包埋-玻璃化法冷冻保存湛江等鞭金藻的研究  

CRYOPRESERVATION OF ISOCHRYSIS ZHANJIANGENSIS BY ENCAPSULATION-VITRIFICATION

作　　者：刘丽娜[1] 王起华[1] 张恩栋[1] 张丽佳[1] 

机构地区：[1]辽宁师范大学生命科学学院,大连116029

出　　处：《水生生物学报》2007年第2期169-173,共5页Acta Hydrobiologica Sinica

基　　金：国家自然科学基金项目(30470184;30170099)资助

摘　　要：采用包埋-玻璃化法冷冻保存湛江等鞭金藻(Isochrysis zhanjiangensis),探讨了装载液成分和浓度、装载时间、脱水时间、洗涤液浓度及洗涤时间对超低温保存后存活率的影响。结果表明在20℃50%PVS(PVS:30%甘油(GLY)+20%乙二醇(EG)+10%二甲基亚砜(DMSO),用f/2培养基定容)装载4.5h,0℃100%PVS脱水50min,冻存24h后取出冻存管并迅速投入40℃恒温水浴中快速化冻约3min,1.0mol/L山梨醇洗涤40min条件下,湛江等鞭金藻的存活率最高,为54%。与常规的两步法和包埋脱水法相比,包埋-玻璃化法简单、快速且存活率高,在藻类种质保存中有广阔的应用前景。lsochrysis zhanfiangensis was preserved in liquid nitrogen by encapsulation-vitrification. The following factors affect the viability of the alga such as the composition and the concentration of loading solution and loading time, the time of dehydration, the concentration of washing solution and washing time were studied. The result showed that the high viability of 54% was obtained when the beads containing the algal cells were loaded with 50% PVS（ PVS：30% （W/V）glycerol, 20% （W/V）ethylene glycol and 10% （W/V） dimethyl sulfoxide in f/2 medium）for 4.5h at 20℃, then dehydrated with 100% PVS for 50 min at 0℃ prior to a direct plunge into liquid nitrogen and washed by lmol/L sorbitol for 40min after warming in 40℃ water bath. The method of encapsulation-vitrification do not need special equipments, simplifies the operation procedure of cryopreservation, so it is a potential method for preservation of I. zhanfiangensis and other algae.

关 键 词：湛江等鞭金藻 包埋-玻璃化 装载 脱水 洗涤 

分 类 号：S984.4[农业科学—捕捞与储运]