海南南渡江唇染色體核型和銀染分析*

李 安 李高俊 王世鋒 柳淑芳 周永燦 申志新 薛獻(xiàn)坵 蔡 巖

李 安1李高俊2王世鋒1柳淑芳3周永燦1申志新2薛獻(xiàn)坵1蔡 巖1①

(1. 海南大學(xué)海洋學(xué)院 海南省熱帶水生生物技術(shù)重點實驗室 ?？?570228；2. 海南省海洋與漁業(yè)科學(xué)院 ?？?570100；3. 中國水產(chǎn)科學(xué)研究院黃海水產(chǎn)研究所 青島 266071)

1 材料與方法

1.1 實驗材料

1.2 實驗方法

1.2.1 染色體標(biāo)本的制備 染色體標(biāo)本的制備參考布氏石斑魚()的染色體標(biāo)本制備方法(蔡巖等, 2012)，每尾魚按10 μl/g魚體重靜脈抽血，再按10 μg/g魚體重劑量腹腔注射植物血凝素(PHA)；3.5 h后，按照1 μg/g魚體重腹腔注射秋水仙素，15 min后，剪鰓放血，取頭腎制備細(xì)胞懸液，經(jīng)500 r/min離心收集細(xì)胞，加入0.075 mol/L KCl溶液，37℃水浴低滲30 min，離心后加入新配制的卡諾氏固定液，室溫固定20 min，之后重復(fù)固定2次。再次離心后取沉淀，加入適量新配制的卡諾氏固定液重懸細(xì)胞，用吸管吸取細(xì)胞懸液滴在玻片上，過酒精燈火焰3次，自然涼干，備用。

1.2.3 Ag-NORs顯帶 參考Howell等(1980)的方法，將濾紙鋪于培養(yǎng)皿底部，用蒸餾水潤濕，上面放2根牙簽，置于60℃水浴鍋中保溫；將制備好的新鮮玻片標(biāo)本樣品面朝上平放在培養(yǎng)皿中，玻片上加20 μl明膠顯影液，再加40 μl 50%硝酸銀溶液，蓋上蓋玻片，在水浴鍋中溫浴至標(biāo)本呈金褐色為止；用蒸餾水快速沖洗，去除蓋玻片，干燥后顯微鏡觀察和拍照。

2 結(jié)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)

圖1 唇中期分裂相及染色體核型

圖2 唇中期分裂相Ag-NORs

實心箭頭和空心箭頭所示分別為第5對和第11對染色體的Ag-NORs

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

3 討論

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 [蔡巖, 周永燦, 謝瑞敏, 等. 布氏石斑魚的染色體核型、銀染和C-帶. 水產(chǎn)學(xué)報, 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 [顧若波, 徐鋼春, 聞海波, 等. 花骨魚染色體組型分析及細(xì)胞核DNA含量的測定. 廣東海洋大學(xué)學(xué)報, 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 [樂佩琦. 魚骨屬魚類的分類整理(鯉形目: 鯉科). 動物分類學(xué)報, 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 [練青平, 宓國強, 胡廷尖, 等. 唇魚骨、花魚骨及其雜交F1的形態(tài)差異分析. 大連海洋大學(xué)學(xué)報, 2011, 26(6): 493499]

Liang SZ, Song W, Jiang KJ,Karyotype analysis ofProgress in Fishery Sciences, 2019, 40(4): 172177 [梁述章, 宋煒, 蔣科技, 等. 棘頭梅童魚染色體核型分析. 漁業(yè)科學(xué)進(jìn)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 [李軍, 駱小年, 金廣海, 等. 網(wǎng)箱養(yǎng)殖唇魚骨的生長性能與生長模型. 廣東海洋大學(xué)學(xué)報, 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. 花魚骨染色體核型的研究. 安徽農(nóng)業(yè)科學(xué), 2008, 36(31): 1365813659]

Li SS, Wang RF, Liu GZ,A karyotype study of eight species of Teleostie fish. Hereditas, 1983(4): 2528, 5152 [李樹深, 王蕊芳, 劉光佐, 等. 八種淡水真骨魚類的核型研究.遺傳, 1983(4): 2528, 5152]

Lou YD. Progress of fish karyotype studies in China. Journal of Fisheries of China, 1997(s1): 8296 [樓允東. 中國魚類染色體組型研究的進(jìn)展. 水產(chǎn)學(xué)報, 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 [寧博, 吳旋, 胡玉花, 等. pH對唇魚骨消化組織消化酶活性的影響. 天津農(nóng)學(xué)院學(xué)報, 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, 崔建勛, 余其興. 中國鯉科魚類染色體核仁組織區(qū)研究. 武漢大學(xué)學(xué)報(自然科學(xué)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, 崔建勛, 余其興. 六種鯉科魚類核仁組織者區(qū)的研究. 遺傳, 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, 劉遠(yuǎn)波, 魏秋蘭, 等. 珠江野生草魚、赤眼鱒的核型、銀染和C帶比較研究. 廣州大學(xué)學(xué)報(自然科學(xué)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, 蘇永全, 王世鋒, 等. 不同地理種群大黃魚染色體核型的比較研究. 海洋學(xué)報, 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 [吳旋, 賈志超, 張家全, 等. 溫度對唇魚骨消化組織消化酶活性的影響. 天津農(nóng)學(xué)院學(xué)報, 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 [伍獻(xiàn)文, 曹文宣, 易伯魯, 等. 中國鯉科魚類志(上下卷). 上海: 上?？茖W(xué)技術(shù)出版, 1982, 443446]

Xu L, Wang HJ, Ouyang S,. Studies on the karyotype ofbleeker. Jiangxi Science, 2007, 25(2): 156157 [徐亮, 王海軍, 歐陽珊, 等. 花魚骨染色體的組型分析. 江西科學(xué), 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 [徐偉, 李池陶, 耿龍武, 等. 烏蘇里江唇魚骨的全人工繁育. 中國水產(chǎn)科學(xué), 2009, 16(4): 550556]

Yang JQ, He SP, J?rg 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 [余先覺, 周暾, 李渝成, 等.中國淡水魚類染色體. 北京: 科學(xué)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 巖，高級實驗師，E-mail: azsure_caiyan@163.com

2019-01-23,

2019-02-22

* 農(nóng)業(yè)基礎(chǔ)性長期性科技工作(ZX08S1623)、中央級科研院所基本科研業(yè)務(wù)費專項(20603022018009)和海南省省屬科研院所技術(shù)開發(fā)研究專項(R100030.304; R100263.3)共同資助 [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

(編輯 馮小花)