Biology subjects

Voxeur, A.

Publications and source records attributed to Voxeur, A..

16 recordsLinked to original sources

bioRxiv2025

APETALA2 controls seed growth by regulating outer integument mechanical identity during seed coat differentiation

Organ morphogenesis requires differentiating cells to acquire mechanical properties that precisely regulate growth. In Arabidopsis thaliana, seed size and shape are largely controlled by the outer integument of the seed coat, whose two layers develop distinct mechanical behaviors critical for growth regulation. Here, we show that the cell identity factor APETALA2 (AP2) both promotes and restricts seed growth by coordinating the post-fertilization differentiation of these layers into distinct seed coat tissues. AP2 drives layer-specific remodeling of cell wall composition, notably affecting pectin and xyloglucan, thereby establishing contrasting mechanical properties between the two layers. Unexpectedly, these changes in wall composition and mechanics do not alter mechanosensitive responses, revealing a surprising uncoupling between cell wall remodeling and mechanotransduction. Our findings identify AP2 as a key coordinator of tissue-specific mechanical differentiation during organ morphogenesis.

plant biology↗

bioRxiv2024

Long-term high temperatures affect seed maturation and seed coat integrity in Brassica napus

Temperatures above the optimum growth threshold affect seed development, accelerating embryo development and producing seeds with ruptured seed coats. However, the underlying mechanisms of this effect remain unclear. To investigate temperature-induced seed coat rupture, we used a multidisciplinary approach combining detailed phenotyping, transcriptomics, histology, immunolabelling, cell wall profiling and nanoindentation-based mechanics of the seed coat in oilseed rape (Brassica napus). Our data reveal that high temperatures accelerate embryo growth, resulting in larger embryos without a compensatory increase in overall seed size. This rapid embryo expansion may exert elevated mechanical stress on the seed coat cells, significantly thinning the seed coat layers. Concurrently, prolonged exposure to high temperatures drives premature biochemical maturation of the seed coat, characterized by accumulation of demethylesterified pectin. Nanoindentation analysis demonstrated that these changes may have compromised the structural integrity of seed coat cell walls. Ultimately, the weakened seed coat may not withstand the internal tension imposed by the fast-growing embryo, leading to seed coat rupture and reduced seed quality. Our work on seed maturation and the mechanism of heat-induced seed coat rupture provides valuable insights for future research into breeding thermotolerant Brassica napus, crucial in the context of a changing climate. HIGHLIGHTSO_LIGrowth of Brassica napus under long-term high temperatures accelerated embryo growth and led to seed coat rupture in approximately half of the seeds. C_LIO_LIPathways linked to cell wall modification were found to be significantly differentially regulated at high temperatures, accompanied by increased pectin demethylesterification in the seed coat. C_LIO_LIA lower reduced modulus and hardness indicate that the surface of HT seeds is mechanically more compliant and softer. Together with the reduced thickness of the seed coat layers, such a seed coat, as a whole structure, is mechanically weaker and less able to resist deformation imposed by embryo growth, resulting in seed coat rupture. C_LI

plant biology↗

bioRxiv2024

Arabidopsis AtGELP53 modulates polysaccharide acetylation and defense response through oligosaccharide-mediated signaling

O-acetylation is a crucial substitution found in hemicelluloses and pectin which are necessary for maintaining the flexibility and structural integrity of the cell. A balanced polysaccharide O-acetylation level is maintained by cell wall acetyl transferases and esterases present in different cell organelles. Specifically, the role of esterases in cell wall acetylation metabolism is less explored. Therefore, we investigated the role of AtGELP53 from the GELP family of esterases or lipases. Here, we show that AtGELP53 is localized in the plasma membrane. Analysis of AtGELP53 overexpressing independent transgenic lines revealed a decrease in xyloglucan acetylation, changes in other polysaccharide acetylation and alteration in cell wall composition. The series of elicitor-based, transcriptomic and proteomic analyses in AtGELP53 overexpressing lines suggested that oligosaccharide-mediated signalling activates the cell wall and defence-related genes primarily because of xyloglucan deacetylation. Furthermore, AtGELP53 overexpression plants showed resistance against Pseudomonas syringae and Ralstonia solanacearum through activation of elicitor-mediated defense response. Overall, our findings outline the role of AtGELP53 in polysaccharide acetylation, cell wall remodelling and defense through the activation of plant cell wall integrity.

