| 1. |
- Blaschek, Leonard, et al.
(author)
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Different combinations of laccase paralogs nonredundantly control the amount and composition of lignin in specific cell types and cell wall layers in Arabidopsis
- 2023
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In: The Plant Cell. - : Oxford University Press (OUP). - 1040-4651 .- 1532-298X. ; 35:2, s. 889-909
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Journal article (peer-reviewed)abstract
- Vascular plants reinforce the cell walls of the different xylem cell types with lignin phenolic polymers. Distinct lignin chemistries differ between each cell wall layer and each cell type to support their specific functions. Yet the mechanisms controlling the tight spatial localization of specific lignin chemistries remain unclear. Current hypotheses focus on control by monomer biosynthesis and/or export, while cell wall polymerization is viewed as random and nonlimiting. Here, we show that combinations of multiple individual laccases (LACs) are nonredundantly and specifically required to set the lignin chemistry in different cell types and their distinct cell wall layers. We dissected the roles of Arabidopsis thaliana LAC4, 5, 10, 12, and 17 by generating quadruple and quintuple loss-of-function mutants. Loss of these LACs in different combinations led to specific changes in lignin chemistry affecting both residue ring structures and/or aliphatic tails in specific cell types and cell wall layers. Moreover, we showed that LAC-mediated lignification has distinct functions in specific cell types, waterproofing fibers, and strengthening vessels. Altogether, we propose that the spatial control of lignin chemistry depends on different combinations of LACs with nonredundant activities immobilized in specific cell types and cell wall layers.
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| 2. |
- Blaschek, Leonard, et al.
(author)
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Functional complexity on a cellular scale : why in situ analyses are indispensable for our understanding of lignified tissues
- 2024
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In: Journal of Agricultural and Food Chemistry. - : American Chemical Society (ACS). - 0021-8561 .- 1520-5118. ; 72:24, s. 13552-13560
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Research review (peer-reviewed)abstract
- Lignins are a key adaptation that enables vascular plants to thrive in terrestrial habitats. Lignin is heterogeneous, containing upward of 30 different monomers, and its function is multifarious: It provides structural support, predetermined breaking points, ultraviolet protection, diffusion barriers, pathogen resistance, and drought resilience. Recent studies, carefully characterizing lignin in situ, have started to identify specific lignin compositions and ultrastructures with distinct cellular functions, but our understanding remains fractional. We summarize recent works and highlight where further in situ lignin analysis could provide valuable insights into plant growth and adaptation. We also summarize strengths and weaknesses of lignin in situ analysis methods.
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| 3. |
- Decou, Raphaël, et al.
(author)
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Analysis of lignin composition and distribution using fluorescence laser confocal microspectroscopy
- 2017. - 1
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In: Xylem. - New York : Humana Press. - 9781493967209 - 9781493967223 ; , s. 233-247
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Book chapter (peer-reviewed)abstract
- Lignin is a polyphenolic polymer specifically accumulating in the cell walls of xylem cells in higher vascular plants. Far from being homogeneous, the lignification of xylem cell walls varies in deposition site, quantity, composition and macromolecular conformation depending on the cell wall compartment, cell type, cell developmental stage and plant species. Here, we describe how confocal microspectroscopy methods using lignin autofluorescence can be used to evaluate the relative lignin amounts, its spatial distribution and composition at the cellular and sub-cellular levels in both isolated cells and histological cross-sections of plant tissues.
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| 4. |
- Menard, Delphine, et al.
(author)
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Establishment and utilization of habituated cell suspension cultures for hormone-inducible xylogenesis
- 2017. - 1
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In: Xylem. - New York : Humana Press. - 9781493967209 - 9781493967223 ; , s. 37-57
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Book chapter (peer-reviewed)abstract
- The development of inducible cell differentiation in suspension cultures led to multiple breakthroughs. It enabled the understanding of the chronology, duration, regulation and interdependency of the multiple events leading to fully functional specialized cells. The most studied cell differentiation in plants using inducible suspension cultures is the formation of tracheary elements (TEs) - the hydro-mineral sap conducting cells. Several in vitro systems established in different plant species have been developed to trigger TE formation on-demand. Here, we describe the establishment, harvesting and analysis of Arabidopsis thaliana stable habituated cell lines inducible by hormones to differentiate into TEs on-demand. Moreover, we explain the means to monitor and modify the chronology, duration and regulation of the progression of TE formation.
