Matrix assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MSI) for direct visualization of plant metabolites in situ

Sturtevant, Drew; Lee, Young Jin; Chapman, Kent D.

doi:10.1016/j.copbio.2015.10.004

Cited by 132 publications

(101 citation statements)

References 51 publications

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“…Recently, MALDI-MS imaging made it possible to study lipid spatial distributions in situ directly from tissue sections (Sturtevant et al, 2016). Previous studies utilizing MALDI-MS imaging in oilseeds of cotton, C. sativa, and Arabidopsis demonstrated that lipid molecular species are not distributed equally between different seed tissues (Horn et al, 2012Sturtevant et al, 2017).…”

Section: Spatial Distribution Of Lipidsmentioning

confidence: 99%

Two Acyltransferases Contribute Differently to Linolenic Acid Levels in Seed Oil

et al. 2017

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ORCID IDs: 0000-0001-6929-7859 (S.M.); 0000-0003-3152-0394 (D.S.); 0000-0001-8255-3255 (C.H.); 0000-0003-0489-3072 (K.C.); 0000-0002-9888-7003 (I.F.).Acyltransferases are key contributors to triacylglycerol (TAG) synthesis and, thus, are of great importance for seed oil quality. The effects of increased or decreased expression of ACYL-COENZYME A:DIACYLGLYCEROL ACYLTRANSFERASE1 (DGAT1) or PHOSPHOLIPID:DIACYLGLYCEROL ACYLTRANSFERASE (PDAT) on seed lipid composition were assessed in several Camelina sativa lines. Furthermore, in vitro assays of acyltransferases in microsomal fractions prepared from developing seeds of some of these lines were performed. Decreased expression of DGAT1 led to an increased percentage of 18:3n-3 without any change in total lipid content of the seed. The tri-18:3 TAG increase occurred predominantly in the cotyledon, as determined with matrix-assisted laser desorption/ionization-mass spectrometry, whereas species with two 18:3n-3 acyl groups were elevated in both cotyledon and embryonal axis. PDAT overexpression led to a relative increase of 18:2n-6 at the expense of 18:3n-3, also without affecting the total lipid content. Differential distributions of TAG species also were observed in different parts of the seed. The microsomal assays revealed that C. sativa seeds have very high activity of diacylglycerol-phosphatidylcholine interconversion. The combination of analytical and biochemical data suggests that the higher 18:2n-6 content in the seed oil of the PDAT overexpressors is due to the channeling of fatty acids from phosphatidylcholine into TAG before being desaturated to 18:3n-3, caused by the high activity of PDAT in general and by PDAT specificity for 18:2n-6. The higher levels of 18:3n-3 in DGAT1-silencing lines are likely due to the compensatory activity of a TAG-synthesizing enzyme with specificity for this acyl group and more desaturation of acyl groups occurring on phosphatidylcholine.

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Section: Spatial Distribution Of Lipidsmentioning

confidence: 99%

Two Acyltransferases Contribute Differently to Linolenic Acid Levels in Seed Oil

et al. 2017

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“…These mass spectra can then be searched for the massto-charge ratio (m/z) of ions of interest, and a heat map or matrix-assisted laser/desorption ionization-mass spectrometry (MALDI-MS) image is generated representing the spatial localization of the ions in question. A summary of the MALDI-MS imaging for the in situ visualization of TAG and PC is shown in Figure 2, but for more detailed descriptions, see Horn and Chapman (2014) and Sturtevant et al (2016).…”

Section: Spatial Mapping Of Pc and Tag At The Midpoint Of Seed Lipid mentioning

confidence: 99%

“…Imaging-based mass spectrometry (MS) approaches have emerged as an alternative, or as an addition to these extraction based methods, to provide an additional spatial dimension to metabolomic studies. Matrix-assisted laser/desorption ionization-mass spectrometry imaging (MALDI-MSI) is a powerful analytical technique that enables the spatial mapping of metabolites within intact biological tissues or directly off tissue surfaces (Cornett et al, 2007;Horn and Chapman, 2014;Sturtevant et al, 2016). This analytical technique was pioneered by the Caprioli group in the 1990s in animal systems (Caprioli et al, 1997) and, over the last decade, has been applied to plant systems to great effect (Horn and Chapman, 2014;Korte et al, 2015;Boughton et al, 2016).…”