plant biology↗

bioRxiv2024

Parallel Evolution of Salinity Tolerance in Arabidopsis thaliana Accessions from Cape Verde Islands

Soil salinization poses a significant threat to crop production impacting one fifth of all cultivated land. The Cape Verde Islands are located 600 km from the coast of Africa and are characterized by high salinity soils and inland water sources. In this study we find that Arabidopsis thaliana plants native to these islands accumulate a metabolite that protects them from salt stress. We partially characterized this metabolite as glucuronyl-mannose. We find that the ability to produce glucuronyl-mannose evolved independently in two different islands from the same archipelago through mutations in the same gene, an alpha glycosidase protein that we named GH38cv. These cases of parallel evolution suggest positive selection towards the increase of salt tolerance with low fitness costs. Indeed, plants carrying derived alleles of GH38cv do not present growth defects or low defenses under normal conditions, but show better germination rates, longer roots and better hydric status than non-mutated plants when exposed to salt stress. These findings provide a knowledge-based method to develop salt resilient crops using natural mechanisms, which could be attractive both to conventional and organic agriculture.

plant biology↗

bioRxiv2024

Galacturonic acid oxidation: a radical way to stick together

The middle lamellae (ML) of plant cells, enriched in Homogalacturonan (HG) is considered to function as a crucial glue responsible for cell-cell cohesion (for review see 1). Mutants with defective HG content exhibit cell adhesion defects2,3. Despite advances, the mechanisms governing cell adhesion during plant development remain elusive. We previously hypothesized that cell-cell cohesion relies on cell wall integrity signaling, yet the specifics remain undefined4. OligoGalacturonans (OG), degradation products of HG, are prime candidates for informing cells about ML status and thereby influencing cell adhesion. This integrity signal is crucial for adhesion homeostasis 4. OGs serve as signaling molecules, recognized by membrane-bound cell wall receptors. Notably, restoring adhesion in qua2-1 mutants by modulating pectin response gene expression in esmd/qua2 mutants underscores potential OGs importance4-6. Deciphering the diversity and role of endogenous OGs is imperative for understanding cell adhesion modulation. Our study aims to identify compounds in this signalling pathway that regulate cell adhesion. We focused on characterizing HG degradation products in dark-grown hypocotyls. Our findings highlight various oligomers, along with two key monomers: galacturonic acid and its oxidized form, galactaric acid. These monomers appear to play a pivotal role in controlling cell adhesion by indirectly enhancing the crosslinking of extensin, a cell wall structural protein. This crosslinking leads to the densification of extensin-based cell wall networks, ultimately restoring cell adhesion in defective mutants. Our research sheds light on the intricate interplay between HG degradation product monomer and cell adhesion mechanisms.

plant biology↗

bioRxiv2024

A change in the cell wall status initiates the elimination of the nucellus in Arabidopsis

The evolution of the seed habit coincides with a change in the cell fate of the nucellus, the sporophytic tissue responsible for female meiosis. Seeds arose when the nucellus retained the female spores instead of releasing them in the environment. As a consequence, the nucellus was partially eliminated to accommodate the growth of the female gametophyte inside the sporophyte. With the evolution of angiosperm seeds, the process of nucellus elimination was requisitioned to allow the growth of the endosperm, the second fertilization product devoted to store nutrients. Cell elimination differs from most known cell death programs as it leads to the apparent dismantling of the cell wall. Here, we show that nucellus elimination in Arabidopsis is initiated by the lysis of the pectic polysaccharides in the cell wall. This process exposes other cell wall components to possible further degradation and precedes a cell death program that leads to nuclear DNA fragmentation. Both pathways are regulated by TRANSPARENT TESTA 16, a MADS-domain transcription factor that evolved with seed plants. Finally, the causal effect of cell wall modification on nucellus development is demonstrated by inhibiting pectin degradation, thus suggesting that a change in the cell wall status might have driven seed evolution.

plant biology↗

bioRxiv2024

Unravelling the Interplay of Nitrogen Nutrition and the Botrytis cinerea pectin lyase BcPNL1 in Modulating Arabidopsis thaliana Susceptibility

