高通量二代测序基因条码技术在油料作物种类鉴别中的应用.pdfVIP

348 2014, Vol.35, No.24 食品科学 ※技术应用 高通量二代测序基因条码技术在油料作物 种类鉴别中的应用 吴亚君，杨艳歌，李 莉，王 斌，刘鸣畅，陈 颖* （中国检验检疫科学研究院，北京 100123） 摘  要：对高通量二代测序基因条码技术在油料作物种类鉴别中的应用进行探索。采用Ion Torrent PGMTM测序技 术，分别对橄榄、花生、大豆、油葵、玉米等14 种油料作物和小麦、大米的叶绿体核酮糖二磷酸羧化酶/氧化酶大 亚基（ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit ，rbcL ）基因246 bp聚合酶链式反应（polymerase chain reaction ，PCR ）产物混合物，及上述作物DNA混合液扩增得到的rbcL基因片段进行测序。结果表明，除小麦 外，所有其他作物成分均获得了鉴定。其中，PCR产物混合物中每种作物成分的测序读数数量比例和该成分PCR产 物在总产物中的比例接近，说明该技术可以用于定量分析。实验结果初步证实了该技术对混合样品中各作物种类鉴 别的准确性，并具有检测效率、流程简便的特点，有望在未来成为食用油标识符合性查验的有力工具。 关键词：二代测序；基因条码；油料；检测 Application of High Throughput Next-Generation Sequencing Based on DNA Barcoding Technology in Species Identification of Edible Oils * WU Ya-jun, YANG Yan-ge, LI Li, WANG Bin, LIU Ming-chang, CHEN Ying (Chinese Academy of Inspection and Quarantine, Beijing 100123, China) Abstract: This study aimed to testify the application of second-generation sequencing based on DNA barcoding technology in the species identification of edible oils. Totally 14 species of oil crops including olive, peanut, soybean, oil sunflower, maize, sesame, camellia, rapeseed, palm and pine nut as well as rice and wheat were selected. Mixtures of the polymerase chain reaction (PCR) products of the 240 bp ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit (rbcL) genes and the rbcL genes amplified from DNA mixtures of these 16 crop species were sequenced by Ion Torrent PGMTM respectively. The results demonstrated that all crop species except wheat were successfully identified. As for two mixture samples, the readi