preliminary results of cryopreservation experiments aimed to long … · 2016-04-21 · national...
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National Agricultural Research and Innovation Centre
Fruitculture Research Institute, Research Station at Újfehértó Vadastag 2., H-4244 Újfehértó, Hungary, [email protected]
Preliminary results of cryopreservation experiments aimed to long-term maintenance of Hungarian sour cherry germplasm
Tamás Lakatos – Tímea Tóth
Introduction: The total area of Hungarian fruit orchards is about 56 000 ha, the most important species is apple with 26 000 ha. In the second place is sour cherry with 14 800 ha. In Hungary, only Hungarian sour cherry varieties are in the production, and more than half of the production give varieties from breeding program of the Research Station at Újfehértó.
The horticultural germplasm collection of the Research Station at Újfehértó contains more than 2 000 accessions, mainly apple, pear and sour cherry varieties. The series of local sour cherry clones is the most valuable part of the collection, because of a successful breeding work based on it. The whole germplasm is located in a 20 ha orchard. The management cost of the collection is significant and increasing. The aim of this present cryopreservation work is to find safe and labour effective alternative of the classical field germplasm collection.
Methods: According to Zhao et al. (2008) five weeks old micropropagated sour cherry plants cv. ‘Csegöldi’ were cold adapted by hold them in an incubator programmed to 16/8 h day/night cycle with 23/4 °C. Dissected CA shoot tips were precultured for 48 h in MS medium containing DMSO (5%) and proline (2%). In 2 ml cryotubes shoot tips were equilibrate with PVS2 or modified PVS2 (containing 3% polyethylene glycol) solution for 30 minutes, and cooled at 1 °C/min rate to -40 °C and submerged into LN2.
Results and Conclusion: The recovery rate after six weeks growing period was relatively low, 27% (5/18 growing shoot tips/total shoot tips) and 38% (8/21 growing shoot tips/total) treated with PVS2 or modified PVS2, respectively.
In our Research Station there is a well-equipped micropropagated lab, however, cryopreservation of the micropropagated shoot tips is relatively expensive, and the recovery of a fruit tree is really slow. Therefore, we would like to apply dormant bud technique, but any of available protocols have been successful in our lab until this time. However, the result of present work suggests that cryopreservation would be a useful technique to maintain our local sour cherry clones.