In this study, we investigated the intricate interplay between nitrogen nutrition, and the dynamics of pectin degradation during plant-pathogen interactions, using Arabidopsis thaliana and Botrytis cinerea as a model pathosystem. Our findings revealed a noteworthy impact of nitrogen availability on the pectin degrading activity of the B. cinerea pectin lyase 1 (PNL) for which the mutant strains presented a reduced pathogenicity restored by complementation. More precisely, infected nitrogen-sufficient (high N) plants exhibited an increased release of PNL-derived oligogalacturonides compared to infected nitrogen-deficient (low N) plants. This correlated with an elevated expression of jasmonic acid repressor genes in high N plants, rendering them more susceptible to B. cinerea. Using{Delta} Bcpnl1 deletion mutants, we demonstrated that the increased production of BcPNL1 -derived oligogalacturonides under high N conditions was responsible for the increased expression of jasmonic acid repressor genes, significantly contributing to the higher susceptibility of high N plantsto B. cinerea. In conclusion, we demonstrated that BcPNL1 is a major pathogenicity factor during B. cinerea interaction that is affected by the plants N nutrition conditions.

plant biology↗

bioRxiv2023

Inositolphosphate glycans accumulate and suppress plant defense during Arabidopsis/Botrytis interaction

This study investigates the presence and significance of previously undiscovered oligosaccharides that accumulate during the interaction between Arabidopsis thaliana and Botrytis cinerea, a pathogenic fungus. Initially focused on characterizing cell wall-derived oligosaccharides, the research uncovered inositol phosphate glycans (IPGs) originating from plant sphingolipids, specifically glycosylinositol phosphorylceramides. Advanced chromatography, mass spectrometry techniques and molecular biology were employed to identify these IPGs, determine their origins, and study their role in the A. thaliana-B. cinerea interaction. Contrary to the conventional belief that oligosaccharides trigger plant defense, this research suggests that B. cinerea releases IPGs identical to those generated by host plant to actually downregulate plant defense mechanisms. This discovery offers insight into the dynamic strategies used by B. cinerea to evade plant defenses and establish successful infections. One-Sentence SummaryA plant pathogen releases products identical to those generated by host plant that aids in evasion of the plant defense.

plant biology↗

bioRxiv2023

Fine-Tuning and Remodelling of Pectins Play a Key Role in the Maintenance of Cell Adhesion

Plant cell adhesion is essential for development and stress response, mediated by pectin-rich middle lamella deposition between cell walls. However, the precise control mechanism of cell adhesion remains unclear. The qua2-1 and esmd1-1 mutants provide a better understanding of this process and suggest a signaling pathway triggering the loss and restoration of adhesion via cell wall modifications. This study attempts to characterize the potential regulatory role of endogenous oligogalacturonides (OGs) and pectin modifications in the control of cell adhesion in Arabidopsis. From dark-grown hypocotyls, our extraction revealed seven distinct endogenous OGs with varying polymerization and modifications. Abundance variations of OGs were observed among wild type, qua2-1, esmd1-1, and qua2-1/esmd1-1 mutants. The structure of homogalacturonans was analyzed by enzymatic fingerprint, in order to identify changes in esterification patterns. Expression analysis of pectin-modifying enzymes showed significant variations in PME, PMEI, and PAE genes. Gene expressions correlate with homogalacturonans modifications and cell adhesion phenotypes. This study enhances our understanding of a feedback loop between the endogenous OGs, homogalacturonans esterification fine tuning, and pectin remodeling enzymes expression in controlling cell adhesion.

plant biology↗

bioRxiv2023

Chitosan stimulates root hair callose deposition and inhibits root hair growth

Although angiosperm plants have a general capacity to react after the immunity elicitor chitin or chitosan treatment by the cell wall callose deposition, this response in particular cell types and its evolutionary conservation is not understood. Here we show that also the growing root hairs (RHs) of Arabidopsis can respond to a mild (0.001%) chitosan treatment by the callose deposition and by a deceleration of the RH growth. We demonstrate that the glucan synthase-like 5 (GSL5)/PMR4 is vital for chitosan-induced callose deposition but not for RH growth inhibition. Upon the higher chitosan concentration (0.01%) treatment, RHs do not deposit callose, while growth inhibition is prominent. To understand the specificities of the low and high concentration chitosan treatments, we analysed the corresponding PTI signalling components, gene expression, and RH cellular endomembrane and cytoskeleton modifications. Importantly, chitosan-induced callose deposition is also present in the functionally analogous and evolutionarily only distantly related RH-like structures rhizophores (lycophytes) and rhizoids (bryophytes). Our results point to the RH callose deposition as a conserved strategy of soil-anchoring plant cells (rhizoids/rhizophores/RHs) to deal with mild biotic stress. At the same time, high chitosan concentration prominently disturbs intracellular dynamics, tip-localised endomembrane compartments and RH growth, precluding callose deposition.

