【作者】 柳学周； 宁鑫； 徐永江； 赵明； 李春广； 李军；

【Author】 LIU Xue-zhou1~* NING Xin1,2 XU Yong-jiang1 ZHAO-Ming1 LI Chun-guang1.2 LI Jun3 (1.Key Laboratory for Sustainable Utilization of Marine Fisheries Resource,Ministry of Agriculture,Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao,266071;2.College of Fisheries and Life Science,ShangHai Ocean University,Shang Hai,201300;3.Institute of Oceanology,Chinese Ac)

【机构】 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； 上海海洋大学水产与生命学院； 中国科学院海洋研究所；

【摘要】 采用紫外线(UV)灭活的冷冻真鲷(Pagrosomus major)精子激发漠斑牙鲆(Paralichthyslethostigma)卵子发育。利用冷休克法抑制卵子第二极体排出,成功获得雌核发育二倍体仔鱼。冷冻真鲷精子适宜的UV灭活剂量为72 mJ/cm2,随着处理时间延长,胚胎的孵化率呈现哈特维希效应,其中40s灭活组胚胎孵化率达到最高29±1.9%,单倍体率100%。冷休克处理温度为0～2℃,处理起始时间3min、持续时间45min时,雌核发育二倍体胚胎的孵化率最高,达9.4±0.71%,获得批量雌核发育仔鱼。雌核发育仔鱼经流式细胞仪和染色体倍性鉴定,均为二倍体(2n=48),未发现单倍体和非整倍体现象。雌核发育二倍体组和单倍体组与正常二倍体组组孵化时间差异显著,分别历时64h59min和55h49min。雌核发育二倍体仔鱼与正常二倍体仔鱼形态特征差异不显著(P>0.05),单倍体仔鱼明显畸形。本研究表明,采用适宜的紫外线剂量灭活的冷冻真鲷精子可成功诱导漠斑牙鲆雌核发育,获得雌核发育二倍体后代,试验结果为漠斑牙鲆全雌化苗种生产提供了技术和理论依据。更多还原

【Abstract】 Female southern flounder(Paralichthys lethostigma) grow faster than males. Therefore,farming all-female progenies could maximize culture profit for farmers.In the present study,diploid gynogenesis was induced by activating flounder egg development with UV irradiated(72 mJ/cm2) Pagrosomus mojor sperm for 3 min in sea water,and then subjecting the eggs to cold shock(0-2℃) treatment for 45 min.Cold shock was employed to prevent extrusion of the second polar body.Control treatments omitted one or more of these steps to separately assess the effectiveness of UV irradiation and cold shock.When initial treatment moment was 3min post hatcing,treatment duration was 45min,the fertilization rate of eggs activated by UV-irradiated heterologous sperm was 37.2±5.1%and the malformation rate of newly hatched larvae was 8.3±2.5%,whilst hatching rate was up to 9.4±0.71%.Flow cytometry and chromosome karyotype were used to examine the ploidy of larvae experienced cold shock treatment.Results showed that all obtained gynogenetic larvae were diploid(2n = 48),neither haploid nor aneuploid phenomena was found.The hatching duration of haploid embryos and gynogenetic embryos更多还原