ISHS Acta Horticulturae 725: V International Symposium on In Vitro Culture and Horticultural Breeding
PROLIFERATION AND COLD HARDENING OF IN VITRO GROWN APPLE SHOOT TIPS |
| Authors: | J. Sedlak, F. Paprstein, A. Bilavcik, J. Zamecnik |
| Keywords: | micropropagation, germplasm, encapsulation, cryopreservation, freezing |
| Abstract:
Shoot tips of apple cultivar ‘Rubin’ were successfully established in vitro. Five proliferation MS (Murashige and Skoog) media containing different concentrations of BAP (0.25, 0.5, 1, 2 mg L-1) or a combination of 1 mg L-1 BAP and 0.1 mg L-1 IBA were tested. The highest proliferation rate was obtained on MS medium with 1 mg L-1 BAP, which produced 4.2 0.3 shoots/explant after one month of cultivation. In vitro plants of cv. ‘Rubin’ were cultivated for 4 weeks at low temperature (4°C) and short photoperiod (8/16-h – light/dark). The assessment of frost resistance of cold hardened and non-hardened in vitro apple plants was done by freezing tests. In vitro plants were exposed to the controlled cooling rate 4°C/h. The lethal temperature (LT) was -13.0°C for cold hardened and 6.5°C for non-hardened in vitro plants. Cold hardened in vitro cultures were used for subsequent cryopreservation. Cryopreservation was made by encapsulation-dehydration vitrification method. The survival and regeneration of cv. ‘Rubin’ was 78% after thawing in 40°C water. | |
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