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Monoclonal antibody-based analysis of cell wall remodeling during xylogenesis

Abstract

Xylogenesis, a process by which woody tissues are formed, entails qualitative and quantitative changes in the cell wall. However, the molecular events that underlie these changes are not completely understood. Previously, we have isolated two monoclonal antibodies, referred to as XD3 and XD27, by subtractive screening of a phage-display library of antibodies raised against a wall fraction of Zinnia elegans xylogenic culture cells. Here we report the biochemical and immunohistochemical characterization of those antibodies. The antibody XD3 recognized (1→4)-β-d-galactan in pectin fraction. During xylogenesis, the XD3 epitope was localized to the primary wall of tracheary-element precursor cells, which undergo substantial cell elongation, and was absent from mature tracheary elements. XD27 recognized an arabinogalactan protein that was bound strongly to a germin-like protein. The XD27 epitope was localized to pre-lignified secondary walls of tracheary elements. Thus these cell-wall-directed monoclonal antibodies revealed two molecular events during xylogenesis. The biological significance of these events is discussed in relation to current views of the plant cell wall.

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Acknowledgments

We thank Tadashi Ishii for help with sugar-linkage analysis; Jin Nakashima for help with cell sectioning; Kuninori Iwamoto for searching ZeGLP1 cDNA sequence; and Maaike de Jong for critical reading of the manuscript. This work was supported partly by Grants-in-Aid from the Ministry of Education, Science, Sports and Culture of Japan (NC-CARP project) and from the Japan Society for the Promotion of Science (23227001) to HF.

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Correspondence to Naoki Shinohara.

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The authors declare that they have no conflict of interest.

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Shinohara, N., Kakegawa, K. & Fukuda, H. Monoclonal antibody-based analysis of cell wall remodeling during xylogenesis. J Plant Res 128, 975–986 (2015). https://doi.org/10.1007/s10265-015-0758-z

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  • DOI: https://doi.org/10.1007/s10265-015-0758-z

Keywords

  • Arabinogalactan protein
  • Cell wall
  • Germin-like protein
  • Pectic galactan
  • Monoclonal antibody
  • Xylogenesis