大菱鲆雌核发育二倍体的真鲷冷冻精子诱导及其生长评价  

Induction and Growth of Meiogynogenetic Diploids of Turbot (Scophthalmus maximus) with Cryopreserved Sperm of Red Sea Bream(Pagrus major)

作　　者：孟振[1,2] 刘新富[1] 雷霁霖[1] 刘滨[1] 张和森 王国栋[1] 

机构地区：[1]中国水产科学研究院黄海水产研究所/青岛市海水鱼类种子工程与生物技术重点实验室,山东青岛266071 [2]中国海洋大学水产学院,山东青岛266003 [3]青岛通用水产养殖有限公司,山东青岛266404

出　　处：《武汉大学学报（理学版）》2013年第4期343-350,共8页Journal of Wuhan University:Natural Science Edition

基　　金：农业部国家鲆鲽类产业技术体系建设专项资金(CARS-50-G02);农业部公益性行业科研专项经费(nyhyzx07-046)资助项目

摘　　要：根据Hertwig效应,采用正交设计实验方法对真鲷(Pagrus major)冷冻精子诱导大菱鲆(Scophthal-mus maximus)减数分裂雌核发育的最佳条件进行了研究.结果表明,未经紫外线照射处理的真鲷精子与大菱鲆卵子受精后形成的胚胎,无论是否经过冷休克处理都不能孵化出膜;经剂量为64.8～72.0mJ.cm-2紫外线照射处理后的真鲷精子与大菱鲆卵子受精后形成的胚胎,初孵仔鱼表现出典型的单倍体综合征;受精和孵化水温为(14.5±0.5)℃时,大菱鲆雌核发育单倍体的冷休克二倍体化的最佳处理起始时间、处理水温和持续时间分别为受精后6.5min,-2℃和45min.至孵化后60日龄,雌核发育苗种的成活率为0.61%,仅相当于其半同胞对照的10%左右,60～180日龄两者的成活率差异不显著,均达90%以上;14月龄前,雌核发育苗种的全长和体重均比半同胞对照要低,此后雌核发育苗种由于雌性比例较高而生长速度加快,至17月龄时其全长和体重与半同胞对照基本相当.Hertwig effect and orthogonal experiment design were employed to determine the optimal protocol for the induction of meiogynogenetic diploids of turbot（Scophthalmus maximus） with cryopreserved sperm of red sea bream（Pagrus major）.With or without cold shock treatment neither of the turbot embryos fertilized with untreated sperm of red sea bream could survive beyond hatching.All of the turbot larvae hatched from eggs activated by UV irradiated sperm of red sea bream at the dosage of 64.8-72.0 mJ·cm-2 were typical haploids.Under the fertilization and hatchery temperature of（14.5±0.5）℃,the maximal diploidization of the meiogynogenetic haploids of turbot could be achieved in-2 ℃ cold water shock treatment for 45 min to the eggs 6.5 min after fertilization with the UV-irradiated sperm of red sea bream.Two batches of meiogynogenetic diploids of turbot were produced by the above UV and cold shock treatments,and a comparison of the survival and growth between one batch of the meiogynogenetic diploids and their half-sib normal diploids had been lasted for 17 months.Up to the 60 days after hatching,the survival rate of the meiogynogenetic dipoloids was only 0.61%,which was less than a tenth of that of their half-sib＇s.Thereafter,survival rates of both group were similar and well above 90%.The growth rate of meiogynogenetic diploids had been lower than that of their half-sib＇s upto the 14th months after the hatching,and became equal to that of their half-sib＇s by the end of 17th months after the hatching,which was possible the result of growth advantage of more females in the meiogynogenetic dipoids.The present study optimizes a procedure for the induction of meiogynogenetic diploids at large scale with heterologous sperm in turbot,thus laying the basis for further applications towards producing monosex and defining the sex determination mechanism.

关 键 词：大菱鲆 真鲷 冷冻精子 雌核发育 

分 类 号：Q78[生物学—分子生物学] S917[农业科学—水产科学]