A correlative light electron microscopy approach reveals plasmodesmata ultrastructure at the graft interface
| Autor: | Sarah J. Cookson, Clément Chambaud, Nathalie Ollat, Lysiane Brocard, Emmanuelle Bayer |
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| Přispěvatelé: | Laboratoire de biogenèse membranaire (LBM), Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique (CNRS) |
| Rok vydání: | 2021 |
| Předmět: |
0106 biological sciences
Physiology Arabidopsis Plant Science Plasmodesma 01 natural sciences law.invention Cell wall 03 medical and health sciences law Genetics [SDV.BV]Life Sciences [q-bio]/Vegetal Biology 030304 developmental biology Microscopy 0303 health sciences Chemistry Endoplasmic reticulum Resolution (electron density) Plasmodesmata Organ Transplantation Hypocotyl Microscopy Electron Electron tomography Ultrastructure Biophysics Breakthrough Technologies Tools and Resources Electron microscope Biogenesis 010606 plant biology & botany |
| Zdroj: | Plant Physiol Plant Physiology Plant Physiology, American Society of Plant Biologists, 2021, ⟨10.1093/plphys/kiab485⟩ |
| ISSN: | 1532-2548 0032-0889 |
| Popis: | Despite recent progress in our understanding of graft union formation, we still know little about the cellular events underlying the grafting process. This is partially due to the difficulty of reliably targeting the graft interface in electron microscopy to study its ultrastructure and three-dimensional architecture. To overcome this technological bottleneck, we developed a correlative light electron microscopy (CLEM) approach to study the graft interface with high ultrastructural resolution. Grafting hypocotyls of Arabidopsis thaliana lines expressing yellow FP or monomeric red FP in the endoplasmic reticulum (ER) allowed efficient targeting of the grafting interface for examination under light and electron microscopy. To explore the potential of our method to study sub-cellular events at the graft interface, we focused on the formation of secondary plasmodesmata (PD) between the grafted partners. We showed that four classes of PD were formed at the interface and that PD introgression into the cell wall was initiated equally by both partners. Moreover, the success of PD formation appeared not systematic with a third of PD not spanning the cell wall entirely. Characterizing the ultrastructural characteristics of these incomplete PD gives us insights into the process of secondary PD biogenesis. We found that the establishment of successful symplastic connections between the scion and rootstock occurred predominantly in the presence of thin cell walls and ER–plasma membrane tethering. The resolution reached in this work shows that our CLEM method advances the study of biological processes requiring the combination of light and electron microscopy. |
| Databáze: | OpenAIRE |
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