青石斑鱼(♀)×蓝身大斑石斑鱼(♂)杂交后代及其父母本群体的染色体核型分析  

Karyotype analysis of hybrid Epinephelus awoara(♀) and Epinephelus tukula(♂) progenies and their parents

作　　者：陈帅 田永胜[1,2] 成美玲 李振通 李子奇 黎琳琳[1,4] 王林娜 王晓梅 CHEN Shuai;TIAN Yongsheng;CHENG Meiling;LI Zhentong;LI Ziqi;LI Linlin;WANG Linna;WANG Xiaomei(Key Laboratory for Sustainable Development of Marine Fisheries,Ministry of Agriculture and Rural Affairs,Yellow Sea Fisheries Research Institute,Chinese Acadenry of Fishery Sciences,Qingdao 266071,China;Laboratory for Marine Fisheries Science and Food Production Processes,Qingdao National Laboratory for Marine Science and Technology,Qingdao 266071,China;College of Fisheries and Life Science,Shanghai Ocean University,Shanghai 201306,China;Chinese Academy of Agricultural Sciences,Beijing 100081,China;Mingbo Aquatic Co.Ltd.,Laizhou 261400,China)

机构地区：[1]中国水产科学研究院黄海水产研究所,农业农村部海洋渔业可持续发展重点实验室,山东青岛266071 [2]青岛海洋科学与技术试点国家实验室,海洋渔业科学与食物产出过程功能实验室,山东青岛266071 [3]上海海洋大学水产与生命学院,上海201306 [4]中国农业科学院,北京100081 [5]莱州明波水产有限公司,山东莱州261400

出　　处：《中国水产科学》2021年第8期988-1000,共13页Journal of Fishery Sciences of China

基　　金：山东省重点研发计划项目(2019GHY112063);山东省良种工程项目(2019LZGC020);山东省泰山产业领军人才工程,烟台市高端人才引进“双百计划”项目;中国水产科学研究院基本科研业务费项目(2020XT0601);中国水产科学研究院黄海水产研究所基本科研业务费项目(20603022019002,20603022018019).

摘　　要：为了探究杂交后代与双亲染色体组型的差异,证明青石斑鱼(Epinephelusawoara)和蓝身大斑石斑鱼(Epinephelustukula)杂交在染色体水平上的可行性,以及建立石斑鱼外周血淋巴细胞培养制备染色体制片技术,本研究以4月龄和16月龄青石斑鱼(♀)×蓝身大斑石斑鱼(♂)杂交后代、4月龄母本后代和父本为实验材料,通过头肾-秋水仙素法和外周血细胞培养法制备染色体制片,经油镜观察和分裂相统计,结果表明青石斑鱼的染色体数目为2n=48,占比91.00%,核型公式为2n=48t,NF=48;蓝身大斑石斑鱼具两个亚中部着丝粒染色体,染色体数目为2n=48,占比82%,核型公式为2n=2sm+46t,NF=50,臂比为1.76±0.11;杂交后代(E.AT)具一个亚中部着丝粒染色体,染色体数目为2n=48,占比78%,核型公式为2n=1sm+47t,NF=49,臂比为1.75±0.29。同时,本研究对4尾16月龄杂交后代鱼的性腺进行组织切片,观察发现所有个体性腺中具有大量卵母细胞,杂交后代均为雌性,初步说明杂交后代所具有的异形染色体与性别无关,推断异形染色体形成的原因是父本提供了23条端部着丝粒染色体和1条亚中部着丝粒染色体,母本提供了24条端部着丝粒染色体。研究结果为石斑鱼杂交后代异形染色体发生、遗传变异,以及杂交后代性状选育和种质改良提供了丰富的资料。The observation of fish chromosomes is of great significance not only in taxonomy and phylogeny, but also in the study of fish breeding, genetic variation, and sex determination. The traditional method of colchicine injection in vivo requires the dissection of fish to obtain the head kidney, which means high costs for parents and rare fish. In contrast, the method of peripheral blood lymphocyte culture can be used to prepare chromosome specimens with minimal damage to the fish. However, the chromosome specimen of grouper has not been prepared by this method owing to its difficulty. In this study, we prepared the chromosome specimen of grouper successfully by optimizing the experimental conditions: the concentration of colchicine was 0.12 mg/mL, the concentration of PHA was 0.1 mg/mL, the rotational speed was 4000 r/h, and the hypotonic temperature is 34 ℃. In addition, hybrid grouper showed heterosis in terms of faster growth and stronger stress resistance, and chromosome research of hybrid offspring is one of the key methods to explore heterosis. To explore the difference in karyotypes between hybrid progeny and parents, we prepared chromosome specimens of the hybrid progeny(E. AT) and maternal progeny by colchicine injection in vivo. The results showed that the chromosome number of E. tukula was 2 n=48, the karyotype formula was 2 n=2 sm+46 t, NF=50, and the arm ratio was 1.76±0.11;the chromosome number of E. awoara was 2 n=48, the karyotype formula was 2 n=48 t, NF=48;the chromosome number of E. AT was 2 n=48, the karyotype formula was 2 n=1 sm+47 t, NF=49, and the arm ratio was 1.75±0.29. Furthermore, we prepared gonadal sections of 16-month-old E. AT to investigate whether the heterochromosome of E. AT was related to sex. The results showed that there were a large number of oocytes in the gonads, which indicated that the heterochromosomes of E. AT had no relationship with sex. Based on the above results, it is speculated that the cause of heterochromosomes in hybrid progenies is that the male parent prov

关 键 词：青石斑鱼 蓝身大斑石斑鱼 杂交后代 染色体核型 异形染色体 性腺切片 

分 类 号：S961[农业科学—水产养殖]