摘要
2～3mm的石蒜茎尖放在MS+0.4mol·L-1蔗糖的培养基上预培养5d,在25℃下用预处理液处理20min,接着用冰浴的玻璃化保护剂PVS2在冰浴中处理80min后,换新鲜PVS2并迅速投入液氮。液氮保存24h后,于40℃水浴中快速解冻2min,用MS+1.2mol·L-1蔗糖的液体培养基洗涤20min,滤纸吸干后接种到恢复培养基中,在25℃下暗培养7d后,转入光照强度为36μmol·m-2·s-1和光暗周期12/12h条件下培养。2周后的成活率最高可达90%,植株再生率达53%。
An efficient cryopreservation procedure by vitrification was developed for long-term conservation of shoot tips of Lycoris radiata Herb in vitro. The procedure included 4 steps： at first, the shoot tips, 2-3 mm in length, were pre-cultured on MS medium enriched with 0.4 mol·L^-1 sucrose for 5 d; and second, pre-cultured shoot tips were treated for 20 min with a loading solution at 25 ℃; and then they were subjected to ice-cooled vitrification solution for 80 min and transferred to 2 mL cryotubes with fresh vitrification solution （PVS2） and plunged into liquid nitrogen; at last rapid thawing took place in 40℃ water bath for 2 min followed by the deloading step for 20 min in a deloading solution consisted of 1.2 mol·L^-1 sucrose liquid MS medium. Further recovery and growth took place on regeneration medium in the dark at 25℃ for 7 d and then with 36 μmol·m^2·s^-1 irradiance and light/dark cycle of 12/12 h. The highest survival rates of the shoot tips reached 90% after 2 weeks incubation and the regeneration rate reached 53%.
出处
《植物生理学通讯》
CSCD
北大核心
2006年第6期1063-1066,共4页
Plant Physiology Communications
基金
上海市科技兴农重点攻关项目[沪农科攻字(2005)第1-6号]
关键词
红花石蒜
茎尖培养
玻璃化法
超低温保存
Lycoris radiata Herb
in vitro-grown shoot tips
vitrification
cryopreservation
