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A quantitative method to measure geranylgeranyl diphosphate (GGPP) and geranylgeranyl monophosphate (GGP) in tomato (Solanum lycopersicum) fruit

BACKGROUND: Isoprenoids are a very large class of metabolites playing a key role in plant physiological processes such as growth, stress resistance, fruit flavor, and color. In chloroplasts and chromoplasts, the diterpene compound geranylgeranyl diphosphate (GGPP) is the metabolic precursor required...

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Autores principales: Zita, Wayne, Shanmugabalaji, Venkatasalam, Ezquerro, Miguel, Rodriguez-Concepcion, Manuel, Kessler, Felix, Glauser, Gaetan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10249317/
https://www.ncbi.nlm.nih.gov/pubmed/37287006
http://dx.doi.org/10.1186/s13007-023-01034-w
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author Zita, Wayne
Shanmugabalaji, Venkatasalam
Ezquerro, Miguel
Rodriguez-Concepcion, Manuel
Kessler, Felix
Glauser, Gaetan
author_facet Zita, Wayne
Shanmugabalaji, Venkatasalam
Ezquerro, Miguel
Rodriguez-Concepcion, Manuel
Kessler, Felix
Glauser, Gaetan
author_sort Zita, Wayne
collection PubMed
description BACKGROUND: Isoprenoids are a very large class of metabolites playing a key role in plant physiological processes such as growth, stress resistance, fruit flavor, and color. In chloroplasts and chromoplasts, the diterpene compound geranylgeranyl diphosphate (GGPP) is the metabolic precursor required for the biosynthesis of tocopherols, plastoquinones, phylloquinone, chlorophylls, and carotenoids. Despite its key role for the plant metabolism, reports on GGPP physiological concentrations in planta have been extremely scarce. RESULTS: In this study, we developed a method to quantify GGPP and its hydrolysis product geranylgeranyl monophosphate (GGP) from tomato fruit, using ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC–MS/MS). Quantification was done by external calibration and the method was validated in terms of specificity, precision, accuracy, and detection and quantitation limits. We further demonstrate the validity of our approach by analysing GGPP contents in the ripe fruits of wild-type tomatoes and mutants defective in GGPP production. Finally, we also show that the sample preparation is key to prevent GGPP hydrolysis and mitigate its conversion to GGP. CONCLUSION: Our study provides an efficient tool to investigate the metabolic fluxes required for GGPP supply and consumption in tomato fruit. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13007-023-01034-w.
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spelling pubmed-102493172023-06-09 A quantitative method to measure geranylgeranyl diphosphate (GGPP) and geranylgeranyl monophosphate (GGP) in tomato (Solanum lycopersicum) fruit Zita, Wayne Shanmugabalaji, Venkatasalam Ezquerro, Miguel Rodriguez-Concepcion, Manuel Kessler, Felix Glauser, Gaetan Plant Methods Methodology BACKGROUND: Isoprenoids are a very large class of metabolites playing a key role in plant physiological processes such as growth, stress resistance, fruit flavor, and color. In chloroplasts and chromoplasts, the diterpene compound geranylgeranyl diphosphate (GGPP) is the metabolic precursor required for the biosynthesis of tocopherols, plastoquinones, phylloquinone, chlorophylls, and carotenoids. Despite its key role for the plant metabolism, reports on GGPP physiological concentrations in planta have been extremely scarce. RESULTS: In this study, we developed a method to quantify GGPP and its hydrolysis product geranylgeranyl monophosphate (GGP) from tomato fruit, using ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC–MS/MS). Quantification was done by external calibration and the method was validated in terms of specificity, precision, accuracy, and detection and quantitation limits. We further demonstrate the validity of our approach by analysing GGPP contents in the ripe fruits of wild-type tomatoes and mutants defective in GGPP production. Finally, we also show that the sample preparation is key to prevent GGPP hydrolysis and mitigate its conversion to GGP. CONCLUSION: Our study provides an efficient tool to investigate the metabolic fluxes required for GGPP supply and consumption in tomato fruit. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13007-023-01034-w. BioMed Central 2023-06-07 /pmc/articles/PMC10249317/ /pubmed/37287006 http://dx.doi.org/10.1186/s13007-023-01034-w Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Methodology
Zita, Wayne
Shanmugabalaji, Venkatasalam
Ezquerro, Miguel
Rodriguez-Concepcion, Manuel
Kessler, Felix
Glauser, Gaetan
A quantitative method to measure geranylgeranyl diphosphate (GGPP) and geranylgeranyl monophosphate (GGP) in tomato (Solanum lycopersicum) fruit
title A quantitative method to measure geranylgeranyl diphosphate (GGPP) and geranylgeranyl monophosphate (GGP) in tomato (Solanum lycopersicum) fruit
title_full A quantitative method to measure geranylgeranyl diphosphate (GGPP) and geranylgeranyl monophosphate (GGP) in tomato (Solanum lycopersicum) fruit
title_fullStr A quantitative method to measure geranylgeranyl diphosphate (GGPP) and geranylgeranyl monophosphate (GGP) in tomato (Solanum lycopersicum) fruit
title_full_unstemmed A quantitative method to measure geranylgeranyl diphosphate (GGPP) and geranylgeranyl monophosphate (GGP) in tomato (Solanum lycopersicum) fruit
title_short A quantitative method to measure geranylgeranyl diphosphate (GGPP) and geranylgeranyl monophosphate (GGP) in tomato (Solanum lycopersicum) fruit
title_sort quantitative method to measure geranylgeranyl diphosphate (ggpp) and geranylgeranyl monophosphate (ggp) in tomato (solanum lycopersicum) fruit
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10249317/
https://www.ncbi.nlm.nih.gov/pubmed/37287006
http://dx.doi.org/10.1186/s13007-023-01034-w
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