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| 5. |
- Ménard, Delphine, et al.
(author)
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Inducible pluripotent suspension cell cultures (iPSCs) to study plant cell differentiation
- 2024. - 2
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In: Xylem. - New York : Humana Press. - 9781071634769 - 9781071634790 - 9781071634776 ; , s. 171-200
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Book chapter (peer-reviewed)abstract
- Inducing the differentiation of specific cell type(s) synchronously and on-demand is a great experimental system to understand the sequential progression of the cellular processes, their timing and their resulting properties for distinct isolated plant cells independently of their tissue context. The inducible differentiation in cell suspension cultures, moreover, enables to obtain large quantities of distinct cell types at specific development stage, which is not possible when using whole plants. The differentiation of tracheary elements (TEs) - the cell type responsible for the hydro-mineral sap conduction and skeletal support of plants in xylem tissues - has been the most studied using inducible cell suspension cultures. We herein describe how to establish and use inducible pluripotent suspension cell cultures (iPSCs) in Arabidopsis thaliana to trigger on-demand different cell types, such as TEs or mesophyll cells. We, moreover, describe the methods to establish, monitor, and modify the sequence, duration, and properties of differentiated cells using iPSCs.
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| 6. |
- Ménard, Delphine, et al.
(author)
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Plant biomechanics and resilience to environmental changes are controlled by specific lignin chemistries in each vascular cell type and morphotype
- 2022
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In: The Plant Cell. - : Oxford University Press. - 1040-4651 .- 1532-298X. ; 34:12, s. 4877-4896
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Journal article (peer-reviewed)abstract
- The biopolymer lignin is deposited in the cell walls of vascular cells and is essential for long-distance water conduction and structural support in plants. Different vascular cell types contain distinct and conserved lignin chemistries, each with specific aromatic and aliphatic substitutions. Yet, the biological role of this conserved and specific lignin chemistry in each cell type remains unclear. Here, we investigated the roles of this lignin biochemical specificity for cellular functions by producing single cell analyses for three cell morphotypes of tracheary elements, which all allow sap conduction but differ in their morphology. We determined that specific lignin chemistries accumulate in each cell type. Moreover, lignin accumulated dynamically, increasing in quantity and changing in composition, to alter the cell wall biomechanics during cell maturation. For similar aromatic substitutions, residues with alcohol aliphatic functions increased stiffness whereas aldehydes increased flexibility of the cell wall. Modifying this lignin biochemical specificity and the sequence of its formation impaired the cell wall biomechanics of each morphotype and consequently hindered sap conduction and drought recovery. Together, our results demonstrate that each sap-conducting vascular cell type distinctly controls their lignin biochemistry to adjust their biomechanics and hydraulic properties to face developmental and environmental constraints.
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| 7. |
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| 8. |
- Sasaki, Takema, et al.
(author)
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Confined-microtubule assembly shapes three-dimensional cell wall structures in xylem vessels
- 2023
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In: Nature Communications. - : Springer Nature. - 2041-1723. ; 14:1
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Journal article (peer-reviewed)abstract
- Properly patterned deposition of cell wall polymers is prerequisite for the morphogenesis of plant cells. A cortical microtubule array guides the two-dimensional pattern of cell wall deposition. Yet, the mechanism underlying the three-dimensional patterning of cell wall deposition is poorly understood. In metaxylem vessels, cell wall arches are formed over numerous pit membranes, forming highly organized three-dimensional cell wall structures. Here, we show that the microtubule-associated proteins, MAP70-5 and MAP70-1, regulate arch development. The map70-1 map70-5 plants formed oblique arches in an abnormal orientation in pits. Microtubules fit the aperture of developing arches in wild-type cells, whereas microtubules in map70-1 map70-5 cells extended over the boundaries of pit arches. MAP70 caused the bending and bundling of microtubules. These results suggest that MAP70 confines microtubules within the pit apertures by altering the physical properties of microtubules, thereby directing the growth of pit arches in the proper orientation. This study provides clues to understanding how plants develop three-dimensional structure of cell walls.
