Lignans in Knotwood of Norway Spruce: Localisation with Soft X-ray Microscopy and Scanning Transmission Electron Microscopy with Energy Dispersive X-ray Spectroscopy

Mansikkala, Tuomas; Patanen, Minna; Kärkönen, Anna; Korpinen, Risto; Pranovich, Andrey; Ohigashi, Takuji; Swaraj, Sufal; Seitsonen, Jani; Ruokolainen, Janne; Huttula, Marko; Saranpää, Pekka; Piispanen, Riikka

doi:10.3390/molecules25132997

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Lignans in Knotwood of Norway Spruce: Localisation with Soft X-ray Microscopy and Scanning Transmission Electron Microscopy with Energy Dispersive X-ray Spectroscopy

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Nano and Molecular Systems Research Unit, Faculty of Science, University of Oulu, P.O. Box 8000, FI-90014 Oulu, Finland

Biocenter Oulu, P.O. Box 5000, University of Oulu, FI-90014 Oulu, Finland

Production Systems, Natural Resources Institute Finland, Latokartanonkaari 9, FI-00790 Helsinki, Finland

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Viikki Plant Science Centre, Department of Agricultural Sciences, University of Helsinki, FI-00014 Helsinki, Finland

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Wood and Paper Chemistry Research Group, Laboratory of Natural Materials Technology, Åbo Akademi University, Porthansgatan 3, FI-20500 Turku, Finland

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UVSOR Facility, Institute for Molecular Science, 38 Nishigo-naka, Myodaiji, Okazaki, Aichi 444-8585, Japan

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SOLEIL Synchrotron, L’Orme des Merisiers, Saint-Aubin, P.O. Box 48, CEDEX, FR-91192 Gif-Sur-Yvette, France

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Nanomicroscopy Center, Department of Applied Physics, Aalto University, P.O. Box 15100, FI-00076 Aalto, Finland

Authors to whom correspondence should be addressed.

Academic Editor: Daniel Granato

Molecules 2020, 25(13), 2997; https://doi.org/10.3390/molecules25132997

Received: 8 June 2020 / Revised: 27 June 2020 / Accepted: 29 June 2020 / Published: 30 June 2020

(This article belongs to the Special Issue Recovery of Phytochemicals from Forest Materials)

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Abstract

Lignans are bioactive compounds that are especially abundant in the Norway spruce (Picea abies L. Karst.) knotwood. By combining a variety of chromatographic, spectroscopic and imaging techniques, we were able to quantify, qualify and localise the easily extractable lignans in the xylem tissue. The knotwood samples contained 15 different lignans according to the gas chromatography-mass spectrometry analysis. They comprised 16% of the knotwood dry weight and 82% of the acetone extract. The main lignans were found to be hydroxymatairesinols HMR1 and HMR2. Cryosectioned and resin-embedded ultrathin sections of the knotwood were analysed with scanning transmission X-ray microscopy (STXM). Cryosectioning was found to retain only lignan residues inside the cell lumina. In the resin-embedded samples, lignan was interpreted to be unevenly distributed inside the cell lumina, and partially confined in deposits which were either readily present in the lumina or formed when OsO₄ used in staining reacted with the lignans. Furthermore, the multi-technique characterisation enabled us to obtain information on the chemical composition of the structural components of knotwood. A simple spectral analysis of the STXM data gave consistent results with the gas chromatographic methods about the relative amounts of cell wall components (lignin and polysaccharides). The STXM analysis also indicated that a torus of a bordered pit contained aromatic compounds, possibly lignin. View Full-Text

Keywords: Picea abies; knotwood; extractives; hydroxymatairesinol; lignin; resin-embedding; cryosectioning; STXM; STEM-EDS; osmium tetroxide

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Mansikkala, T.; Patanen, M.; Kärkönen, A.; Korpinen, R.; Pranovich, A.; Ohigashi, T.; Swaraj, S.; Seitsonen, J.; Ruokolainen, J.; Huttula, M.; Saranpää, P.; Piispanen, R. Lignans in Knotwood of Norway Spruce: Localisation with Soft X-ray Microscopy and Scanning Transmission Electron Microscopy with Energy Dispersive X-ray Spectroscopy. Molecules 2020, 25, 2997. https://doi.org/10.3390/molecules25132997

AMA Style

Mansikkala T, Patanen M, Kärkönen A, Korpinen R, Pranovich A, Ohigashi T, Swaraj S, Seitsonen J, Ruokolainen J, Huttula M, Saranpää P, Piispanen R. Lignans in Knotwood of Norway Spruce: Localisation with Soft X-ray Microscopy and Scanning Transmission Electron Microscopy with Energy Dispersive X-ray Spectroscopy. Molecules. 2020; 25(13):2997. https://doi.org/10.3390/molecules25132997

Chicago/Turabian Style

Mansikkala, Tuomas, Minna Patanen, Anna Kärkönen, Risto Korpinen, Andrey Pranovich, Takuji Ohigashi, Sufal Swaraj, Jani Seitsonen, Janne Ruokolainen, Marko Huttula, Pekka Saranpää, and Riikka Piispanen. 2020. "Lignans in Knotwood of Norway Spruce: Localisation with Soft X-ray Microscopy and Scanning Transmission Electron Microscopy with Energy Dispersive X-ray Spectroscopy" Molecules 25, no. 13: 2997. https://doi.org/10.3390/molecules25132997

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Lignans in Knotwood of Norway Spruce: Localisation with Soft X-ray Microscopy and Scanning Transmission Electron Microscopy with Energy Dispersive X-ray Spectroscopy

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