鳗鲡幼苗种类的DNA指纹鉴定

林能锋; 陈强; 杨金先; 许斌福; 曾红

doi:10.19303/j.issn.1008-0384.2017.03.006

鳗鲡幼苗种类的DNA指纹鉴定

doi: 10.19303/j.issn.1008-0384.2017.03.006

林能锋^1,,
陈强¹,
杨金先¹,
许斌福¹,
曾红^2, ,

1.
福建省农业科学院生物技术研究所，福建福州 350003
2.
福建师范大学生命科学学院，福建福州 350108

基金项目:

福建省科技计划项目——省属公益类科研院所基本科研专项 2015R1019-11

详细信息

作者简介:
林能锋 (1972-)，男，博士，副研究员，主要从事海洋动物遗传育种与水产动物病害防治研究 (E-mail:lnfeng@hotmail.com)

通讯作者:
曾红 (1972-)，女，博士，副教授，主要从事动物细胞生物学研究 (E-mail:zhbil@fjne.edu.cn)

中图分类号: Q935
计量
- 文章访问数: 1634
- HTML全文浏览量: 128
- PDF下载量: 211
- 被引次数: 0
出版历程
- 收稿日期: 2016-08-22
- 修回日期: 2017-01-10
- 刊出日期: 2017-03-01

Molecular Identification of Juvenile Eels by DNA Fingerprints

1.
Institute of Biotechnology, Fujian Academy of Agriculture Sciences, Fuzhou, Fujian 350003, China
2.
College of Life Sciences, Fujian Normal University, Fuzhou, Fujian 350117, China

摘要

摘要: 养殖鳗鲡种苗来源日趋多样，确定鳗苗种类是决定养殖成败的关键因素之一。应用鳗鲡属鱼类mtDNA COI基因的保守序列设计一对特异性引物，对21批次，共109个白苗期鳗鲡样品进行种类鉴定。结果显示，应用该引物可以在所采集到的鳗鲡样品中扩增出652 bp特异性基因片段，将样品COI基因序列与BOLD数据库中鳗鲡COI基因标准序列进行比对，共鉴定得7个种类归属，分别为日本鳗鲡A.japonica、美洲鳗鲡A.rostrata、欧洲鳗鲡A.anguilla、花鳗鲡A.marmorata、莫桑比克鳗鲡A.mossambica、吕宋鳗鲡A.luzonensis和太平洋双色鳗A.bicolor pacifica。对COI基因序列进行聚类分析，得到7个有很高节点支持率的主要类群。结果证实mtDNA COI基因在鳗鲡种类鉴定上具有很好的实用性。此外，在种类的鉴定中发现其中1批次的样品定种错误，有1个批次的鳗鲡样品中存在有2个种类的混杂，提示在实际的苗种养殖及交易中，对鳗鲡的种类鉴定上仍存在一定的错误和混杂，有必要引入DNA分子标记技术进行更加方便和准确的鉴定。
- 鳗鲡 /
- 幼苗 /
- COI基因 /
- 分子鉴定
Abstract: Accurate species identification is crucial for eel aquaculture, especially when increasing taxa of the fish were raised in the eel farms in China. This study applied a pair of specific primers from the conservative fragment of mtDNA COI gene in Anguilla to amplify 109 DNA samples extracted from 21 batches of eel juveniles for taxon identification. It was found that a specific 652bp fragment could be amplified from all of the samples. The BOLD Identification System (Barcoding of Life Data Systems: www.boldsystems.org) was used for sequence similarity search on the COI genes. Seven species, including A. japonica, A. rostrata, A.anguilla, A.marmorata, A.mossambica, A.luzonensis and A.bicolor pacifica, were identified. The cluster analysis showed 7 main clades with high rate of support at the nodes. It suggested that the COI identification method could be satisfactorily employed for eel juveniles identification. Incidentally, during the course of this study a former mistaken identification on a batch of juvenile fish was discovered. Therefore, the DNA barcode-based identification method was highly recommended for implementation to benefit the commercial trade and eel aquaculture.
- eel /
- juveniles /
- COI /
- molecular identification

