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Instruments / Technical Area	Publication Details	Published
I12-JEEP: Joint Engineering, Environmental and Processing	Preferential wheat (Triticum aestivum. L cv. Fielder) root growth in different sized aggregates Tinashe Mawodza , Manoj Menon , Harriet Brooks , Oxana V. Magdysyuk , Genoveva Burca , Stuart Casson Soil And Tillage Research 212 DOI: 10.1016/j.still.2021.105054 Diamond Proposal Number(s): [22992] Show Abstract ▼ Abstract: Soil structure is one of the most important environmental factors affecting root architectural development and consequently plant yield. Understanding how plant roots respond to soils with variable soil structure is important as it enables soil management practices that promote optimal root growth. Many contemporary, non-invasive experiments investigating how plant root architecture responds to soil structural variations have often focused on compaction, often neglecting the role of soil aggregate size in determining root configuration. To better understand this, in this study, we used non-invasive neutron and X-Ray imaging to investigate how variable aggregate size affects the early root architectural establishment in wheat plants. Sandy loam soil derived macro-aggregates of two distinct sizes (0.25−0.5 and 2−4 mm) were used to infer the suitability of each aggregate size for use in wheat seedbeds. We also grew wheat seedlings in partitioned containers with the two different aggregate size classes filled side by side to establish whether there would be preferential growth of roots in either aggregate size class. Our results showed significantly increased root growth in the smaller 0.25−0.5 mm aggregates as compared to the larger 2−4 mm aggregates. This was mainly as a result of enhanced lateral root growth when the wheat plants were grown in the finer aggregates. On the other hand, coarser aggregates induced significantly increased seminal root axes which partially offset the differences in total root length between the two aggregate sizes. Plants growing in partitioned containers similarly indicated preferential root growth in smaller aggregate with an even more pronounced difference in root growth in the smaller aggregates. As inferred from our results, seedbeds dominated by smaller macro-aggregates (finer soil tilth) may be optimal to enhance wheat seedling root growth in sandy loam soils.	Aug 2021
B18-Core EXAFS	Evaluation of cadmium and arsenic effects on wild and cultivated cardoon genotypes selected for metal phytoremediation and bioenergy purposes Chiara Leonardi , Valeria Toscano , Claudia Genovese , Julian Frederick Willem Mosselmans , Bryne Tendolo Ngwenya , Salvatore Antonino Raccuia Environmental Science And Pollution Research 15 DOI: 10.1007/s11356-021-14705-9 Diamond Proposal Number(s): [15231] Open Access Show Abstract ▼ Abstract: Cynara cardunculus L. is a multipurpose crop, characterized by high production of biomass suitable for energy purposes and green chemistry. Taking advantage of its already demonstrated ability to grow in polluted environments that characterize many world marginal lands, the aim of this work was to investigate the response of different cardoon genotypes to exposure to cadmium (Cd) and arsenic (As) pollution, in order to use this crop for rehabilitation of contaminated sites and its biomass for energy production. In this study, seeds of two wild cardoon accessions harvested in rural and industrial Sicilian areas and of a selected line of domestic cardoon were used, and the grown plants were spiked with As and Cd, alone or in combination, at two different concentrations (500 and 2000 μM) and monitored for 45 days. The growth parameters showed that all the plants survived until the end of experiment, with growth stimulation in the presence of low concentrations of As and Cd, relative to metal-free controls. Biomass production was mostly allocated in the roots in As treatment and in the shoots in Cd treatment. Cd EXAFS analysis showed that tolerance to high concentrations of both metals was likely linked to complexation of Cd with oxygen-containing ligands, possibly organic acids, in both root and leaf biomass with differences in behaviour among genotypes. Under As+Cd contamination, the ability of the plants to translocate As to aboveground system increased also showing that, for both metal(loid)s, there were significant differences between genotypes studied. Moreover, the results showed that Cynara cardunculus var. sylvestris collected in an industrial area is the genotype that, among those studied, had the best phytoextraction capability for each metal(loid).	Jun 2021
