Matrix-free UV-laser desorption/ionization (LDI) mass spectrometric imaging at the single-cell level: distribution of secondary metabolites of Arabidopsis thaliana and Hypericum species
Matrix-free UV-laser desorption/ionization (LDI) mass spectrometric imaging at the single-cell level: distribution of secondary metabolites of Arabidopsis thaliana and Hypericum species
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DOI：
10.1111/j.1365-313x.2009.04012.x
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发表时间：
2009-12-01
期刊：
影响因子：
7.2
通讯作者：
Svatos, Ales
中科院分区：
文献类型：
--
作者：
Hoelscher, Dirk;Shroff, Rohit;Svatos, Ales
本文介绍了无基质激光解吸/电离质谱仪(LDI-MSI)在单细胞分辨率下对植物组织中高度局域的紫外吸收次生代谢物的成像。对于金丝桃属植物，讨论了该方法的范围和局限性。在金丝桃叶、胎座、雄蕊和花柱上的暗腺中，可以找到金丝桃素和假金丝桃素等萘二蒽酮；在这种重要的植物医学植物的花粉中也可以找到双黄酮类化合物。获得的最高空间分辨率为10微米，远远高于常用的基质辅助激光解吸/电离(MALDI)成像方案。来自成像实验的数据得到了对冷冻切片、激光显微解剖和新鲜切割的植物材料的独立LDI-TOF/MS分析的支持。结果证实了激光显微切割(LMD)与LDI-TOF/MS或LDI-MSI联用分析植物次生代谢物的可行性。此外，还对拟南芥进行了分析，以证明LDI-MSI用于其他常见化合物，如黄酮类化合物的可行性。山奈酚、栎素和异鼠李素及其糖苷在细胞水平上的器官特异性分布被成像。
P>The present paper describes matrix-free laser desorption/ionisation mass spectrometric imaging (LDI-MSI) of highly localized UV-absorbing secondary metabolites in plant tissues at single-cell resolution. The scope and limitations of the method are discussed with regard to plants of the genus Hypericum. Naphthodianthrones such as hypericin and pseudohypericin are traceable in dark glands on Hypericum leaves, placenta, stamens and styli; biflavonoids are also traceable in the pollen of this important phytomedical plant. The highest spatial resolution achieved, 10 mu m, was much higher than that achieved by commonly used matrix-assisted laser desorption/ionization (MALDI) imaging protocols. The data from imaging experiments were supported by independent LDI-TOF/MS analysis of cryo-sectioned, laser-microdissected and freshly cut plant material. The results confirmed the suitability of combining laser microdissection (LMD) and LDI-TOF/MS or LDI-MSI to analyse localized plant secondary metabolites. Furthermore, Arabidopsis thaliana was analysed to demonstrate the feasibility of LDI-MSI for other commonly occurring compounds such as flavonoids. The organ-specific distribution of kaempferol, quercetin and isorhamnetin, and their glycosides, was imaged at the cellular level.