异源精子诱导犬齿牙鲆的雌核发育  

Gynogenesis induced by heterogenous sperms in summer flounder Paralichthys dentatus

作　　者：杨景峰[1,2,3] 陈松林[1,4] 徐亘博[1] 苏鹏志[1] 罗心慧[1] 邵长伟[1] 翟介明 

机构地区：[1]中国水产科学研究院黄海水产研究所农业部海洋渔业资源可持续利用重点开放实验室,山东青岛266071 [2]中国海洋大学海洋生命学院,山东青岛266003 [3]内蒙古民族大学动物科技学院,内蒙古通辽028042 [4]上海高校水产养殖学E-研究院,上海海洋大学水产与生命学院,上海201306 [5]山东莱州明波水产有限公司,山东莱州266031

出　　处：《水产学报》2009年第4期533-541,共9页Journal of Fisheries of China

基　　金：国家"八六三"高技术研究发展计划(2006AA10A403);山东省泰山学者工程专项;上海高校水产养殖学E-研究院项目(E030090)

摘　　要：用冷冻保存的花鲈精子作为异源精子，采用紫外线（uV）对精子进行照射使其遗传物质失活，然后与卵子进行“授精”，可以刺激犬齿牙鲆鱼卵进行雌核发育，在受精后一定时间采用冷休克处理“授精”卵，抑制第二极体排出成功获得了犬齿牙鲆雌核发育二倍体鱼苗。实验表明：犬齿牙鲆同源精子经80mJ／cm^2的紫外线照射可以完全失活，冻存花鲈精子经紫外线照射后也同样具有诱导犬齿牙鲆雌核发育的能力。无论同源精子还是异源精子，最佳诱导条件为在18℃条件下，精子经80mJ／cm^2的紫外线照射，受精后4～5min将受精卵放在3℃海水中进行冷休克处理，持续时间为45min，同源精子和异源精子二倍体诱导分别为32．66％±7．03％和28．00％±6．48％。采用形态学、流式细胞仪DNA含量分析和微卫星标记技术对雌核发育鱼苗进行了分析，证明了雌核发育鱼苗为雌核发育二倍体。Effective methods for induction of diploid gynogenesis of the summer flounder Paralichthys dentatus are needed to initiate monosex culture, which will allow farmers to take advantage of the more rapid growth and larger size attained by females and will maximize profit potential for the culture of this species. To induce diploid gynogenesis in summer flounder, eggs were fertilized with heterogenous cryopreservative sperm of Lateolabrax japonicas, which was irradiated with UV ray to be genetically inactivated, and cold shock was used to prevent extrusion of the second polar body. Results indicated that, just like the homologous sperm of summer flounder, the cryopreservative sperm of Lateolabrax japonicas could induce diploid gynogenesis of summer flounder. Diploid gynogenesis was induced by activating egg development with UV irradiated sperm （80 mJ/cm^2, the homologous or the heterogenous sperm） for 5 rain in seawater at 18 ℃, and then subjecting the eggs to cold shock in 3℃ seawater for 45 rain. And the dipoid gynogenesis induction rates of the homologous sperm and the heterogenous sperm were 32.66% ±7.03% and 28. 00% ±6. 48%, respectively. It was certified that the larvae were gynogenetic diploid by morphology, flow cytometry and microsatellite DNA analysis. It is the first time to report effective methods for induction diploid gynogenesis of the summer flounder by heterogenous cryopreservative sperm of Lateolabrax japonicas, which provides technological measures for sex control and genetic improvement of summer flounder.

关 键 词：犬齿牙鲆 花鲈 雌核发育 异源精子 冷休克 微卫星 

分 类 号：S917[农业科学—水产科学]