Condello E, Caboni E, Andre E, Piette B, Druart R, Swennen R, Panis B
CRA - Fruit Tree Research Institute, Rome, Italy.
Cryo Letters. 2011 Mar-Apr;32(2):175-85.
In vitro axillary buds of two apple cultivars, Pinova and Jonagold, were successfully cryopreserved by droplet-vitrification. In vitro axillary buds of cv. Pinova were subjected to PVS2 for 15, 30, 45, 60, 80 or 100 min, while Jonagold buds were treated with PVS2 for 15, 30, 45 or 60 min. In addition, the effect of age of in vitro mother-plants on recovery after cryopreservation was evaluated. Recovery was performed on medium with various combinations of BA, IBA and GA3. Regrowth percentages for cv. Pinova increased in line with increasing PVS2 exposure durations, from 15 to 60 min. Cv. Jonagold showed a similar trend with an increase in regrowth from 30 to 60 min PVS2 exposure. Improved regrowth was observed when axillary buds were excised from aged mother-plants in comparison to those excised from plantlets that were regularly subcultured. The highest shoot regrowth was obtained when applying a 60 min PVS2 treatment to axillary buds excised from non-preconditioned 4-month old in vitro shoots and performing regrowth on recovery medium containing 4.50 microM BA and 0.50 microM IBA. This optimal protocol was also successfully applied to apple rootstocks M26 and Jork 9.
通过玻璃化法成功对两个苹果品种皮诺娃（Pinova）和乔纳金（Jonagold）的离体腋芽进行了超低温保存。皮诺娃品种的离体腋芽用PVS2处理15、30、45、60、80或100分钟，而乔纳金品种的芽用PVS2处理15、30、45或60分钟。此外，还评估了离体母株年龄对超低温保存后恢复情况的影响。在含有不同组合的苄氨基嘌呤（BA）、吲哚丁酸（IBA）和赤霉素（GA3）的培养基上进行恢复培养。皮诺娃品种的再生率随着PVS2处理时间从15分钟增加到60分钟而增加。乔纳金品种也呈现类似趋势，PVS2处理时间从30分钟增加到60分钟时再生率增加。与从定期继代培养的组培苗上切取的腋芽相比，从老龄母株上切取的腋芽再生情况有所改善。对从未经预处理的4个月龄离体苗上切取的腋芽进行60分钟PVS2处理，并在含有4.50微摩尔BA和0.50微摩尔IBA的恢复培养基上进行再生培养时，获得了最高的芽再生率。该优化方案也成功应用于苹果砧木M26和约克9号。
