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| 1 | 豫北两种虾虎鱼DNA条形码研究显示文摘虾虎鱼是鲈形目一类常见鱼类,广泛分布在我国各个水系,本文以线粒体细胞色素C亚基I(COI)DNA序列为分子标记,通过研究豫北四个采样点虾虎鱼DNA条形码来揭示物种多样性及分子系统发育关系,从而全面了解我省虾虎鱼遗传多样性现状。采用苯酚/氯仿抽提法提取虾虎鱼的基因组,凝胶电泳检测提取效果,采用常用的鱼类COI基因扩增引物扩增目的片段,商业公司送样测序获得其DNA条形码序列。研究共获得2种虾虎鱼(子陵吻虾虎Rhinogobius giurinus和褐吻虾虎Rhinogobius Brunneus)共39条序列,MEGA5.0软件分析结果表明:第三密码子G含量最低,COI基因各位点平均A+T含量要高于G+C含量。以臀斑颌鳞虾虎鱼(Gnatholepis anjerensis)为外类群,本研究涉及的两个物种分别聚成高支持率的单系类群。两种虾虎鱼种间平均遗传距离是0.19,种内平均遗传距离为0.0037,种间平均遗传距离是种内平均遗传距离的54倍,说明采用COI作为分子标记能识别出豫北虾虎鱼物种,为有效的分子标记,同时系统发育分析显示COI可作为分子标记建立虾虎鱼科的系统发育关系。 | 周传江 伍振兴 顾钱洪 孟晓林 聂国兴 | 2015 | 河南水产2015，，4： | 0 |
| 2 | Phylogenetic Analysis of Cynoglossidae in the Yangtze Estuary Based on Partial Sequence of Mitochondrial CO Ⅰ显示文摘To determine the role of mitochondrial COⅠgene in classification and identification of species,a total of 39 single individuals from nine species pertaining to two genera of Cynoglossidae in the Yangtze Estuary were barcoded by COⅠ,sequenced and compared with that of other Cynoglossidae species recorded in the Gen Bank. Total genomic DNA was extracted from each scale sample using the classic phenol / chloroform extraction method. Six hundred and fifty base pairs( bp)COⅠfragments were amplified using the primers 'i. e. ' F1: 5'- TCA ACC AAC CAC AAA GAC ATT GGC AC- 3',R1: 5'- TAG ACT TCT GGG TGG CCA AAG AAT CA- 3'. Every PCR amplification was performed in a total volume of 50 μL of PCR mixture. PCR products were purified and then sequenced in both forward and reverse directions using an ABI PRISMTM 3730 XL Automated Sequencer. DNA sequences were aligned with clustal W using default parameters. Base composition,variable and parsimony informative sites were determined using MEGA 5. 0. Neighbor- joining( NJ) and Maximum parsimony( MP) phylogenetic trees were constructed for COⅠhaplotypes( Kimura 2 Parameter substitution model,K2P; 1 000 bootstraps pseudoreplications) using MEGA 5. 0. Using the MEGA5. 0 software for statistical analysis,the averaged AT content was greater than the GC content( Tab. 2). The GC content of codon position 1 averaged 53. 8%( 51. 8%-57. 3%),which of position 2 for 42. 0%,and that of position 3 ranged from 28. 1% to 37. 8% in average of 32. 4%( Tab. 4). The transitional pairs( si) was slightly more than the transversional pairs( sv),and the ratio( R = si/sv) was 1. 45( Tab. 3). Analysis of the frequency of amino acids in COⅠgene encoding protein showed that the highest frequency of amino acid was leucine,and the lowest frequency of amino acid was tryptophan( Tab. 5). The average K2 P distances pairwise-species and within-species were 0. 191 and 0. 003,respectively( Tab. 6). The K2 P distance pairwise-species was 63. 7 times of that within-species. According to the MP and NJ trees for all 39 sequences,it demonstrated that Cynoglossinae in the Yangtze Estuary was a monophyletic group( Fig. 1). However,the phylogenetic relationships revealed by the COⅠsequence analyses were not consistent with those inferred from morphological classification. In contrast to morphological classification,Paraplagusis japonica of Rhinoplagusia and Cynoglossus robustus of Cynoglossoides were placed in a same clade with support of a high bootstrap value( Fig. 1). The species in the sub-genera of Areliscus clustered on an independent branch. But synonymic phenomena existed in the following two groups of species: C. abbreviatus and C. purpureomaculatus,C. lighti and C. joyner,with the pairwise-species of 0. 002 and 0. 007,respectively( Tab. 6). Our results highlight that the information from COⅠsequences not only can filter out the synonym of the same species,but also be able to be used to carry out effective identification for Cynoglossidae species,which further shows that mitochondrial COⅠis feasible as the classification barcode. | Song Chao Yu Yanan Zhang Tao Yang Gang Zhang Longzhen | 2015 | Animal Husbandry and Feed Science2015，7，6： | 0 |
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