Imaging Mass Spectrometry, Metabolism, and New Views of the Microbial World

Hoefler, B. Christopher; Straight, Paul D.

doi:10.1002/9781118876015.ch10

Cited by 4 publications

(5 citation statements)

References 235 publications

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“…However, for MALDI-TOF-IMS bacterial cultures are initially grown on an agar plate, which is subsequently covered in MALDI matrix and scanned over a two-dimensional area by programming a mass spectrometer to collect mass spectra at each X,Y with defined raster distance. The mass spectra are composited together into a spatially organized dataset such that the distribution of molecules produced by each organism can be mapped ( Yang et al, 2009 ; Hoefler and Straight, 2014 ). MALDI-TOF-IMS has been used to investigate bacterial competition in a number of systems.…”

Section: Culture-dependent Models For Bacterial Competitionmentioning

confidence: 99%

Bacterial Communities: Interactions to Scale

Vargas-Bautista²,

2016

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In the environment, bacteria live in complex multispecies communities. These communities span in scale from small, multicellular aggregates to billions or trillions of cells within the gastrointestinal tract of animals. The dynamics of bacterial communities are determined by pairwise interactions that occur between different species in the community. Though interactions occur between a few cells at a time, the outcomes of these interchanges have ramifications that ripple through many orders of magnitude, and ultimately affect the macroscopic world including the health of host organisms. In this review we cover how bacterial competition influences the structures of bacterial communities. We also emphasize methods and insights garnered from culture-dependent pairwise interaction studies, metagenomic analyses, and modeling experiments. Finally, we argue that the integration of multiple approaches will be instrumental to future understanding of the underlying dynamics of bacterial communities.

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Section: Culture-dependent Models For Bacterial Competitionmentioning

confidence: 99%

Bacterial Communities: Interactions to Scale

Vargas-Bautista²,

2016

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“…Data analysis using FlexImaging: The spectra were internally calibrated by matrix ions and the datasets were analyzed using FlexImaging 4.0 (Bruker Daltonics, Bremen, Germany), in which the ion of interest was manually filtered from the mean or individual spectra using 0.5-1.0 Da increments. The images were normalized using TIC mode, and a hypothetical color was assigned to each specific ion [15,17,23].…”

Section: Maldi-msi Analysismentioning

confidence: 99%

“…Mass spectrometry (MS) techniques have been widely applied in this area [12]. Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) has also been applied in metabolomics studies, offering the opportunity to monitor the spatial and temporal distributions of metabolites [13][14][15]. Recently, MALDI-MSI was successful in revealing secondary metabolites involved in intracellular metabolism [16], as well as monitoring metabolic exchanges in interspecies relationships [17,18].…”

Section: Introductionmentioning

confidence: 99%

Influence of Citrus sunki and Poncirus trifoliata Root Extracts on Metabolome of Phytophthora parasitica

Maximo,

Araújo,

Pagotto

et al. 2024

Metabolites

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Phytophthora parasitica is an oomycete pathogen that infects a broad range of crops of worldwide economic interest; among them are citrus species. In general, some Citrus and the rootstocks of related genera offer considerable resistance against P. parasitica; therefore, understanding the mechanisms involved in the virulence of this pathogen is crucial. In this work, P. parasitica secondary metabolite production was studied using matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) and ultrahigh-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (UHPLC/ESI-Q-TOF-MS) combined with chemometric tools, and its metabolic profile was evaluated under the influence of Citrus sunki (a highly susceptible host) and Poncirus trifoliata (a resistant genotype) extracts. The root extracts of Citrus sunki had an influence on the growth and hyphae morphology, and the root extracts of P. trifoliata had an influence on the zoospore behavior. In parallel, the spatial distribution of several metabolites was revealed in P. parasitica colonies using MALDI-MSI, and the metabolite ion of m/z 246 was identified as the protonated molecule of Arg-Ala. The MALDI-MSI showed variations in the surface metabolite profile of P. parasitica under the influence of the P. trifoliata extract. The P. parasitica metabolome analysis using UHPLC-ESI-Q-TOF-MS resulted in the detection of Arg-Gln (m/z 303.1775), as well as L-arginine (m/z 175.1191) and other unidentified metabolites. Significant variations in this metabolome were detected under the influence of the plant extracts when evaluated using UHPLC-ESI-Q-TOF-MS. Both techniques proved to be complementary, offering valuable insights at the molecular level when used to assess the impact of the plant extracts on microbial physiology in vitro. The metabolites identified in this study may play significant roles in the interaction or virulence of P. parasitica, but their functional characterization remains to be analyzed. Overall, these data confirm our initial hypotheses, demonstrating that P. parasitica has the capabilities of (i) recognizing host signals and altering its reproductive programing and (ii) distinguishing between hosts with varying responses in terms of reproduction and the production of secondary metabolites.

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“…To investigate interactions in detail, matrix assisted laser desorption/ionization time-of-flight imaging mass spectrometry (MALDI-TOF-IMS) can be used. This approach provides detailed information on the composition and distribution of small molecules and peptides produced by interacting microbial colonies with high spatial resolution, and has been used in multiple studies of bacterial interactions to characterize the mechanisms of competition [12][13][14][15] . However, MALDI-TOF-IMS often requires laborious sample preparation, specialized expertise to operate the equipment, and expensive, specialized mass spectrometers.…”

Section: Introductionmentioning

confidence: 99%

High Throughput Co-culture Assays for the Investigation of Microbial Interactions

Carlson

Stubbendieck

Currie

et al. 2019

JoVE

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The study of interactions between microorganisms has led to numerous discoveries, from novel antimicrobials to insights in microbial ecology. Many approaches used for the study of microbial interactions require specialized equipment, are expensive, and/or are time intensive. This paper presents a protocol for co-culture interaction assays that are inexpensive, scalable to large sample numbers, and easily adaptable to numerous experimental designs. Microorganisms are cultured together on 12-well plates, with each well representing one pairwise combination of microorganisms. A test organism is cultured on one side of each individual well, and first incubated in monoculture. Subsequently, target organisms are simultaneously inoculated onto the opposite side of each of the wells using a 3D printed inoculation stamp. After co-culture, the completed assays are scored for visual phenotypes, such as growth or inhibition. These assays can be used to confirm phenotypes or identify patterns among isolates of interest. Using this simple and effective method, users can analyze many combinations of microorganisms rapidly and efficiently. This co-culture approach is applicable to antibiotic discovery, as well as, culture-based microbiome research and has already been successfully applied to both applications.

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Imaging Mass Spectrometry, Metabolism, and New Views of the Microbial World

Cited by 4 publications

References 235 publications

Bacterial Communities: Interactions to Scale

Bacterial Communities: Interactions to Scale

Influence of Citrus sunki and Poncirus trifoliata Root Extracts on Metabolome of Phytophthora parasitica

High Throughput Co-culture Assays for the Investigation of Microbial Interactions

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