino', Thidiazuron