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(diǎn)實(shí)驗(yàn)室 海口 570228；2. 海南省海洋與漁業(yè)科學(xué)院 海口 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(shí)驗(yàn)材料

1.2 實(shí)驗(yàn)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(rùn)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?zèng)_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(shí)心箭頭和空心箭頭所示分別為第5對(duì)和第11對(duì)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

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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(jí)實(shí)驗(yàn)師，E-mail: azsure_caiyan@163.com

2019-01-23,

2019-02-22

* 農(nóng)業(yè)基礎(chǔ)性長(zhǎng)期性科技工作(ZX08S1623)、中央級(jí)科研院所基本科研業(yè)務(wù)費(fèi)專項(xiàng)(20603022018009)和海南省省屬科研院所技術(shù)開發(fā)研究專項(xiàng)(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)