Bacete, Laura - Plant cell wall dynamics

@article{fernandez-calvo_leucine_2024,
	title = {Leucine rich repeat-malectin receptor kinases {IGP1}/{CORK1}, {IGP3} and {IGP4} are required for arabidopsis immune responses triggered by β-1,4-{D}-{Xylo}-oligosaccharides from plant cell walls},
	volume = {11},
	issn = {2468-2330},
	url = {https://www.sciencedirect.com/science/article/pii/S2468233024000069},
	doi = {10.1016/j.tcsw.2024.100124},
	abstract = {Pattern-Triggered Immunity (PTI) in plants is activated upon recognition by Pattern Recognition Receptors (PRRs) of Damage- and Microbe-Associated Molecular Patterns (DAMPs and MAMPs) from plants or microorganisms, respectively. An increasing number of identified DAMPs/MAMPs are carbohydrates from plant cell walls and microbial extracellular layers, which are perceived by plant PRRs, such as LysM and Leucine Rich Repeat-Malectin (LRR-MAL) receptor kinases (RKs). LysM-RKs (e.g. CERK1, LYK4 and LYK5) are needed for recognition of fungal MAMP chitohexaose (β-1,4-D-(GlcNAc)6, CHI6), whereas IGP1/CORK1, IGP3 and IGP4 LRR-MAL RKs are required for perception of β-glucans, like cellotriose (β-1,4-D-(Glc)3, CEL3) and mixed-linked glucans. We have explored the diversity of carbohydrates perceived by Arabidopsis thaliana seedlings by determining PTI responses upon treatment with different oligosaccharides and polysaccharides. These analyses revealed that plant oligosaccharides from xylans [β-1,4-D-(xylose)4 (XYL4)], glucuronoxylans and α-1,4-glucans, and polysaccharides from plants and seaweeds activate PTI. Cross-elicitation experiments of XYL4 with other glycans showed that the mechanism of recognition of XYL4 and the DAMP 33-α-L-arabinofuranosyl-xylotetraose (XA3XX) shares some features with that of CEL3 but differs from that of CHI6. Notably, XYL4 and XA3XX perception is impaired in igp1/cork1, igp3 and igp4 mutants, and almost not affected in cerk1 lyk4 lyk5 triple mutant. XYL4 perception is conserved in different plant species since XYL4 pre-treatment triggers enhanced disease resistance in tomato to Pseudomonas syringae pv tomato DC3000 and PTI responses in wheat. These results expand the number of glycans triggering plant immunity and support IGP1/CORK1, IGP3 and IGP4 relevance in Arabidopsis thaliana glycans perception and PTI activation.
Significance Statement
The characterization of plant immune mechanisms involved in the perception of carbohydrate-based structures recognized as DAMPs/MAMPs is needed to further understand plant disease resistance modulation. We show here that IGP1/CORK1, IGP3 and IGP4 LRR-MAL RKs are required for the perception of carbohydrate-based DAMPs β-1,4-D-(xylose)4 (XYL4) and 33-α-L-arabinofuranosyl-xylotetraose (XA3XX), further expanding the function of these LRR-MAL RKs in plant glycan perception and immune activation.},
	urldate = {2024-08-09},
	journal = {The Cell Surface},
	author = {Fernández-Calvo, Patricia and López, Gemma and Martín-Dacal, Marina and Aitouguinane, Meriem and Carrasco-López, Cristian and González-Bodí, Sara and Bacete, Laura and Mélida, Hugo and Sánchez-Vallet, Andrea and Molina, Antonio},
	month = jun,
	year = {2024},
	keywords = {Cell wall, Disease resistance, Leucine rich repeat-malectin receptor kinases, Pattern triggered immunity, Xylans, Xylotetraose},
	pages = {100124},
}

Pattern-Triggered Immunity (PTI) in plants is activated upon recognition by Pattern Recognition Receptors (PRRs) of Damage- and Microbe-Associated Molecular Patterns (DAMPs and MAMPs) from plants or microorganisms, respectively. An increasing number of identified DAMPs/MAMPs are carbohydrates from plant cell walls and microbial extracellular layers, which are perceived by plant PRRs, such as LysM and Leucine Rich Repeat-Malectin (LRR-MAL) receptor kinases (RKs). LysM-RKs (e.g. CERK1, LYK4 and LYK5) are needed for recognition of fungal MAMP chitohexaose (β-1,4-D-(GlcNAc)6, CHI6), whereas IGP1/CORK1, IGP3 and IGP4 LRR-MAL RKs are required for perception of β-glucans, like cellotriose (β-1,4-D-(Glc)3, CEL3) and mixed-linked glucans. We have explored the diversity of carbohydrates perceived by Arabidopsis thaliana seedlings by determining PTI responses upon treatment with different oligosaccharides and polysaccharides. These analyses revealed that plant oligosaccharides from xylans [β-1,4-D-(xylose)4 (XYL4)], glucuronoxylans and α-1,4-glucans, and polysaccharides from plants and seaweeds activate PTI. Cross-elicitation experiments of XYL4 with other glycans showed that the mechanism of recognition of XYL4 and the DAMP 33-α-L-arabinofuranosyl-xylotetraose (XA3XX) shares some features with that of CEL3 but differs from that of CHI6. Notably, XYL4 and XA3XX perception is impaired in igp1/cork1, igp3 and igp4 mutants, and almost not affected in cerk1 lyk4 lyk5 triple mutant. XYL4 perception is conserved in different plant species since XYL4 pre-treatment triggers enhanced disease resistance in tomato to Pseudomonas syringae pv tomato DC3000 and PTI responses in wheat. These results expand the number of glycans triggering plant immunity and support IGP1/CORK1, IGP3 and IGP4 relevance in Arabidopsis thaliana glycans perception and PTI activation. Significance Statement The characterization of plant immune mechanisms involved in the perception of carbohydrate-based structures recognized as DAMPs/MAMPs is needed to further understand plant disease resistance modulation. We show here that IGP1/CORK1, IGP3 and IGP4 LRR-MAL RKs are required for the perception of carbohydrate-based DAMPs β-1,4-D-(xylose)4 (XYL4) and 33-α-L-arabinofuranosyl-xylotetraose (XA3XX), further expanding the function of these LRR-MAL RKs in plant glycan perception and immune activation.

