ISHS Acta Horticulturae 356: II International Symposium on Olive Growing
ISOLATION AND CULTURE OF OLIVE (OLEA EUROPAEA L.) CULTIVAR PROTOPLASTS. |
| Authors: | E. Perri, M.V. Parlati, E. Rugini |
| Keywords: | In vitro culture, petiole, leaf, shoot, 'Dolce Agogia', 'Canino', Thidiazuron |
| Abstract:
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. | |
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