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.
作者
王玉英
王琴
李枝林
魏全涛
商正蕊
凌青
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)
出处
《北方园艺》
CAS
北大核心
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)。
关键词
虎雪兰
原球茎
玻璃化
超低温
脱毒
Cymbidium tracyanum var.huanghua×Cymbidium mastersii
protocorm
vitrification
cryopreservation
virus elimination
作者简介
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.