@article{alonso_baez_cell_2023,
	title = {Cell wall dynamics: novel tools and research questions},
	volume = {74},
	issn = {0022-0957},
	shorttitle = {Cell wall dynamics},
	url = {https://doi.org/10.1093/jxb/erad310},
	doi = {10.1093/jxb/erad310},
	abstract = {Years ago, a classic textbook would define plant cell walls based on passive features. For instance, a sort of plant exoskeleton of invariable polysaccharide composition, and probably painted in green. However, currently, this view has been expanded to consider plant cell walls as active, heterogeneous, and dynamic structures with a high degree of complexity. However, what do we mean when we refer to a cell wall as a dynamic structure? How can we investigate the different implications of this dynamism? While the first question has been the subject of several recent publications, defining the ideal strategies and tools needed to address the second question has proven to be challenging due to the myriad of techniques available. In this review, we will describe the capacities of several methodologies to study cell wall composition, structure, and other aspects developed or optimized in recent years. Keeping in mind cell wall dynamism and plasticity, the advantages of performing long-term non-invasive live-imaging methods will be emphasized. We specifically focus on techniques developed for Arabidopsis thaliana primary cell walls, but the techniques could be applied to both secondary cell walls and other plant species. We believe this toolset will help researchers in expanding knowledge of these dynamic/evolving structures.},
	number = {21},
	urldate = {2023-11-24},
	journal = {Journal of Experimental Botany},
	author = {Alonso Baez, Luis and Bacete, Laura},
	month = nov,
	year = {2023},
	pages = {6448--6467},
}

Years ago, a classic textbook would define plant cell walls based on passive features. For instance, a sort of plant exoskeleton of invariable polysaccharide composition, and probably painted in green. However, currently, this view has been expanded to consider plant cell walls as active, heterogeneous, and dynamic structures with a high degree of complexity. However, what do we mean when we refer to a cell wall as a dynamic structure? How can we investigate the different implications of this dynamism? While the first question has been the subject of several recent publications, defining the ideal strategies and tools needed to address the second question has proven to be challenging due to the myriad of techniques available. In this review, we will describe the capacities of several methodologies to study cell wall composition, structure, and other aspects developed or optimized in recent years. Keeping in mind cell wall dynamism and plasticity, the advantages of performing long-term non-invasive live-imaging methods will be emphasized. We specifically focus on techniques developed for Arabidopsis thaliana primary cell walls, but the techniques could be applied to both secondary cell walls and other plant species. We believe this toolset will help researchers in expanding knowledge of these dynamic/evolving structures.

@article{vukasinovic_cracking_2023,
	title = {Cracking the green wall code: insights into cell wall integrity across organisms},
	volume = {1},
	issn = {2813-821X},
	shorttitle = {Cracking the green wall code},
	url = {https://www.frontiersin.org/journals/plant-physiology/articles/10.3389/fphgy.2023.1323899/full},
	doi = {10.3389/fphgy.2023.1323899},
	abstract = {{\textless}p{\textgreater}Cell walls are not just passive barriers; they are dynamic and adaptable structures that are actively remodeled in response to both internal and external cues. They are crucial in defining cellular identity, ensuring structural integrity, and mediating interactions with the environment. The concept of cell wall integrity (CWI) encompasses the mechanisms by which cells monitor and maintain their walls, ensuring proper function and response to challenges. While significant knowledge has been accumulated on CWI in certain model organisms, there remains a vast landscape of uncharted territory in others. In this review, we aim to bridge this gap, offering a comparative perspective on CWI across different evolutionary lineages, from the well-studied yeasts to the diverse world of plants. We focus especially on the green lineage –the group of green algae and land plants, hence the {\textless}italic{\textgreater}green wall{\textless}/italic{\textgreater}–, but also consider some insights from organisms with radically different lifestyles and cell wall arrangements, which serves as a base to some intriguing questions about the role of CWI across evolution and environmental adaptation.{\textless}/p{\textgreater}},
	language = {English},
	urldate = {2024-10-02},
	journal = {Frontiers in Plant Physiology},
	author = {Vukašinović, Nemanja and Serif, Manuel and Bacete, Laura},
	month = nov,
	year = {2023},
	note = {Publisher: Frontiers},
	keywords = {Bacillariophyceae, Cell wall physicochemical properties, Cell wall plasticity, Charophyceae, Mechanosensing, Streptophyta, Zygnematophyceae, cell wall dynamics},
}

\textlessp\textgreaterCell walls are not just passive barriers; they are dynamic and adaptable structures that are actively remodeled in response to both internal and external cues. They are crucial in defining cellular identity, ensuring structural integrity, and mediating interactions with the environment. The concept of cell wall integrity (CWI) encompasses the mechanisms by which cells monitor and maintain their walls, ensuring proper function and response to challenges. While significant knowledge has been accumulated on CWI in certain model organisms, there remains a vast landscape of uncharted territory in others. In this review, we aim to bridge this gap, offering a comparative perspective on CWI across different evolutionary lineages, from the well-studied yeasts to the diverse world of plants. We focus especially on the green lineage –the group of green algae and land plants, hence the \textlessitalic\textgreatergreen wall\textless/italic\textgreater–, but also consider some insights from organisms with radically different lifestyles and cell wall arrangements, which serves as a base to some intriguing questions about the role of CWI across evolution and environmental adaptation.\textless/p\textgreater

@article{bacete_dynamics_2023,
	title = {Dynamics and mechanics of plant cell walls: insights into plant growth, defence, and stress response},
	volume = {113},
	issn = {1573-5028},
	shorttitle = {Dynamics and mechanics of plant cell walls},
	url = {https://doi.org/10.1007/s11103-023-01395-9},
	doi = {10.1007/s11103-023-01395-9},
	language = {en},
	number = {6},
	urldate = {2024-01-04},
	journal = {Plant Molecular Biology},
	author = {Bacete, Laura and Mélida, Hugo},
	month = dec,
	year = {2023},
	pages = {329--330},
}

