鲈鱼胚胎程序化冷冻保存的研究  

Study on programmed cryopreservation of sea perch embryos

作　　者：于过才[1] 陈松林[1] 孔晓瑜[1] 田永胜[1] 季相山[1] 

机构地区：[1]中国水产科学研究院黄海水产研究所

出　　处：《海洋水产研究》2004年第1期1-8,共8页Marine Fisheries Research

基　　金：国家863高技术研究发展项目(2001AA621100)资助

摘　　要：对鲈鱼(Lateolabraxjaponicus)胚胎的程序降温冷冻保存进行了研究。通过稀释液的筛选实验 ,选择了一种培养效果好的稀释液DS1液。研究了不同发育阶段胚胎对抗冻液的毒性耐受力 ,表明肌肉效应期胚胎的耐受力最强。测定了心跳期胚胎在抗冻液A1 、A2 、B1 、B2 中的平衡处理时间 ,为进行鲈鱼胚胎冻前平衡提供了参考时间。比较了植冰和不植冰对鲈鱼胚胎冷冻保存结果的影响及不同洗脱方法洗脱胚胎的效果。结果表明 ,在冷冻过程中诱导植冰的胚胎成活率高于未植冰组 ,两步法洗脱效果优于一步法和三步法。用不同降温速率进行了鲈鱼胚胎的超低温冷冻保存实验 ,结果表明 ,采用 1 .5℃ /min的降温速率降温 ,在液氮中冷冻保存 30min的 72粒心跳期胚胎中有 3粒成活 ,成活率为 4.2 %。Programmed freezingof sea perch embryos was studied in this paper.We selected an effective extender-DS1 by treating closure stage of blastopore in 8 extenders for 10 h. The toxicity tolerance of embryos at vari ous stages of development towards the solution containing 10% DMSO was examined. The result showed that embryos at muscular contraction stage tolerated the high er toxicity of the cryoprotectant than the others. Equilibration time that embry os were treated in solution A 1 、 A 2 、 B 1 、 B 2 was determ ined and this providedevidence for treating embryos in cryoprotective ag ents before cryopreservation.Comparing effects of seeding and no-seedin g on cryopreservation and evaluating effectsof different elution methods , we foundthat survival rate of seeding group was higher than th at of no-seeding group and that two-step method (Embryos were first placed in 0.25 mol/L sucrose for 10min and then in sea water) was better than one -step method and three-step method.Cryopreservation of sea perch embryos was done with different cooling rates of 0.4 ℃/min、 1.0 ℃/min、 1 .2 ℃/min、 1.5 ℃/min、 2.0 ℃/min, respectively. The result showed that 3 e mbryos survived from 72 embryos at heartbeat stage that were cryopreservated wit h cooling rate of 1.5 ℃/min and heldin liquid nitrogen for 30 min befor e thawing,the survival rate was 4.2%.

关 键 词：鲈鱼 胚胎 程序冷冻 

分 类 号：Q813[生物学—生物工程] Q959.483