Live Single-Cell Plant Hormone Analysis by Video-Mass Spectrometry

Shimizu, Takafumi; Miyakawa, Shinya; Esaki, Tsuyoshi; Mizuno, Hajime; Masujima, Tsutomu; Koshiba, Tomokazu; Seo, Mitsunori

doi:10.1093/pcp/pcv042

Cited by 40 publications

(33 citation statements)

References 30 publications

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“…It will be interesting in future work to investigate the question of synthesis versus sensitivity. Recently developed single cell approaches [ 50–52 ,] should make it possible to investigate whether elevated [CO 2 ] induces an increase in guard cell ABA (either through de novo synthesis or release from conjugated forms). Similarly, the question of sensitivity could be probed in the nced3 nced5 background by investigating whether there is a level of exogenously applied ABA that does not promote stomatal closure on its own but that imparts sensitivity of this mutant to [CO 2 ].…”

Section: Resultsmentioning

confidence: 99%

Elevated CO 2 -Induced Responses in Stomata Require ABA and ABA Signaling

et al. 2015

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SummaryAn integral part of global environment change is an increase in the atmospheric concentration of CO2 ([CO2]) [1]. Increased [CO2] reduces leaf stomatal apertures and density of stomata that plays out as reductions in evapotranspiration [2–4]. Surprisingly, given the importance of transpiration to the control of terrestrial water fluxes [5] and plant nutrient acquisition [6], we know comparatively little about the molecular components involved in the intracellular signaling pathways by which [CO2] controls stomatal development and function [7]. Here, we report that elevated [CO2]-induced closure and reductions in stomatal density require the generation of reactive oxygen species (ROS), thereby adding a new common element to these signaling pathways. We also show that the PYR/RCAR family of ABA receptors [8, 9] and ABA itself are required in both responses. Using genetic approaches, we show that ABA in guard cells or their precursors is sufficient to mediate the [CO2]-induced stomatal density response. Taken together, our results suggest that stomatal responses to increased [CO2] operate through the intermediacy of ABA. In the case of [CO2]-induced reductions in stomatal aperture, this occurs by accessing the guard cell ABA signaling pathway. In both [CO2]-mediated responses, our data are consistent with a mechanism in which ABA increases the sensitivity of the system to [CO2] but could also be explained by requirement for a CO2-induced increase in ABA biosynthesis specifically in the guard cell lineage. Furthermore, the dependency of stomatal [CO2] signaling on ABA suggests that the ABA pathway is, in evolutionary terms, likely to be ancestral.

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Section: Resultsmentioning

confidence: 99%

Elevated CO 2 -Induced Responses in Stomata Require ABA and ABA Signaling

et al. 2015

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“…[90][91][92] Fore xample,t his technique was applied to the identification and quantitation of plant hormones in single plant cells by introducing stable isotope-labeled standards in the electrospray solution. [93] Recently,this technique was also applied to detect the localization of terpenoid indole alkaloids in specific cell types in Catharanthus roseus stems. [94] Capillary microsampling ESI-IMS-MS utilizes custom tailored capillaries and micromanipulators to extract 5-10 % of the contents from cells of interest.…”

Section: Ambient Ionization Methods Offer Low Perturbationmentioning

confidence: 99%

Single‐Cell Mass Spectrometry Approaches to Explore Cellular Heterogeneity

2018

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Compositional diversity is a fundamental property in cell populations. Single-cell analysis promises new insight into this cellular heterogeneity on the genomic, transcriptomic, proteomic, and metabolomic levels. Mass spectrometry (MS) is a label-free technique that enables the multiplexed analysis of proteins, peptides, lipids, and metabolites in individual cells. The abundances of these molecular classes are correlated with the physiological states and environmental responses of the cells. In this Minireview, we discuss recent advances in single-cell MS techniques with an emphasis on sampling and ionization methods developed for volume-limited samples. Strategies for sample treatment, separation methods, and data analysis require special considerations for single cells. Ongoing analytical challenges include subcellular heterogeneity, non-normal statistical distributions of cellular properties, and the need for high-throughput, high molecular coverage and minimal perturbation.

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“…Diese Methode wurde auf die Stoffwechselanalyse einzelner Säugetier‐ und Pflanzenzellen angewendet . So wurden mit dieser Technik Pflanzenhormone in pflanzlichen Einzelzellen identifiziert und quantifiziert, indem die Elektrospray‐Lösung mit stabilen isotopenmarkierten Standardsubstanzen versetzt wurde . Diese Methode diente auch dazu, die Lokalisation von Terpenoid‐Indolalkaloiden in bestimmten Zellarten von Catharanthus‐roseus ‐Stämmen zu ermitteln …”

Section: Einzelzellproteomik Und ‐Metabolomikunclassified

Einzelzell‐Massenspektrometrie zur Untersuchung zellulärer Heterogenität

Zhang

Vertes

2018

Angewandte Chemie

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Die Diversität der Zusammensetzung ist eine fundamentale Eigenschaft von Zellpopulationen. Neue Einblicke in diese zelluläre Heterogenität auf Genom‐, Transkriptom‐, Proteom‐ und Metabolomniveau verspricht die Einzelzellanalyse. Die Massenspektrometrie (MS) ist ein markierungsfreies Verfahren, das die Multiplex‐Analyse von Proteinen, Peptiden, Lipiden und Metaboliten in Einzelzellen ermöglicht. Die Häufigkeiten dieser Verbindungsklassen werden mit den physiologischen Zuständen und den Reaktionen der Zellen auf Umgebungsänderungen korreliert. Dieser Kurzaufsatz beschreibt neue Fortschritte bei den Einzelzell‐MS‐Techniken mit dem Schwerpunkt auf Entnahme‐ und Ionisationsmethoden, die für volumenbegrenzte Proben entwickelt wurden. Bei Einzelzellen erfordern die Strategien zur Probenbehandlung, die Trennmethoden und die Datenanalyse besondere Überlegungen. Aktuelle analytische Herausforderungen betreffen unter anderem die subzelluläre Heterogenität, nicht normale statistische Verteilungen von Zelleigenschaften sowie die Notwendigkeit von Hochdurchsatzanwendungen, einem breiten Molekülspektrum und minimaler Störung.

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Live Single-Cell Plant Hormone Analysis by Video-Mass Spectrometry

Cited by 40 publications

References 30 publications

Elevated CO 2 -Induced Responses in Stomata Require ABA and ABA Signaling

Elevated CO 2 -Induced Responses in Stomata Require ABA and ABA Signaling

Single‐Cell Mass Spectrometry Approaches to Explore Cellular Heterogeneity

Einzelzell‐Massenspektrometrie zur Untersuchung zellulärer Heterogenität

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