@article{soni_interplay_2023,
	title = {The interplay between cell wall integrity and cell cycle progression in plants},
	volume = {113},
	issn = {1573-5028},
	url = {https://doi.org/10.1007/s11103-023-01394-w},
	doi = {10.1007/s11103-023-01394-w},
	abstract = {Plant cell walls are dynamic structures that play crucial roles in growth, development, and stress responses. Despite our growing understanding of cell wall biology, the connections between cell wall integrity (CWI) and cell cycle progression in plants remain poorly understood. This review aims to explore the intricate relationship between CWI and cell cycle progression in plants, drawing insights from studies in yeast and mammals. We provide an overview of the plant cell cycle, highlight the role of endoreplication in cell wall composition, and discuss recent findings on the molecular mechanisms linking CWI perception to cell wall biosynthesis and gene expression regulation. Furthermore, we address future perspectives and unanswered questions in the field, such as the identification of specific CWI sensing mechanisms and the role of CWI maintenance in the growth-defense trade-off. Elucidating these connections could have significant implications for crop improvement and sustainable agriculture.},
	language = {en},
	number = {6},
	urldate = {2023-12-22},
	journal = {Plant Molecular Biology},
	author = {Soni, Nancy and Bacete, Laura},
	month = dec,
	year = {2023},
	keywords = {Auxin, Cell cycle progression, Cell wall sensing, Cytokinin, Endoreplication, Growth-defense trade-off, Plant cell wall integrity},
	pages = {367--382},
}

Plant cell walls are dynamic structures that play crucial roles in growth, development, and stress responses. Despite our growing understanding of cell wall biology, the connections between cell wall integrity (CWI) and cell cycle progression in plants remain poorly understood. This review aims to explore the intricate relationship between CWI and cell cycle progression in plants, drawing insights from studies in yeast and mammals. We provide an overview of the plant cell cycle, highlight the role of endoreplication in cell wall composition, and discuss recent findings on the molecular mechanisms linking CWI perception to cell wall biosynthesis and gene expression regulation. Furthermore, we address future perspectives and unanswered questions in the field, such as the identification of specific CWI sensing mechanisms and the role of CWI maintenance in the growth-defense trade-off. Elucidating these connections could have significant implications for crop improvement and sustainable agriculture.

@article{bacete_theseus1_2022,
	title = {{THESEUS1} modulates cell wall stiffness and abscisic acid production in {Arabidopsis} thaliana},
	volume = {119},
	url = {https://www.pnas.org/doi/full/10.1073/pnas.2119258119},
	doi = {10.1073/pnas.2119258119},
	abstract = {Plant cells can be distinguished from animal cells by their cell walls and high-turgor pressure. Although changes in turgor and the stiffness of cell walls seem coordinated, we know little about the mechanism responsible for coordination. Evidence has accumulated that plants, like yeast, have a dedicated cell wall integrity maintenance mechanism. It monitors the functional integrity of the wall and maintains integrity through adaptive responses induced by cell wall damage arising during growth, development, and interactions with the environment. These adaptive responses include osmosensitive induction of phytohormone production, defense responses, as well as changes in cell wall composition and structure. Here, we investigate how the cell wall integrity maintenance mechanism coordinates changes in cell wall stiffness and turgor in Arabidopsis thaliana. We show that the production of abscisic acid (ABA), the phytohormone-modulating turgor pressure, and responses to drought depend on the presence of a functional cell wall. We find that the cell wall integrity sensor THESEUS1 modulates mechanical properties of walls, turgor loss point, ABA biosynthesis, and ABA-controlled processes. We identify RECEPTOR-LIKE PROTEIN 12 as a component of cell wall integrity maintenance–controlling, cell wall damage–induced jasmonic acid (JA) production. We propose that THE1 is responsible for coordinating changes in turgor pressure and cell wall stiffness.},
	number = {1},
	urldate = {2023-03-10},
	journal = {Proceedings of the National Academy of Sciences},
	author = {Bacete, Laura and Schulz, Julia and Engelsdorf, Timo and Bartosova, Zdenka and Vaahtera, Lauri and Yan, Guqi and Gerhold, Joachim Matthias and Tichá, Tereza and Øvstebø, Camilla and Gigli-Bisceglia, Nora and Johannessen-Starheim, Svanhild and Margueritat, Jeremie and Kollist, Hannes and Dehoux, Thomas and McAdam, Scott A. M. and Hamann, Thorsten},
	month = jan,
	year = {2022},
	note = {Publisher: Proceedings of the National Academy of Sciences},
	pages = {e2119258119},
}

Plant cells can be distinguished from animal cells by their cell walls and high-turgor pressure. Although changes in turgor and the stiffness of cell walls seem coordinated, we know little about the mechanism responsible for coordination. Evidence has accumulated that plants, like yeast, have a dedicated cell wall integrity maintenance mechanism. It monitors the functional integrity of the wall and maintains integrity through adaptive responses induced by cell wall damage arising during growth, development, and interactions with the environment. These adaptive responses include osmosensitive induction of phytohormone production, defense responses, as well as changes in cell wall composition and structure. Here, we investigate how the cell wall integrity maintenance mechanism coordinates changes in cell wall stiffness and turgor in Arabidopsis thaliana. We show that the production of abscisic acid (ABA), the phytohormone-modulating turgor pressure, and responses to drought depend on the presence of a functional cell wall. We find that the cell wall integrity sensor THESEUS1 modulates mechanical properties of walls, turgor loss point, ABA biosynthesis, and ABA-controlled processes. We identify RECEPTOR-LIKE PROTEIN 12 as a component of cell wall integrity maintenance–controlling, cell wall damage–induced jasmonic acid (JA) production. We propose that THE1 is responsible for coordinating changes in turgor pressure and cell wall stiffness.

@article{molina_arabidopsis_2021,
	title = {Arabidopsis cell wall composition determines disease resistance specificity and fitness},
	volume = {118},
	url = {https://www.pnas.org/doi/abs/10.1073/pnas.2010243118},
	doi = {10.1073/pnas.2010243118},
	abstract = {Plant cell walls are complex structures subject to dynamic remodeling in response to developmental and environmental cues and play essential functions in disease resistance responses. We tested the specific contribution of plant cell walls to immunity by determining the susceptibility of a set of Arabidopsis cell wall mutants (cwm) to pathogens with different parasitic styles: a vascular bacterium, a necrotrophic fungus, and a biotrophic oomycete. Remarkably, most cwm mutants tested (29/34; 85.3\%) showed alterations in their resistance responses to at least one of these pathogens in comparison to wild-type plants, illustrating the relevance of wall composition in determining disease-resistance phenotypes. We found that the enhanced resistance of cwm plants to the necrotrophic and vascular pathogens negatively impacted cwm fitness traits, such as biomass and seed yield. Enhanced resistance of cwm plants is not only mediated by canonical immune pathways, like those modulated by phytohormones or microbe-associated molecular patterns, which are not deregulated in the cwm tested. Pectin-enriched wall fractions isolated from cwm plants triggered immune responses in wild-type plants, suggesting that wall-mediated defensive pathways might contribute to cwm resistance. Cell walls of cwm plants show a high diversity of composition alterations as revealed by glycome profiling that detect specific wall carbohydrate moieties. Mathematical analysis of glycome profiling data identified correlations between the amounts of specific wall carbohydrate moieties and disease resistance phenotypes of cwm plants. These data support the relevant and specific function of plant wall composition in plant immune response modulation and in balancing disease resistance/development trade-offs.},
	number = {5},
	urldate = {2023-03-10},
	journal = {Proceedings of the National Academy of Sciences},
	author = {Molina, Antonio and Miedes, Eva and Bacete, Laura and Rodríguez, Tinguaro and Mélida, Hugo and Denancé, Nicolas and Sánchez-Vallet, Andrea and Rivière, Marie-Pierre and López, Gemma and Freydier, Amandine and Barlet, Xavier and Pattathil, Sivakumar and Hahn, Michael and Goffner, Deborah},
	month = feb,
	year = {2021},
	note = {Publisher: Proceedings of the National Academy of Sciences},
	pages = {e2010243118},
}

