13 种河豚鱼CO I基因部分DNA序列分析及在物种鉴定中的应用

应用CO I基因聚合酶链式反应（polymerase chain reaction，PCR）扩增通用引物，对福建省和海南省搜集的3?属13?种河豚鱼样品与9?个未知种名的河豚鱼样品的CO?I基因靶序列片段进行PCR扩增和测序，13?个已知物种样品的DNA序列提交基因库（GenBank），取得相应的登录号。各物种CO I基因靶序列长度均为681 bp。应用DNAMAN V6软件对样品的DNA序列进行同源性比对分析，建立样品间系统关系同源树。基于CO I基因序列，供试13 个样品被划分为4?个类群组。群间的同源率为84%，群内同源率为90%～100%。根据序列同源性分析结果，9?个未知种名的样品被归类到2?个类群组中，判定这9?个样品为东方鲀属或腹刺鲀属，其中1?个样品（HNW2）为月腹刺鲀，4?个样品（HNW3、HNW4、FJW2、FJW5）为棕斑腹刺鲀，1?个样品（HNW1）为暗鳍腹刺鲀；3?个样品（FJW1、FJW3、FJW4）为横纹东方鲀。探讨基于DNA条形码技术的CO I基因靶序列片段在河豚鱼种属鉴别中应用的可能性。

Analysis of Partial DNA Sequence of CO I Gene and Its Application for Species Identification of Thirteen Species of Puffer Fish

Partial fragments of the cytochrome oxidase subunit I (CO I) genes from13 species in 3 genera of puffer fish and 9 samples from unknown species collected in Fujian province and Hainan province were amplified by polymerase chain reaction (PCR) using universal primers and the PCR products were sequenced. The DNA sequences of 13 known species were submitted to the GenBank with accession numbers. The target gene sequence was 681 bp in length for all species. Sequence alignment was carried out using DNAMAN V6 software, and a phylogenetic tree among species was established. Thirteen known species were divided into 4 groups, with 84% homology between the groups and 90%–100% within the groups. Nine samples of unknown puffer fish species were classed into 2 groups: the Takifugu and Gastrophysus genera; HNW2 was identified as Gastrophysus lunaris, HNW3, HNW4, FJW2, and FJW5 as G. spadiceus, HNW1 as G. gloveri and FJW1, FJW3 and FJW4 as T. oblongus.