生物技术通报 ›› 2020, Vol. 36 ›› Issue (11): 222-229.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0198

• 技术与方法 • 上一篇    下一篇

基于代谢组学的转基因水稻生物安全评价方法研究

兰青阔1(), 赵新1, 沈晓玲2, 魏静娜1, 刘双2, 陈锐1, 檀建新2, 王永1()   

  1. 1.天津市农业科学院,天津 300381
    2.河北农业大学,保定 071001
  • 收稿日期:2020-02-28 出版日期:2020-11-26 发布日期:2020-11-20
  • 作者简介:兰青阔,男,研究员,研究方向:转基因生物安全评价技术;E-mail: lanqingkuo@163.com
  • 基金资助:
    转基因重大专项课题(2019ZX08010-002);转基因重大专项课题(2019ZX08015-002);天津市农业科学院青年科研人员创新研究与实验项目(2018005)

Biosafety Assessment Technology Research for Genetically Modified Rice Based on Metabolomics

LAN Qing-kuo1(), ZHAO Xin1, SHEN Xiao-ling2, WEI Jing-na1, LIU Shuang2, CHEN Rui1, TAN Jian-xin2, WANG Yong1()   

  1. 1. Tianjin Academy of Agricultural Sciences,Tianjin 300192
    2. Hebei Agricultural University,Baoding 071001
  • Received:2020-02-28 Published:2020-11-26 Online:2020-11-20

摘要:

代谢组学因其全面无偏向性、非靶标轮廓分析的技术优势,成为目前转基因生物安全评价技术研究的热点。以转基因抗虫水稻及其非转基因受体为研究对象,采用基于超高效液相色谱-四级杆-飞行时间串联质谱(UPLC-QTOF/MS)分析的转基因作物代谢组评价技术流程,对代谢组提取方法、数据处理方法、分析平台等进行了探索。结果表明,应用80%甲醇水能有效提取水稻苗期叶片中的代谢物;经UPLC分离、QTOF/MS分析、正交校正的偏最小二乘辨别分析(OPLS-DA)等计算方法,能够有效发现转基因水稻及其受体之间的差异代谢物。本研究建立的完整的转基因水稻代谢组分析方法具有分辨率高、易掌握的优点。本研究丰富了现有的转基因生物安全评价的内容和方法,为转基因生物安全管理提供技术支撑。

关键词: 转基因抗虫水稻, 代谢组学与代谢物, 转基因生物安全评价, UPLC-QTOF/MS分析

Abstract:

Currently metabolomics is becoming the hotspot in safety assessment research of genetically modified organism(GMO)because of its unbiased and untargeted analysis. Here metabolome extraction method,multivariate processing methods,and data analysis platform were explored while transgenic insect-resistant rice and its non-transgenic receptor were taken as the research object and technical process of metabolomics evaluation of genetically modified crops was used based on the ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-QTOF/MS). The results showed that metabolites in the leaves of rice seedlings were effectively extracted with extraction buffer containing 80% methanol. By UPLC separation,QTOF/MS analysis,and orthogonal partial least squares-discriminate analysis(OPLS-DA),the differential metabolites between genetically modified rice and its non-genetically modified rice were effectively discovered. The systematic analysis method of transgenic rice metabolome established in this study demonstrates the advantages of high resolution and easy to operate. This study enriches the current contents and methods of safe assessment for GMO as well as provides technical support for the safety management of GMO.

Key words: transgenic insect-resistant rice, metabolomics and metabolites, GMO safety assessment, UPLC-QTOF/MS analysis