Plant cell walls are complex structures subject to dynamic remodeling in response to developmental and environmental cues and play essential functions in disease resistance responses. We tested the specific contribution of plant cell walls to immunity by determining the susceptibility of a set of Arabidopsis cell wall mutants (cwm) to pathogens with different parasitic styles: a vascular bacterium, a necrotrophic fungus, and a biotrophic oomycete. Remarkably, most cwm mutants tested (29/34; 85.3%) showed alterations in their resistance responses to at least one of these pathogens in comparison to wild-type plants, illustrating the relevance of wall composition in determining disease-resistance phenotypes. We found that the enhanced resistance of cwm plants to the necrotrophic and vascular pathogens negatively impacted cwm fitness traits, such as biomass and seed yield. Enhanced resistance of cwm plants is not only mediated by canonical immune pathways, like those modulated by phytohormones or microbe-associated molecular patterns, which are not deregulated in the cwm tested. Pectin-enriched wall fractions isolated from cwm plants triggered immune responses in wild-type plants, suggesting that wall-mediated defensive pathways might contribute to cwm resistance. Cell walls of cwm plants show a high diversity of composition alterations as revealed by glycome profiling that detect specific wall carbohydrate moieties. Mathematical analysis of glycome profiling data identified correlations between the amounts of specific wall carbohydrate moieties and disease resistance phenotypes of cwm plants. These data support the relevant and specific function of plant wall composition in plant immune response modulation and in balancing disease resistance/development trade-offs.

@article{rebaque_cell_2021,
	title = {Cell wall-derived mixed-linked β-1,3/1,4-glucans trigger immune responses and disease resistance in plants},
	volume = {106},
	issn = {1365-313X},
	url = {https://onlinelibrary.wiley.com/doi/abs/10.1111/tpj.15185},
	doi = {10.1111/tpj.15185},
	abstract = {Pattern-triggered immunity (PTI) is activated in plants upon recognition by pattern recognition receptors (PRRs) of damage- and microbe-associated molecular patterns (DAMPs and MAMPs) derived from plants or microorganisms, respectively. To understand better the plant mechanisms involved in the perception of carbohydrate-based structures recognized as DAMPs/MAMPs, we have studied the ability of mixed-linked β-1,3/1,4-glucans (MLGs), present in some plant and microbial cell walls, to trigger immune responses and disease resistance in plants. A range of MLG structures were tested for their capacity to induce PTI hallmarks, such as cytoplasmic Ca2+ elevations, reactive oxygen species production, phosphorylation of mitogen-activated protein kinases and gene transcriptional reprogramming. These analyses revealed that MLG oligosaccharides are perceived by Arabidopsis thaliana and identified a trisaccharide, β-d-cellobiosyl-(1,3)-β-d-glucose (MLG43), as the smallest MLG structure triggering strong PTI responses. These MLG43-mediated PTI responses are partially dependent on LysM PRRs CERK1, LYK4 and LYK5, as they were weaker in cerk1 and lyk4 lyk5 mutants than in wild-type plants. Cross-elicitation experiments between MLG43 and the carbohydrate MAMP chitohexaose [β-1,4-d-(GlcNAc)6], which is also perceived by these LysM PRRs, indicated that the mechanism of MLG43 recognition could differ from that of chitohexaose, which is fully impaired in cerk1 and lyk4 lyk5 plants. MLG43 treatment confers enhanced disease resistance in A. thaliana to the oomycete Hyaloperonospora arabidopsidis and in tomato and pepper to different bacterial and fungal pathogens. Our data support the classification of MLGs as a group of carbohydrate-based molecular patterns that are perceived by plants and trigger immune responses and disease resistance.},
	language = {en},
	number = {3},
	urldate = {2023-03-10},
	journal = {The Plant Journal},
	author = {Rebaque, Diego and del Hierro, Irene and López, Gemma and Bacete, Laura and Vilaplana, Francisco and Dallabernardina, Pietro and Pfrengle, Fabian and Jordá, Lucía and Sánchez-Vallet, Andrea and Pérez, Rosa and Brunner, Frédéric and Molina, Antonio and Mélida, Hugo},
	year = {2021},
	note = {\_eprint: https://onlinelibrary.wiley.com/doi/pdf/10.1111/tpj.15185},
	keywords = {Arabidopsis thaliana, Capsicum annuum, Hyaloperonospora arabidopsidis, Solanum lycopersicum, cell wall, disease resistance, mixed-linked glucan, pattern triggered immunity, plant immunity},
	pages = {601--615},
}

Pattern-triggered immunity (PTI) is activated in plants upon recognition by pattern recognition receptors (PRRs) of damage- and microbe-associated molecular patterns (DAMPs and MAMPs) derived from plants or microorganisms, respectively. To understand better the plant mechanisms involved in the perception of carbohydrate-based structures recognized as DAMPs/MAMPs, we have studied the ability of mixed-linked β-1,3/1,4-glucans (MLGs), present in some plant and microbial cell walls, to trigger immune responses and disease resistance in plants. A range of MLG structures were tested for their capacity to induce PTI hallmarks, such as cytoplasmic Ca2+ elevations, reactive oxygen species production, phosphorylation of mitogen-activated protein kinases and gene transcriptional reprogramming. These analyses revealed that MLG oligosaccharides are perceived by Arabidopsis thaliana and identified a trisaccharide, β-d-cellobiosyl-(1,3)-β-d-glucose (MLG43), as the smallest MLG structure triggering strong PTI responses. These MLG43-mediated PTI responses are partially dependent on LysM PRRs CERK1, LYK4 and LYK5, as they were weaker in cerk1 and lyk4 lyk5 mutants than in wild-type plants. Cross-elicitation experiments between MLG43 and the carbohydrate MAMP chitohexaose [β-1,4-d-(GlcNAc)6], which is also perceived by these LysM PRRs, indicated that the mechanism of MLG43 recognition could differ from that of chitohexaose, which is fully impaired in cerk1 and lyk4 lyk5 plants. MLG43 treatment confers enhanced disease resistance in A. thaliana to the oomycete Hyaloperonospora arabidopsidis and in tomato and pepper to different bacterial and fungal pathogens. Our data support the classification of MLGs as a group of carbohydrate-based molecular patterns that are perceived by plants and trigger immune responses and disease resistance.

