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Alternative methods for sampling and preservation of photosynthetic pigments and tocopherols in plant material from remote locations

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Abstract

Current methods for the study of pigments involve freezing in liquid nitrogen and storage at −80°C or lyophilization until HPLC analysis. These requirements greatly restrict ecophysiological research in remote areas where such resources are hardly available. We aimed to overcome such limitations by developing several techniques not requiring freezing or lyophilization. Two species with contrasting foliar characteristics (Olea europaea and Taraxacum officinale) were chosen. Seven preservation methods were designed, optimized and tested in a field trial. These protocols were compared with a control immediately frozen after collection. Pigments and tocopherols were analysed by HPLC. Main artefacts were chlorophyll epimerization or phaeophytinization, carotenoid isomerization, altered de-epoxidation index and tocopherol degradation. Among all methods, sample desiccation in silica gel provides robust samples (pigment composition was unaffected by storage time or temperature) and almost unaltered pigment profiles, except for a shift in epoxidation state. Although liquid nitrogen freezing and subsequent lyophilization or freezer storage were preferred, when these facilities are either not available or not suitable for long-distance transport, desiccation with silica gel, passive extraction in acetone and/or storage of fresh samples in water vapour saturated atmospheres enable a complete pigment characterization. Silica gel is advisable for long-term sample conservation.

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Acknowledgements

We thank Beatriz Fernandez-Marín and Noé Velazquez for help during analyses. R. E. received a doctoral grant from the Basque Government and M. P.-M. from the University of Barcelona. Funding for this research was provided by the Spanish Ministry of Education and Science (projects BFU 2007-62637, CGL 2005-03062/BOS, CGL2005-03998/BOS) and by the Basque Government (project UPV/EHU-GV IT-299-07).

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Authors and Affiliations

  1. Departamento de Biología Vegetal y Ecología, Universidad del País Vasco/EHU, Apdo 644, 48080, Bilbao, Spain

    Raquel Esteban, Unai Artetxe, José María Becerril & José Ignacio García-Plazaola

  2. Departamento de Biología Vegetal I, Facultad de Biología, Universidad Complutense de Madrid, 28040, Madrid, Spain

    Luis Balaguer & Rafael Rubio de Casas

  3. Departamento de Fisiología y Ecología Vegetal, CCMA-CSIC, Serrano 15 dpdo, 28006, Madrid, Spain

    Esteban Manrique & Raúl Ochoa

  4. Departament de Biologia Vegetal, Facultat de Biologia, Universitat de Barcelona, Diagonal 645, 08028, Barcelona, Spain

    Isabel Fleck & Marta Pintó-Marijuan

  5. Serveis Científico-Tècnics, Universitat de Barcelona, 08028, Barcelona, Spain

    Isidre Casals

  6. Departamento de Biología Vegetal, Universidad de La Laguna, 38207, La Laguna, Tenerife, Spain

    Domingo Morales, María Soledad Jiménez & Roberto Lorenzo

Authors
  1. Raquel Esteban
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  2. Luis Balaguer
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  3. Esteban Manrique
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  4. Rafael Rubio de Casas
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  6. Isabel Fleck
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  7. Marta Pintó-Marijuan
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  8. Isidre Casals
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  9. Domingo Morales
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  10. María Soledad Jiménez
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  11. Roberto Lorenzo
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  12. Unai Artetxe
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  13. José María Becerril
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  14. José Ignacio García-Plazaola
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Corresponding author

Correspondence to José Ignacio García-Plazaola.

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Esteban, R., Balaguer, L., Manrique, E. et al. Alternative methods for sampling and preservation of photosynthetic pigments and tocopherols in plant material from remote locations. Photosynth Res 101, 77–88 (2009). https://doi.org/10.1007/s11120-009-9468-5

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  • DOI: https://doi.org/10.1007/s11120-009-9468-5

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