澳大利亚本土植物茎尖新的真空渗透玻璃化（VIV）冷冻保存技术的评估
Evaluation of the new vacuum infiltration vitrification (viv) cryopreservation technique for native Australian plant shoot tips.
作者信息
Funnekotter Bryn, Whiteley Susan E, Turner Shane R, Bunn Eric, Mancera Ricardo L
机构信息
School of Biomedical Sciences, CHIRI Biosciences, Curtin University, GPO Box U1987, Perth, WA 6845.Australia. Botanic Gardens and Parks Authority, Fraser Avenue, West Perth WA 6005.Australia.
Botanic Gardens and Parks Authority, Fraser Avenue, West Perth WA 6005.Australia. School of Plant Biology, Faculty of Natural and Agricultural Sciences, University of Western Australia, Crawley WA 6009, Australia.
出版信息
Cryo Letters. 2015 Mar-Apr;36(2):104-13.
BACKGROUND
The application of a vacuum during the incubation in cryoprotective agents such as PVS2 allows for increased penetration, reducing total incubation times required before vitrification and post-cryopreservation regeneration is achieved.
OBJECTIVE
This study compared a conventional droplet-vitrification protocol to the new vacuum infiltration vitrification protocol in four Australian plant species.
MATERIALS AND METHODS
The new vacuum infiltration vitrification applied an 80 kPa vacuum during incubations in loading solution and PVS2. Infiltration of the cryoprotective agents into shoot tips was determined by differential scanning calorimetry measuring ice formation in the thermographs comparing a range of loading solution and PVS2 incubation times.
RESULTS AND CONCLUSION
The application of the vacuum infiltration vitrification technique resulted in a significantly reduced PVS2 incubation time for cryogenic survival and regeneration for all four species, reducing the time needed to adequately protect shoot tips by half to a quarter when compared to a conventional droplet-vitrification technique.
背景
在诸如PVS2等冷冻保护剂孵育过程中施加真空，可提高其渗透性，减少实现玻璃化和冷冻保存后再生所需的总孵育时间。
目的
本研究比较了四种澳大利亚植物物种的传统液滴玻璃化方案与新的真空渗透玻璃化方案。
材料与方法
新的真空渗透玻璃化在装载溶液和PVS2孵育期间施加80 kPa的真空。通过差示扫描量热法测量热成像图中的冰形成，比较一系列装载溶液和PVS2孵育时间，来确定冷冻保护剂渗入茎尖的情况。
结果与结论
真空渗透玻璃化技术的应用使所有四个物种的低温存活和再生所需的PVS2孵育时间显著减少，与传统液滴玻璃化技术相比，将充分保护茎尖所需的时间减少了一半至四分之一。
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