Wang Min-Rui, Chen Long, Zhang Zhibo, Blystad Dag-Ragnar, Wang Qiao-Chun
State Key Laboratory of Crop Stress Biology for Arid Areas, College of Horticulture, Northwest A&F University, Yangling, Shaanxi, China.
Division of Biotechnology and Plant Health, Norwegian Institute of Bioeconomy Research, Ås, Norway.
Methods Mol Biol. 2018;1815:257-268. doi: 10.1007/978-1-4939-8594-4_17.
Virus diseases have been a great threat to production of economically important crops. In practice, the use of virus-free planting material is an effective strategy to control viral diseases. Cryotherapy, developed based on cryopreservation, is a novel plant biotechnology tool for virus eradication. Comparing to the traditional meristem culture for virus elimination, cryotherapy resulted in high efficiency of pathogen eradication. In general, cryotherapy includes seven major steps: (1) introduction of infected plant materials into in vitro cultures, (2) shoot tip excision, (3) tolerance induction of explants to dehydration and subsequent freezing in liquid nitrogen (LN), (4) a short-time treatment of explants in LN, (5) warming and post-culture for regeneration, (6) re-establishment of regenerated plants in greenhouse conditions, and (7) virus indexing.
病毒病害一直对经济作物的生产构成巨大威胁。在实际生产中，使用无病毒种植材料是控制病毒病害的有效策略。基于冷冻保存技术发展而来的低温疗法，是一种用于根除病毒的新型植物生物技术工具。与传统的用于消除病毒的茎尖培养相比，低温疗法能高效根除病原体。一般来说，低温疗法包括七个主要步骤：（1）将受感染的植物材料引入离体培养；（2）切取茎尖；（3）诱导外植体耐受脱水并随后在液氮中冷冻；（4）外植体在液氮中进行短时间处理；（5）解冻并进行继代培养以实现再生；（6）在温室条件下使再生植株重新定植；（7）病毒检测。
