植物学报 ›› 2018, Vol. 53 ›› Issue (6): 812-828.DOI: 10.11983/CBB17179

• 技术方法 • 上一篇    下一篇

基于液质联用的莱茵衣藻极性甘油酯组定性定量分析

杨淼1,2,3, 孟迎迎1,3, 褚亚东1, 薛松1,*()   

  1. 1中国科学院大连化学物理研究所, 大连 116023
    2中国科学院大学, 北京 100049
    3大连理工大学生命科学与技术学院, 大连 116024
  • 收稿日期:2017-09-24 出版日期:2018-11-01 发布日期:2018-12-05
  • 通讯作者: 薛松
  • 作者简介:

    作者简介:白克智, 1959年开始在中国科学院植物研究所工作, 先后任助理研究员、研究员, 长期从事植物生长发育及其调控的研究。1986年,其主持的“满江红生物学特性研究”荣获中国科学院科技进步二等奖。曾任《植物生理学报》编委、《植物学报》常务编委、中国植物生长调节剂协会主任等职。

  • 基金资助:
    国家自然科学基金(No.21576253)

Qualitative and Quantitative Analyses of Polar Glycerolipids in Chlamydomonas reinhardtii based on LC-MS Techniques

Yang Miao1,2,3, Meng Yingying1,3, Chu Yadong1, Xue Song1,*()   

  1. 1Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China
    2University of Chinese Academy of Sciences, Beijing 100049, China
    3School of Life Sciences and Biotechnology, Dalian University of Technology, Dalian 116024, China
  • Received:2017-09-24 Online:2018-11-01 Published:2018-12-05
  • Contact: Xue Song

摘要:

以模式藻株莱茵衣藻(Chlamydomonas reinhardtii)为材料, 基于液质联用技术对其极性甘油酯组进行定性定量分析。通过综合利用UPLC-ESI-Q-Trap/MS的一级质谱扫描(中性丢失或母离子扫描)及UPLC-ESI-Orbitrap/MS2的二级碎片信息扫描, 共鉴定出109种极性甘油酯分子; 再通过外标法利用UPLC-ESI-Q-Trap/MS在多级反应监测模式下对各分子进行靶向定量分析。结果表明, 莱茵衣藻的极性脂以糖脂MGDG、DGDG及甜菜碱脂DGTS为主, 所有极性脂的分子组成表明, DGDG、SQDG、DGTS及PI是C18脂肪酸的去饱和载体。该研究利用液质联用技术建立了莱茵衣藻极性甘油酯组的结构图谱及定量分析技术平台, 为微藻极性脂生物学功能及脂质代谢研究奠定了基础。

关键词: 莱茵衣藻, 液质联用技术, 极性甘油酯组, 定性定量分析

Abstract:

The polar glycerolipidome of the model algal strain Chlamydomonas reinhardtii was analyzed qualitatively and quantitatively by using liquid chromatography coupled with mass spectrometry (LC-MS). Based on UPLC-ESI-Q-Trap/MS and UPLC-ESI-Orbitrap/MS2, MS scanning, including neutral loss or precursor ion scanning, and MS/MS scanning were used to identify 109 molecular species possessing distinct mass-to-charge ratios and acyl chains. Targeted quantitative analyses based on external standards with UPLC-ESI-Q-Trap/MS in multiple reactions monitoring (MRM) mode was used to analyze individual molecular species. The results indicated that the glycerolipids of C. reinhardtii mainly included glycolipids (i.e., MGDG and DGDG) and betaine lipid, DGTS. In addition, analysis of fatty acyl constituents of all polar membrane lipids revealed that DGDG, SQDG, DGTS and PI acted as carriers for desaturation of C18 fatty acids. This study established a platform for characterizing the structure spectra and quantitative analysis with LC-MS techniques for C. reinhardtii, which provides a useful tool for studies of the biological function of polar membrane lipids and lipid metabolism in microalgae.

Key words: Chlamydomonas reinhardtii, LC-MS technique, polar glycerolipids, qualitative and quantitative analyses