杂交兰种苗超低温脱毒技术研究

doi:10.13304/j.nykjdb.2017.0122

中国农业科技导报 ›› 2018, Vol. 20 ›› Issue (1): 147-153.DOI: 10.13304/j.nykjdb.2017.0122

• 方法与技术创新 • 上一篇

杂交兰种苗超低温脱毒技术研究

李涵,陆琳,瞿素萍,田敏,邹凌,李慧敏,黎霞,曹桦*

云南省农业科学院花卉研究所, 国家观赏园艺工程技术研究中心, 昆明 650100

收稿日期:2017-03-08 出版日期:2018-01-15 发布日期:2017-07-11
通讯作者: 曹桦，副研究员，硕士，主要从事观赏植物遗传育种研究。E-mail: lyxhope@163.com
作者简介:李涵，副研究员，博士，主要从事观赏植物遗传育种研究。E-mail: lihan528@126.com。
基金资助:
云南省科技厅重点新产品开发计划项目（2014BB009）资助。

Study of Hybrid Orchid Seedlings on Virus Elimination Using Cryopreservation Technology

LI Han, LU Lin, QU Suping, TIAN Min, ZOU Ling, LI Huimin, LI Xia, CAO Hua*

National Engineering Research Center For Ornamental Horticulture, Flower Research Institute, Yunnan Agricultural Academy Science, Kunming 650100, China

Received:2017-03-08 Online:2018-01-15 Published:2017-07-11

摘要/Abstract

摘要： 以携带建兰花叶病毒(CyMV)和齿兰环斑病毒(ORSV)的杂交兰根状茎为试材，采用包埋玻璃化超低温保存法对杂交兰病毒脱除进行研究。结果表明：低温炼苗21 d的茎尖，在0.5 mol/L的蔗糖浓度预培养基中培养3 d，然后浸入0.4 mol/L高糖液体培养基中加载90 min，之后转入玻璃化溶液PVS2(protect vitrification solution 2)中冰上处理6 h，再液氮保存1 h，最后于37℃水浴中解冻2~3 min，1.2 mol/L高糖液体培养基中卸载20 min，待恢复培养后，杂交兰茎尖成活率为64%以上，随机检测30个样品，两种病毒脱毒率均为100%。研究为杂交兰种苗脱毒奠定了一定的理论基础。

关键词: 杂交兰, 超低温保存, 脱毒, CyMV, ORSV

Abstract: Taking rhizomes of hybrid Cymbidium with CyMV and ORSV as test material, the paper studied virus elimination of hybrid orchid by encapsulation vitrification cryopreservation method. The results showed that treating shoot tips under low temperature for 21 d then cultured in 0.5 mol/L sucrose of preculture medium for 3 d，and immersed them in the liquid culture medium with 0.4 mol/L high sucrose for 90 min, and put into PVS2 for 6 h on ice, then preserved in liquid nitrogen for 1 h and cultured in liquid medium with 1.2 mol/L high sucrose for 20 min afterwards treated at 37℃ for 2~3 min. Their survival rate was over 64% after being cultured for recovery. 30 samples were selected randomly for virus detection, 2 of them were virus-free seedlings. Their virus detoxification rates were 100%. This study laid certain theoretical basis for virus detoxification of hybrid orchid seedlings.

Key words: hybrid orchid, cryopreservation, virus-free, CyMV, ORSV

李涵,陆琳,瞿素萍,田敏,邹凌,李慧敏,黎霞,曹桦*. 杂交兰种苗超低温脱毒技术研究[J]. 中国农业科技导报, 2018, 20(1): 147-153.

LI Han, LU Lin, QU Suping, TIAN Min, ZOU Ling, LI Huimin, LI Xia, CAO Hua*. Study of Hybrid Orchid Seedlings on Virus Elimination Using Cryopreservation Technology[J]. Journal of Agricultural Science and Technology, 2018, 20(1): 147-153.

[1]	王婧莹1,王琢2,闫培生1*. 黄曲霉毒素的脱毒研究进展[J]. 中国农业科技导报, 2019, 21(4): 42-51.
[2]	黎霞1,曹桦1,闻永慧2,杨春梅1,赵培飞1,2,阮继伟1,李涵1*. 大花蕙兰与朱砂兰杂种根状茎增殖和分化的研究[J]. 中国农业科技导报, 2017, 19(2): 28-34.
[3]	印南日1,2,李培武1,3*,周海燕2,4,白艺珍2,3,丁小霞1,2. 我国食用棉籽油质量安全[J]. , 2013, 15(4): 20-24.

杂交兰种苗超低温脱毒技术研究

Study of Hybrid Orchid Seedlings on Virus Elimination Using Cryopreservation Technology

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