Population structure of pilchards using sequence of mitochondrial DNA

本研究是为了阐明在日本沿海地区采集的沙丁鱼种群结构。方法采集日本沿海地区沙丁鱼（Sardinops melanosttictus）、日本沙丁鱼（Engraulis japonicus）和特鲁米乌鱼（Etrumeus teres）的肌肉进行总DNA提取。用苯酚-氯仿从乙醇保存标本的肌肉中提取DNA。cyt。采用聚合酶链反应（PCR）从总DNA中扩增出gnome线粒体DNA （mtDNA）的b基因、d环和ITS基因。采用直接测序法对纯化的双链PCR产物进行测序。采用Kimra的2参数法估计遗传距离值。结果d环区突变率最高。由于对D-loop进行测序是为了确定当地沙丁鱼的群体结构，因此在收集的地点之间没有发现明显的遗传差异。因此，日本沿海沙丁鱼的种群结构可能是单一的。

This study was perform to elucidate the population structure of pilchard collected form coastal area of Japan.MethodsFish muscle (Sardinops melanostictus, Engraulis japonicus and Etrumeus teres) for total DNA extraction were collected from the coastal area of Japan. DNA was extracted from muscle of ethanol-preserved specimens using phenol-chloroform. The cyt.b gene and D-loop of the mitochondrial DNA (mtDNA) and ITS gene of gnome were amplified from total DNA using the polymerase chain reaction (PCR). The double strand PCR product purified were sequenced using direct sequencing method. Genetic distance values were estimated using the method of Kimra's 2-parameter.ResultsHighest mutation rate was observed in the D-loop among the analyzed regions of pilchards. Since D-loop was sequenced in order to identify the local population structure of pilchard, no obvious genetic differences were observed among the collected sites. Thus, single population structure is suggested in pilchard from coast of Japan.