海南南渡江唇染色体核型和银染分析*

李 安 李高俊 王世锋 柳淑芳 周永灿 申志新 薛献坵 蔡 岩

李 安1李高俊2王世锋1柳淑芳3周永灿1申志新2薛献坵1蔡 岩1①

(1. 海南大学海洋学院 海南省热带水生生物技术重点实验室 海口 570228；2. 海南省海洋与渔业科学院 海口 570100；3. 中国水产科学研究院黄海水产研究所 青岛 266071)

1 材料与方法

1.1 实验材料

1.2 实验方法

1.2.1 染色体标本的制备 染色体标本的制备参考布氏石斑鱼()的染色体标本制备方法(蔡岩等, 2012)，每尾鱼按10 μl/g鱼体重静脉抽血，再按10 μg/g鱼体重剂量腹腔注射植物血凝素(PHA)；3.5 h后，按照1 μg/g鱼体重腹腔注射秋水仙素，15 min后，剪鳃放血，取头肾制备细胞悬液，经500 r/min离心收集细胞，加入0.075 mol/L KCl溶液，37℃水浴低渗30 min，离心后加入新配制的卡诺氏固定液，室温固定20 min，之后重复固定2次。再次离心后取沉淀，加入适量新配制的卡诺氏固定液重悬细胞，用吸管吸取细胞悬液滴在玻片上，过酒精灯火焰3次，自然凉干，备用。

1.2.3 Ag-NORs显带 参考Howell等(1980)的方法，将滤纸铺于培养皿底部，用蒸馏水润湿，上面放2根牙签，置于60℃水浴锅中保温；将制备好的新鲜玻片标本样品面朝上平放在培养皿中，玻片上加20 μl明胶显影液，再加40 μl 50%硝酸银溶液，盖上盖玻片，在水浴锅中温浴至标本呈金褐色为止；用蒸馏水快速冲洗，去除盖玻片，干燥后显微镜观察和拍照。

2 结果

Tab.1 The results of the metaphase chromosome number of H. labeo

Tab.2 The relative length and arm ratio of chromosome of H. labeo (Mean±SD)

图1 唇中期分裂相及染色体核型

图2 唇中期分裂相Ag-NORs

实心箭头和空心箭头所示分别为第5对和第11对染色体的Ag-NORs

Solid arrows and hollow arrows indicated the Ag-NORs on chromosome No.5 and No.11, respectively

3 讨论

Arai R. Fish Karyotypes: A check list. Springer, 2011, 5559

Cai Y, Zhou YC, Xie RM,. A study on the karyotype, Ag-NORs and C-banding inJournal of Fisheries of China, 2012, 36(5): 647–651 [蔡岩, 周永灿, 谢瑞敏, 等. 布氏石斑鱼的染色体核型、银染和C-带. 水产学报, 2012, 36(5): 647651]

Cui J, Ren X, Yu Q. Nuclear DNA content variation in fishes. Cytologia, 1991, 56(3): 425429

Gan GM, Zhang YG. Cytological study on fertilization in of. Acta Hydrobiologica Sinica, 2006, 30(3): 284291

Gu RB, Xu GC, Wen HB,. Chromosome karyotype and nuclear DNA contents of the spotted steed,Journal of Guangdong Ocean University, 2008, 28(1): 12–14 [顾若波, 徐钢春, 闻海波, 等. 花骨鱼染色体组型分析及细胞核DNA含量的测定. 广东海洋大学学报, 2008, 28(1): 12–14]

Howell WM, Black DA. Controlled silver-staining of nucleolus organizer regions with a protective colloidal developer: A 1-step method. Expenertia, 1980, 36(8): 10141015

Le PQ. Taxonomic revision of the Hemibarbus fishes (Cypriniformes: Cyprinidae). Zoological Systematics, 1995, 20(1): 116123 [乐佩琦. 鱼骨属鱼类的分类整理(鲤形目: 鲤科). 动物分类学报, 1995, 20(1): 116123]

Levan A, Fredga K, Sandberg AA. Nomenclature for centromeric position on chromosomes. Hereditas, 1964, 52(2): 201220

Lian QP, Fu GQ, Hu TJ,. Morphological variations in skin carp,and their hybrid F1(♀´♂). Journal of Dalian Ocean University, 2011, 26(6): 493499 [练青平, 宓国强, 胡廷尖, 等. 唇鱼骨、花鱼骨及其杂交F1的形态差异分析. 大连海洋大学学报, 2011, 26(6): 493499]

