WANG Yuying,WANG Qin,LI Zhilin,et al.Study on Cymbidium tracyanum var. huanghua×Cymbidium mastersii on Virus Elimination Using Vitrification-cryopreservation Technology[J].Northern Horticulture,2020,44(05):61-66.[doi:10.11937/bfyy.20191600]
虎雪兰玻璃化超低温法脱毒技术的研究
- Title:
- Study on Cymbidium tracyanum var. huanghua×Cymbidium mastersii on Virus Elimination Using Vitrification-cryopreservation Technology
- Keywords:
- Cymbidium tracyanum var.huanghua×Cymbidium mastersii; protocorm; vitrification; cryopreservation; virus elimination
- 文献标志码:
- A
- 摘要:
- 以兰属花卉虎雪兰组培原球茎为试材，采用玻璃化超低温法对兰花病毒脱除进行了研究，以期为虎雪兰玻璃化超低温法脱毒体系的建立提供参考依据。结果表明：在蔗糖浓度0.5 mol·L-1预培养4 d，然后在蔗糖0.6 mol·L-1加载液冰上处理50 min，之后转入PVS2溶液冰上玻璃化处理120 min，再液氮冷冻40 min，37 ℃水浴解冻3 min，最后卸载液（1/2MS+1.2 mol·L-1蔗糖）卸载20 min，待恢复培养后，原球茎成活率可达到65%以上，随机检测不同处理样品的脱毒率可达97%。
- Abstract:
- The original bulbs of the Cymbidium tracyanum var.huanghua×Cymbidium mastersii tissue culture were used as the test material.The vitrification ultra-low temperature method was used to study the removal of Cymbidium virus.It would provide reference for virus elimination technology of orchids.This experiment was a study about virus-elimicaton of orchids using vitrification-cryopreservation technology,and the tissue culture protocorm of Cymbidium tracyanum var.huanghua×Cymbidium mastersii was used as the experiment material.The results showed that pre-culture for 4 days in 0.5 mol·L-1sucrose concentration,then treat 50 minutes in loading liquid that with sucrose concentration of 0.6 mol·L-1 on ice,the next step was shift into PVS-2 liquid and vitrification treatment 120 minutes on ice,then freeze 40 minutes in liquid nitrogen,after that thaw 3 minutes in 37 ℃ centigrade for 3 minutes,finally load-off 20 minutes in unloading liquid (1/2MS+1.2 mol·L-1 sucrose).After resuming culture,the survive rate of protocorm could reach more than 65%,and the detoxification rate could reach 97% by random detection of different samples.
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备注/Memo
第一作者简介：王玉英（1980-），女，云南大理人，博士，副教授，现主要从事植物资源的利用和创新等研究工作。E-mail:wyysxp@126.com.责任作者：凌青（1968-），女，云南昆明人，硕士，副教授，现主要从事植物的利用和创新等研究工作。E-mail:wyysxp@126.com.基金项目：云南省重点新产品开发计划资助项目（2012BB008）；云南省重大专项资助项目（2018BB010）；云南农业大学自然科学青年科研基金资助项目（2016ZR10）；云南省大学生创业创新资助项目（201710676067）；云南省昆明市科学技术局重点资助项目（2015-1-N-00984）。收稿日期：2019-07-15