@article{bacete_arabidopsis_2020,
	title = {Arabidopsis {Response} {Regulator} 6 ({ARR6}) {Modulates} {Plant} {Cell}-{Wall} {Composition} and {Disease} {Resistance}},
	volume = {33},
	issn = {0894-0282},
	url = {https://apsjournals.apsnet.org/doi/10.1094/MPMI-12-19-0341-R},
	doi = {10.1094/MPMI-12-19-0341-R},
	abstract = {The cytokinin signaling pathway, which is mediated by Arabidopsis response regulator (ARR) proteins, has been involved in the modulation of some disease-resistance responses. Here, we describe novel functions of ARR6 in the control of plant disease-resistance and cell-wall composition. Plants impaired in ARR6 function (arr6) were more resistant and susceptible, respectively, to the necrotrophic fungus Plectosphaerella cucumerina and to the vascular bacterium Ralstonia solanacearum, whereas Arabidopsis plants that overexpress ARR6 showed the opposite phenotypes, which further support a role of ARR6 in the modulation of disease-resistance responses against these pathogens. Transcriptomics and metabolomics analyses revealed that, in arr6 plants, canonical disease-resistance pathways, like those activated by defensive phytohormones, were not altered, whereas immune responses triggered by microbe-associated molecular patterns were slightly enhanced. Cell-wall composition of arr6 plants was found to be severely altered compared with that of wild-type plants. Remarkably, pectin-enriched cell-wall fractions extracted from arr6 walls triggered more intense immune responses than those activated by similar wall fractions from wild-type plants, suggesting that arr6 pectin fraction is enriched in wall-related damage-associated molecular patterns, which trigger immune responses. This work supports a novel function of ARR6 in the control of cell-wall composition and disease resistance and reinforces the role of the plant cell wall in the modulation of specific immune responses.},
	number = {5},
	urldate = {2023-03-10},
	journal = {Molecular Plant-Microbe Interactions®},
	author = {Bacete, Laura and Mélida, Hugo and López, Gemma and Dabos, Patrick and Tremousaygue, Dominique and Denancé, Nicolas and Miedes, Eva and Bulone, Vincent and Goffner, Deborah and Molina, Antonio},
	month = may,
	year = {2020},
	note = {Publisher: Scientific Societies},
	keywords = {Arabidopsis response regulators (ARR), Plectosphaerella cucumerina, Ralstonia solanacearum, cell wall, cytokinin, damage-associated molecular patterns (DAMPs), disease resistance, immunity},
	pages = {767--780},
}

The cytokinin signaling pathway, which is mediated by Arabidopsis response regulator (ARR) proteins, has been involved in the modulation of some disease-resistance responses. Here, we describe novel functions of ARR6 in the control of plant disease-resistance and cell-wall composition. Plants impaired in ARR6 function (arr6) were more resistant and susceptible, respectively, to the necrotrophic fungus Plectosphaerella cucumerina and to the vascular bacterium Ralstonia solanacearum, whereas Arabidopsis plants that overexpress ARR6 showed the opposite phenotypes, which further support a role of ARR6 in the modulation of disease-resistance responses against these pathogens. Transcriptomics and metabolomics analyses revealed that, in arr6 plants, canonical disease-resistance pathways, like those activated by defensive phytohormones, were not altered, whereas immune responses triggered by microbe-associated molecular patterns were slightly enhanced. Cell-wall composition of arr6 plants was found to be severely altered compared with that of wild-type plants. Remarkably, pectin-enriched cell-wall fractions extracted from arr6 walls triggered more intense immune responses than those activated by similar wall fractions from wild-type plants, suggesting that arr6 pectin fraction is enriched in wall-related damage-associated molecular patterns, which trigger immune responses. This work supports a novel function of ARR6 in the control of cell-wall composition and disease resistance and reinforces the role of the plant cell wall in the modulation of specific immune responses.

@article{melida_arabinoxylan-oligosaccharides_2020,
	title = {Arabinoxylan-{Oligosaccharides} {Act} as {Damage} {Associated} {Molecular} {Patterns} in {Plants} {Regulating} {Disease} {Resistance}},
	volume = {11},
	issn = {1664-462X},
	url = {https://www.frontiersin.org/articles/10.3389/fpls.2020.01210},
	doi = {10.3389/fpls.2020.01210},
	abstract = {Immune responses in plants can be triggered by damage/microbe-associated molecular patterns (DAMPs/MAMPs) upon recognition by plant pattern recognition receptors (PRRs). DAMPs are signaling molecules synthesized by plants or released from host cellular structures (e.g., plant cell walls) upon pathogen infection or wounding. Despite the hypothesized important role of plant cell wall-derived DAMPs in plant-pathogen interactions, a very limited number of these DAMPs are well characterized. Recent work demonstrated that pectin-enriched cell wall fractions extracted from the cell wall mutant impaired in Arabidopsis Response Regulator 6 (arr6), that showed altered disease resistance to several pathogens, triggered more intense immune responses than those activated by similar cell wall fractions from wild-type plants. It was hypothesized that arr6 cell wall fractions could be differentially enriched in DAMPs. In this work, we describe the characterization of the previous immune-active fractions of arr6 showing the highest triggering capacities upon further fractionation by chromatographic means. These analyses pointed to a role of pentose-based oligosaccharides triggering plant immune responses. The characterization of several pentose-based oligosaccharide structures revealed that β-1,4-xylooligosaccharides of specific degrees of polymerization and carrying arabinose decorations are sensed as DAMPs by plants. Moreover, the pentasaccharide 33-α-L-arabinofuranosyl-xylotetraose (XA3XX) was found as a highly active DAMP structure triggering strong immune responses in Arabidopsis thaliana and enhancing crop disease resistance.},
	urldate = {2023-03-10},
	journal = {Frontiers in Plant Science},
	author = {Mélida, Hugo and Bacete, Laura and Ruprecht, Colin and Rebaque, Diego and del Hierro, Irene and López, Gemma and Brunner, Frédéric and Pfrengle, Fabian and Molina, Antonio},
	year = {2020},
}

