牙鲆同质雌核发育人工诱导研究  

Study on artificial induction of mitogynogenetic diploid in Paralichthys olivaceus

作　　者：尤锋[1] 许建和[1,2] 倪静[1] 孙威[1] 朱香萍[1,3] 徐冬冬[1] 徐永立[1] 张培军[1] 

机构地区：[1]中国科学院海洋研究所,青岛266071 [2]淮海工学院,连云港222005 [3]青岛农业大学,青岛266109

出　　处：《高技术通讯》2008年第8期874-880,共7页Chinese High Technology Letters

基　　金：863计划(2006AA10A404、2002AA629160);国家自然科学基金(30571445)资助项目

摘　　要：采用紫外线照射对牙鲆(Paralichthys olivaceus)精子遗传物质进行完全灭活,灭活精子与牙鲆卵子授精获得雌核发育单倍体受精卵,再通过冷休克或静水压法对其进行倍性恢复,成功诱导出同质雌核发育牙鲆。牙鲆精子遗传灭活的最适紫外照射剂量为3.6×10^(-3)J/mm^2(照射时间为180s);(15±0.2)℃水温下冷休克法诱导的最适条件为受精后85min,0～2℃水温处理45min;静水压法的最适诱导参数为受精后85min,600kg/cm^2压力处理6min,诱导效率静水压法明显高于冷休克法。两种方法诱导组胚胎发育较对照组在原肠期前都有明显滞后现象,开口前同质雌核发育仔鱼比异质雌核发育仔鱼存活率低,但二者全长生长无显著差异(P<0.05)。这些结果为牙鲆纯系建立及其应用提供了重要依据。Gynogenetic haploids of Paralichthys olivaceus were obtained by activating eggs development with the ultraviolet light （UV）-irradiated homologous sperm and then mitogynogenetic diploids were induced successfully by inhibiting the early egg cleavage with the application of cold shock or pressure shock. The results of UV irradiation experiments revealed that the optimal UV dosage for genetic inactivation of P. olivaceus sperm was 3.6 × 10^-3 J/mm^2（180 s）. At （15 ± 0.2）℃, the treatment optima were 1 - 2℃ for 45min at 85min after fertilization for cold shock and 600kg/cm^2 for 6min at the same treatment timing for pressure shocks. As far as the treatment methods were concerned, pressure shock was more effective than cold shock to inhibit the egg cleavage. Compared with diploid controls, embryonic developments in both shock groups were suppressed before the end of blastula stage. Du.ring 1 - 4d after hatching, survival rates of mitogynogenetic diploid larvae were markedly lower than those of their diploid control and meiogynogenetic counterparts even though no significant differences among them were observed in total length of larvae before first feeding （ P 〈 0.05）. These above results provide the basic data for the establishment and usage of pure lines in the cultured P. olivaceus.

关 键 词：牙鲆 同质雌核发育二倍体 人工诱导 UV 冷休克和静水压 

分 类 号：S917.4[农业科学—水产科学]