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基于线粒体COI基因序列探讨广西钦州湾牡蛎的遗传分化
李咏梅1, 陈秀荔1, 彭 敏1
广西壮族自治区水产研究所 遗传育种与健康养殖重点实验室
摘要:
[目的]初步了解我国广西钦州湾牡蛎的遗传多样性及其分类和系统进化情况.[方法]以采自广西钦州湾的37个白肉牡蛎和29个红肉牡蛎个体为材料,对其线粒体细胞色素氧化酶亚基Ⅰ(COI)的部分序列进行PCR扩增和序列比对.检测牡蛎的遗传多样性;计算牡蛎不同单倍型及GenBank中已登载的4种牡蛎间的遗传距离,采用聚类分析方法,构建其NJ和UPGMA系统进化树.[结果]66个白肉牡蛎和红肉牡蛎线粒体COI基因片段的PCR产物大小约为600 bp,扩增产物纯化并去除引物序列后获得约535 bp的核苷酸序列;碱基组成平均为:T 37.5%.C18.2%,A 23.8%,G 20.5%,A+T 60.5%,C+G 39.5%,A+T含量高于G+C含量.运用DNAStar软件对66个钦州湾牡蛎序列进行比对,白肉牡蛎中共检测到4个多态位点,包括3个转换位点和1个颠换位点,有4种单倍型;红肉牡蛎中共检测到26个多态位点,转换位点和颠换位点各13个,有3种单倍型.GenBank中香港牡蛎COI序列与白肉牡蛎的遗传距离为0.000,有明巨牡蛎COI序列与红肉牡蛎的遗传距离为0.011,白肉牡蛎与红肉牡蛎的遗传距离为0.146;NJ和UPGMA系统进化树表明,白肉牡蛎与红肉牡蛎亲缘关系较远,白肉牡蛎与香港牡蛎聚为一支,红肉牡蛎与有明巨牡蛎聚为一支.[结论]序列特征、遗传距离和系统进化树分析结果表明,COI基因可用于牡蛎的种类鉴定和系统发育分析.
关键词:  牡蛎  COI基因  线粒体  广西
DOI:
分类号:
基金项目:广西壮族自治区自然科学基金,广西壮族自治区科技创新能力建设项目
Genetic differentiation of oyster from Qinzhou Bay based on mitochondrial cytochrome oxidase subunit I gene
Abstract:
【Objective】 Preliminary understanding of genetic diversity,its classification and condition of systemic evolution of oyster from Qinzhou Bay of Guangxi was made.【Method】 37 white meat oyster and 29 red meat oyster were used as experimental materials from Qinzhou Bay of Guangxi.Mitochondrial cytochrome oxidase subunit I(COI)gene fragment were amplified using PCR technique and aligned of sequence,detecting genetic diversity of oyster,and calculating genetic distance between different haplotypes of oyster and 4 kinds of oyster in GenBank.Clustering analysis could build NJ and UPGMA phylogenetic tree.【Result】 The mitochondrial cytochrome oxidase subunit COI gene fragment PCR products of 66 white meat oyster and red meat oyster were about 600 bp long,535 bp nucleotide sequences were determined for oysters after being purified and excluded the primer sequence; The average base compositions were T 37.5%,C 18.2%,A 23.8%,G 20.5%,A+T 60.5%,C+G 39.5% respectively,A+T content were higher than G+C content.By using DNAStar to align and compare the sequences of the 66 individuals with each other,4 polypeptide sites were detected in white meat oyster,including 3 transition sites and 1 transversion sites.There were 4 haplotypes.26 polypeptide sites were detected in red meat oyster,including 13 transition sites and transversion sites each,and there were 3 haplotypes.Genetic distances between COI sequence of C.hongkongsis and white meat oysters were 0.000;Genetic distances between COI sequence of C.ariakensis and red meat oysters were 0.011;Genetic distances between white meat oysters and red meat oysters were 0.146.NJ and UPGMA phylogenetic trees showed that the genetic relationship between white meat oysters and red meat oysters were far,white meat oysters and C.hongkongsis assembled a branch,the same to red meat oysters and C.ariakensis.【Conclusion】 The sequence character,genetic distance and phylogenetic trees indicated that COI gene is useful in species identification and phylogenetic analysis of oyster.
Key words:  oyster  COI gene  mitochondrial  Guangxi