食品科学

• 技术应用 • 上一篇    

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*   

  1. 中国检验检疫科学研究院,北京 100123
  • 出版日期:2014-12-25 发布日期:2014-12-29
  • 通讯作者: 陈 颖
  • 基金资助:

    国家高技术研究发展计划(863计划)项目(2011AA100807);公益性(质检)行业科研专项(201110015)

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*   

  1. Chinese Academy of Inspection and Quarantine, Beijing 100123, China
  • Online:2014-12-25 Published:2014-12-29
  • Contact: CHEN Ying

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(polymerasechain 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

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
reading ratio of each plant was consistent with the proportion of its PCR product. This study preliminarily proved that the
combined technique is capable of identifying the plant species in blended oils accurately and efficiently. The procedure could
be streamlined to be a promising detection method for oil product supervision in the near future.

Key words: next-generation sequencing, DNA barcoding, edible oil crops, detection

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