HTML全文

图 1 鳗鲡mtDNA COI基因序列的聚类分析

Figure 1. Cluster analysis on mtDNA COI sequences of Anguilla

下载: 全尺寸图片幻灯片

表 1 基于mtDNA COI基因序列的7种鳗鲡种内及种间遗传距离

Table 1. Genetic distances among 7 eel species according to partial mtDNA COI gene sequence

eel	A.rostrata	A.anguilla	A.mossambica	A.marmorata	A.luzonensis	A.bicolor pacifica	A.japonica
A.rostrata	0.004
A.anguilla	0.033	0.007
A.mossambica	0.075	0.081	0.003
A.marmorata	0.073	0.082	0.070	0.004
A.luzonensis	0.078	0.082	0.076	0.035	0.005
A.bicolor pacifica	0.087	0.090	0.081	0.053	0.061	0.019
A.japonica	0.087	0.094	0.086	0.058	0.062	0.087	0.003

下载: 导出CSV

参考文献(15)

[1]	HEBERT P D N, RATNASINGHAM S, DEWAARD J R. Barcoding animal life:cytochrome c oxidase subunit 1 divergences among closely related species[J]. Proceedings:Biological Sciences, 2003, 270:S96-S99. doi: 10.1098/rsbl.2003.0025
[2]	WARD R D, HANNER R, HEBERT P D N. The campaign to DNA barcode all fishes, FISH-BOL[J].Journal of Fish Biology, 2009, 74(2):329-356. doi: 10.1111/jfb.2009.74.issue-2
[3]	林能锋, 苏永全, 丁少雄, 等.大黄鱼微卫星标记引物在石首鱼科几个近缘种中的通用性研究[J].中国水产科学, 2008, 15(2):237-243. http://www.cnki.com.cn/Article/CJFDTOTAL-ZSCK200802007.htm
[4]	UNTERGASSER A, CUTCUTACHE I, KORESSAAR T, et al.Primer3-new capabilities and interfaces[J]. Nucleic Acids Research, 2012, 40(15):e115. doi: 10.1093/nar/gks596
[5]	KORESSAAR T, REMM M.Enhancements and modifications of primer design program Primer3[J].Bioinformatics, 2007, 23(10):1289-1291. doi: 10.1093/bioinformatics/btm091
[6]	RATNASINGHAM S and HEBERT P D N. BOLD:The barcode of life data system (www.barcodinglife.org)[J]. Molecular Ecology Note, 2007, 7:355-364. doi: 10.1111/j.1471-8286.2007.01678.x
[7]	KUMAR S, STECHER G and TAMURA K. MEGA7:Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets[J]. Molecular Biology and Evolution, 2016, 33(7):1870-1874. doi: 10.1093/molbev/msw054
[8]	BASTROP R, STREHLOW B, JURSS K, STURMBAUER C. A new molecular phylogenetic hypothesis for the evolution of freshwater eels[J]. Molecular Phylogenetics and Evolution, 2000, 14(2):250-258. doi: 10.1006/mpev.1999.0694
[9]	AOYAMA J, NISHIDA M, TSUKAMOTO K. Molecular phylogeny and evolution of the freshwater eel, genus Anguilla[J]. Molecular Phylogenetics and Evolution, 2001, 20(3):450-459. doi: 10.1006/mpev.2001.0959
[10]	罗土炎, 饶小珍, 林岗, 等. 4种鳗鲡同工酶的比较研究[J].福建农业学报, 2009, 24(5):421-428. http://www.fjnyxb.cn/CN/abstract/abstract385.shtml
[11]	杨弘, 王希道, 吴婷婷, 等.用RAPD技术研究3种鳗鱼的种质鉴定[J].中国水产科学, 2002, 9(3):269-272. http://www.cnki.com.cn/Article/CJFDTOTAL-ZSCK200203018.htm
[12]	张新艳, 樊海平, 范斯敏, 等.应用RAPD技术对6种玻璃鳗的种质鉴定和遗传多样性分析[J].江西农业大学学报, 2010, 32(4):683-688. http://www.cnki.com.cn/Article/CJFDTOTAL-JXND201004013.htm
[13]	梁俊, 李道季, 卢莉琼.日本鳗鲡 (Anguilla japonica) 和欧洲鳗鲡 (A.anguilla) 的微卫星差异[J].海洋与湖沼, 2003, 34(4):414-421. doi: 10.11693/hyhz200304008008
[14]	HEBERT P D N, CYWINSKA A, BALL S L, et al. Biological identifications through DNA barcodes[J]. Proc.R. Soc. Lond.B, 2003, 70:313-321.
[15]	吴宁, 黎中宝, 林小云, 等. 6种鳗鲡 (Anguilla) 线粒体DNA COI序列的比较研究[J].海洋与湖沼, 2010, 41(6):930-934. doi: 10.11693/hyhz201006020020