Molecular Phylogeny Determined by Mitochondrial Genes 16S rRNA, COX1 and Cytb of Eleven Anemonefish Species
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摘要: 采用线粒体16S rRNA、COX1和Cytb基因测定11种小丑鱼的序列，分析其基因序列差异和遗传距离关系，并用邻接法、最大似然法和贝叶斯法构建系统进化树，探讨了11种小丑鱼的系统发育关系。序列分析结果显示，经比对处理后的3个基因总长度4 388 bp，其中多态位点1 028个，简约信息位点616个，转换与颠换比为2.12，A+T的平均含量为54.7%，高于C+G的平均含量45.3%。系统发育关系表明：（1）双锯鱼亚科为单系发生，但双锯鱼属不为单系发生，建议对棘颊雀鲷属重新定位；在双锯鱼属中，公子小丑鱼Amphiprion ocellaris和黑边公子小丑鱼Amphiprion percula聚为一支，为双锯鱼亚科分子亲缘关系基础；（2）透红小丑鱼Premnas biaculeatus在双锯鱼属的定位较为模糊，贝叶斯树显示透红小丑鱼P.biaculeatus与公子小丑鱼A.ocellaris和黑边公子小丑鱼A.percula聚成一支并位于该支的底端，而NJ树和ML树中透红小丑鱼却没能和公子小丑鱼A.ocellaris和黑边公子小丑鱼A.percula聚为一支，而是和其他小丑鱼汇聚成一支；（3）透红小丑鱼P.biaculeatus与金透红小丑鱼Premnas epigrammata是同一种。Abstract: Sequences of mitochondrial 16S rRNA, COX1 and Cytb genes were determined to establish the phylogenetic relationship among 11 species of anemonefish using the neighbor-joining (NJ), maximum-likelihood (ML) and Bayesian methods with the divergence and genetic distances determined as well. The 3 genes had a combined length of 4 388 bp that consisted of 1 028 polymorphic and 616 parsimonious-information sites. The ratio of transition to transversion was 2.12, and the average A+T content was 54.7%, which was higher than 45.3% for C+G in the genes. It was found that (1) the subfamily Amphiprioninae was monophyly, but the genus Amphiprion was not monophyletic, implying Premnas should be reclassified; (2) the phylogenetic trees grouped the percula complex into a clade that formed the ancestral taxon of anemonefish, but the phylogenetic relationship was uncertain in regard to Premnas biaculeatus; and, while the Bayesian tree placed the percula complex and P. biaculeatus in a same clade, NJ and ML tree classified P. biaculeatus in a clade with subgenera Amphiprion, Paramphiprion and Phalerebus; and, (3) the genetic distance between species suggested a possibility of P. biaculeatus and Premnas epigrammata being identical.
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Key words:
- 16S rRNA gene /
- COX1 gene /
- Cytb gene /
- anemonefish /
- genetic differences /
- phylogenetic evolution
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图 1 基于16S rRNA、COX1 和Cytb基因序列构建的NJ树
Figure 1. NJ phylogenetic tree of anemonefish based on mitochondrial 16S rRNA, COX1 and Cytb genes
图 2 基于16S rRNA、COX1和Cytb基因序列构建的ML树
Figure 2. ML phylogenetic tree of anemonefish based on mitochondrial 16S rRNA, COX1 and Cytb genes
图 3 基于16S rRNA、COX1 和Cytb基因序列构建的贝叶斯树
Figure 3. Bayesian phylogenetic tree of anemonefish based on mitochondrial 16S rRNA, COX1 and Cytb genes
表 1 小丑鱼种类及采集信息
Table 1. Sampling and species of anemonefish under study
种类 拉丁名 亚属 采集位置 鞍背小丑鱼 Amphiprion polymnus Paramphiprion 菲律宾海域 公子小丑鱼 Amphiprion ocellaris Actinicola 菲律宾海域 黑边公子小丑鱼 Amphiprion percula Actinicola 印尼巴东海域 黑红小丑鱼 Amphiprion melanopus Amphiprion 菲律宾海域 印度红番茄小丑鱼 Amphiprion ephippium Amphiprion 菲律宾海域 红小丑鱼 Amphiprion frenatus Amphiprion 菲律宾海域 双带小丑鱼 Amphiprion clarkii Amphiprion 菲律宾海域 宽带小丑鱼 Amphiprion latezonatus Paramphiprion 菲律宾海域 银线小丑鱼 Amphiprion akallopisos Phalerebus 菲律宾海域 金透红小丑鱼 Premnas epigrammata* premnas 印尼巴东海域 透红小丑鱼 Premnas Biaculeatus premnas 菲律宾海域 注：*表示对学名说法不一，有人称之为The Gold Stripe Maroon Clownfish。 
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表 2 11种小丑鱼间的遗传距离
Table 2. Genetic distances among 11 species of anemonefish according to kimura-2-parameters model
项目 银线小丑 双带小丑 印度红番茄小丑 红小丑 宽带小丑 黑红小丑 公子小丑 黑边公子小丑 鞍背小丑 透红小丑 金透红小丑 银线小丑 双带小丑 0.046 印度红番茄小丑 0.029 0.048 红小丑 0.029 0.048 0.009 宽带小丑 0.039 0.052 0.032 0.031 黑红小丑 0.030 0.047 0.010 0.002 0.030 公子小丑 0.098 0.100 0.094 0.094 0.098 0.094 黑边公子小丑 0.098 0.099 0.093 0.093 0.098 0.093 0.002 鞍背小丑 0.038 0.052 0.030 0.028 0.020 0.028 0.094 0.093 透红小丑 0.096 0.098 0.096 0.095 0.101 0.094 0.093 0.093 0.098 金透红小丑 0.093 0.097 0.093 0.091 0.098 0.091 0.091 0.091 0.095 0.010
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