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云纹石斑鱼(♀)×鞍带石斑鱼()杂交后代染色体核型分析
成美玲1,2, 田永胜1,3, 吴玉萍1,2, 李振通1,4, 张晶晶1,4, 王林娜1,3, 李文升5, 马文辉5, 王晓梅5, 翟介明5, 李 波5
1.农业部海洋渔业可持续发展重点实验室 中国水产科学研究院黄海水产研究所 青岛 266071;2.大连海洋大学水产与生命学院 大连 116023;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 青岛 266071;4.上海海洋大学水产与生命学院 上海 201306;5.莱州明波水产有限公司 烟台 261400
摘要:
本研究对云纹石斑鱼(Epinephelus moara♀)×鞍带石斑鱼(Epinephelus lanceolatus♂)杂交后代利用植物血球凝集素(PHA)及秋水仙素通过活体注射法制作染色体标本。选用头肾和鳃组织制作细胞悬浊液,并对冷滴片和热滴片法进行比较,结合空气干燥法制作染色体分裂相玻片,经吉姆萨染色后,在显微镜下观察筛选清晰完整的分裂相。杂交子代(俗称云龙斑)选取75个分裂相,统计染色体数目并进行核型分析。结果显示,云龙斑二倍体染色体数为48,其中,19对染色体为端部着丝粒染色体,2对为亚中部着丝粒染色体,3对为亚端部着丝粒染色体,其染色体臂数(NF)为58,核型公式为2n=48,4sm+6st+38t。杂交后代云龙斑与父母本染色体数目相同,都为2n=48,但云龙斑染色体与其父母本组型不同,在遗传过程中发生了复杂的染色体重新配对和变异。本研究为杂交后代“云龙斑”的种质鉴定、亲缘关系分析、杂交育种等提供了重要的科学根据。
关键词:  云纹石斑鱼  鞍带石斑鱼  云龙斑  染色体PHA注射法  核型分析
DOI:
分类号:
基金项目:
Chromosome Karyotype Analysis of Hybrids of Epinephelus moara♀×Epinephelus lanceolatus♂
CHENG Meiling1,2, TIAN Yongsheng1,3, WU Yuping1,2, LI Zhentong1,4, ZHANG Jingjing1,4, WANG Linna1,3, LI Wensheng5, MA Wenhui5, WANG Xiaomei5, ZHAI Jieming5, LI Bo5
1.Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071;2.College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023;3.National Laboratory of Marine Science and Technology, Laboratory of Marine Fisheries Science and Food Production, Qingdao 266071;4.College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306;5.Laizhou Mingbo Aquatic Co., Ltd., Yantai 261400
Abstract:
In this research, the chromosomal preparation and karyotype of Epinephelus moara♀ × Epinephelus lanceolatus♂ hybrid progeny (commonly known as Yunlong grouper) were studied. The juvenile fish weighing 60~100 g were injected subcutaneously with 6 μg/g phytohemagglutinin (PHA) and 0.1‰ colchicine. After 21 h, the head kidney and gill were dissected for cell suspensions, slices of their chromosome split phase were made by cold and hot drop method combing with air-drying. After being stained with 10% Giemsa, a clear and complete split phase was screened under a microscope. Chromosomes in 75 mitotic phases were selected and the karyotype analysis was performed. The results showed that the karyotype formula was determined to be 2n=48, 4sm+6st+38t, NF=58 in Yunlong grouper. Hybrids Yunlong grouper had the same chromosome number with their parents (2n=48), but their karyotype was different, indicating the complex chromosomal recombination and mutation occurred in the genetic process. This study provided an important scientific basis for the germplasm identification genetic relationship analysis and crossbreeding of Yunlong grouper.
Key words:  Epinephelus moara  Epinephelus lanceolatus  Yunlong grouper  Chromosome PHA injection method  Karyotype analysis