| COⅠ和16S rRNA基因在高原裂腹鱼物种鉴定中的应用 |
| Application of COⅠand 16S rRNA Gene for Identification of Tibetan Plateau Schizothorax Species |
| 投稿时间：2015-02-09 修订日期：2015-09-21 |
| DOI：10.15928/j.1674-3075.2015.04.014 |
| 中文关键词:COⅠ 16S rRNA 高原裂腹鱼 物种鉴定 |
| 英文关键词:COⅠ 16S rRNA plateau Schizothorax fish species identification |
| 基金项目:国家自然科学基金重点项目（31130049）；教育部科学技术研究项目（213013A）；上海市教委科研创新项目曙光计划（13SG51）；上海市教委水产学一流学科项目。 |
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| 中文摘要: |
| 针对裂腹鱼种类繁多、种间形态学差异不明显的特点，利用分子标记对高原裂腹鱼进行准确的物种鉴定具有重要价值。对采集的60尾高原裂腹鱼进行了COⅠ和16S rRNA基因的部分序列测定，并通过GenBank数据库进行鱼种鉴定；运用邻接法（NJ）构建系统发育树，对实验鱼进行群系划分；通过DNASP软件分析比较实验鱼COⅠ和16S rRNA基因的序列变异；依据MEGA 6.0的Kimura-2-parameter模型计算实验鱼的种内遗传距离和种间遗传距离，并对COⅠ和16S rRNA基因在高原裂腹鱼物种鉴定中的适用性进行探讨。结果表明，采集的样本包含3属、5种裂腹鱼，分别为尖裸鲤属的尖裸鲤（Oxygymnocypris stewartii）、裸鲤属的软刺裸鲤（Gymnocypris dobula）、裂腹鱼属的拉萨裂腹鱼（Schizothorax waltoni）、巨须裂腹鱼（Schizothorax macropogon）和异齿裂腹鱼（Schizothorax oconnori）。在COⅠ基因构建的系统发育树中，5种高原裂腹鱼均形成独立的一支，而16S rRNA基因不能准确地对裂腹鱼属的3个鱼种实现区分。研究显示，5种高原裂腹鱼COⅠ基因核苷酸的变异水平（Pi）和单倍型多样性指数（Hd）均高于16S rRNA基因；COⅠ基因计算得到实验鱼种间遗传距离和种内遗传距离平均值分别为0.025和0.002，种内和种间的最大遗传差异约为13倍；16S rRNA基因得出结果为0.026和0.005，而种内和种间的最大遗传差异约为6倍。综合COⅠ和16S rRNA基因能有效鉴定高原裂腹鱼物种，而在单个基因的选择上，COⅠ基因具有更高的适用性。 |
| 英文摘要: |
| The plateau Schizothorax species are distributed primarily on the Tibetan plateau and adjacent waters. Schizothorax species vary widely and possess high scientific and economic value, but the morphological differences between species are not obvious. Therefore, the ability to identify plateau Schizothorax species using molecular markers would be very useful. In this study, 60 Schizothorax species collected from the Tibetan plateau were identified based on their 16S rRNA and COⅠ base sequences. Based on our results, the effectiveness of using the two genes for identifying plateau Schizothorax species was examined. The 60 Schizothorax samples for the study were selected from 200 Schizothorax collected in August, 2012 and May and September, 2013 and labeled S1- S60. The 16S rRNA and COⅠ base sequences of the 60 fish were extracted and amplified using PCR. The PCR products of 16S rRNA and COⅠ genes were then sequenced and the sequences were used for species identification by comparison with the GenBank database. Results indicate that the 60 Schizothorax were from 3 genera and 5 species; Oxygymnocypris stewartii, Gymnocypris dobula, Schizothorax waltoni, Schizothorax macropogon and Schizothorax oconnori. Phylogenetic trees of the 60 Schizothorax, based on COⅠ and 16S rRNA base sequences, were constructed using the neighbor-joining method. The phylogenetic trees show that the five plateau Schizothorax species form an independent clade based on COⅠ base sequences, but the 16S rRNA sequence failed to separate three Schizothorax species. The nucleotide diversity (Pi) and haploid type diversity (Hd) of the COⅠ gene, measured by DNASP software, were higher than those of the 16S rRNA gene. In addition, the interspecific and intraspecific genetic distances of the two genes were calculated by Kimura-2-parameter model from MEGA 6.0. The average interspecific and intraspecific genetic distances of COⅠ gene for the five Schizothorax species were 0.025 and 0.002 and the largest genetic difference between them was about a factor of 13. However, the average interspecific and intraspecific genetic distances of 16S rRNA gene were 0.026 and 0.005 and the largest genetic difference was about a factor of 6. In conclusion, species identification of plateau Schizothorax can be accomplished effectively by combining the COⅠ and 16S rRNA genes and the COⅠ gene is more sensitive than the 16S rRNA gene for discriminating among the closely related plateau Schizothorax species. |
| 产久林,姜华鹏,刘一萌,胡星星,王丛丛,许强华.2015.COⅠ和16S rRNA基因在高原裂腹鱼物种鉴定中的应用[J].水生态学杂志,36(4):98-104. |
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