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| 9. |
- Segura, Javier, et al.
(author)
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Boreal tree species affect soil organic matter composition and saprotrophic mineralization rates
- 2019
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In: Plant and Soil. - : Springer. - 0032-079X .- 1573-5036. ; 441:1-2, s. 173-190
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Journal article (peer-reviewed)abstract
- Aims: To investigate how different tree species affect the composition of SOM and its mineralization in boreal forest ecosystems.Methods: We used pyrolysis GC-MS for molecular-level characterization of the SOM formed under five common boreal tree species at a replicated field experiment similar to 50years after plantation. We incubated soil samples at 4, 9, 14 and 19 degrees C and measured inherent CO2 production and substrate-induced respiration. We then evaluated if the saprotrophic microbial activity and its temperature sensitivity was controlled by the SOM composition.Results: The molecular composition of the SOM emerged as key factor influencing SOM properties in plots with different tree species. Most of the variance in the SOM content was explained by the organo-chemical composition of the SOM. More importantly, the fraction of the microbial community able to utilize the native SOM was largely controlled by the SOM organo-chemical composition. Temperature sensitivity of CO2 production (Q(10)) was not explained by SOM composition. However, the microbial access to different SOM pools varied with temperature.Conclusions: These results bridge the gap between the paradigms of short-term litter and long-term SOM decomposition showing that, on an intermediate timescale (similar to 50 years), boreal tree species affect SOM molecular composition and saprotrophic mineralization rates.
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| 10. |
- Serk, Henrik, 1980-
(author)
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Cellular Aspects of Lignin Biosynthesis in Xylem Vessels of Zinnia and Arabidopsis
- 2015
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Doctoral thesis (other academic/artistic)abstract
- Lignin is the second most abundant biopolymer on earth and is found in the wood (xylem) of vascular land plants. To transport the hydro-mineral sap, xylem forms specialized conduit cells, called tracheary elements (TEs), which are hollow dead cylinders reinforced with lateral secondary cell walls (SCW). These SCWs incorporate lignin to gain mechanical strength, water impermeability and resistance against pathogens. The aim of this thesis is to understand the spatio-temporal deposition of lignin during TE differentiation and the relationship with its neighbouring cells. In vitro TE differentiating cell cultures of Zinnia elegans and Arabidopsis thaliana are ideal tools to study this process: cells differentiate simultaneously into 30-50% TEs while the rest remain parenchymatic (non-TEs). Live-cell imaging of such TEs indicated that lignification occurs after programmed cell death (PCD), in a non-cell autonomous manner, in which the non-TEs provide the lignin monomers.This thesis confirms that lignification occurs and continues long after TE PCD in both in vitro TE cultures and whole plants using biochemical, pharmacological and cytological methods. The cooperative supply of lignin monomers by the non-TEs was demonstrated by using Zinnia and Arabidopsis in vitro TE cultures. Inhibitor experiments revealed further that the non-TEs supply reactive oxygen species (ROS) to TEs and that ROS are required for TE post-mortem lignification. Characterization of the non-TEs showed an enlarged nucleus with increased DNA content, thus indicating that non-TEs are in fact endoreplicated xylem parenchyma cells (XP). The cooperative lignification was confirmed in whole plants by using knock-out mutants in a lignin monomer synthesis gene, which exhibit reduced TE lignification. The XP specific complementation of these mutants led to nearly completely rescuing the TE lignin reduction. Using microscopic techniques, the spatial distribution of lignin was analyzed in TEs from in vitro cultures and whole plants, revealing that lignification is restricted to TE SCWs in both protoxylem and metaxylem. These specific deposition domains were established by phenoloxidases, i.e. laccases localized to SCWs and peroxidases, present in SCWs and the apoplastic space. Laccases were cell-autonomously produced by developing TEs, indicating that the deposition domains are defined before PCD.Altogether, these results highlight that the hydro-mineral sap transport through TEs is enabled by the spatially and temporally controlled lignification of the SCW. Lignification occurs post-mortem by the supply of monomers and ROS from neighbouring XP cells and is restricted to specific deposition domains, defined by the pre-mortem sequestration of phenoloxidases.
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