I04-Macromolecular Crystallography	The allelic rice immune receptor Pikh confers extended resistance to strains of the blast fungus through a single polymorphism in the effector binding interface Juan Carlos De La Concepcion , Josephine H. R. Maidment , Apinya Longya , Gui Xiao , Marina Franceschetti , Mark J. Banfield Plos Pathogens 17 DOI: 10.1371/journal.ppat.1009368 Diamond Proposal Number(s): [13467] Open Access Show Abstract ▼ Abstract: Arms race co-evolution drives rapid adaptive changes in pathogens and in the immune systems of their hosts. Plant intracellular NLR immune receptors detect effectors delivered by pathogens to promote susceptibility, activating an immune response that halts colonization. As a consequence, pathogen effectors evolve to escape immune recognition and are highly variable. In turn, NLR receptors are one of the most diverse protein families in plants, and this variability underpins differential recognition of effector variants. The molecular mechanisms underlying natural variation in effector recognition by NLRs are starting to be elucidated. The rice NLR pair Pik-1/Pik-2 recognizes AVR-Pik effectors from the blast fungus Magnaporthe oryzae, triggering immune responses that limit rice blast infection. Allelic variation in a heavy metal associated (HMA) domain integrated in the receptor Pik-1 confers differential binding to AVR-Pik variants, determining resistance specificity. Previous mechanistic studies uncovered how a Pik allele, Pikm, has extended recognition to effector variants through a specialized HMA/AVR-Pik binding interface. Here, we reveal the mechanistic basis of extended recognition specificity conferred by another Pik allele, Pikh. A single residue in Pikh-HMA increases binding to AVR-Pik variants, leading to an extended effector response in planta. The crystal structure of Pikh-HMA in complex with an AVR-Pik variant confirmed that Pikh and Pikm use a similar molecular mechanism to extend their pathogen recognition profile. This study shows how different NLR receptor alleles functionally converge to extend recognition specificity to pathogen effectors.	Mar 2021
B18-Core EXAFS	X-ray absorption spectroscopy evidence of sulfur-bound Cadmium in the Cd-hyperaccumulator Solanum nigrum and the non-accumulator Solanum melongena Marie-Laure Pons , Blanche Collin , Emmanuel Doelsch , Perrine Chaurand , Till Fehlauer , Clement Levard , Catherine Keller , Jérôme Rose Environmental Pollution 108 DOI: 10.1016/j.envpol.2021.116897 Diamond Proposal Number(s): [23312] Show Abstract ▼ Abstract: It has been proposed that non-protein thiols and organic acids play a major role in cadmium phytoavailability and distribution in plants. In the Cd-hyperaccumulator Solanum nigrum and non-accumulator Solanum melongena, the role of these organic ligands in the accumulation and detoxification mechanisms of Cd are debated. In this study, we used X-ray absorption spectroscopy to investigate Cd speciation in these plants (roots, stem, leaves) and in the soils used for their culture to unravel the plants responses to Cd exposure. The results show that Cd in the 100 mg.kg-1 Cd-doped clayey loam soil is sorbed onto iron oxyhydroxides. In both S. nigrum and S. melongena, Cd in roots and fresh leaves is mainly bound to thiol ligands, with a small contribution of inorganic S ligands in S. nigrum leaves. We interpret the Cd binding to sulfur ligands as detoxification mechanisms, possibly involving the sequestration of Cd complexed with glutathione or phytochelatins in the plant vacuoles. In the stems, results show an increase binding of Cd to -O ligands (>50% for S. nigrum). We suggest that Cd is partly complexed by organic acids for transportation in the sap.	Mar 2021
B18-Core EXAFS	Role of plant growth promoting bacteria in driving speciation gradients across soil-rhizosphere-plant interfaces in zinc-contaminated soils Nyekachi C. Adele , Bryne T. Ngwenya , Kate V. Heal , J. Frederick W. Mosselmans Environmental Pollution 283 DOI: 10.1016/j.envpol.2021.116909 Diamond Proposal Number(s): [10429] Show Abstract ▼ Abstract: Inoculation of soil or seeds with plant growth promoting bacteria ameliorates metal toxicity to plants by changing metal speciation in plant tissues but the exact location of these changes remains unknown. Knowing where the changes occur is a critical first step to establish whether metal speciation changes are driven by microbial metabolism or by plant responses. Since bacteria concentrate in the rhizosphere, we hypothesised steep changes in metal speciation across the rhizosphere. We tested this by comparing speciation of zinc (Zn) in roots of Brassica juncea plants grown in soil contaminated with 600 mg kg-1 of Zn with that of bulk and rhizospheric soil using synchrotron X-ray absorption spectroscopy (XAS). Seeds were either uninoculated or inoculated with Rhizobium leguminosarum bv. trifolii and Zn was supplied in the form of sulfide (ZnS nanoparticles) and sulfate (ZnSO4). Consistent with previous studies, Zn toxicity, as assessed by plant growth parameters, was alleviated in B. juncea inoculated with Rhizobium leguminosarum. XAS results showed that in both ZnS and ZnSO4 treatments, the most significant changes in speciation occurred between the rhizosphere and the root, and involved an increase in the proportion of organic acids and thiol complexes. In ZnS treatments, Zn phytate and Zn citrate were the dominant organic acid complexes, whilst Zn histidine also appeared in roots exposed to ZnSO4. Inoculation with bacteria was associated with the appearance of Zn cysteine and Zn formate in roots, suggesting that these two forms are driven by bacterial metabolism. In contrast, Zn complexation with phytate, citrate and histidine is attributed to plant responses, perhaps in the form of exudates, some with long range influence into the bulk soil, leading to shallower speciation gradients.	Mar 2021
