Simultaneous Determination of Sulphur metabolites in <i>Arabidopsis thaliana</i> via LC‐ESI‐MS/MS and <sup>34</sup>S‐metabolic Labelling

Hsieh, Chin‐Lin; Yeh, Kai‐Wun; Kok, Luit J. De; Pan, Ryh‐Nan; Kuo, Yueh‐Hsiung; Tseng, Mei‐Hui

doi:10.1002/pca.1361

Cited by 5 publications

(5 citation statements)

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“…The cysteine autotrophy of plants allows labeling of sulfur‐containing metabolites by feeding of stable sulfur isotope anions (Giavalisco et al ., ; Hsieh et al ., ). In order to determine the degree of isotope labeling and to optimize the strategy for labeling sulfur‐containing compounds in Arabidopsis, seeds were germinated and seedlings were grown in the presence of various concentrations of 34 S‐sulfate.…”

Section: Resultsmentioning

confidence: 97%

“…The reproducibility and comparability of the analysis are critical for identification of metabolites that are selectively enriched in samples by using an untargeted approach (Saito and Matsuda, ). Recently, 34 S labeling was introduced for metabolome analysis using online liquid chromatography and high‐resolution MS (Giavalisco et al ., ; Hsieh et al ., ). The studies identified sulfur‐containing metabolites in Arabidopsis based on the labeling‐specific isotopic pattern.…”

Section: Discussionmentioning

confidence: 97%

“…Hsieh et al . () used LC‐MS and positive ionization for quantification of prominent sulfur compounds such as GSH in plant extracts. However, the capability for sulfur metabolite detection in a targeted or untargeted manner was not explored.…”

Section: Discussionmentioning

confidence: 99%

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Exploring the Arabidopsis sulfur metabolome

et al. 2013

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SUMMARYSulfur plays a crucial role in protein structure and function, redox status and plant biotic stress responses. However, our understanding of sulfur metabolism is limited to identified pathways. In this study, we used a high-resolution Fourier transform mass spectrometric approach in combination with stable isotope labeling to describe the sulfur metabolome of Arabidopsis thaliana. Databases contain roughly 300 sulfur compounds assigned to Arabidopsis. In comparative analyses, we showed that the overlap of the expected sulfur metabolome and the mass spectrometric data was surprisingly low, and we were able to assign only 37 of the 300 predicted compounds. By contrast, we identified approximately 140 sulfur metabolites that have not been assigned to the databases to date. We used our method to characterize the c-glutamyl transferase mutant ggt4-1, which is involved in the vacuolar breakdown of glutathione conjugates in detoxification reactions. Although xenobiotic substrates are well known, only a few endogenous substrates have been described. Among the specifically altered sulfur-containing masses in the ggt4-1 mutant, we characterized one endogenous glutathione conjugate and a number of further candidates for endogenous substrates. The small percentage of predicted compounds and the high proportion of unassigned sulfur compounds identified in this study emphasize the need to re-evaluate our understanding of the sulfur metabolome.

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Section: Resultsmentioning

confidence: 97%

Section: Discussionmentioning

confidence: 97%

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Exploring the Arabidopsis sulfur metabolome

et al. 2013

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“…Perchloric acid was used to protect metabolites from enzymatic degradation [ 41 ], and to avoid sulfur metabolites oxidation and degradation [ 14 , 42 , 43 ] under basic and neutral condition. Formic acid was used for enhancing electrospray ionization.…”

Section: Methodsmentioning

confidence: 99%

Simultaneous untargeted and targeted metabolomics profiling of underivatized primary metabolites in sulfur-deficient barley by ultra-high performance liquid chromatography-quadrupole/time-of-flight mass spectrometry