Immune responses in plants can be triggered by damage/microbe-associated molecular patterns (DAMPs/MAMPs) upon recognition by plant pattern recognition receptors (PRRs). DAMPs are signaling molecules synthesized by plants or released from host cellular structures (e.g., plant cell walls) upon pathogen infection or wounding. Despite the hypothesized important role of plant cell wall-derived DAMPs in plant-pathogen interactions, a very limited number of these DAMPs are well characterized. Recent work demonstrated that pectin-enriched cell wall fractions extracted from the cell wall mutant impaired in Arabidopsis Response Regulator 6 (arr6), that showed altered disease resistance to several pathogens, triggered more intense immune responses than those activated by similar cell wall fractions from wild-type plants. It was hypothesized that arr6 cell wall fractions could be differentially enriched in DAMPs. In this work, we describe the characterization of the previous immune-active fractions of arr6 showing the highest triggering capacities upon further fractionation by chromatographic means. These analyses pointed to a role of pentose-based oligosaccharides triggering plant immune responses. The characterization of several pentose-based oligosaccharide structures revealed that β-1,4-xylooligosaccharides of specific degrees of polymerization and carrying arabinose decorations are sensed as DAMPs by plants. Moreover, the pentasaccharide 33-α-L-arabinofuranosyl-xylotetraose (XA3XX) was found as a highly active DAMP structure triggering strong immune responses in Arabidopsis thaliana and enhancing crop disease resistance.

@article{bacete_plant_2020,
	title = {Plant {Biology}: {Plants} {Turn} {Down} the {Volume} to {Respond} to {Cell} {Swelling}},
	volume = {30},
	issn = {0960-9822},
	shorttitle = {Plant {Biology}},
	url = {https://www.sciencedirect.com/science/article/pii/S0960982220307806},
	doi = {10.1016/j.cub.2020.06.001},
	abstract = {Turgor manipulation to induce plant cell swelling is one of the classic experiments undertaken in biology courses in schools and at universities. However, only now do we start to understand the molecular mechanisms responsible for detecting plant cell swelling.},
	language = {en},
	number = {14},
	urldate = {2023-03-10},
	journal = {Current Biology},
	author = {Bacete, Laura and Hamann, Thorsten},
	month = jul,
	year = {2020},
	pages = {R804--R806},
}

Turgor manipulation to induce plant cell swelling is one of the classic experiments undertaken in biology courses in schools and at universities. However, only now do we start to understand the molecular mechanisms responsible for detecting plant cell swelling.

@article{bacete_role_2020,
	title = {The {Role} of {Mechanoperception} in {Plant} {Cell} {Wall} {Integrity} {Maintenance}},
	volume = {9},
	copyright = {http://creativecommons.org/licenses/by/3.0/},
	issn = {2223-7747},
	url = {https://www.mdpi.com/2223-7747/9/5/574},
	doi = {10.3390/plants9050574},
	abstract = {The plant cell walls surrounding all plant cells are highly dynamic structures, which change their composition and organization in response to chemical and physical stimuli originating both in the environment and in plants themselves. They are intricately involved in all interactions between plants and their environment while also providing adaptive structural support during plant growth and development. A key mechanism contributing to these adaptive changes is the cell wall integrity (CWI) maintenance mechanism. It monitors and maintains the functional integrity of cell walls by initiating adaptive changes in cellular and cell wall metabolism. Despite its importance, both our understanding of its mode of action and knowledge regarding the molecular components that form it are limited. Intriguingly, the available evidence implicates mechanosensing in the mechanism. Here, we provide an overview of the knowledge available regarding the molecular mechanisms involved in and discuss how mechanoperception and signal transduction may contribute to plant CWI maintenance.},
	language = {en},
	number = {5},
	urldate = {2023-03-10},
	journal = {Plants},
	author = {Bacete, Laura and Hamann, Thorsten},
	month = may,
	year = {2020},
	note = {Number: 5
Publisher: Multidisciplinary Digital Publishing Institute},
	keywords = {cell wall, cell wall integrity, mechanoperception, mechanosensing, plant defense, plant environment interaction},
	pages = {574},
}

The plant cell walls surrounding all plant cells are highly dynamic structures, which change their composition and organization in response to chemical and physical stimuli originating both in the environment and in plants themselves. They are intricately involved in all interactions between plants and their environment while also providing adaptive structural support during plant growth and development. A key mechanism contributing to these adaptive changes is the cell wall integrity (CWI) maintenance mechanism. It monitors and maintains the functional integrity of cell walls by initiating adaptive changes in cellular and cell wall metabolism. Despite its importance, both our understanding of its mode of action and knowledge regarding the molecular components that form it are limited. Intriguingly, the available evidence implicates mechanosensing in the mechanism. Here, we provide an overview of the knowledge available regarding the molecular mechanisms involved in and discuss how mechanoperception and signal transduction may contribute to plant CWI maintenance.

