超低温冷冻保存对七带石斑鱼胚胎酶活性的影响  

Effects of Cryopreservation on Enzyme Activity in Seven-band Grouper Epinephelus septemfasciatus Embryos

作　　者：姜静[1,2] 田永胜[1] 翟介明 陈松林[1] 

机构地区：[1]农业部海洋渔业可持续发展重点实验室,中国水产科学研究院黄海水产研究所,青岛266071 [2]上海海洋大学,水产与生命学院,上海201306 [3]山东莱州明波水产有限公司,莱州261418

出　　处：《农业生物技术学报》2014年第4期476-485,共10页Journal of Agricultural Biotechnology

基　　金：国家高技术研究发展计划(863)(No.2012AA10A402和No.2012AA10A408);国家自然科学基金项目(No.31372510);山东省泰山学者建设工程专项

摘　　要：七带石斑鱼(Epinephelus septemfasciatus)是一种生长速度快、低温下耐受性强具有较高经济价值的海水鱼类,在自然环境中种群数量较少,对其种质资源进行长期保存迫在眉睫。本研究以七带石斑鱼胚胎为实验材料进行种质冷冻保存,探讨超低温冷冻(-196℃)对七带石斑鱼胚胎内超氧化物歧化酶(superoxide dismutase,SOD)、肌酸激酶(creatine kinase,CK)、Na+/K+-ATPase、乳酸脱氢酶(lactate dehydrogenase,LDH)、丙二醛(malondialdehyde,MDA)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性的影响,以检测抗冻剂在冷冻保存中对胚胎的影响。本研究分别测定了经过PM(24%1,2-丙二醇(PG)+16%甲醇(MeOH))、PMG(15.75%1,2-丙二醇+8.75%甲醇(MeOH)+8.75%甘油(Gly))和PMGT(15.75%1,2-丙二醇+8.75%甲醇(MeOH)+8.75%甘油(Gly)+5%海藻糖(trehalose))3种玻璃化液处理冷冻前后七带石斑鱼胚胎内6种酶活性的变化。结果表明,胚胎分别经过以上3种玻璃化液处理后,6种酶活性均发生显著性变化,与冷冻前相比,冷冻后胚胎内MDA酶活性显著升高(P<0.05);与之相反,冷冻后胚胎内的SOD、CK、Na+/K+-ATPase、LDH和GSH-Px酶活性与冷冻前相比,均显著下降,且差异显著(P<0.05)。而在PM玻璃化液处理后的未冷冻胚胎中Na+/K+-ATPase的表达量与对照组无显著性差异(P>0.05)。冷冻使MDA酶活性显著升高,而使其他5种酶的表达量水平降低。PM玻璃化液对CK和Na+/K+-ATPase的活性影响较小,说明对胚胎具有一定的保护作用。本研究为七带石斑鱼胚胎超低温冷冻保存提供理论依据。Seven-band grouper （Epinephelus septemfasciatus） is a seawater fish with high economic values that grow fast and tolerate strong with low temperature. Population quantity is less in the natural environment, and the germplasm resources for long-term preservation are imminent. Therefore, based on the cryopreservation of seven-band grouper embryos, the effects of cryopreservation （-196 ℃） on the enzyme activity （superoxide dismutase （SOD）, creatine kinase （CK）, Na＋/K＋-ATPase, lactate dehydrogenase （LDH）, malondialdehyde （MDA）, glutathione peroxidase （GSH-Px）） in Epinephelus septemfasciatus embryos were studied. In the present study, the changes of enzyme activity in embryos were analyzed by kits to clarify damage mechanism of embryos. The results indicated that the MDA enzyme activity of embryos after cryopreservation treated with the vitrifications solution of PM（24% 1,2 - propylene glycol（PG）＋16% methanol（MeOH））、PMG（15.75% pylene glycol（PG）＋8.75% methanol（MeOH）＋8.75% glycerol（Gly））和PMGT（15.75% pylene glycol（PG）＋8.75% methanol（MeOH）＋8.75%glycerol（Gly）＋5% Trehalose） significantly increased compared with no cryopreservation groups. However, the activity of some other enzymes descended significantly before and after cryopreservation compared with control group （P〈0.05）, and the latter was significantly lower than the former. However, there was no significant difference between Na＋/K＋-ATPase enzyme activity of no-frozen embryos treated with PM vitrification group and the control group. From various enzymes activity change from seven-band grouper before and after cryopreservation, cryopreservation and cryoprotectants have significant effect on the enzymes activity of seven-band grouper. Therefore, through the detection and analysis of the concentration of enzymes activity change from frozen embryos, freeze made MDA enzymes activity increase, other 5 enzymes activity decreased. PM vitrification solution had less ef

关 键 词：七带石斑鱼 超低温冷冻保存 胚胎 酶活性 

分 类 号：S961.6[农业科学—水产养殖]