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A comprehensive comparison of four methods for extracting lipids from Arabidopsis tissues

BACKGROUND: The plant lipidome is highly complex, and the composition of lipids in different tissues as well as their specific functions in plant development, growth and stress responses have yet to be fully elucidated. To do this, efficient lipid extraction protocols which deliver target compounds...

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Autores principales: Kehelpannala, Cheka, Rupasinghe, Thusitha W. T., Hennessy, Thomas, Bradley, David, Ebert, Berit, Roessner, Ute
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7713330/
https://www.ncbi.nlm.nih.gov/pubmed/33292337
http://dx.doi.org/10.1186/s13007-020-00697-z
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author Kehelpannala, Cheka
Rupasinghe, Thusitha W. T.
Hennessy, Thomas
Bradley, David
Ebert, Berit
Roessner, Ute
author_facet Kehelpannala, Cheka
Rupasinghe, Thusitha W. T.
Hennessy, Thomas
Bradley, David
Ebert, Berit
Roessner, Ute
author_sort Kehelpannala, Cheka
collection PubMed
description BACKGROUND: The plant lipidome is highly complex, and the composition of lipids in different tissues as well as their specific functions in plant development, growth and stress responses have yet to be fully elucidated. To do this, efficient lipid extraction protocols which deliver target compounds in solution at concentrations adequate for subsequent detection, quantitation and analysis through spectroscopic methods are required. To date, numerous methods are used to extract lipids from plant tissues. However, a comprehensive analysis of the efficiency and reproducibility of these methods to extract multiple lipid classes from diverse tissues of a plant has not been undertaken. RESULTS: In this study, we report the comparison of four different lipid extraction procedures in order to determine the most effective lipid extraction protocol to extract lipids from different tissues of the model plant Arabidopsis thaliana. CONCLUSION: While particular methods were best suited to extract different lipid classes from diverse Arabidopsis tissues, overall a single-step extraction method with a 24 h extraction period, which uses a mixture of chloroform, isopropanol, methanol and water, was the most efficient, reproducible and the least labor-intensive to extract a broad range of lipids for untargeted lipidomic analysis of Arabidopsis tissues. This method extracted a broad range of lipids from leaves, stems, siliques, roots, seeds, seedlings and flowers of Arabidopsis. In addition, appropriate methods for targeted lipid analysis of specific lipids from particular Arabidopsis tissues were also identified.
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spelling pubmed-77133302020-12-03 A comprehensive comparison of four methods for extracting lipids from Arabidopsis tissues Kehelpannala, Cheka Rupasinghe, Thusitha W. T. Hennessy, Thomas Bradley, David Ebert, Berit Roessner, Ute Plant Methods Methodology BACKGROUND: The plant lipidome is highly complex, and the composition of lipids in different tissues as well as their specific functions in plant development, growth and stress responses have yet to be fully elucidated. To do this, efficient lipid extraction protocols which deliver target compounds in solution at concentrations adequate for subsequent detection, quantitation and analysis through spectroscopic methods are required. To date, numerous methods are used to extract lipids from plant tissues. However, a comprehensive analysis of the efficiency and reproducibility of these methods to extract multiple lipid classes from diverse tissues of a plant has not been undertaken. RESULTS: In this study, we report the comparison of four different lipid extraction procedures in order to determine the most effective lipid extraction protocol to extract lipids from different tissues of the model plant Arabidopsis thaliana. CONCLUSION: While particular methods were best suited to extract different lipid classes from diverse Arabidopsis tissues, overall a single-step extraction method with a 24 h extraction period, which uses a mixture of chloroform, isopropanol, methanol and water, was the most efficient, reproducible and the least labor-intensive to extract a broad range of lipids for untargeted lipidomic analysis of Arabidopsis tissues. This method extracted a broad range of lipids from leaves, stems, siliques, roots, seeds, seedlings and flowers of Arabidopsis. In addition, appropriate methods for targeted lipid analysis of specific lipids from particular Arabidopsis tissues were also identified. BioMed Central 2020-12-03 /pmc/articles/PMC7713330/ /pubmed/33292337 http://dx.doi.org/10.1186/s13007-020-00697-z Text en © The Author(s) 2020 Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://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
Kehelpannala, Cheka
Rupasinghe, Thusitha W. T.
Hennessy, Thomas
Bradley, David
Ebert, Berit
Roessner, Ute
A comprehensive comparison of four methods for extracting lipids from Arabidopsis tissues
title A comprehensive comparison of four methods for extracting lipids from Arabidopsis tissues
title_full A comprehensive comparison of four methods for extracting lipids from Arabidopsis tissues
title_fullStr A comprehensive comparison of four methods for extracting lipids from Arabidopsis tissues
title_full_unstemmed A comprehensive comparison of four methods for extracting lipids from Arabidopsis tissues
title_short A comprehensive comparison of four methods for extracting lipids from Arabidopsis tissues
title_sort comprehensive comparison of four methods for extracting lipids from arabidopsis tissues
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7713330/
https://www.ncbi.nlm.nih.gov/pubmed/33292337
http://dx.doi.org/10.1186/s13007-020-00697-z
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