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Vibrational microspectroscopy enables chemical characterization of single pollen grains as well as comparative analysis of plant species based on pollen ultrastructure

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Chemical imaging of pollen by vibrational microspectroscopy enables characterization of pollen ultrastructure, in particular phenylpropanoid components in grain wall for comparative study of extant and extinct plant species.

A detailed characterization of conifer (Pinales) pollen by vibrational microspectroscopy is presented. The main problems that arise during vibrational measurements were scatter and saturation issues in Fourier transform infrared (FTIR), and fluorescence and penetration depth issues in Raman. Single pollen grains larger than approx. 15 µm can be measured by FTIR microspectroscopy using conventional light sources, while smaller grains may be measured by employing synchrotron light sources. Pollen grains that were larger than 50 µm were too thick for FTIR imaging since the grain constituents absorbed almost all infrared light. Chemical images of pollen were obtained on sectioned samples, unveiling the distribution and concentration of proteins, carbohydrates, sporopollenins and lipids within pollen substructures. The comparative analysis of pollen species revealed that, compared with other Pinales pollens, Cedrus atlantica has a higher relative amount of lipid nutrients, as well as different chemical composition of grain wall sporopollenin. The pre-processing and data analysis, namely extended multiplicative signal correction and principal component analysis, offer simple estimate of imaging spectral data and indirect estimation of physical properties of pollen. The vibrational microspectroscopy study demonstrates that detailed chemical characterization of pollen can be obtained by measurement of an individual grain and pollen ultrastructure. Measurement of phenylpropanoid components in pollen grain wall could be used, not only for the reconstruction of past environments, but for assessment of diversity of plant species as well. Therefore, analysis of extant and extinct pollen species by vibrational spectroscopies is suggested as a valuable tool in biology, ecology and palaeosciences.

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Acknowledgments

We thank Elin Ørmen from the Microscopy lab, Imaging Centre—NMBU, for the help with the SEM measurements, and Paul Dumas from the SMIS beamline, Synchrotron SOLEIL, for the help with the FTIR synchrotron measurements. The research was supported by the Norwegian Research Council (ISP project No. 216687), by the SOLEIL, French national synchrotron facility (project No. 20120345), and by the European Commission through the Seventh Framework Programme (FP7-PEOPLE-2012-IEF project No. 328289).

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Authors and Affiliations

  1. Department of Mathematical Sciences and Technology, Faculty of Environmental Science and Technology, Norwegian University of Life Sciences, Drøbakveien 31, 1430, Ås, Norway

    Boris Zimmermann, Murat Bağcıoğlu & Achim Kohler

  2. Synchrotron SOLEIL, L’Orme des Merisiers, Saint-Aubin, BP 48, 91192, Gif-sur-Yvette, France

    Christophe Sandt

  3. Nofima AS, Osloveien 1, 1430, Ås, Norway

    Achim Kohler

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  1. Boris Zimmermann
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  4. Achim Kohler
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Correspondence to Boris Zimmermann.

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Zimmermann, B., Bağcıoğlu, M., Sandt, C. et al. Vibrational microspectroscopy enables chemical characterization of single pollen grains as well as comparative analysis of plant species based on pollen ultrastructure. Planta 242, 1237–1250 (2015). https://doi.org/10.1007/s00425-015-2380-7

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