Subcellular-level resolution MALDI-MS imaging of maize leaf metabolites by MALDI-linear ion trap-Orbitrap mass spectrometer

Korte, Andrew R.; Yandeau‐Nelson, Marna D.; Nikolau, Basil J.; Lee, Young Jin

doi:10.1007/s00216-015-8460-5

Cited by 127 publications

(115 citation statements)

References 37 publications

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“…MALDI-MSI can obtain high-spatial-resolution images of ions, allowing for the visualization of fine structures. Spatial resolution of ;20 mm has become routine in MALDI-MSI, and 2-to 5-mm spatial resolution has been demonstrated by several groups, with the latter providing a means to locate metabolites within subcellular compartments (Zavalin et al, 2013;Korte et al, 2015).…”

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Spatial Mapping and Profiling of Metabolite Distributions during Germination

et al. 2017

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Germination is a highly complex process by which seeds begin to develop and establish themselves as viable organisms. In this study, we utilize a combination of gas chromatography-mass spectrometry, liquid chromatography-fluorescence, and mass spectrometry imaging approaches to profile and visualize the metabolic distributions of germinating seeds from two different inbreds of maize (Zea mays) seeds, B73 and Mo17. Gas chromatography and liquid chromatography analyses demonstrate that the two inbreds are highly differentiated in their metabolite profiles throughout the course of germination, especially with regard to amino acids, sugar alcohols, and small organic acids. Crude dissection of the seed followed by gas chromatography-mass spectrometry analysis of polar metabolites also revealed that many compounds were highly sequestered among the various seed tissue types. To further localize compounds, matrix-assisted laser desorption/ionization mass spectrometry imaging was utilized to visualize compounds in fine detail in their native environments over the course of germination. Most notably, the fatty acyl chain-dependent differential localization of phospholipids and triacylglycerols was observed within the embryo and radicle, showing correlation with the heterogeneous distribution of fatty acids. Other interesting observations include unusual localization of ceramides on the endosperm/scutellum boundary and subcellular localization of ferulate in the aleurone.Plants use seeds as the propagule to ensure reproduction to the next generation, and over the past 10,000 years, human civilizations have established agricultural practices to ensure a seed-based food supply (Larson et al., 2014). Therefore, deciphering the processes that enable seeds to perform their biological functions is of importance in understanding how plants are propagated and also is of practical importance to improve agriculture. Seeds are designed to survive long periods of dormancy in a relatively dry state, and the process of germination is initiated by the imbibition of water. During this germination process, many metabolic changes occur, most of which are associated with the catabolism of seed storage products (proteins, polysaccharides, and lipids) into metabolically usable, simpler chemical forms that are either used as precursors to assemble the growing seedling or are oxidized via energy-producing biochemical pathways to thermodynamically support growth (Bewley, 1997(Bewley, , 2001.The specific metabolic processes that support seed germination are somewhat dependent on the taxonomic clade of the plant, which determines seed tissue/ organ organization and the nature of the seed storage compounds. Specifically, the seeds of the Poaceae family of monocots are characterized by a starch-and protein-filled endosperm, and their catabolism by digestive enzymes produced in the outermost layer of the endosperm (i.e. the aleurone) provides the carbonbased and nitrogenous precursors for the growth of the embryo, which is composed of the embryonic ax...

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Spatial Mapping and Profiling of Metabolite Distributions during Germination

et al. 2017

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“…Procedures for corn seed germination and plant growth are described in detail elsewhere [11,12], and briefly described below. A maize seed was imbibed for 10 min and placed with the embryo side facing down on top of moist filter paper in a Petri dish that was placed in a climate-controlled greenhouse.…”

Section: Plant Growth and Harvestmentioning

confidence: 99%

“…The instrument was modified to use an external 355 nm Nd:YAG laser (UVFQ; Elforlight Ltd., Daventry, UK) and a custom optical set-up [12]. TunePlus and Xcalibur software (Thermo Scientific) were used to define imaging acquisition parameters and for data acquisition.…”

Section: Mass Spectrometry Analysismentioning

confidence: 99%

Matrix Recrystallization for MALDI-MS Imaging of Maize Lipids at High-Spatial Resolution

Dueñas

Carlucci

Lee

2016

J. Am. Soc. Mass Spectrom.

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Abstract. Matrix recrystallization is optimized and applied to improve lipid ion signals in maize embryos and leaves. A systematic study was performed varying solvent and incubation time. During this study, unexpected side reactions were found when methanol was used as a recrystallization solvent, resulting in the formation of a methyl ester of phosphatidic acid. Using an optimum recrystallization condition with isopropanol, there is no apparent delocalization demonstrated with a transmission electron microscopy (TEM) pattern and maize leaf images obtained at 10 μm spatial resolution.

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“…The technique enables simultaneous label-free visualization of the lateral distribution of numerous biomolecules in the sections (e.g., peptides, lipids, further metabolites). With current MALDI-MSI instruments, lateral resolutions in the range of a few 10 μm are routinely achieved, and several reports even demonstrated MALDI-MSI analysis with pixel sizes in the low micrometer range [3][4][5][6][7].…”

Section: Introductionmentioning

confidence: 99%

Influence of the Laser Spot Size, Focal Beam Profile, and Tissue Type on the Lipid Signals Obtained by MALDI-MS Imaging in Oversampling Mode

Wiegelmann

Dreisewerd

Soltwisch

2016

J. Am. Soc. Mass Spectrom.

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Abstract. To improve the lateral resolution in matrix-assisted laser desorption/ ionization mass spectrometry imaging (MALDI-MSI) beyond the dimensions of the focal laser spot oversampling techniques are employed. However, few data are available on the effect of the laser spot size and its focal beam profile on the ion signals recorded in oversampling mode. To investigate these dependencies, we produced 2 times six spots with dimensions between~30 and 200 μm. By optional use of a fundamental beam shaper, square flat-top and Gaussian beam profiles were compared. MALDI-MSI data were collected using a fixed pixel size of 20 μm and both pixel-by-pixel and continuous raster oversampling modes on a QSTAR mass spectrometer. Coronal mouse brain sections coated with 2,5-dihydroxybenzoic acid matrix were used as primary test systems. Sizably higher phospholipid ion signals were produced with laser spots exceeding a dimension of~100 μm, although the same amount of material was essentially ablated from the 20 μm-wide oversampling pixel at all spot size settings. Only on white matter areas of the brain these effects were less apparent to absent. Scanning electron microscopy images showed that these findings can presumably be attributed to different matrix morphologies depending on tissue type. We propose that a transition in the material ejection mechanisms from a molecular desorption at large to ablation at smaller spot sizes and a concomitant reduction in ion yields may be responsible for the observed spot size effects. The combined results indicate a complex interplay between tissue type, matrix crystallization, and laser-derived desorption/ablation and finally analyte ionization.

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Subcellular-level resolution MALDI-MS imaging of maize leaf metabolites by MALDI-linear ion trap-Orbitrap mass spectrometer

Cited by 127 publications

References 37 publications

Spatial Mapping and Profiling of Metabolite Distributions during Germination

Spatial Mapping and Profiling of Metabolite Distributions during Germination

Matrix Recrystallization for MALDI-MS Imaging of Maize Lipids at High-Spatial Resolution

Influence of the Laser Spot Size, Focal Beam Profile, and Tissue Type on the Lipid Signals Obtained by MALDI-MS Imaging in Oversampling Mode

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