@article{melida_non-branched_2018,
	title = {Non-branched β-1,3-glucan oligosaccharides trigger immune responses in {Arabidopsis}},
	volume = {93},
	issn = {1365-313X},
	url = {https://onlinelibrary.wiley.com/doi/abs/10.1111/tpj.13755},
	doi = {10.1111/tpj.13755},
	abstract = {Fungal cell walls, which are essential for environmental adaptation and host colonization by the fungus, have been evolutionarily selected by plants and animals as a source of microbe-associated molecular patterns (MAMPs) that, upon recognition by host pattern recognition receptors (PRRs), trigger immune responses conferring disease resistance. Chito-oligosaccharides [β-1,4-N-acetylglucosamine oligomers, (GlcNAc)n] are the only glycosidic structures from fungal walls that have been well-demonstrated to function as MAMPs in plants. Perception of (GlcNAc)4–8 by Arabidopsis involves CERK1, LYK4 and LYK5, three of the eight members of the LysM PRR family. We found that a glucan-enriched wall fraction from the pathogenic fungus Plectosphaerella cucumerina which was devoid of GlcNAc activated immune responses in Arabidopsis wild-type plants but not in the cerk1 mutant. Using this differential response, we identified the non-branched 1,3-β-d-(Glc) hexasaccharide as a major fungal MAMP. Recognition of 1,3-β-d-(Glc)6 was impaired in cerk1 but not in mutants defective in either each of the LysM PRR family members or in the PRR-co-receptor BAK1. Transcriptomic analyses of Arabidopsis plants treated with 1,3-β-d-(Glc)6 further demonstrated that this fungal MAMP triggers the expression of immunity-associated genes. In silico docking analyses with molecular mechanics and solvation energy calculations corroborated that CERK1 can bind 1,3-β-d-(Glc)6 at effective concentrations similar to those of (GlcNAc)4. These data support that plants, like animals, have selected as MAMPs the linear 1,3-β-d-glucans present in the walls of fungi and oomycetes. Our data also suggest that CERK1 functions as an immune co-receptor for linear 1,3-β-d-glucans in a similar way to its proposed function in the recognition of fungal chito-oligosaccharides and bacterial peptidoglycan MAMPs.},
	language = {en},
	number = {1},
	urldate = {2023-03-10},
	journal = {The Plant Journal},
	author = {Mélida, Hugo and Sopeña-Torres, Sara and Bacete, Laura and Garrido-Arandia, María and Jordá, Lucía and López, Gemma and Muñoz-Barrios, Antonio and Pacios, Luis F. and Molina, Antonio},
	year = {2018},
	note = {\_eprint: https://onlinelibrary.wiley.com/doi/pdf/10.1111/tpj.13755},
	keywords = {BAK1, CERK, cell wall, chitin, glucan, necrotrophic fungi, plant immunity},
	pages = {34--49},
}

Fungal cell walls, which are essential for environmental adaptation and host colonization by the fungus, have been evolutionarily selected by plants and animals as a source of microbe-associated molecular patterns (MAMPs) that, upon recognition by host pattern recognition receptors (PRRs), trigger immune responses conferring disease resistance. Chito-oligosaccharides [β-1,4-N-acetylglucosamine oligomers, (GlcNAc)n] are the only glycosidic structures from fungal walls that have been well-demonstrated to function as MAMPs in plants. Perception of (GlcNAc)4–8 by Arabidopsis involves CERK1, LYK4 and LYK5, three of the eight members of the LysM PRR family. We found that a glucan-enriched wall fraction from the pathogenic fungus Plectosphaerella cucumerina which was devoid of GlcNAc activated immune responses in Arabidopsis wild-type plants but not in the cerk1 mutant. Using this differential response, we identified the non-branched 1,3-β-d-(Glc) hexasaccharide as a major fungal MAMP. Recognition of 1,3-β-d-(Glc)6 was impaired in cerk1 but not in mutants defective in either each of the LysM PRR family members or in the PRR-co-receptor BAK1. Transcriptomic analyses of Arabidopsis plants treated with 1,3-β-d-(Glc)6 further demonstrated that this fungal MAMP triggers the expression of immunity-associated genes. In silico docking analyses with molecular mechanics and solvation energy calculations corroborated that CERK1 can bind 1,3-β-d-(Glc)6 at effective concentrations similar to those of (GlcNAc)4. These data support that plants, like animals, have selected as MAMPs the linear 1,3-β-d-glucans present in the walls of fungi and oomycetes. Our data also suggest that CERK1 functions as an immune co-receptor for linear 1,3-β-d-glucans in a similar way to its proposed function in the recognition of fungal chito-oligosaccharides and bacterial peptidoglycan MAMPs.

@article{bacete_plant_2018,
	title = {Plant cell wall-mediated immunity: cell wall changes trigger disease resistance responses},
	volume = {93},
	issn = {1365-313X},
	shorttitle = {Plant cell wall-mediated immunity},
	url = {https://onlinelibrary.wiley.com/doi/abs/10.1111/tpj.13807},
	doi = {10.1111/tpj.13807},
	abstract = {Plants have evolved a repertoire of monitoring systems to sense plant morphogenesis and to face environmental changes and threats caused by different attackers. These systems integrate different signals into overreaching triggering pathways which coordinate developmental and defence-associated responses. The plant cell wall, a dynamic and complex structure surrounding every plant cell, has emerged recently as an essential component of plant monitoring systems, thus expanding its function as a passive defensive barrier. Plants have a dedicated mechanism for maintaining cell wall integrity (CWI) which comprises a diverse set of plasma membrane-resident sensors and pattern recognition receptors (PRRs). The PRRs perceive plant-derived ligands, such as peptides or wall glycans, known as damage-associated molecular patterns (DAMPs). These DAMPs function as ‘danger’ alert signals activating DAMP-triggered immunity (DTI), which shares signalling components and responses with the immune pathways triggered by non-self microbe-associated molecular patterns that mediate disease resistance. Alteration of CWI by impairment of the expression or activity of proteins involved in cell wall biosynthesis and/or remodelling, as occurs in some plant cell wall mutants, or by wall damage due to colonization by pathogens/pests, activates specific defensive and growth responses. Our current understanding of how these alterations of CWI are perceived by the wall monitoring systems is scarce and few plant sensors/PRRs and DAMPs have been characterized. The identification of these CWI sensors and PRR–DAMP pairs will help us to understand the immune functions of the wall monitoring system, and might allow the breeding of crop varieties and the design of agricultural strategies that would enhance crop disease resistance.},
	language = {en},
	number = {4},
	urldate = {2023-03-10},
	journal = {The Plant Journal},
	author = {Bacete, Laura and Mélida, Hugo and Miedes, Eva and Molina, Antonio},
	year = {2018},
	note = {\_eprint: https://onlinelibrary.wiley.com/doi/pdf/10.1111/tpj.13807},
	keywords = {Arabidopsis, DAMP, PRR, cell wall, cell wall integrity, cell wall mutant, disease resistance, immunity, wall sensor},
	pages = {614--636},
}

Plants have evolved a repertoire of monitoring systems to sense plant morphogenesis and to face environmental changes and threats caused by different attackers. These systems integrate different signals into overreaching triggering pathways which coordinate developmental and defence-associated responses. The plant cell wall, a dynamic and complex structure surrounding every plant cell, has emerged recently as an essential component of plant monitoring systems, thus expanding its function as a passive defensive barrier. Plants have a dedicated mechanism for maintaining cell wall integrity (CWI) which comprises a diverse set of plasma membrane-resident sensors and pattern recognition receptors (PRRs). The PRRs perceive plant-derived ligands, such as peptides or wall glycans, known as damage-associated molecular patterns (DAMPs). These DAMPs function as ‘danger’ alert signals activating DAMP-triggered immunity (DTI), which shares signalling components and responses with the immune pathways triggered by non-self microbe-associated molecular patterns that mediate disease resistance. Alteration of CWI by impairment of the expression or activity of proteins involved in cell wall biosynthesis and/or remodelling, as occurs in some plant cell wall mutants, or by wall damage due to colonization by pathogens/pests, activates specific defensive and growth responses. Our current understanding of how these alterations of CWI are perceived by the wall monitoring systems is scarce and few plant sensors/PRRs and DAMPs have been characterized. The identification of these CWI sensors and PRR–DAMP pairs will help us to understand the immune functions of the wall monitoring system, and might allow the breeding of crop varieties and the design of agricultural strategies that would enhance crop disease resistance.

