泰山螭霖鱼线粒体COⅠ基因序列的遗传多样性分析

摘要

[目的]通过克隆泰山螭霖鱼的COⅠ基因序列，研究其与鲤科几属鱼类的进化关系，确定泰山螭霖鱼的分类地位。[方法]以泰山螭霖鱼30个个体为材料，对其线粒体细胞色素氧化酶亚基Ⅰ（ COⅠ）的部分序列进行PCR扩增和序列对比，检测泰山螭霖鱼的遗传多样性；计算泰山螭霖鱼不同单倍型与GenBank中鲤科5个属鱼类的遗传距离，采用聚类分析方法，构建NJ和UPGMA系统进化树。[结果]30个个体泰山螭霖鱼线粒体COⅠ基因片段长度约1600 bp，无碱基的插入和缺失，纯化并去除引物序列后获得约1493 bp的核苷酸序列，存在6种单倍型，共检测到6个多态位点，包括4个转化、2个颠换，其单倍型多样性（Hd）为0．460，核苷酸多样性（Pi）为0．00078，平均核苷酸差异数（K）1．163。遗传距离和NJ和UPGMA系统进化树表明，泰山螭霖鱼与多鳞白甲鱼的COⅠ序列一致，泰山螭霖鱼所在的突吻鱼属与光唇鱼属的亲缘关系最近，与金线鲃属、倒刺鲃属、华鲮属的亲缘关系较远。[结论]该研究为其种质资源的管理与保护及遗传育种提供分子生物学依据。%Objective] Through cloning COⅠ gene sequence of Varicorhinus macrolepis, its evolutionary relationship with other genus fish in Cyprinidae was studied, the classification status of V.macrolepis was determined.[ Method] 30 V.macrolepis were used as experimental mate-rials, mitochondrial cytochrome oxidase subunitⅠ( COⅠ) gene fragment was amplified using PCR technique and aligned of sequence, genetic diversity of V.macrolepis was detected,and genetic distance between different haplotypes of V.macrolepi and 5 genus of Cyprinidae in Gen-Bank was calculated.Clustering analysis was used to build NJ and UPGMA phylogenetic tree.[ Result] The results showed that the COⅠgene fragment PCR products of 30 V.macrolepis were about 1 600 bp, and no insertion/deletionwas observed, 1 493 bp nucleotide sequences were determined after being purified and excluded the primer sequence.There were 6 haplotypes, and 6 polymorphic sites were detected, including 4 transition sites and 2 transversion sites.Haplotype diversity (Hd) of V.macrolepis was 0.460, nucleotide diversity (Pi) was 0.000 78, av-erage number of nucleotide differences ( K) was 1.163.Genetic distance and NJ UPGMA phylogenetic trees showed that the sequences of V.macrolepis and O.macrolepis are the same.The genetic relationship between Varicorhinus and Acrossocheilus was the closest, while the ge-netic relationship between Varicorhinus and Spinibarbus, Sinilabeo,Sinocyclocheilus were far.[ Conclusion] The study can provide molecular biology basis for management, protection and genetic breeding of germplasm resources.