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云纹石斑鱼精子冷冻保存 被引量:5

Sperm cryopreservation of kelp grouper Epinephelus moara
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摘要 以云纹石斑鱼精液为实验材料,对精子稀释液、抗冻剂种类和适宜浓度、冷冻保存液进行了筛选.结果表明,利用9 g/L NaC1、10 g/L KHCO3和10%小牛血清配制而成的稀释液EM1-2适宜于云纹石斑鱼精子冷冻保存,以2ml冷冻管为精子容器,在60L液氮生物保存罐中冷冻保存精子,冷冻解冻精子活力可达56.67%±5.77%,要优于TS-2、ES1-3和其他EM系列稀释液冷冻保存精子活力.利用EM1-2为基础液对抗冻保护剂进行筛选,结果显示,10%~20%的二甲基亚砜(DMSO)和1-2-丙二醇(PG)冷冻保存后精子活力无显著差异(P>0.05),其中15%的DMSO和10% PG冷冻保存精子效果最优,解冻后精子活力分别可达54.52%±7.81%和57.24%±3.69%.利用冷冻保存1年的精液与云纹石斑鱼卵进行受精,受精率和孵化率均达到80%以上,与新鲜精子无显著性差异(P>0.05).本研究表明,利用EM1-2配制15%的DMSO或10%的PG可用于冷冻保存云纹石斑鱼精液.在此基础上,建立了精子冷冻库,保存精子130ml,为人工繁育和杂交育种提供了丰富的精子源. To study the cryopreservation of Epinephelus rnoara sperm, mature E. moara semen was used as experimental material, and concentration of various semen diluents, and con centration of cryoprotectant were screened. We used 60L liquid nitrogen jars and 2ml freezing tubes to preserve sperms by "three-step cooling procedure". The result showed that, EM1 2 semen diluent prepared with μg/L NaC1, 10g/L KHCO3 and 10% FBS(fetal bovine serum) was better than TS-2, ES1-3 and other EM semen diluents, which preserved 56.67%+5. 77% sperm motility after unfrozen. Using EM1-2 as the liquid foundation to prepare cryoprotectant, no significant difference was observed in sperm motility by 10%-20% DMSO and PG cryopr- eservation(P〉0.05). The best of all is 15% DMSO and 10% PG, which preserved 54.52%± 7.81% and 57.24 % ± 3.69%sperm motility respectively after unfrozen. Using 1-year eryopre- served semen to fertilize E. moara eggs, the rates of fertilization and hatching reached 80 % or above, without significant difference with fresh sperms(P〉0.05). Our study showed that 15 DMSO or 10% PG using EM1-2 as the liquid foundation could cryopreserve E. moara semen. We established a frozen sperm library based on this tificial breeding and cross breeding. study, and it will provide a basis for the artificial breeding and cross breeding.
出处 《渔业科学进展》 CSCD 北大核心 2014年第1期26-33,共8页 Progress in Fishery Sciences
基金 “863”高技术研究发展计划(2012AA10A402 2012AA10A408) 国家自然科学基金项目(30972244) 山东省泰山学者建设工程专项共同资助
关键词 云纹石斑鱼 精子 冷冻保存 精子稀释液 Epinephelus moara Sperm Cryopreservation Diluents
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