Abstract
The development of dormancy, frost resistance and cryotolerance of in vitro apple plants (Malus domestica Borkh.), cv. Greensleeves during their exposure to cold hardening was studied. In vitro cultures were cold hardened at 4°C under a short photoperiod up to 25 weeks. The dormancy status, non-structural saccharides, proline, water content and frost resistance were evaluated for optimization of cryopreservation. According to regrowth tests, in vitro cultures exhibited endogenous dormancy after the maximal frost resistance was reached. The highest regeneration ability of shoot tips after cryopreservation by encapsulation–dehydration method coincided with the period of the plant’s dormant state and maximum of frost resistance. All studied saccharides and proline exhibited the maximal values at the beginning of cold hardening and/or the dormancy phase. Contrary to the accumulation of saccharides and proline, water content showed the inverse time behaviour. According to these results, the cold hardening-induced endodormancy, high frost resistance and accumulation of saccharides and proline are the important prerequisites for the successful cryopreservation of shoot tips of in vitro grown apple plants.
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Acknowledgments
This work was supported by 0002700604 project of the Ministry of Agriculture of the Czech Republic and the OC08062 project financed by the Ministry of Education, Youth and Sport of the Czech Republic. Authors are thankful to Naďa Kantnerová, student of Czech Agricultural University, Prague who assisted in experimental part of in vitro cultures, to Dr. Helena Lipavská and Dr. Hana Konrádová, Charles University, Prague for estimations of saccharides content in plantlets and to Prof. Paul T. Lynch, University Derby, UK providing us by Greensleeves apple cultivar.
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Bilavčík, A., Zámečník, J., Grospietsch, M. et al. Dormancy development during cold hardening of in vitro cultured Malus domestica Borkh. plants in relation to their frost resistance and cryotolerance. Trees 26, 1181–1192 (2012). https://doi.org/10.1007/s00468-012-0694-7
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DOI: https://doi.org/10.1007/s00468-012-0694-7