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confidence: 99%

“…Moreover, observing the spatial context of lipid distribution gives new insights into how associated metabolic networks might be differentially regulated between different tissue types. MALDI-MSI has been used previously with great success to show the spatial heterogeneity of lipid distribution in various plant seeds, such as cotton (Gossypium hirsutum; Horn et al, 2012;Sturtevant et al, 2017b), maize (Zea mays; Feenstra et al, 2015), camelina (Camelina sativa; , and Arabidopsis (Sturtevant et al, 2016(Sturtevant et al, , 2017a, all of which show a certain degree of tissue-specific lipid accumulation across several different lipid classes.…”

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confidence: 99%

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Spatial and Temporal Mapping of Key Lipid Species in Brassica napus Seeds

et al. 2017

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The regulation of lipid synthesis in oil seeds is still not fully understood. Oilseed rape (Brassica napus) is the third most productive vegetable oil crop on the global market; therefore, increasing our understanding of lipid accumulation in oilseed rape seeds is of great economic, as well as intellectual, importance. Matrix-assisted laser/desorption ionization-mass spectrometry imaging (MALDI-MSI) is a technique that allows the mapping of metabolites directly onto intact biological tissues, giving a spatial context to metabolism. We have used MALDI-MSI to study the spatial distribution of two major lipid species, triacylglycerols and phosphatidylcholines. A dramatic, heterogenous landscape of molecular species was revealed, demonstrating significantly different lipid compositions between the various tissue types within the seed. The embryonic axis was found to be particularly enriched in palmitic acid, while the seed coat/aleurone layer accumulated vaccenic, linoleic, and a-linoleic acids. Furthermore, the lipid composition of the inner and outer cotyledons differed from each other, a remarkable discovery given the supposed identical functionality of these two tissues. Triacylglycerol and phosphatidylcholine molecular species distribution was analyzed through a developmental time series covering early seed lipid accumulation to seed maturity. The spatial patterning of lipid molecular species did not vary significantly during seed development. Data gathered using MALDI-MSI was verified through gas chromatography analysis of dissected seeds. The distinct lipid distribution profiles observed imply differential regulation of lipid metabolism between the different tissue types of the seed. Further understanding of this differential regulation will enhance efforts to improve oilseed rape productivity and quality.

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“…However, achieving such high resolution requires careful matrix embedding and sample preparation, which may affect tissue structure and localization of biomolecules. Furthermore, the spatial resolution is insufficient to interrogate individual oil bodies (0.05–3 µm diameter) at this time [24,27]. …”

Section: Introductionmentioning

confidence: 99%

Label-freein situimaging of oil body dynamics and chemistry in germination

Waschatko

Billecke

Schwendy

et al. 2016

J. R. Soc. Interface.

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Plant oleosomes are uniquely emulsified lipid reservoirs that serve as the primary energy source during seed germination. These oil bodies undergo significant changes regarding their size, composition and structure during normal seedling development; however, a detailed characterization of these oil body dynamics, which critically affect oil body extractability and nutritional value, has remained challenging because of a limited ability to monitor oil body location and composition during germination in situ. Here, we demonstrate via in situ, label-free imaging that oil bodies are highly dynamic intracellular organelles that are morphologically and biochemically remodelled extensively during germination. Label-free, coherent Raman microscopy (CRM) combined with bulk biochemical measurements revealed the temporal and spatial regulation of oil bodies in native soya bean cotyledons during the first eight days of germination. Oil bodies undergo a cycle of growth and shrinkage that is paralleled by lipid and protein compositional changes. Specifically, the total protein concentration associated with oil bodies increases in the first phase of germination and subsequently decreases. Lipids contained within the oil bodies change in saturation and chain length during germination. Our results show that CRM is a well-suited platform to monitor in situ lipid dynamics and local chemistry and that oil bodies are actively remodelled during germination. This underscores the dynamic role of lipid reservoirs in plant development.

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Matrix assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MSI) for direct visualization of plant metabolites in situ

Cited by 132 publications

References 51 publications

Two Acyltransferases Contribute Differently to Linolenic Acid Levels in Seed Oil

Two Acyltransferases Contribute Differently to Linolenic Acid Levels in Seed Oil

Spatial and Temporal Mapping of Key Lipid Species in Brassica napus Seeds

Label-freein situimaging of oil body dynamics and chemistry in germination

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