plant biology↗

bioRxiv2022

The structural, dynamical and biochemical characterizations of Verticillium dahliae pectate lyase, VdPelB, highlight its specificities

Pectins, complex polysaccharides and major components of the plant primary cell wall, can be degraded by pectate lyases (PLs). PLs cleave glycosidic bonds of homogalacturonans (HG), the main pectic domain, by {beta}-elimination, releasing unsaturated oligogalacturonides (OGs). To understand the catalytic mechanism and structure/function of these enzymes, we characterized VdPelB from Verticillium dahliae, a plant pathogen. We first solved the crystal structure of VdPelB at 1.2[A] resolution showing that it is a right-handed parallel {beta}-helix structure. Molecular dynamics (MD) simulations further highlighted the dynamics of the enzyme in complex with substrates that vary in their degree of methylesterification, identifying amino acids involved in substrate binding and cleavage of non-methylesterified pectins. We then biochemically characterized wild type and mutated forms of VdPelB. VdPelB was most active on non-methylesterified pectins, at pH 8 in presence of Ca2+ ions. VdPelB-G125R mutant was most active at pH 9 and showed higher relative activity compared to native enzyme. The OGs released by VdPelB differed to that of previously characterized PLs, showing its peculiar specificity in relation to its structure. OGs released from Verticillium-partially tolerant and sensitive flax cultivars differed which could facilitate the identification VdPelB-mediated elicitors of defence responses.

biochemistry↗

bioRxiv2022

Differences in the structure of plant polygalacturonases specify enzymes dynamics and processivities to fine-tune pectins and root development

The fine-tuning of pectins by polygalacturonases (PGs) plays a key role in modulating plant cell wall chemistry and mechanics, impacting plant development. In plants, the high number of PGs encoded in the genome questions the regulation of pectin depolymerization and the roles of distinct isozymes in the control of development. Here we report the first crystal structures of two PGs from Arabidopsis, PGLR and ADPG2 whose expression overlap in roots. Albeit having overall conserved folds and active sites, PGLR and ADPG2 differed in the structure of their binding grooves and in the amino-acids of the subsites. We determined the structural features that explain the absence of inhibition of the plant PGs by endogenous PG-Inhibiting Proteins (PGIPs). By combining molecular dynamic simulations, analysis of enzymes kinetics and hydrolysis products, we showed that subtle differences in PGLR and ADPG2 structures translated into distinct enzyme-substrate dynamics and enzymes processivities. Using the plant root as a developmental model, exogenous application of purified enzymes showed that these distinct PGLR/ADPG2 processivities ultimately translated into different impacts on development. The highly processive ADPG2 had major effects on both root cell elongation and cell adhesion. Our study suggests that, in plants, gene redundancy is unlikely to reflect redundant biochemical specificities. Isozymes of distinct specificities and processivities are likely to be of major importance for the fine spatial and temporal regulation of pectin structure. Significance StatementPlant polygalacturonases (PG) are enzymes that play a key role in the regulation of cell wall pectin chemistry by controlling the degree of polymerization of the HG chains. The high number of genes encoding PG in Arabidopsis questions the rationale for such abundance. We solved the crystal structure of two PG (PGLR and ADPG2) whose expression overlap in roots and showed, using combined computational and experimental approaches, that they differ in their enzyme-substrate dynamics, leading to distinct processivities. The highly processive ADPG2 can generate digestion products of shorter degree of polymerization, and upon exogenous application on developing roots, induced drastic developmental defects. Our study suggests that gene redundancy is unlikely to reflect redundant biochemical specificities of isozymes.

biochemistry↗

bioRxiv2021

Impairment of the cellulose degradation machinery enhances fungal virulence but limits reproductive fitness

