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An efficient method for medium throughput screening of cuticular wax composition in different plant species

Abstract

Introduction

Most aerial plant organs are covered by a cuticle, which largely consists of cutin and wax. Cuticular waxes are mixtures of dozens of compounds, mostly very-long-chain aliphatics that are easily extracted by solvents. Over the last four decades, diverse cuticular wax analysis protocols have been developed, most of which are complex and time-consuming, and need to be adapted for each plant species or organ. Plant genomics and breeding programs often require mid-throughput metabolic phenotyping approaches to screen large numbers of individuals and obtain relevant biological information.

Objectives

To generate a fast, simple and user-friendly methodology able to capture most wax complexity independently of the plant, cultivar and organ.

Methods

Here we present a simple GC–MS method for screening relatively small wax amounts, sampled by short extraction with a versatile, uniform solvent. The method will be tested and validated in leaves and fruits from three different crop species: tomato (Solanum lycopersicum), apple (Malus domestica) and hybrid aspen (Populus tremula × tremuloides).

Results

Consistent results were obtained in tomato cultivar M82 across three consecutive years (2010–2012), two organs (leaf and fruit), and also in two different tomato (M82 and MicroTom) and apple (Golden Delicious and Granny Smith) cultivars. Our results on tomato wax composition match those reported previously, while our apple and hybrid aspen analyses provide the first comprehensive cuticular wax profile of these species.

Conclusion

This protocol allows standardized identification and quantification of most cuticular wax components in a range of species.

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Acknowledgments

We thank the Adelis Foundation, the Leona M. and Harry B. Helmsley Charitable Trust, the Jeanne and Joseph Nissim Foundation for Life Sciences, the Tom and Sondra Rykoff Family Foundation Research and the Raymond Burton Plant Genome Research Fund for supporting the A. A. lab activity. A. A. is the incumbent of the Peter J. Cohn Professorial Chair. A. A., A. G. and J. P. F. M. thank COST FA1106 Quality Fruit for STSM and networking activities.

Funding

Research at the IBMCP was supported by MINECO Grant BIO2013-42193-R and from EC H2020 TRADITOM SFS7a-2014- (contract 634561) to Antonio Granell and by FPU-MECD personal Grant to Josefina Patricia Fernandez Moreno (AP-2007-01905). Research at the Weizmann Institute of Sciences was supported by the Israel Science Foundation (ISF) personal Grant to Asaph Aharoni (ISF Grant No. 646/11). We also thank COST FA1106 Quality Fruit for funding networking activities.

Author information

Correspondence to Asaph Aharoni or Antonio Granell.

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Authors declare that they have not conflict of interest.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Fernandez-Moreno, J., Malitsky, S., Lashbrooke, J. et al. An efficient method for medium throughput screening of cuticular wax composition in different plant species. Metabolomics 12, 73 (2016). https://doi.org/10.1007/s11306-016-0982-0

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Keywords

  • Metabolic profiling
  • Cuticular waxes
  • Fruit surface
  • Fleshy fruit