Research Group EnVOC, Department of Green Chemistry and Technology, Faculty of Bioscience Engineering, Ghent University, Coupure links 653, 9000, Ghent, Belgium.
Research Group Epigenetics and Defence, Department of Biotechnology, Faculty of Bioscience Engineering, Ghent University, Coupure links 653, 9000, Ghent, Belgium.
Anal Bioanal Chem. 2018 Jul;410(18):4527-4539. doi: 10.1007/s00216-018-1112-9. Epub 2018 May 23.
Phytohormones are signaling and regulating metabolites involved in numerous plant processes, including growth, development, and responses to stress. Currently, the focus is on the analysis of multiple phytohormones in order to characterize crosstalk and hormone signaling networks. In this paper, representative phytohormones of the major classes are simultaneously determined in rice tissues by a generic solid-liquid extraction, followed by liquid chromatography and electrospray ionization high-resolution tandem mass spectrometry using a Q-Exactive™ instrument. After a thorough optimization of the sample preparation, the analytical method was fully validated toward the ultra-trace quantification of six a priori selected plant hormones using three scan modes of the quadrupole-Orbitrap instrument: full-scan high-resolution mass spectrometry, targeted single ion monitoring (t-SIM), and t-SIM followed by data-dependent tandem mass spectrometry. Overall, a similar quantitative performance was noticed for the different scan modes. The analytical method was successfully applied to measure basal phytohormone levels in six different rice accessions, comprising Oryza sativa ssp. japonica, indica, and Oryza glaberrima. Hormone concentrations were higher in shoots than in roots or at least similar. Except for a lower level of salicylic acid in shoots of O. glaberrima versus O. sativa, no other differences in hormone levels could be noticed that were dependent of the (sub)species assignment of the analyzed accessions. Making use of the benefits of full-scan high-resolution mass spectrometry, a first post-run suspect screening was performed, suggesting - based on accurate mass measurements and isotopic patterns - the possible presence of about 50 additional plant hormones in the rice tissues. Graphical abstract ᅟ.
植物激素是参与植物生长、发育和应激反应等多种过程的信号和调节代谢物。目前，重点是分析多种植物激素，以描述串扰和激素信号网络。在本文中，通过通用的固液萃取法同时在水稻组织中同时测定了主要类别的代表性植物激素，然后通过液相色谱和电喷雾电离高分辨串联质谱仪（Q-Exactive™仪器）进行分析。在对样品制备进行了彻底优化后，使用四极杆-Orbitrap 仪器的三种扫描模式：全扫描高分辨质谱、靶向单离子监测（t-SIM）和 t-SIM 后进行数据依赖性串联质谱，对超痕量定量分析六种预先选择的植物激素的分析方法进行了全面验证。总体而言，不同扫描模式的定量性能相似。该分析方法成功应用于测量六种不同水稻品系（包括粳稻、籼稻和非洲稻）中基础植物激素的水平。与根或至少相似相比，芽中的激素浓度更高。除了非洲稻的芽中水杨酸水平较低外，根据分析品系的（亚种）归属，未观察到其他激素水平的差异。利用全扫描高分辨质谱的优势，进行了一次初步的运行后嫌疑筛查，基于精确质量测量和同位素模式，提示水稻组织中可能存在约 50 种其他植物激素。
