Biology subjects

Pelloux, J.

Publications and source records attributed to Pelloux, J..

7 recordsLinked to original sources

bioRxiv2024

Oligogalacturonides, produced by enzymatic degradation of sugar beet by-products, confer partial protection against wheat powdery mildew

Nowadays, it becomes urgent to use new alternative strategies for a more sustainable agriculture, by developing eco-friendly biocontrol compounds leading to an induction of plant resistance against pathogens. In recent years, oligogalacturonides (OGs), fragments of pectin resulting from the degradation of the plant cell wall, have been particularly under focus for their ability to induce plant defense, leading to plant protection against pathogens. In the current work, 2 mixtures of OGs, bio-sourced from sugar beet by-products, were produced and assessed for their protective efficacy on wheat against Blumeria graminis f. sp. tritici (Bgt), responsible for powdery mildew. These OGs pools were obtained by hydrolysing pectins with two A. thaliana polygalacturonases (PGs), ADPG2 and PGLR (EC 3.2.1.15), produced in heterologous system Pichia pastoris. No direct effect of OGs generated by either ADPG2, named OGs-ADPG2 (degree of polymerisation/DP 1-10 and degree of esterification/DE 41%) or PGLR, named OGs-PGLR (DP 1-16, DE 34 %), was observed on Bgt spore germination in vitro. After a preventive foliar spraying of wheat by OGs, only OGs-PGLR provided a 31 % partial protection against Bgt. Both pools of OGs triggered an increase in the expression of some plant defense-related genes (peroxidase or PR proteins encoding genes), before inoculation with the pathogen. However, only OGs-PGLR induced the expression of a gene encoding a lipoxygenase (LOX), which is the first enzyme involved in the octadecanoic pathway, leading to oxylipins production in plant defense. Taking together, these findings revealed that some OGs, produced from sugar beet by-products by enzymatic hydrolysis, could be used as an alternative of phytochemicals.

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↗

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

TBL38 is an atypical homogalacturonan acetylesterase with a peculiar cell wall microdomain localization in Arabidopsis seed mucilage secretory cells

Plant cell walls are made of complex polysaccharidic/proteinaceous network whose biosynthesis and dynamics implicate several cell compartments and impact plant development. The synthesis and remodeling of homogalacturonan pectins is associated with multiple developmental processes ranging from growth to response to biotic/abiotic stress. It encompasses Golgi-localized methylation and acetylation and subsequent demethylation and deacetylation in the cell wall. In the last decade, our comprehension of plant polysaccharides acetylation has increased significantly thanks to the study of the TRICHOME BIREFRINGENCE-LIKE (TBL) protein family. TBLs are mostly described as Golgi-localized acetyltransferases specifically targeting diverse hemicelluloses or pectins. Various tbl mutants showed altered wall mechanical properties and dynamics. Here, we study TBL38 that is co-expressed with PECTIN METHYLESTERASE INHIBITOR6 (PMEI6) and PEROXIDASE 36 (PRX36) during the development of Arabidopsis seed mucilage secretory cells (MSCs). We demonstrate the atypical TBL38 cell wall localization restricted to the PMEI6/PRX36 MSC cell wall microdomain. A tbl38 mutant displays an intriguing homogalacturonan immunological phenotype in this cell wall microdomain and in a MSC surface-enriched abrasion powder. This fraction was further characterized by mass spectrometry oligosaccharide profiling revealing an increased homogalacturonan acetylation phenotype. Finally, a recombinant TBL38 is shown to display pectin acetylesterase activity in vitro. These results indicate that TBL38 is an atypical cell wall-localized TBL that displays a homogalacturonan acetylesterase activity rather than a Golgi-localized acetyltransferase activity as observed in previously studied TBLs. TBL38 function during seed development is discussed.

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↗

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

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↗