Liang SZ, Song W, Jiang KJ,Karyotype analysis ofProgress in Fishery Sciences, 2019, 40(4): 172177 [梁述章, 宋炜, 蒋科技, 等. 棘头梅童鱼染色体核型分析. 渔业科学进展, 2019, 40(4): 172177]

Li J, Luo XN, Jin GH,Growth performance and models ofcultured in net cages. Journal of Guangdong Ocean University, 2015, 35(1): 99103 [李军, 骆小年, 金广海, 等. 网箱养殖唇鱼骨的生长性能与生长模型. 广东海洋大学学报, 2015, 35(1): 99103]

Li JX. Study on chromosome karyotype of spotted steed. Journal of Anhui Agricultural Science, 2008, 36(31): 1365813659 [李均祥. 花鱼骨染色体核型的研究. 安徽农业科学, 2008, 36(31): 1365813659]

Li SS, Wang RF, Liu GZ,A karyotype study of eight species of Teleostie fish. Hereditas, 1983(4): 2528, 5152 [李树深, 王蕊芳, 刘光佐, 等. 八种淡水真骨鱼类的核型研究.遗传, 1983(4): 2528, 5152]

Lou YD. Progress of fish karyotype studies in China. Journal of Fisheries of China, 1997(s1): 8296 [楼允东. 中国鱼类染色体组型研究的进展. 水产学报, 1997(s1): 8296]

Martínez-Rodríguez G, Thode G, Álvarez MDC,C-banding and Ag-NOR reveal heterogeneity among karyotypes of serranids (Perciformes). Cytobios, 1989, 58(2): 5360

Medrano L, Bernardi G, Couturier J,. Chromosome banding and genome compartmentalization in fishes .Chromosome, 1988, 96(2): 178183

Ning B, Wu X, Hu YH,. Effects of pH on digestive enzymes in digestive tissues ofJournal of Tianjin Agricultural University, 2013, 20(1): 3436 [宁博, 吴旋, 胡玉花, 等. pH对唇鱼骨消化组织消化酶活性的影响. 天津农学院学报, 2013, 20(1): 3436]

Natarajan R, Subrahmanyam K. A karyotype study of some teleosts from portonovo waters. Proceedings of the Indian Academy of Sciences - Section B, 1974, 79(5): 173196

Nelson JS. Fishes of the world. Quarterly Review of Biology, 1976, 7(1995): 1–16

Perazzo G, Noleto RB, Vicari MR,. Chromosomal studies in, and, (Cichlidae, Perciformes) from extreme Southern Brazil. Reviews in Fish Biology & Fisheries, 2011, 21(3): 509515

Ren X, Cui J, Yu Q. Chromosomal heterochromatin differentiation of cyprinid fishes. Genetica, 1992, 87(1): 4751

Ren X, Yu X. Characterization of nucleolar organizer regions of twelve species of Chinese cyprinid fishes. Caryologia, 2014, 46(23): 201207

Ren XH, Cui JX, Yu QX. Chromosomal nucleolar organizer regions differentiations in Chinese Cyprinid Fishes. Wuhan University Journal of Natural Sciences, 1996, 42(4): 475480 [任修海, 崔建勋, 余其兴. 中国鲤科鱼类染色体核仁组织区研究. 武汉大学学报(自然科学版), 1996, 42(4): 475480]

Ren XH, Cui JX, Yu QX. Studies on NORs of 6S Peeisin Cyprinidae (Pisees). Hereditas, 1993, 15(4): 1113 [任修海, 崔建勋, 余其兴. 六种鲤科鱼类核仁组织者区的研究. 遗传, 1993, 15(4): 1113]

Sharma AK, Shar A. Chromosomes in evolution of eukaryotic groups. CRC Press, 1983, 434(4): 32833292

Shu H, Liu YB, Wei QL,. Studies on chromosome karyotype, Ag-NORs and C-banding patterns of wildands in the Pearl River. Journal of Guangzhou University( Natural Science), 2014, 13(2): 5359 [舒琥, 刘远波, 魏秋兰, 等. 珠江野生草鱼、赤眼鳟的核型、银染和C带比较研究. 广州大学学报(自然科学版), 2014, 13(2): 5359]

Sola L, Innocentiis SD, Gornung E,Cytogenetic analysis of(Pisces: Serranidae), with the chromosome localization of the 18S and 5S rRNA genes and of the (TTAGGG)n telomeric sequence. Marine Biology, 2000, 137(1): 4751

Tan X, Qin JG, Chen B,. Karyological analyses on redclaw crayfish, (Decapoda: Parastacidae). Aquaculture, 2004, 234(1): 6576

