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Polygalacturonases (PGs) fine-tune pectins to modulate cell wall chemistry and mechanics, impacting plant development. The large number of PGs encoded in plant genomes leads to questions on the diversity and specificity of distinct isozymes. Herein, we report the crystal structures of 2 Arabidopsis thaliana PGs, POLYGALACTURONASE LATERAL ROOT (PGLR), and ARABIDOPSIS DEHISCENCE ZONE POLYGALACTURONASE2 (ADPG2), which are coexpressed during root development. We first determined the amino acid variations and steric clashes that explain the absence of inhibition of the plant PGs by endogenous PG-inhibiting proteins (PGIPs). Although their beta helix folds are highly similar, PGLR and ADPG2 subsites in the substrate binding groove are occupied by divergent amino acids. By combining molecular dynamic simulations, analysis of enzyme kinetics, and hydrolysis products, we showed that these structural differences translated into distinct enzyme-substrate dynamics and enzyme processivities: ADPG2 showed greater substrate fluctuations with hydrolysis products, oligogalacturonides (OGs), with a degree of polymerization (DP) of ≤4, while the DP of OGs generated by PGLR was between 5 and 9. Using the Arabidopsis root as a developmental model, exogenous application of purified enzymes showed that the highly processive ADPG2 had major effects on both root cell elongation and cell adhesion. This work highlights the importance of PG processivity on pectin degradation regulating plant development., Competing Interests: Conflict of interest statement. None declared., (© American Society of Plant Biologists 2023. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Cadmium (Cd) was shown to co-localise with calcium (Ca) in oxalate crystals in the stems and leaves of Cd tolerant Gomphrena claussenii, but Cd binding remained unresolved. Using synchrotron radiation X-ray absorption near edge spectroscopy we demonstrate that in oxalate crystals of hydroponically grown G. claussenii the vast majority of Cd is bound to oxygen ligands in oxalate crystals (>88%; Cd-O-C coordination) and the remaining Cd is bound to sulphur ligands (Cd-S-C coordination). Cadmium binding to oxalate does not depend on the amount of Ca supplied or from which organs the crystals originate (stems and mature leaves). By contrast, roots contain no oxalate crystals and therein Cd is bound predominantly by S ligands. The potential to remove Cd by extraction of Cd-rich oxalate crystals from plant material should be tested in phytoextraction or phytomining strategies.

Typha latifolia is a plant species widely used for phytoremediation. Accumulation, localization and distribution of Pb and mineral nutrients were investigated in roots, rhizomes and leaves of Typha latifolia grown at 0, 50, 100 and 250 μM Pb concentrations in a pot experiment under controlled conditions. Bulk elemental concentrations were determined by X-ray fluorescence (XRF) spectroscopy whereas micro-proton-induced X-ray emission (micro-PIXE) was used for element localization in root and rhizome tissues. Gradual increase in bulk Pb concentrations was observed in Typha latifolia roots and rhizomes treated with increasing Pb concentrations, however in rhizomes Pb concentrations were an order of magnitude lower than in roots. In leaves Pb concentrations were around the limit of detection for XRF (~20 μg g(-1)). An increase in concentration of K and Ca in roots, rhizomes and leaves, of iron and zinc in roots and leaves, and of Mn in rhizomes was observed either at 50 and/or 100 μM Pb treatments, whereas for K and Ca in roots, rhizomes and leaves, Fe and Zn in roots and leaves and Mn in rhizomes, or at 250 μM Pb treatment the increase was seen for concentrations of Fe and Zn in rhizomes and Cu in roots. Mn concentrations decreased with Pb treatments in roots and leaves. Element localization using micro-PIXE analysis demonstrated Pb accumulation in epidermal and cortical tissues of treated roots and rhizomes, while in endodermis and vascular tissues Pb was not detected. A displacement of Ca from epidermal to cortical tissues was observed in Pb treated roots and rhizomes, pointing to cell wall immobilization of Pb as one of the tolerance mechanisms in Typha latifolia. High level of colocalization of Pb with P (r = 0.60), S (r = 0.37) and Zn (r = 0.70) was observed in Pb treated roots, while in rhizomes colocalization with the mentioned elements was still positive, but not that prominent. These results indicate that Pb may form complexes with phosphorus and sulfur compounds in roots and rhizomes, which may also represent attraction sites for binding Zn. Because of its large root and rhizome surface area acting as main sites for Pb adsorption, Typha latifolia may represent potentially efficient plant species for phytoremediation of Pb contaminated soils and waters., (This journal is © The Royal Society of Chemistry 2012)

Mature developed seeds are physiologically and biochemically committed to store nutrients, principally as starch, protein, oils, and minerals. The composition and distribution of elements inside the aleurone cell layer reflect their biogenesis, structural characteristics, and physiological functions. It is therefore of primary importance to understand the mechanisms underlying metal ion accumulation, distribution, storage, and bioavailability in aleurone subcellular organelles for seed fortification purposes. Synchrotron radiation soft X-ray full-field imaging mode (FFIM) and low-energy X-ray fluorescence (LEXRF) spectromicroscopy were applied to characterize major structural features and the subcellular distribution of physiologically important elements (Zn, Fe, Na, Mg, Al, Si, and P). These direct imaging methods reveal the accumulation patterns between the apoplast and symplast, and highlight the importance of globoids with phytic acid mineral salts and walls as preferential storage structures. C, N, and O chemical topographies are directly linked to the structural backbone of plant substructures. Zn, Fe, Na, Mg, Al, and P were linked to globoid structures within protein storage vacuoles with variable levels of co-localization. Si distribution was atypical, being contained in the aleurone apoplast and symplast, supporting a physiological role for Si in addition to its structural function. These results reveal that the immobilization of metals within the observed endomembrane structures presents a structural and functional barrier and affects bioavailability. The combination of high spatial and chemical X-ray microscopy techniques highlights how in situ analysis can yield new insights into the complexity of the wheat aleurone layer, whose precise biochemical composition, morphology, and structural characteristics are still not unequivocally resolved.