Matrix assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MSI) for direct visualization of plant metabolites in situ Drew Sturtevant 1 , Young-Jin Lee 2,3 and Kent D Chapman 1 Direct visualization of plant tissues by matrix assisted laser desorption ionization-mass spectrometry imaging (MALDI- MSI) has revealed key insights into the localization of metabolites in situ. Recent efforts have determined the spatial distribution of primary and secondary metabolites in plant tissues and cells. Strategies have been applied in many areas of metabolism including isotope flux analyses, plant interactions, and transcriptional regulation of metabolite accumulation. Technological advances have pushed achievable spatial resolution to subcellular levels and increased instrument sensitivity by several orders of magnitude. It is anticipated that MALDI-MSI and other MSI approaches will bring a new level of understanding to metabolomics as scientists will be encouraged to consider spatial heterogeneity of metabolites in descriptions of metabolic pathway regulation. Addresses 1 Center for Plant Lipid Research and Department of Biological Sciences, University of North Texas, 1155 Union Circle #305220, Denton, TX 76203, United States 2 Department of Chemistry, Iowa State University, 35A Roy J. Carver Co-Laboratory, Ames, IA 50011, United States 3 Ames Laboratory, US Department of Energy, Ames, IA 50011, United States Corresponding author: Chapman, Kent D ([email protected]) Current Opinion in Biotechnology 2016, 37:53–60 This review comes from a themed issue on Plant biotechnology Edited by John A Morgan and Anne Osbourn For a complete overview see the Issue and the Editorial Available online 22nd November 2015 http://dx.doi.org/10.1016/j.copbio.2015.10.004 0958-1669/# 2015 Elsevier Ltd. All rights reserved. Introduction Plant based agricultural products and chemical byproducts contribute hundreds of billions dollars annually to the global economy (www.faostat.fao.org), and originate from a diversity of grain, oilseed, fiber, and numerous other crops worldwide. Optimal production of chemicals derived from these crops requires an underlying understanding of the synthesis and metabolism of these compounds, especially in support of widespread metabolic engineering efforts to generate sustainable, bio-based products. An improved understanding of plant metabolism relies on analytical capabilities for the accurate identification and quantifica- tion of metabolites, which metabolomics has largely addressed through the development of sophisticated sep- aration techniques, mass spectrometry approaches, and computational tools [1]. Analysis of chemical/tissue extracts by these methodologies provides excellent quali- tative and quantitative information about chemical com- position, but provides little or no data regarding the original spatial distribution of metabolites in situ. A number of years ago, the Caprioli group pioneered the development of mass spectrometry imaging for localizing molecules of interest directly on the sections of mammalian tissues [2,3]. This methodology, termed matrix assisted laser desorption/ion- ization-mass spectrometry imaging (MALDI-MSI, also referred to as imaging mass spectrometry (IMS)), has since been expanded to visualize molecules directly in plant tissues and surfaces for the localization of lipids [4,5 ,6,7], proteins [8], secondary metabolites [9,10,11 ,12 ,13], and various small molecules [14 ,15–17] at unprecedented spatial and chemical resolution. The last several years have seen increasing applications of MALDI-MSI to plant tissues which have been enabled, in part, by novel instruments and matrices to improve spatial resolution and chemical coverage [18 ,19 ], streamlined matrix and sample preparation [20], and easily accessible open-source image processing free-ware [21–23]. These and other advances have enabled scientists to not only generate high resolution chemical images of plant tissues, but also to combine spatial metabolite data with further quantitative MS studies and gene expression analysis. In addition to MALDI-MSI there are other MSI platforms being utilized for MS imaging of plant tissues in situ, including desorption electrospray ionization (DESI-MS), laser ablation electrospray ionization (LAESI-MS) and secondary ion mass spectrometry (SIMS), and several salient reviews of these platforms in the context of plant MSI are available [6,24–26]. Here we highlight the most recent applications of MALDI-MS imaging in plant tissues (Table 1) for metabolite localization in situ. Efforts exam- ined here have also begun to address next-level biological questions in biochemical and metabolic contexts. It is anticipated that this new dimension of spatial analysis will provide important insights into the function, regulation and manipulation of plant metabolism. MALDI-MSI: a procedural overview A typical experimental workflow for MALDI-MSI is provided in Figure 1. First, plant tissues are flash-frozen Available online at www.sciencedirect.com ScienceDirect www.sciencedirect.com Current Opinion in Biotechnology 2016, 37:53–60
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Matrix assisted laser desorption/ionization-massspectrometry imaging (MALDI-MSI) for directvisualization of plant metabolites in situDrew Sturtevant1, Young-Jin Lee2,3 and Kent D Chapman1
Available online at www.sciencedirect.com
ScienceDirect
Direct visualization of plant tissues by matrix assisted laser
Imaging plant metabolites by mass spectrometry Sturtevant, Lee and Chapman 59
differences in distribution established and can this be
further addressed with complementary techniques such
as in situ hybridization of mRNA or MALDI-MSI of
metabolic pathway enzymes? Can three-dimensional
MALDI-MS images be developed to improve the overall
perspective of metabolite heterogeneity? What are the
technical advances that need to be made to visualize more
small molecules in situ with confidence? What ways can
MALDI-MSI be used to trace metabolism over time? Can
quantification be routinely achieved with MALDI-MSI?
Answers to these and other questions will help move
MALDI-MSI, and other approaches to localize metabo-
lites directly in plant cells and tissues, to the forefront in
biochemical research, and will continue to promise a visual
perspective of metabolism with profound and far reaching
implications.
AcknowledgementsSupport for the authors research efforts in MSI is from Cotton Incorporated(Agreement #08-395) and U.S. Department of Energy, Office of Science,Basic Energy Sciences program (DE-FG02-14ER15647) to KDC. YL wassupported by the US Department of Energy, Office of Basic EnergySciences, Division of Chemical Sciences, Geosciences, andBiosciences. The Ames Laboratory is operated by Iowa State Universityunder Contract DE-AC02-07CH11358. We thank Maria Duenas forobtaining MS images of Arabidopsis seed cross-sections shown in Figure 1.
References and recommended readingPapers of particular interest, published within the period of review,have been highlighted as:
� of special interest�� of outstanding interest
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5.�
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