Wang DX, Su YQ, Wang SF,. The karyotypes and their polymorphisms of thefrom different populations. Acta Oceanologica Sinica, 2006, 28(6): 176–178 [王德祥, 苏永全, 王世锋, 等. 不同地理种群大黄鱼染色体核型的比较研究. 海洋学报, 2006, 28(6): 176– 178]

Wu X, Jia ZC, Zhang JQ,. Effects of temperature on activity of digestive enzymes in. Journal of Tianjin Agricultural University, 2012, 19(4): 3235 [吴旋, 贾志超, 张家全, 等. 温度对唇鱼骨消化组织消化酶活性的影响. 天津农学院学报, 2012, 19(4): 3235]

Wu XW, Cao WX, Yi BL,. Cyprinidae fish of China (Up and down volume). Shanghai: Shanghai Scientific & Technical Publishers, 1982, 443446 [伍献文, 曹文宣, 易伯鲁, 等. 中国鲤科鱼类志(上下卷). 上海: 上海科学技术出版, 1982, 443446]

Xu L, Wang HJ, Ouyang S,. Studies on the karyotype ofbleeker. Jiangxi Science, 2007, 25(2): 156157 [徐亮, 王海军, 欧阳珊, 等. 花鱼骨染色体的组型分析. 江西科学, 2007, 25(2): 156157]

Xu W, Li CT, Geng LW,. Growth and reproduction of rearedin the Wusuli River. Journal of Fishery Sciences of China, 2009, 16(4): 550556 [徐伟, 李池陶, 耿龙武, 等. 乌苏里江唇鱼骨的全人工繁育. 中国水产科学, 2009, 16(4): 550556]

Yang JQ, He SP, Jörg F,. The phylogenetic relationships of the Gobioninae (Teleostei: Cyprinidae) inferred from mitochondrial cytochromeb, gene sequences. Hydrobiologia, 2006, 553(1): 255266

Yu XJ, Zhou D, Li YC,. Chinese freshwater fish chromosome. Beijing: Science Press, 1989 [余先觉, 周暾, 李渝成, 等.中国淡水鱼类染色体. 北京: 科学出版社, 1989]

The Karyotype and Ag-NORs offrom Nandujiang River of Hainan

LI An1, LI Gaojun2, WANG Shifeng1, LIU Shufang3, ZHOU Yongcan1, SHEN Zhixin2, XUE Xianqiu1, CAI Yan1①

(1. Ocean College, Hainan University, Key Laboratory of Tropical Aquatic Biotechnology of Hainan Province, Haikou 570228; 2. Hainan Academy of Ocean and Fisheries Sciences, Haikou 570100; 3. Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071)

In this study, the karyotype and Ag-NORs of wildfrom Nandujiang River of Hainan were analyzed using the methods of intraperitoneal injection of phytohemagglutinin (PHA) and colchicine, and direct preparation of head kidney cells. Results showed thathad a diploid chromosome number of 50, the karyotype formula was 2n = 18m + 12sm + 12st + 8t, the number of chromosome arms (NF) was 80, no heterotypic chromosome was observed, and Ag-NORs were located on the ends of the short arms of pair No.5 and pair No.11. Comprehensive analyses of the chromosomal evolution in subfamily Gobioninae indicated thathad the primitive karyotype of Gobioninae fish. Our study will not only supplement the cytogenetic data ofand provide information on the taxonomy and evolution of the subfamily Gobioninae, genus, but will also lay the theoretical foundation for germplasm protection and germplasm improvement of.

; Chromosome; Karyotype; Ag-NORs; Gobioninae

Q343.2

A

2095-9869(2020)02-0095-08

蔡 岩，高级实验师，E-mail: azsure_caiyan@163.com

2019-01-23,

2019-02-22

* 农业基础性长期性科技工作(ZX08S1623)、中央级科研院所基本科研业务费专项(20603022018009)和海南省省属科研院所技术开发研究专项(R100030.304; R100263.3)共同资助 [This work was supported by Scientific and Technological Work Information of Agricultural (ZX08S1623), Central Public-Interest Scientific Institution Basal Research Fund (20603022018009), and Hainan Province Scientific Institution Basal Research Fund (R100030.304; R100263.3)]. 李 安，E-mail: 2541431771@qq.com

10.19663/j.issn2095-9869.20190123002

http://www.yykxjz.cn/

Li A, Li GJ, Wang SF, Liu SF, Zhou YC, Shen ZX, Xue XQ, Cai Y. The karyotype and Ag-NORs offrom Nandujiang River of Hainan. Progress in Fishery Sciences, 2020, 41(2): 95–102

CAI Yan, E-mail: azsure_caiyan@163.com

(编辑 冯小花)