研究目的
Understanding rhizospheric processes by imaging complex molecular transformations at relevant spatial scales within the root soil continuum.
研究成果
The findings demonstrate the potential to elucidate the spatial distribution of multiple classes of organic compounds within rhizosphere systems. The approach offers a path for moving beyond bulk-omic analyses, providing a deeper understanding of rhizosphere processes on the submillimeter scale.
研究不足
The current limitation to detect certain molecules in situ by MSI is reflected in the incompatibility of the surface chemistry of the blotting membrane with negative ionization mode analysis, which is generally better for detecting sugars and organic acids.
1：Experimental Design and Method Selection:
The method involves extracting organic compounds from the rhizosphere onto a PVDF membrane while maintaining their 2D distribution and then imaging the distribution of chemical compounds using MALDI-MS.
2：Sample Selection and Data Sources:
Switchgrass was grown in rhizoboxes with removable side panels containing soil from Kellogg Biological Station, Michigan, USA.
3：List of Experimental Equipment and Materials:
High-density polyethylene rhizoboxes, PVDF membrane, MALDI target plate, double-sided copper tape, MALDI matrix (2,5-dihydroxybenzoic acid), 15 Tesla Fourier transform ion cyclotron resonance (FTICR)-MS.
4：Experimental Procedures and Operational Workflow:
After 8 weeks of plant growth, a side panel was removed from the rhizobox and a prewetted PVDF membrane was installed against the soil/root interface for 48h. The membrane was then mounted on a MALDI target plate, and MALDI matrix was applied. MSI was performed in positive ion mode.
5：8h. The membrane was then mounted on a MALDI target plate, and MALDI matrix was applied. MSI was performed in positive ion mode. Data Analysis Methods:
5. Data Analysis Methods: The ultrahigh mass resolution of the 15T-FTICR-MS enabled annotation of exact molecular formulas of ions ablated from the PVDF membrane, and orthogonal LC-MS analysis assisted in confident molecular identification.
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