Skip to main content

ORIGINAL RESEARCH article

Front. Plant Sci.
Sec. Technical Advances in Plant Science
Volume 15 - 2024 | doi: 10.3389/fpls.2024.1440885
This article is part of the Research Topic Cutting Edge Microscopy and Imaging Techniques in Plant and Algal Research View all 7 articles

Colocalising proteins and polysaccharides in plants for cell wall and trafficking studies

Provisionally accepted
Edwin R. Lampugnani Edwin R. Lampugnani 1Staffan Persson Staffan Persson 2Allison Van De Meene Allison Van De Meene 3,4*
  • 1 Menzies Institute for Medical Research, College of Health and Medicine, University of Tasmania, Hobart, Tasmania, Australia
  • 2 Department of Plant and Environmental Sciences, Faculty of Natural and Life Sciences, University of Copenhagen, Frederiksberg, Capital Region of Denmark, Denmark
  • 3 School of Biosciences, Faculty of Science, University of Melbourne, Parkville, Victoria, Australia
  • 4 Ian Holmes Imaging Centre, Bio21, The University of Melbourne, Parkville, Australia

The final, formatted version of the article will be published soon.

    Plant cell walls (PCWs) are intricate structures with complex polysaccharides delivered by distinct trafficking routes. Unravelling the intricate trafficking pathways of polysaccharides and proteins involved in PCW biosynthesis is a crucial first step towards understanding the complexities of plant growth and development. This study investigated the feasibility of employing a multi-modal approach that combines transmission electron microscopy (TEM) with molecular-genetic tagging and antibody labelling techniques to differentiate these pathways at the nanoscale. The genetically encoded electron microscopy (EM) tag APEX2 was fused to Arabidopsis thaliana cellulose synthase 6 (AtCESA6) and Nicotiana alata ARABINAN DEFICIENT LIKE 1 (NaARADL1), and these were transiently expressed in Nicotiana benthamiana leaves. APEX2 localization was then combined with immunolabeling using pectin-specific antibodies (JIM5 and JIM7). Our results demonstrate distinct trafficking patterns for CESA6 and NaARADL, with CESA6 localized primarily to the plasma membrane and vesicles, while NaARADL1 was found in the trans-Golgi network and cytoplasmic vesicles. Pectin epitopes were observed near the plasma membrane, in Golgi-associated vesicles, and in secretory vesicle clusters (SVCs) with both APEX2 constructs. Notably, JIM7 labelling was found in vesicles adjacent to APEX2-AtCESA6 vesicles, suggesting potential co-trafficking. This integrative approach offers a powerful tool for elucidating the dynamic interactions between PCW components at the nanoscale level. The methodology presented here facilitates the precise mapping of protein and polysaccharide trafficking pathways, advancing our understanding of PCW biosynthesis and providing avenues for future research aimed at engineering plant cell walls for various applications.

    Keywords: cell walls, APEX2, Antibodies, cellulose synthase, Pectins, Polysaccharides, electron microscopy Plant Cell Walls (PCWs), non-cellulosic polysaccharides (NCPs)

    Received: 30 May 2024; Accepted: 12 Aug 2024.

    Copyright: © 2024 Lampugnani, Persson and Van De Meene. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: Allison Van De Meene, School of Biosciences, Faculty of Science, University of Melbourne, Parkville, 3010, Victoria, Australia

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.