Single-Cell Metabolite Profiling of Stalk and Glandular Cells of Intact Trichomes with Internal Electrode Capillary Pressure Probe Electrospray Ionization Mass Spectrometry
Single-Cell Metabolite Profiling of Stalk and Glandular Cells of Intact Trichomes with Internal Electrode Capillary Pressure Probe Electrospray Ionization Mass Spectrometry
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DOI：
10.1021/acs.analchem.5b03366
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发表时间：
2016-03-15
影响因子：
7.4
通讯作者：
Nonami, Hiroshi
中科院分区：
文献类型：
--
作者：
Nakashima, Taiken;Wada, Hiroshi;Nonami, Hiroshi
在本报告中，我们开发了具有内电极毛细管的压力探针电喷雾电离质谱法（IEC-PPESI-MS），该方法能够实现高空间分辨率的细胞采样，精确的采样后操作和高检测灵敏度。利用该技术，对番茄植物（Solanum lycopersicum L.）中构成毛状体单元的两种相邻细胞类型茎细胞和腺细胞进行了原位比较单细胞代谢物分析，以阐明两种细胞类型之间以及不同类型毛状体之间的代谢分化程度。由于高。该系统的灵敏度，小于一皮升细胞SAP从一个单一的茎细胞充分产生氨基酸，有机酸，碳水化合物和类黄酮的一些峰。从茎细胞中提取最少的细胞液而不严重干扰毛状体结构，这使得对同一毛状体上相邻腺细胞的序列分析成为可能，这表明两种相邻细胞类型之间代谢物组成存在显著差异。不同类型毛状体之间的比较还揭示了代谢物谱的显著差异，特别是黄酮类和酰基糖的组成。一些代谢物仅在特定的细胞类型或特定的毛状体类型中发现。虽然对番茄毛状体腺细胞进行了广泛的代谢组学分析，但这是第一次描述茎状体和腺状细胞以及不同毛状体类型的代谢物组成在细胞间的变化。该技术的进一步应用可能为揭示植物细胞在形状、大小、功能和理化性质上的不同代谢提供新的见解。
In this report, we developed the pressure probe electrospray ionization-mass spectrometry with internal electrode capillary (IEC-PPESI-MS) which enables high spatial-resolution cell sampling, precise postsampling manipulation, and high detection sensitivity. Using this technique, a comparative in situ single-cell metabolite profiling of stalk and glandular cells, the two adjacent cell types comprising a trichome unit in tomato plants (Solanum lycopersicum L.), were performed to clarify the extent of Metabolic differentiation between two cell types as well as among different types of trichomes. Owing to high. sensitivity of the system, less than a picoliter cell sap from a single stalk cell sufficiently yielded a number of peaks of amino acids, organic acids, carbohydrates, and flavonoids. The minimal cell sap removal from a stalk cell without severe disturbance of trichome structure enabled sequential analysis of adjacent glandular cell on the same trichome, which showed the presence of striking differences in metabolite compositions between two adjacent cell types. Comparison among different types of trichome also revealed significant variations in metabolite profiles, particularly in flavonoids and acyl sugars compositions. Some metabolites were found only in specific cell types or particular trichome types. Although extensive metabolomics analysis of glandular cells of tomato trichomes has been previously documented, this is the first report describing cell-to-cell variations in metabolite compositions of stalk and glandular cells as well as in different trichome types. Further application of this technique may provide new insights into distinct metabolism in plant cells displaying variations in shape, size, function and physicochemical properties.