Alternative methods for sampling and preservation of photosynthetic pigments and tocopherols in plant material from remote locations |
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Authors: | Raquel Esteban Luis Balaguer Esteban Manrique Rafael Rubio de Casas Raúl Ochoa Isabel Fleck Marta Pintó-Marijuan Isidre Casals Domingo Morales María Soledad Jiménez Roberto Lorenzo Unai Artetxe José María Becerril José Ignacio García-Plazaola |
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Institution: | 1. Departamento de Biología Vegetal y Ecología, Universidad del País Vasco/EHU, Apdo 644, 48080, Bilbao, Spain 2. Departamento de Biología Vegetal I, Facultad de Biología, Universidad Complutense de Madrid, 28040, Madrid, Spain 3. Departamento de Fisiología y Ecología Vegetal, CCMA-CSIC, Serrano 15 dpdo, 28006, Madrid, Spain 4. Departament de Biologia Vegetal, Facultat de Biologia, Universitat de Barcelona, Diagonal 645, 08028, Barcelona, Spain 5. Serveis Científico-Tècnics, Universitat de Barcelona, 08028, Barcelona, Spain 6. Departamento de Biología Vegetal, Universidad de La Laguna, 38207, La Laguna, Tenerife, Spain
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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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Keywords: | HPLC Liquid nitrogen Lyophilization Silica gel Xanthophylls |
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