线粒体COⅢ基因分析三种常见鲳鱼的亲缘关系

Relationship Analysis and Species Identification of Three Common Pomfrets Using Mitochondrial COⅢ Gene

金鲳(Trachinotusovatus)、银鲳(Pampusargenteus)和中间低鳍鲳(Peprilusmedius)是重要经济鱼种,其中后两种鱼外形较为相似,分类一直存在分歧.本研究随机采集烟台、青岛地区银鲳、中间低鳍鲳和金鲳的生鲜海鱼样本,提取3种鱼的全基因组DNA,设计通用引物对目标基因进行PCR扩增和测序,后对其线粒体COⅢ全序列及ATP6部分序列进行基因多态性分析,探讨3种鱼的亲缘关系;依据测序结果,设计各鱼种的特异引物,用特异引物和通用引物进行复合PCR扩增方法对各鱼种进行种源性鉴定.通过比对分析测序结果,获得了中间低鳍鲳COⅢ基因全序列及ATP6基因部分序列且收录在NCBI核苷酸数据库中,登录号分别为KT210112和KT210113.证明NCBIGenBank中发布的刺鲳(Psenopsis anomala)序列(KP334103.1)实为中间低鳍鲳.遗传距离分析发现银鲳与金鲳的距离最大,与中间低鳍鲳次之,中间低鳍鲳与金鲳的遗传距离最小.采用复合PCR方法,实现了在同一复合PCR体系中有效区分3种鱼的方法,具有更加便捷、准确的优势.

Trachinotus ovatus,Pampus argenteus and Peprilus medius are important economic fish.As Pampus argenteus and Peprilus medius are more similar in shape,there existed much controversy over the classification between the two species.In this study,Trachinotus ovatus,Peprilus medius and Pampus argenteus were randomly collected rom the markets of Yantai and Qingdao,and the whole genomic DNAs of the three kinds of fish were extracted.Universal primer pairs were designed for the amplification of the target genes with PCR,followed by sequencing programs.Then,the genetic polymorphism of mitochondrial COⅢsequences and ATP6 partial sequences were analyzed,and the genetic relationship of the three fish species was discussed.Based on the sequencing results,species-specific primers were designed,and the specific and universal primers were used to identify the species with the method of complex PCR.The full COⅢgene sequences and part of ATP6 sequences of Peprilus medius were obtained and published in NCBI,with the accession numbers KT210112 and KT210113 respectively.It has also been determined that the sequence of Psenopsis anomala(KP334103.1),published in NCBI,is in fact Peprilus medius.Genetic distance analysis reveals that Pampus argenteus and Trachinotus ovatus have comparatively distant relationship,while the genetic distance between Peprilus medius and Trachinotus ovatus is considerably small.With multiplex PCR,three fish species can be distinguished in one multiplex PCR system.This system provides more convenient and accurate solutions.