菊花茎尖的玻璃化超低温保存研究  

Study on Cryopreservation of Shoot-tips of Chrysanthemum through Vitrification

作　　者：刘艳霞[1,2] 刘灶长[1] 林田[1] 李天菲[1] 陈发棣[3] Lee Inho 罗利军[1] 

机构地区：[1]上海市农业生物基因中心,上海201106 [2]华中农业大学,武汉430070 [3]南京农业大学,南京210095 [4]Sunchon National University,Jeonnam 540-742,Korea

出　　处：《植物遗传资源学报》2009年第2期249-254,共6页Journal of Plant Genetic Resources

基　　金：上海市科技兴农重点攻关项目沪农科攻字(2006)第1-8号

摘　　要：本文建立了适合中国菊花种质资源长期保存的玻璃化超低温保存技术体系。在4℃下，把1-2mm的菊花茎尖放在含0，4mol／L蔗糖的MS培养基上暗培养2-3d，用预处理液在25℃下处理30min，再用玻璃化试剂PVS2在冰浴条件下处理15min，换新鲜的PVS2试剂并迅速投入液氮。液氮保存24h后，40％水浴解冻2min，用含蔗糖1．2mol／L的MS液体培养基洗涤20mn，滤纸吸千后接种到恢复培养基中，在25℃条件下弱光培养1-3d转入正常光照培养条件下培养，2周后成活率可达86％以上，成活的茎尖均可再生。An efficient cryopreservation procedure by vitrification was developed for long-term conservation of chrysanthemum. The procedure included 4 steps ： a） The 1-2 mm length in vitro-grown shoot tips were precultured on MS medium enriched with 0.4 mol/L sucrose in darkness at 4℃for 2-3 d. b） Pre-cultured shoot tips were immersed for 20 rain in loading solution （LS） containing 2 mol/L glycerol and 0.4 mol/L sucrose at 25℃. c） Treat the tips with ice-cooled PVS2 solution for 15 min, and then tips were put into cryo-tubes filled with fresh PVS2 and plunged into liquid nitrogen, d） The preserved tips were rapidly thawed in 40℃ water bath for 2 rain and unloaded the PVS2 with liquid MS medium contained 1.2 mol/L sucrose for 20 min. Further recovery and growth took place on regeneration medium in the darkness for less than 3 days. Successfully vitrified shoot tips developed shoots directly within 3- 14d of culture. The survival rate of the shoot tips reached to 86% and the survived tips could all regenerated into new shoots.

关 键 词：菊花 茎尖 玻璃化法 超低温保存 

分 类 号：S682.11[农业科学—观赏园艺]