Molecular Identification of Juvenile Eels by DNA Fingerprints
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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。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.
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Key words:
- eel /
- juveniles /
- COI /
- molecular identification
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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 -
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