The effect of isolation methods of tomato pollen on the results of metabolic profiling

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Introduction: Untargeted metabolomics is a powerful tool to detect hundreds of metabolites within a given tissue and to compare the metabolite composition of samples in a comprehensive manner. However, with regard to pollen research such comprehensive metabolomics approaches are yet not well developed. To enable isolation of pollen that is tightly enclosed within the anthers of the flower, such as immature pollen, the current pollen isolation protocols require the use of a watery solution. These protocols raise a number of concerns for their suitability in metabolomics analyses, in view of possible metabolic activities in the pollen and contamination with anther metabolites. Objectives: We assessed the effect of different sample preparation procedures currently used for pollen isolation for their suitability to perform metabolomics of tomato pollen. Methods: Pollen were isolated using different methods and the metabolic profiles were analysed by liquid chromatography–mass spectrometry (LC–MS). Results: Our results demonstrated that pollen isolation in a watery solution led to (i) rehydration of the pollen grains, inducing marked metabolic changes in flavonoids, phenylpropanoids and amino acids and thus resulting in a metabolite profile that did not reflect the one of mature dry pollen, (ii) hydrolysis of sucrose into glucose and fructose during subsequent metabolite extraction, unless the isolated and rehydrated pollen were lyophilized prior to extraction, and (iii) contamination with anther-specific metabolites, such as alkaloids, thus compromising the metabolic purity of the pollen fraction. Conclusion: We conclude that the current practices used to isolate pollen are suboptimal for metabolomics analyses and provide recommendations on how to improve the pollen isolation protocol, in order to obtain the most reliable metabolic profile from pollen tissue.

Original languageEnglish
Article number11
JournalMetabolomics
Volume15
Issue number1
DOIs
Publication statusPublished - 1 Jan 2019

Fingerprint

Lycopersicon esculentum
Metabolites
Pollen
Metabolomics
Contamination
Tissue
Fructose
Alkaloids
Flavonoids
Spectrometry
Metabolome
Sucrose
Hydrolysis
Amino Acids
Glucose
Liquids
Chemical analysis
Fluid Therapy

Keywords

  • Anther
  • Metabolite
  • Metabolome
  • Metabolomics
  • Pollen
  • Tomato

Cite this

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title = "The effect of isolation methods of tomato pollen on the results of metabolic profiling",
abstract = "Introduction: Untargeted metabolomics is a powerful tool to detect hundreds of metabolites within a given tissue and to compare the metabolite composition of samples in a comprehensive manner. However, with regard to pollen research such comprehensive metabolomics approaches are yet not well developed. To enable isolation of pollen that is tightly enclosed within the anthers of the flower, such as immature pollen, the current pollen isolation protocols require the use of a watery solution. These protocols raise a number of concerns for their suitability in metabolomics analyses, in view of possible metabolic activities in the pollen and contamination with anther metabolites. Objectives: We assessed the effect of different sample preparation procedures currently used for pollen isolation for their suitability to perform metabolomics of tomato pollen. Methods: Pollen were isolated using different methods and the metabolic profiles were analysed by liquid chromatography–mass spectrometry (LC–MS). Results: Our results demonstrated that pollen isolation in a watery solution led to (i) rehydration of the pollen grains, inducing marked metabolic changes in flavonoids, phenylpropanoids and amino acids and thus resulting in a metabolite profile that did not reflect the one of mature dry pollen, (ii) hydrolysis of sucrose into glucose and fructose during subsequent metabolite extraction, unless the isolated and rehydrated pollen were lyophilized prior to extraction, and (iii) contamination with anther-specific metabolites, such as alkaloids, thus compromising the metabolic purity of the pollen fraction. Conclusion: We conclude that the current practices used to isolate pollen are suboptimal for metabolomics analyses and provide recommendations on how to improve the pollen isolation protocol, in order to obtain the most reliable metabolic profile from pollen tissue.",
keywords = "Anther, Metabolite, Metabolome, Metabolomics, Pollen, Tomato",
author = "Paupi{\`e}re, {Marine J.} and Tikunov, {Yury M.} and Nurit Firon and {de Vos}, {Ric C.H.} and Chris Maliepaard and Visser, {Richard G.F.} and Bovy, {Arnaud G.}",
year = "2019",
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day = "1",
doi = "10.1007/s11306-018-1471-4",
language = "English",
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journal = "Metabolomics",
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The effect of isolation methods of tomato pollen on the results of metabolic profiling. / Paupière, Marine J.; Tikunov, Yury M.; Firon, Nurit; de Vos, Ric C.H.; Maliepaard, Chris; Visser, Richard G.F.; Bovy, Arnaud G.