I18-Microfocus Spectroscopy	Co-application of Se and a biostimulant at different wheat growth stages: Influence on grain development Tingting Xiao , Roberto Boada , Mercè Llugany , Manuel Valiente Plant Physiology And Biochemistry 160, 184 - 192 DOI: 10.1016/j.plaphy.2021.01.025 Diamond Proposal Number(s): [18671] Show Abstract ▼ Abstract: An appropriate selenium intake can be beneficial for human health. Se-biofortified food in Se-deficient regions is becoming an increasingly common practice but there are still issues to be addressed regarding the observed Se-induced toxicity to the plant. In this respect, plant biostimulants are used to enhance nutrition efficiency, abiotic stress tolerance and crop quality. In this work, the efficacy of a plant biostimulant to counteract the Se-induced stress in wheat plants is experimentally assessed. The co-application of different Se-biofortification treatments and the biostimulant at different growth stages (tillering or heading stage) was investigated. The use of micro focused X-ray spectroscopy allows us to confirm organic Se species to be the main Se species found in wheat grain and that the proportion of organic Se species is only slightly affected by the Se application stage. Our study proves that the biostimulant had a key role in the enhancement of both the amount of grains produced per spike and their dry biomass without hindering Se enrichment process, neither diminishing the Se concentration nor massively disrupting the Se species present. This information will be useful to minimize both plant toxicity and economic cost towards a more effective and plant healthy selenium supplementation.	Mar 2021
I03-Macromolecular Crystallography	Multiple variants of the fungal effector AVR-Pik bind the HMA domain of the rice protein OsHIPP19, providing a foundation to engineer plant defence Josephine H. R. Maidment , Marina Franceschetti , Abbas Maqbool , Hiromasa Saitoh , Chatchawan Jantasuriyarat , Sophien Kamoun , Ryohei Terauchi , Mark J. Banfield Journal Of Biological Chemistry 7 DOI: 10.1016/j.jbc.2021.100371 Diamond Proposal Number(s): [13467] Open Access Show Abstract ▼ Abstract: Microbial plant pathogens secrete effector proteins which manipulate the host to promote infection. Effectors can be recognised by plant intracellular nucleotide-binding leucine-rich repeat (NLR) receptors, initiating an immune response. The AVR-Pik effector from the rice blast fungus Magnaporthe oryzae is recognised by a pair of rice NLR receptors, Pik-1 and Pik-2. Pik-1 contains a non-canonical integrated heavy metal-associated (HMA) domain, which directly binds AVR-Pik to activate plant defences. The host targets of AVR-Pik are also HMA domain-containing proteins, namely heavy metal-associated isoprenylated plant proteins (HIPPs) and heavy metal-associated plant proteins (HPPs). Here, we demonstrate that one of these targets interacts with a wider set of AVR-Pik variants compared to the Pik-1 HMA domains. We define the biochemical and structural basis of the interaction between AVR-Pik and OsHIPP19, and compare the interaction to that formed with the HMA domain of Pik-1. Using analytical gel filtration and surface plasmon resonance, we show that multiple AVR-Pik variants, including the stealthy variants AVR-PikC and AVR-PikF which do not interact with any characterised Pik-1 alleles, bind to OsHIPP19 with nanomolar affinity. The crystal structure of OsHIPP19 in complex with AVR-PikF reveals differences at the interface that underpin high-affinity binding of OsHIPP19-HMA to a wider set of AVR-Pik variants than achieved by the integrated HMA domain of Pik-1. Our results provide a foundation for engineering the HMA domain of Pik-1 to extend binding to currently unrecognised AVR-Pik variants and expand disease resistance in rice to divergent pathogen strains.	Feb 2021
