星斑川鲽雌核发育二倍体、单倍体与普通二倍体及杂交胚胎发育的比较  

Comparison of gynogenetic diploid and haploid embryonic development and hybridization of starry flounder Platichthys stellatusr

作　　者：段会敏[1,2] 田永胜[1,3] 李文龙[1,3] 李振通[1,2] 陈张帆[1,3] 唐江[1,2] 成美玲[1,4] 郭华[1,2] 孙宗哲 陈松林[1,3] 

机构地区：[1]中国水产科学院黄海水产研究所农业部海洋渔业可持续发展重点实验室,山东青岛266071 [2]上海海洋大学水产与生命学院,上海201306 [3]青岛海洋科学与技术国家实验室海洋渔业科学与食物产出过程功能实验室,山东青岛266273 [4]大连海洋大学水产与生命学院,辽宁大连116023 [5]蓬莱市宗哲水产养殖公司,山东蓬莱265617

出　　处：《中国水产科学》2017年第3期477-487,共11页Journal of Fishery Sciences of China

基　　金：青岛市南区科技发展资金项目(2015-6-023-ZH);青岛海洋科学与技术国家实验室鳌山科技创新计划项目(2015ASKJ02)

摘　　要：通过紫外线(UV)对冷冻的鲈(Lateolabrax japonicus)精子进行灭活,利用冷休克和压力休克方法诱导星斑川鲽(Platichthys stellatus)雌核发育二倍体,同时利用灭活鲈精子制备单倍体胚胎,未灭活鲈精子受精制备杂交胚胎,星斑川鲽精子受精制备正常发育胚胎。对以上几种胚胎发育时序、发育生物学特征进行了观察比较。结果表明,卵裂期单倍体、杂交二倍体和雌核发育二倍体胚胎发育速度与普通二倍体胚胎没有明显差异,从低囊胚期开始各实验组胚胎发育速度均慢于普通二倍体胚胎;杂交胚胎在胚体形成期基本死亡,单倍体胚胎在尾芽期停止发育死亡,均不能正常孵出。雌核发育二倍体与普通二倍体具有相似的发育时序,普通二倍体100 h 10 min孵化出膜,而雌核发育二倍体104 h 50 min孵化出膜。雌核发育胚体畸形率(53.59±0.36)%,孵化率(0.11±0.01)%;普通二倍体胚体畸形率(35.11±6.19)%,孵化率(58.01±5.30)%;与普通二倍体相比,雌核发育二倍体胚体畸形率高,孵化率低,但孵化鱼苗能够正常发育,获得了雌核发育群体。本研究为星斑川鲽雌核发育提供了技术方法,同时为单倍体、杂交和雌核发育胚胎的发育生物学研究提供了细胞生物学证据。Artificially induced gynogenesis is a major technique used for controlling the sex ratio of aquatic animals as well as for culturing selectively bred colonies. In this study, cryopreserved sperm of Japanese seabass Lateolabrax japonicus was UV-irradiated and used to fertilize eggs of starry flounder Platichthys stellatus and diploid gynogenesis was induced using cold and hydrostatic pressure shocks. Meanwhile, P. stellatus eggs fertilized with UV-irradiated L. japonicas sperm but without cold or hydrostatic pressure shock were used as the haploid group; P. stellatus eggs fertilized with L. japonicus sperm were used for hybridization; and P. stellatus eggs fertilized with P. stellatus sperm were used as a control group. The results showed that cell division started at a similar point in time in the control group as in the gynogenetic haploids and hybridized embryos. However, as compared with the controls, embryonic development among each of the three experimental groups Was retarded from the late blastula stage. All the hybridized embryos died before formation of an embryonic body; all the haploid embryos died before the heart started to beat. Thus, none of the embryos in these two groups could be successfully hatched. Embryonic development of a small portion of the gynogenetic diploids followed a pattern resembling that of the controls, although with less success： （58.01±5.30）% of embryos in the control group were hatched out at 100 h 10 min, with （35.11±6.19）% malformed; in comparison, only （0.11±0.01）% of the gynogenetic diploids were hatched out at 104 h 50 min, with （53.59±0.36）% malformed. Although the gynogenetic diploids had a very low hatching rate and a relatively high rate of malformation, we were still able to construct a gynogenetic colony of P. stellatus with normally developing larvae. Hence, we propose that this study uncovers a promising technique for P. stellatus gynogenesis, and the results provide abundant cytobiological evidence contributing to information on the developm

关 键 词：星斑川鲽 雌核发育 单倍体 杂交胚胎 胚胎发育 

分 类 号：S917[农业科学—水产科学]