Population structure of pilchards using sequence of mitochondrial DNA

本研究旨在阐明采自日本沿海地区的沙丁鱼的种群结构。方法从日本沿海地区采集用于提取总DNA的鱼肌肉(Sardinopsolostictus、Engraulis Japan onicus和Etrumeus teres)。用苯酚-氯仿从乙醇保存的肌肉标本中提取DNA。用聚合酶链式反应(PCR)从基因组总DNA中扩增出线粒体DNA(MtDNA)的cyt.b基因和D-loop基因及其基因。用直接测序法对纯化的双链PCR产物进行测序。结果在所分析的沙丁鱼区域中，D-环突变率最高。由于对D-loop进行测序，以确定沙丁鱼的地方种群结构，因此在收集的地点之间没有明显的遗传差异。因此，建议采用日本沿海沙丁鱼的单一种群结构。

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.