Aronen T, Ryynanen L
Finnish Forest Research Institute, Punkaharju, Finland.
Cryo Letters. 2014 Sep-Oct;35(5):385-94.
For the conservation of hybrid aspen germplasm, cryostorage of dormant in vivo buds is a convenient back-up method for field collections. In practice in Finland, bud collection is performed from February to March.
The aim of this study was to assess how this time schedule can be extended without compromising regeneration. In addition, an easily measurable marker for successful cryopreservation was examined.
Timing of cryopreservation was tested from August to February, using dormant buds from both outdoor and indoor plants. To find a marker, water content and gene expression of hydrid aspens, as well as environmental factors such as temperature, temperature sum, and light period were followed.
Cryopreservation was successful from October to February, when, on an average, at least 75 % of the buds regenerated through micropropagation, and there was no difference to non-frozen controls. Significant genotypic variation was observed in October and February, with regeneration rates of 61-100 % and 37-98 %, respectively. No marker for successful cryopreservation was found among the studied factors.
The results provide flexibility for the undertaking of practical work, with a recommendation that cryopreservation can be carried out from November to January - earlier than the current practice.
为了保存杂种山杨种质，对休眠的体内芽进行冷冻保存是田间收集的一种便捷备用方法。在芬兰的实际操作中，芽的采集在2月至3月进行。
本研究的目的是评估如何在不影响再生的情况下延长这个时间安排。此外，还研究了一种易于测量的成功冷冻保存的标志物。
利用室外和室内植物的休眠芽，在8月至2月期间测试冷冻保存的时间。为了找到一个标志物，跟踪了杂种山杨的含水量、基因表达以及温度、积温、光照周期等环境因素。
10月至2月冷冻保存成功，平均至少75%的芽通过微繁殖再生，与未冷冻的对照没有差异。10月和2月观察到显著的基因型差异，再生率分别为61%-100%和37%-98%。在所研究的因素中未发现成功冷冻保存的标志物。
研究结果为实际工作提供了灵活性，建议在11月至1月进行冷冻保存，比目前的做法更早。