@article{gonneau_receptor_2018,
	title = {Receptor {Kinase} {THESEUS1} {Is} a {Rapid} {Alkalinization} {Factor} 34 {Receptor} in {Arabidopsis}},
	volume = {28},
	issn = {0960-9822},
	url = {https://www.sciencedirect.com/science/article/pii/S0960982218307115},
	doi = {10.1016/j.cub.2018.05.075},
	abstract = {The growth of plants, like that of other walled organisms, depends on the ability of the cell wall to yield without losing its integrity. In this context, plant cells can sense the perturbation of their walls and trigger adaptive modifications in cell wall polymer interactions. Catharanthus roseus receptor-like kinase 1-like (CrRLK1L) THESEUS1 (THE1) was previously shown in Arabidopsis to trigger growth inhibition and defense responses upon perturbation of the cell wall, but so far, neither the ligand nor the role of the receptor in normal development was known. Here, we report that THE1 is a receptor for the peptide rapid alkalinization factor (RALF) 34 and that this signaling module has a role in the fine-tuning of lateral root initiation. We also show that RALF34-THE1 signaling depends, at least for some responses, on FERONIA (FER), another RALF receptor involved in a variety of processes, including immune signaling, mechanosensing, and reproduction [1]. Together, the results show that RALF34 and THE1 are part of a signaling network that integrates information on the integrity of the cell wall with the coordination of normal morphogenesis.},
	language = {en},
	number = {15},
	urldate = {2023-03-10},
	journal = {Current Biology},
	author = {Gonneau, Martine and Desprez, Thierry and Martin, Marjolaine and Doblas, Verónica G. and Bacete, Laura and Miart, Fabien and Sormani, Rodnay and Hématy, Kian and Renou, Julien and Landrein, Benoit and Murphy, Evan and Van De Cotte, Brigitte and Vernhettes, Samantha and De Smet, Ive and Höfte, Herman},
	month = aug,
	year = {2018},
	keywords = {FERONIA, cell wall, lateral root initiation, rapid alkalinization factor},
	pages = {2452--2458.e4},
}

The growth of plants, like that of other walled organisms, depends on the ability of the cell wall to yield without losing its integrity. In this context, plant cells can sense the perturbation of their walls and trigger adaptive modifications in cell wall polymer interactions. Catharanthus roseus receptor-like kinase 1-like (CrRLK1L) THESEUS1 (THE1) was previously shown in Arabidopsis to trigger growth inhibition and defense responses upon perturbation of the cell wall, but so far, neither the ligand nor the role of the receptor in normal development was known. Here, we report that THE1 is a receptor for the peptide rapid alkalinization factor (RALF) 34 and that this signaling module has a role in the fine-tuning of lateral root initiation. We also show that RALF34-THE1 signaling depends, at least for some responses, on FERONIA (FER), another RALF receptor involved in a variety of processes, including immune signaling, mechanosensing, and reproduction [1]. Together, the results show that RALF34 and THE1 are part of a signaling network that integrates information on the integrity of the cell wall with the coordination of normal morphogenesis.

@incollection{bacete_characterization_2017,
	address = {New York, NY},
	series = {Methods in {Molecular} {Biology}},
	title = {Characterization of {Plant} {Cell} {Wall} {Damage}-{Associated} {Molecular} {Patterns} {Regulating} {Immune} {Responses}},
	isbn = {978-1-4939-6859-6},
	url = {https://doi.org/10.1007/978-1-4939-6859-6_2},
	abstract = {The plant cell wall is one of the first defensive barriers that pathogens need to overcome to successfully colonize plant tissues. Plant cell wall is considered a dynamic structure that regulates both constitutive and inducible defense mechanisms. The wall is a potential source of a diverse set of Damage-Associated Molecular Patterns (DAMPs), which are signalling molecules that trigger immune responses. However, just a few active wall ligands, such as oligogalacturonic acids (OGs), have been characterized so far. To identify additional wall-derived DAMPs, we obtained different plant wall fractions and tested their capacity to trigger immune responses using a calcium read-out system. To characterize the active DAMPs structures present in these fractions, we applied Glycome Profiling, a technology that uses a large and diverse set of specific monoclonal antibodies against wall carbohydrate ligands. The methods describe here can be used in combination with other biochemical approaches to identify and purify new plant cell wall DAMPs.},
	language = {en},
	urldate = {2023-03-10},
	booktitle = {Plant {Pattern} {Recognition} {Receptors}: {Methods} and {Protocols}},
	publisher = {Springer},
	author = {Bacete, Laura and Mélida, Hugo and Pattathil, Sivakumar and Hahn, Michael G. and Molina, Antonio and Miedes, Eva},
	editor = {Shan, Libo and He, Ping},
	year = {2017},
	doi = {10.1007/978-1-4939-6859-6_2},
	keywords = {Arabidopsis, Cell wall, Hemicellulose, Immunity, Pectin},
	pages = {13--23},
}

The plant cell wall is one of the first defensive barriers that pathogens need to overcome to successfully colonize plant tissues. Plant cell wall is considered a dynamic structure that regulates both constitutive and inducible defense mechanisms. The wall is a potential source of a diverse set of Damage-Associated Molecular Patterns (DAMPs), which are signalling molecules that trigger immune responses. However, just a few active wall ligands, such as oligogalacturonic acids (OGs), have been characterized so far. To identify additional wall-derived DAMPs, we obtained different plant wall fractions and tested their capacity to trigger immune responses using a calcium read-out system. To characterize the active DAMPs structures present in these fractions, we applied Glycome Profiling, a technology that uses a large and diverse set of specific monoclonal antibodies against wall carbohydrate ligands. The methods describe here can be used in combination with other biochemical approaches to identify and purify new plant cell wall DAMPs.