| 20种溪蟹线粒体COI基因比较分析及系统发育研究 |
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| 引用本文： | 刘东东,高佳怡,张璐瑶,龚理.20种溪蟹线粒体COI基因比较分析及系统发育研究[J].南方农业学报,2022,53(10):2736-2747. |
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| 作者姓名： | 刘东东 高佳怡 张璐瑶 龚理 |
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| 作者单位： | 浙江海洋大学海洋科学与技术学院/海洋生物种质发掘与利用国家地方研究中心, 浙江舟山 316022 |
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| 基金项目： | 浙江省自然科学基金项目（LY21C190007）；浙江省大学生科技创新活动计划（新苗人才计划）项目（2021R411003） |
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| 摘 要： | 【目的】比较分析20种溪蟹的线粒体COI基因序列，并基于COI基因序列构建溪蟹科系统发育进化树分析不同种类间的亲缘关系，探究COI基因作为分子标记在溪蟹物种鉴定中的适用性，同时为溪蟹科的物种鉴定及系统发育研究提供理论依据。【方法】测定2种龙溪蟹属（Longpotamon）代表种的线粒体COI基因全序列，结合GenBank中已公布的18种溪蟹科COI基因全序列，利用MEGA X计算其碱基组成、保守位点和遗传距离，采用MatGAT 2.02进行多序列相似性比较分析，并以PhyloSuite构建贝叶斯树（BI）和最大似然树（ML），探究溪蟹科物种内部的亲缘关系。【结果】20种溪蟹的COI基因序列全长1534~1539 bp，连续编码511~512个氨基酸残基，所有物种均以ATG为起始密码子；碱基含量略有不同，分别为35.9%~40.7%（T）、16.4%~20.2%（C）、26.8%~28.9%（A）和14.7%~17.2%（G），呈明显的AT偏向性。COI基因核苷酸序列及其推导氨基酸序列比较分析结果显示分别有577和98个变异位点，表明密码子存在简并性。20种溪蟹的线粒体COI基因遗传距离、序列相似性及系统发育进化分析结果均显示，长安龙溪蟹（Longpotamon changanense）与龙溪蟹未定种（Longpotamon sp.）的亲缘关系最近。虽然采用不同方法基于不同数据集构建的系统发育进化树在拓扑结构上有所不同，但所有树型均显示龙溪蟹属（Longpotamon）的小龙溪蟹（L.parvum）并未与该属其他物种聚类在一起，华溪蟹属（Sinolapotamon）、近溪蟹属（Potamiscus）和小石蟹属（Tenuilapotamon）物种也散布在系统发育进化树不同分支中，暗示这些物种在分类鉴定上为非单系群，还需进一步研究确定。【结论】20种溪蟹的线粒体COI基因序列平均种间遗传距离为0.173，均具有区别于其他种类的特异位点，即线粒体COI基因序列可作为溪蟹科物种鉴定的分子标记。
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| 关 键 词： | 溪蟹 线粒体 COI基因 DNA条形码 系统发育 |
| 收稿时间： | 2022-04-11 |
Comparative analysis of mitochondrial COI gene of 20 Potamidae species and its phylogenetic studies |
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| LIU Dong-dong,GAO Jia-yi,ZHANG Lu-yao,GONG Li.Comparative analysis of mitochondrial COI gene of 20 Potamidae species and its phylogenetic studies[J].Journal of Southern Agriculture,2022,53(10):2736-2747. |
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| Authors: | LIU Dong-dong GAO Jia-yi ZHANG Lu-yao GONG Li |
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| Institution: | School of Marine Science and Technology, Zhejiang Ocean University/National and Provincial Joint Laboratory of Exploration and Utilization of Marine Aquatic Genetic Resources, Zhoushan, Zhejiang 316022, China |
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| Abstract: | 【Objective】 To comparatively analyze the sequence differences of mitochondrial COI gene in 20 Potamidae species. The phylogenetic relationship of Potamidae species was constructed based on the COI gene sequences to clarify the evolution of Potamidae, to explore the applicability of COI gene as molecular marker in species identification of Potamidae, and to provide theoretical basis for species identification and phylogenetic studies of Potamidae.【Method】 The complete mitochondrial COI gene sequences of two representative species of Longpotamon were determined. Combined with the complete mitochondrial COI gene sequences of 18 species of Longpotamon published in GenBank, MEGA X was used to calculate the base composition, conserved loci and genetic distance, and MatGAT 2.02 was used to analyze the multi-sequence similarity. And PhyloSuite constructed Bayesian trees(BI) and maximum likelihood trees(ML) to explore the internal phylogenetic relationships of species in Potamidae.【Result】The COI gene sequence of 20 species of Potamidae was 1534-1539 bp, encoding 511-512 amino acid residues, and all species started with ATG as the codon. Base contents were slightly different, which were 35.9%-40.7%(T), 16.4%-20.2%(C), 26.8%-28.9%(A) and 14.7%-17.2%(G), and it was obvious AT bias. The nucleotide sequences of COI gene and its derived amino acid sequences showed 577 and 98 variation sites, respectively, indicating the existence of codon degeneracy. The genetic distance, sequence similarity and phylogenetic evolution of mitochondrial COI genes of 20 species of Potamidae showed that Longpotamon changanense was the closest relative to Longpotamon sp. Although phylogenetic trees constructed by different methods and based on different data sets differed in topology, all tree types showed that L. parvum of Longpotamon was not clustered with other species in the genus. Sinolapotamon, Potamiscus and Tenuilapotamon species were also scattered in the phylogenetic tree, suggesting that these species were non-monophyletic groups in taxonomic identification and needed further study to confirm.【Conclusion】 The mitochondrial COI gene sequences of 20 Potamidae species have average genetic distance of 0.173, and they all have specific sites differing from each other. Therefore, mitochondrial COI gene sequences can be considered as a molecular marker for classification and identification of Potamidae. |
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