摘要
采用真鲷精子激活牙鲆卵子,经(0±0.5)℃冷休克处理,诱导了牙鲆减数分裂雌核发育。灭活真鲷精子,紫外线照射剂量3.4mJ/cm2时孵化率最低;随着照射剂量的增加,孵化率恢复,照射剂量73mJ/cm2时,孵化率最高,呈现典型的Hertwig效应。用流式细胞仪检测倍性,未经冷休克处理的胚胎全部为单倍体,经冷休克处理的均为二倍体,表明精子灭活有效。冷休克实验结果表明,冷休克起始时间2～5min均有效,以3min为最好;冷休克持续时间30～90min均有效,以45min为最好。综合两项因素,授精3min后,休克时间持续45min组的授精率和孵化率均为最高,与其他处理组差异显著(P<0.05)。RAPD分析结果显示,异源精子的遗传信息未在雌核发育二倍体的电泳图中出现,表明其遗传信息没有传递给子代。
Japanese flounder（Paralichthys olivaceus） is a commercially important marine finfish in China.The growth rate of Japanese flounder is significantly faster in females than in males.Effective methods for gynogenetic inducement in this species are important for all-female population construction.Meiogynogenesis was induced in Japanese flounder（P.olivaceus） by using UV irradiated sperm of red sea bream（Pagrus major）.The sperm of red sea bream exhibited a significant Hertwig effect after irradiating by UV.The embryo had the lowest hatching rate（2.0% ± 0.1%） when UV dosage was 3.4 mJ/cm2,and recovered with the increase of UV dosage.It reached the highest hatching rate（57.7% ± 3.1%） when UV dosage was 73 mJ/ cm2,and ploidy analysis by flow cytometer showed that the embryos were all haploids.After a series of tests for initiation and duration at water temperature of（0 ± 0.5） ℃,the best parameters for meiogynogenesis inducing in Japanese flounder were determined that cold shock begun at 3 min after insemination and continued for 45 min.The fertilization rate and hatching rate at this scenario were 80.7% ± 1.1% and 57.3% ± 2.3%,respectively,which were significantly higher than all the other treatment groups（P 0.05）.Fries of gynogenesis were confirmed to be diploids via flow cytometer.Further analysis of RAPD analysis showed that genetic material of red sea bream sperm was destroyed after UV irradiation and was not transmitted to progenies.This study provided an effective method to induce meiogynogenesis for all female population construction in Japanese flounder.
出处
《水产学报》
CAS
CSCD
北大核心
2010年第4期508-514,共7页
Journal of Fisheries of China
基金
国家现代产业技术体系资助项目(nycytx-50-G02)
关键词
牙鲆
真鲷
减数分裂
雌核发育
异源精子
冷休克
Paralichthys olivaceus
Pagrus major
meiosis
gynogenesis
heterogenenos sperm
cold shock