Fungal endophytes grow in the apoplastic space, in constant contact with the plant cell wall (CW) that hinders microbe progression, while representing a source of nutrients. Although numerous fungal CW modifying proteins have been identified, their role during host colonization remains underexplored. Here we show that the root-infecting plant pathogen Fusarium oxysporum (Fo) does not require its complete arsenal of cellulases to infect the host plant. Quite the opposite, Fo mutants impaired in cellulose degradation become hypervirulent by enhancing the secretion of virulence factors. On the other hand, the reduction on cellulase activity had a severe negative effect on saprophytic growth and microconidia production during the final stages of the Fo infection cycle. These findings enhance our understanding on the function of plant CW degradation on the outcome of host-microbe interactions and reveal an unexpected role of cellulose degradation in a pathogens evolutionary success. TeaserUnexpectedly, fungi compromised in their capacity to degrade plant cellulose are hypervirulent but impaired in sporulation.

microbiology↗

bioRxiv2021

Pectin remodeling belongs to a homeostatic system and triggers transcriptomic and hormonal modulations

O_LIHere, we focused on the biological modifications arisen from a strong and transient variation of the pectin methylesterification status during the seed-to-seedling transition. C_LIO_LIA reverse genetic approach was used to trigger specific reduction of pectin de-methylesterification during the seed maturation stage and the related physiological effects were assessed using a combination of biochemical, transcriptomic and microscopic analyses. C_LIO_LIArabidopsis PME36 is required to implement the characteristic pattern of de-methylesterified pectin in the mature seed. While this pattern is strongly impaired in pme36-1 and pme36-2 mature seed, no phenotypical effect is observed in the knockout mutant during seed germination. By analyzing hormone homeostasis and gene expression regulation, we show a strong and dynamic physiological disorder in the mutant, which reveals the existence of a complex compensatory mechanism overcoming the defect in pectin de-methylesterification. C_LIO_LIOur results reveal that pectin methylesterification status acts as upstream modulator involved in an undescribed homeostatic system in which pectin remodeling, hormone signaling and transcriptomic regulations interact to ensure the maintenance of a normal seed-to-seedling developmental program. C_LI

plant biology↗

bioRxiv2021

Plant inositol-phosphate-glycans and fucosylated xyloglucan oligosaccharide are accumulated upon Arabidopsis thaliana/ Botrytis cinerea infection

In mammals, insulin is involved in controlling blood glucose levels and its role in modulating immunity is being more and more documented. This hormone promotes the release of inositolphosphate glycans (IPG) which act as mediators. In plants, one IG has already been identified in plant culture cells (Smith and Fry, 1999; Smith et al., 1999) but, to our knowledge, no IPG have been yet identified. Here, we discovered 7 IPG that are accumulated upon Arabidopsis thaliana-Botrytis cinerea interaction, concomitantly with oligogalacturonides and a fucosylated xyloglucan oligosaccharide. Further structural characterization showed that they come from the hydrolysis of polar heads of Serie A to H glycosylinositol phosphorylceramides presumably via a phospholipase C activity. Taken together with the emerging role of insulin as immune regulator, these results question the role of IPG as damage associated molecular pattern both in animal and plant kingdoms.

plant biology↗

bioRxiv2021

The pH-dependent processivity of Arabidopsis AtPME2 can control cell wall mechanical properties

Pectin methylesterases (PMEs) modify homogalacturonans (HG) chemistry and play a key role in regulating primary cell wall mechanical properties. How PME activity can fine-tune pectin structure in the growing plant has remained elusive. Here we report on the Arabidopsis AtPME2, which we found to be highly expressed during lateral root emergence and dark-grown hypocotyl elongation. We produced the mature active enzyme using heterologous expression in Pichia pastoris and characterized it through the use of a generic plant PME antiserum suitable for detecting recombinant and native enzyme independent of species source. At neutral pH AtPME2 is preferentially active on pectins with a degree of 55-70% methylesterification and can be inhibited by PME inhibitor protein (PMEI). We show that the mode of action for AtPME2 can switch from full processivity (at pH 8), creating large blocks of unmethylated galacturonic acid, to low processivity (at pH 5) and relate these observations to the differences in electrostatic potential of the protein at acidic and alkaline pH. To assess the role of AtPME2 in development, we characterized two knock-out lines. We show that in the context of acidified apoplast, low-processive demethylesterification by AtPME2 can loosen the cell wall, with consequent increase in cell elongation and etiolated hypocotyl length. Our study brings insights into how the pH-dependent regulation by PME activity could affect pectin structure and associated cell wall mechanical properties in expansion. One sentence summaryThe processivity of AtPME2, a pectin methylesterase that fine-tunes cell wall pectins is modulated by pH in vitro and impacts the mechanical properties of the wall, affecting development in planta.

plant biology↗