Ghosson

Schwarzenberg²,

Jamois³

et al. 2018

Plant Methods

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BackgroundMetabolomics based on mass spectrometry analysis are increasingly applied in diverse scientific domains, notably agronomy and plant biology, in order to understand plants’ behaviors under different stress conditions. In fact, these stress conditions are able to disrupt many biosynthetic pathways that include mainly primary metabolites. Profiling and quantifying primary metabolites remain a challenging task because they are poorly retained in reverse phase columns, due to their high polarity and acid–base properties. The aim of this work is to develop a simultaneous untargeted/targeted profiling of amino acids, organic acids, sulfur metabolites, and other several metabolites. This method will be applied on sulfur depleted barley, in order to study this type of stress, which is difficult to detect at early stage. Also, this method aims to explore the impact of this stress on barley’s metabolome.ResultsUltra-high performance liquid chromatography–high resolution mass spectrometry-based method was successfully applied to real samples allowing to discriminate, detect, and quantify primary metabolites in short-runs without any additional sampling step such as derivatization or ion pairing. The retention of polar metabolites was successfully achieved using modified C18 columns with high reproducibility (relative standard deviation below 10%). The quantification method showed a high sensitivity and robustness. Furthermore, high resolution mass spectrometry detection provided reliable quantification based on exact mass, eliminating potential interferences, and allowing the simultaneous untargeted metabolomics analysis. The untargeted data analysis was conducted using Progenesis QI software, performing alignment, peak picking, normalization and multivariate analysis. The simultaneous analysis provided cumulative information allowing to discriminate between two plant batches. Thus, discriminant biomarkers were identified and validated. Simultaneously, quantification confirmed coherently the relative abundance of these biomarkers.ConclusionsA fast and innovated simultaneous untargeted/targeted method has successfully been developed and applied to sulfur deficiency on barley. This work opens interesting perspectives in both fundamental and applied research. Biomarker discovery give precious indication to understand plant behavior during a nutritional deficiency. Thus, direct or indirect measurement of these compounds allows a real time fertilization management and encounter the challenges of sustainable agriculture.Electronic supplementary materialThe online version of this article (10.1186/s13007-018-0329-0) contains supplementary material, which is available to authorized users.

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“…e number of distinct proteins that can be identi ed from complex samples has been increased by the use of shotgun proteomics, a gel-free liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis, compared to traditional gel-based approaches [36,46,[58][59][60]. However, there is no single standardized procedure for the analysis of all proteins and metabolites because these are highly diverse and biochemically heterogeneous [37].…”

Section: Proteomics Studiesmentioning

confidence: 99%

Arabidopsis thaliana and omics approaches: a review

Carneiro¹,

Madrid²,

Maciel³

et al. 2015

JIOMICS

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Arabidopsis thaliana is a small, owering plant that is widely used as a model organism in plant biology, mainly because it is the rst plant to have its entire genome sequenced. It has since proven to be an ideal organism for studying plant development. Arabidopsis is commonly used as a model plant for genomics, metabolomics and proteomics studies, and more recently it has been utilized in metallomic studies. Because of its widespread applications, many methods for Arabidopsis sample preparation, analytes separation and data quanti cation have been explored. is review brie y describes the Arabidopsis thaliana characteristics, the developed researches and the primary methods using this plant in di erent elds of OMICS. In the future, the availability of Arabidopsis genomic information may result in its continuous development for nanoparticles and metallomics studies.

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Simultaneous Determination of Sulphur metabolites in Arabidopsis thaliana via LC‐ESI‐MS/MS and ³⁴S‐metabolic Labelling

Abstract: The validated method for determination of methionine, reduced glutathione and oxidised glutathione was effectively applied to measure metabolite dynamics of sulphur-containing metabolites at the whole-plant level.

Cited by 5 publications

References 41 publications

Exploring the Arabidopsis sulfur metabolome

Exploring the Arabidopsis sulfur metabolome

Simultaneous untargeted and targeted metabolomics profiling of underivatized primary metabolites in sulfur-deficient barley by ultra-high performance liquid chromatography-quadrupole/time-of-flight mass spectrometry

Arabidopsis thaliana and omics approaches: a review

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Simultaneous Determination of Sulphur metabolites in Arabidopsis thaliana via LC‐ESI‐MS/MS and 34S‐metabolic Labelling

Abstract: The validated method for determination of methionine, reduced glutathione and oxidised glutathione was effectively applied to measure metabolite dynamics of sulphur-containing metabolites at the whole-plant level.

Cited by 5 publications

References 41 publications

Exploring the Arabidopsis sulfur metabolome

Exploring the Arabidopsis sulfur metabolome

Simultaneous untargeted and targeted metabolomics profiling of underivatized primary metabolites in sulfur-deficient barley by ultra-high performance liquid chromatography-quadrupole/time-of-flight mass spectrometry

Arabidopsis thaliana and omics approaches: a review

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Simultaneous Determination of Sulphur metabolites in Arabidopsis thaliana via LC‐ESI‐MS/MS and ³⁴S‐metabolic Labelling