I13-1-Coherence I18-Microfocus Spectroscopy	Developing a system for in vivo imaging of maize roots containing iodinated contrast media in soil using synchrotron XCT and XRF Callum P. Scotson , Arjen Van Veelen , Katherine A. Williams , Nicolai Koebernick , Dan Mckay Fletcher , Tiina Roose Plant And Soil 4 DOI: 10.1007/s11104-020-04784-x Diamond Proposal Number(s): [18762, 19399] Open Access Show Abstract ▼ Abstract: Aims: We sought to develop a novel experimental system which enabled application of iodinated contrast media to in vivo plant roots intact in soil and was compatible with time-resolved synchrotron X-ray computed tomography imaging. The system was developed to overcome issues of low contrast to noise within X-ray computed tomography images of plant roots and soil environments, the latter of which can complicate image processing and result in the loss of anatomical information. Methods: To demonstrate the efficacy of the system we employ the novel use of both synchrotron X-ray computed tomography and synchrotron X-ray fluorescence mapping to capture the translocation of the contrast media through root vasculature into the leaves. Results: With the application of contrast media we identify fluid flow in root vasculature and visualise anatomical features, which are otherwise often only observable in ex vivo microscopy, including: the xylem, metaxylem, pith, fibres in aerenchyma and leaf venation. We are also able to observe interactions between aerenchyma cross sectional area and solute transport in the root vasculature with depth. Conclusions: Our novel system was capable of successfully delivering sufficient contrast media into root and leaf tissues such that anatomical features could be visualised and internal fluid transport observed. We propose that our system could be used in future to study internal plant transport mechanisms and parameterise models for fluid flow in plants.	Dec 2020
Krios III-Titan Krios III at Diamond	High-light versus low-light: Effects on paired photosystem II supercomplex structural rearrangement in pea plants Alessandro Grinzato , Pascal Albanese , Roberto Marotta , Paolo Swuec , Guido Saracco , Martino Bolognesi , Giuseppe Zanotti , Cristina Pagliano International Journal Of Molecular Sciences 21 DOI: 10.3390/ijms21228643 Diamond Proposal Number(s): [19714] Open Access Show Abstract ▼ Abstract: In plant grana thylakoid membranes Photosystem II (PSII) associates with a variable number of antenna proteins (LHCII) to form different types of supercomplexes (PSII-LHCII), whose organization is dynamically adjusted in response to light cues, with the C2S2 more abundant in high-light and the C2S2M2 in low-light. Paired PSII-LHCII supercomplexes interacting at their stromal surface from adjacent thylakoid membranes were previously suggested to mediate grana stacking. Here, we present the cryo-electron microscopy maps of paired C2S2 and C2S2M2 supercomplexes isolated from pea plants grown in high-light and low-light, respectively. These maps show a different rotational offset between the two supercomplexes in the pair, responsible for modifying their reciprocal interaction and energetic connectivity. This evidence reveals a different way by which paired PSII-LHCII supercomplexes can mediate grana stacking at diverse irradiances. Electrostatic stromal interactions between LHCII trimers almost completely overlapping in the paired C2S2 can be the main determinant by which PSII-LHCII supercomplexes mediate grana stacking in plants grown in high-light, whereas the mutual interaction of stromal N-terminal loops of two facing Lhcb4 subunits in the paired C2S2M2 can fulfil this task in plants grown in low-light. The high-light induced accumulation of the Lhcb4.3 protein in PSII-LHCII supercomplexes has been previously reported. Our cryo-electron microscopy map at 3.8 Å resolution of the C2S2 supercomplex isolated from plants grown in high-light suggests the presence of the Lhcb4.3 protein revealing peculiar structural features of this high-light-specific antenna important for photoprotection.	Nov 2020
I02-Macromolecular Crystallography I03-Macromolecular Crystallography I04-Macromolecular Crystallography I24-Microfocus Macromolecular Crystallography	The active site of magnesium chelatase Nathan B. P. Adams , Claudine Bisson , Amanda A. Brindley , David A. Farmer , Paul A. Davison , James D. Reid , C. Neil Hunter Nature Plants 104 DOI: 10.1038/s41477-020-00806-9 Diamond Proposal Number(s): [8987, 12788] Show Abstract ▼ Abstract: The insertion of magnesium into protoporphyrin initiates the biosynthesis of chlorophyll, the pigment that underpins photosynthesis. This reaction, catalysed by the magnesium chelatase complex, couples ATP hydrolysis by a ChlID motor complex to chelation within the ChlH subunit. We probed the structure and catalytic function of ChlH using a combination of X-ray crystallography, computational modelling, mutagenesis and enzymology. Two linked domains of ChlH in an initially open conformation of ChlH bind protoporphyrin IX, and the rearrangement of several loops envelops this substrate, forming an active site cavity. This induced fit brings an essential glutamate (E660), proposed to be the key catalytic residue for magnesium insertion, into proximity with the porphyrin. A buried solvent channel adjacent to E660 connects the exterior bulk solvent to the active site, forming a possible conduit for the delivery of magnesium or abstraction of protons.	Nov 2020

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