In: Metabolomics, Vol. 15, No. 1, 11, 01.01.2019.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - The effect of isolation methods of tomato pollen on the results of metabolic profiling

AU - Paupière, Marine J.

AU - Tikunov, Yury M.

AU - Firon, Nurit

AU - de Vos, Ric C.H.

AU - Maliepaard, Chris

AU - Visser, Richard G.F.

AU - Bovy, Arnaud G.

PY - 2019/1/1

Y1 - 2019/1/1

N2 - Introduction: Untargeted metabolomics is a powerful tool to detect hundreds of metabolites within a given tissue and to compare the metabolite composition of samples in a comprehensive manner. However, with regard to pollen research such comprehensive metabolomics approaches are yet not well developed. To enable isolation of pollen that is tightly enclosed within the anthers of the flower, such as immature pollen, the current pollen isolation protocols require the use of a watery solution. These protocols raise a number of concerns for their suitability in metabolomics analyses, in view of possible metabolic activities in the pollen and contamination with anther metabolites. Objectives: We assessed the effect of different sample preparation procedures currently used for pollen isolation for their suitability to perform metabolomics of tomato pollen. Methods: Pollen were isolated using different methods and the metabolic profiles were analysed by liquid chromatography–mass spectrometry (LC–MS). Results: Our results demonstrated that pollen isolation in a watery solution led to (i) rehydration of the pollen grains, inducing marked metabolic changes in flavonoids, phenylpropanoids and amino acids and thus resulting in a metabolite profile that did not reflect the one of mature dry pollen, (ii) hydrolysis of sucrose into glucose and fructose during subsequent metabolite extraction, unless the isolated and rehydrated pollen were lyophilized prior to extraction, and (iii) contamination with anther-specific metabolites, such as alkaloids, thus compromising the metabolic purity of the pollen fraction. Conclusion: We conclude that the current practices used to isolate pollen are suboptimal for metabolomics analyses and provide recommendations on how to improve the pollen isolation protocol, in order to obtain the most reliable metabolic profile from pollen tissue.

AB - Introduction: Untargeted metabolomics is a powerful tool to detect hundreds of metabolites within a given tissue and to compare the metabolite composition of samples in a comprehensive manner. However, with regard to pollen research such comprehensive metabolomics approaches are yet not well developed. To enable isolation of pollen that is tightly enclosed within the anthers of the flower, such as immature pollen, the current pollen isolation protocols require the use of a watery solution. These protocols raise a number of concerns for their suitability in metabolomics analyses, in view of possible metabolic activities in the pollen and contamination with anther metabolites. Objectives: We assessed the effect of different sample preparation procedures currently used for pollen isolation for their suitability to perform metabolomics of tomato pollen. Methods: Pollen were isolated using different methods and the metabolic profiles were analysed by liquid chromatography–mass spectrometry (LC–MS). Results: Our results demonstrated that pollen isolation in a watery solution led to (i) rehydration of the pollen grains, inducing marked metabolic changes in flavonoids, phenylpropanoids and amino acids and thus resulting in a metabolite profile that did not reflect the one of mature dry pollen, (ii) hydrolysis of sucrose into glucose and fructose during subsequent metabolite extraction, unless the isolated and rehydrated pollen were lyophilized prior to extraction, and (iii) contamination with anther-specific metabolites, such as alkaloids, thus compromising the metabolic purity of the pollen fraction. Conclusion: We conclude that the current practices used to isolate pollen are suboptimal for metabolomics analyses and provide recommendations on how to improve the pollen isolation protocol, in order to obtain the most reliable metabolic profile from pollen tissue.

KW - Anther

KW - Metabolite

KW - Metabolome

KW - Metabolomics

KW - Pollen

KW - Tomato

U2 - 10.1007/s11306-018-1471-4

DO - 10.1007/s11306-018-1471-4

M3 - Article

VL - 15

JO - Metabolomics

JF - Metabolomics

SN - 1573-3882

IS - 1

M1 - 11

ER -