虎雪兰玻璃化超低温法脱毒技术的研究  

Study on Cymbidium tracyanum var. huanghua×Cymbidium mastersii on Virus Elimination Using Vitrification-cryopreservation Technology

作　　者：王玉英[1] 王琴 李枝林[1] 魏全涛 商正蕊 凌青[2] WANG Yuying;WANG Qin;LI Zhilin;WEI Quantao;SHANG Zhengrui;LING Qing(College of Garden and Horticulture,Yunnan Agricultural University,Kunming,Yunnan 650201;Yunnan Agricultural Vocational and Technical College,Kunming,Yunnan 650031)

机构地区：[1]云南农业大学园林园艺学院,云南昆明650201 [2]云南农业职业技术学院,云南昆明650031

出　　处：《北方园艺》2020年第5期61-66,共6页Northern Horticulture

基　　金：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)。

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%。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(1/2 MS+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.

关 键 词：虎雪兰 原球茎 玻璃化 超低温 脱毒 

分 类 号：S682.31[农业科学—观赏园艺]