岱衢族大黄鱼(Pseudosciaena crocea) 线粒体基因组全序列的扩增及其特异鉴别引物的开发

采用PCR扩增、克隆获得了岱衢族和闽-粤东族大黄鱼(Pseudosciaena crocea)线粒体基因全长序列,并扩增了各25条样本的高变区域,测序结果可分成3个操作分类单元(Operational Taxonomic Unit,OTU);闽-粤东族含3个类型:D1-N1、D1-N2和D2型;而岱衢族样品均为D1-N1型;设计出能区分不同分型的鉴别引物J1和J2,且优化了鉴别条件。结果表明:根据凝胶电泳的图片,如果样品分型为D1-N2和D2型,则100%为闽-粤东族;如果样品分型为D1-N1型,则该样品为岱衢族的概率为86.2%,为闽-粤东族的概率为13.8%。研究结果为岱衢族大黄鱼的种质资源保护、良种选育和市场销售等方面提供技术支持。

THE AMPLIFICATION OF THE COMPLETE MITOCHONDRIAL GENOME OF THE DAI-QU STOCK PSEUDOSCIAENA CROCEA AND THE DEVELOPMENT OF ITS DIAGNOSTIC PRIMERS

The complete mitochondrial DNA (mtDNA) genomes of Pseudosciaena crocea sampled from Min-yue and Dai-qu Stock were amplified by PCR and sequenced. The sequences of high variability regions from 25 samples respectively were obtained, which were divided into 3 Operational Taxonomic Units (OTUs): D1-N1, D1-N2 and D2. The results indicated that: all the Dai-qu Stock samples were clustered into D1-N1 type; Min-yue Stock samples were clustered into D1-N1, D1-N2 and D2 type. Primer pairs J1 and J2 were designed to significantly distinguish different OTUs, and PCR conditions were optimizated for identification. The results show that the accurate rate of identification for Min-yue Stock P. crocea is 100%, if the band pattern on photo belongs to D1-N2 or D2 type, and the accurate rate of identification for Dai-qu and Min-yue Stock P. crocea are respectively 86.2% and 13.8%, if it belongs to D1-N1 type, according to gel images of the samples. The research results provide technical support for varieties resources conservation, selective improved variety and market sales of the Dai-qu Stock P. crocea.